GB2368512A - Increasing soluble fibre content in pasta products using enzymes - Google Patents

Abstract

A process of producing pasta or noodle products with increased water soluble fibre content comprises the addition of a fibre-hydrolysing enzyme during pasta production, e.g. added to the raw materials for pasta. Insoluble fibre may also be added. The water soluble fibre in the pasta products preferably does not leach during cooking of the pasta. The enzyme may be added as a microbial organism expressing the enzyme, e.g. Aspergillus niger or Bacillus subtillis and the enzyme may be an endoxylanase. The resulting pasta products may comprise water-extractable arabinoxylan in a specific concentration which consists of molecules of a variable molecular weight distribution. The pasta products may have a health benefit to a consumer, such as lowering cholesterol levels. A machine may be specifically designed for the process of pasta production.

Description

Process for preparing pasta products with increased levels of soluble fiber Field of the invention The present invention is related to a process for the preparation of pasta products with increased levels of non-leachable water soluble (dietary) fiber. This process comprises the addition of endoxylanases (E. C. 3.2. 1. 8) or any other fiber hydrolysing enzymes such as cellulases (such as E. C. 3.2. 1.4), glucanases (such as E. C. 3.2. 1. 6),... or combinations thereof during pasta production. Possible substrates for these enzymes are arabinoxylans (AX), cellulose, P-glucans, glucomannans, lignin and pectins. Most of them are classified as nonstarch-polysaccharides (NSP).

Technological background of the invention In this text, dietary fiber is defined as the remnants of edible plant cell polysaccharides, lignin and associated substances resistant to hydrolysis by human alimentary enzymes. AX, such as e. g. present in cereals is therefore a dietary fiber. Dietary fiber can also be referred to as dietary fibre, fibre or fiber.

Endoxylanases are arabinoxylan hydrolysing enzymes. The term dietary fiber hydrolysing enzymes is used for all enzymes able to hydrolyze dietary fiber.

Dietary fiber can be either water-extractable or water-unextractable. For the purposes of this text, the term soluble fiber is used for the water-extractable fiber or the fiber solubilised as result of enzymic hydrolysis or combinations thereof (unless specified otherwise).

AX consist of a backbone of 1, 4 linked ss-Dxylopyranosyl units partially substituted with a-1-2 and/ or ci-1-3 L-arabinofuranosyl side chains (Perlin 1951).

They can either be water-extractable (WE-AX) or waterunextractable (WU-AX). The reported total AX (TOT-AX) contents in semolina, the milling product of durum wheat (Triticum durum Desf. ), which is commonly used for the production of pasta, vary between 2.25% and 3. 02% respectively (d. b. , dry basis) (Bains and Irvine 1965, Lempereur et al 1997) and 0.28% and 0.36% (d. b. ) for WE- AX (Lintas and D'Appolonia 1973, Roels et al 1999). Durum wheat AX contain a higher proportion of arabinose than common wheat AX, indicating a more branched structure (Medcalf and Gilles 1968, Medcalf et al 1968, Ciaccio et al 1982, Roels et al 1999). In common wheat flour, the percentage of TOT-AX varies between ca. 1.30% and 2.30% and that of WE-AX between ca. 0.40 and 0.70% (Cleemput et al 1993). Rye flour contains between ca. 1.50% and 3.80% WE-AX and between ca. 3.80% and 8.00% TOT-AX (Nyman et al 1984, Meuser and Suckow 1986).

Endoxylanases (EC: 3.2. 1.8) are AX hydrolysing enzymes, able to transform both WU-AX and WE-AX. The use of endoxylanases is widespread in e. g. the breadmaking industry and extensive studies have focussed on their importance in the process. In breadmaking, some endoxylanases have a beneficial effect on bread volume (Kulp 1968, McCleary 1986, ter Haseborg and Himmelstein 1988, Rouau et al 1994, Courtin et al 1999). Courtin et al (1999) demonstrated that WU-AX have a negative effect on bread volume and that their enzymic solubilisation without drastic reduction of the molecular weight (MW) of the solubilised AX is of primary importance for optimal bread quality.

Endoxylanases can change the AX dietary fiber population in five distinct ways (Courtin 2000). In the light of the present invention, the following actions are worthwhile to be mentioned: the endoxylanases can reduce the level of WU-AX by solubilising them, which results in an increase in the level of solubilised AX. They will also reduce the MW of the solubilised AX as well as that of the original WE-AX.

Cereal ss-glucans consist of linear chains of P-D- glucopyranosyl residues joined by (1-3) and (1-4) glycosidic linkages. They represent ca. 25% in wheat and ca. 75% in barley of the cell wall NSP.

p-glucanases are endoglucanase enzymes that hydrolyze the 1-3 and/or 1-4 p-D-glucosidic bonds in ssglucans.

Cellulose consists of linear chains of (1-4) P-D glucopyranosyl residues.

Glucomannans consist of varying proportions of (1-4) -linked R-D-glucopyranosyl and P-D-mannopyranosyl residues.

Lignin is a three-dimensional, highly complex network built up of aromatic p-hydroxyphenyl, guaiacyl, and syringyl units.

Pectins are polymers consisting of D-galacturonic structural units joined by 1-4 linkages. Other components are rhamnose units, D-qalactan, arabinan, fucose and xylose sugars.

Pasta (products) are cereal-based products that are formed from a dough but are not leavened. The processes (e. g. extrusion, sheeting,...) by which they are formed vary, as do the type of raw materials used (see field of the invention) such as (whole meal) durum wheat semolina or bread wheat flour or rice flour or maize

flour or soya flour or buckwheat flour,... or combinations thereof (Hoseney 1994). Ingredients used are water, semolina or flour or combinations thereof and in some instances other ingredients such as ascorbic acid, salt, alkaline salts, fruit or vegetable extracts or both, coloring substances, enzymes such as glucose oxidase, amylases,..., emulsifiers, eggs,.... For some pasta products, a yellow color is desired and oxidation of carotenoid pigments originally present in e. g. durum wheat semolina is to be avoided. For this reason, mixing of the dough may be carried out under partial or full vacuum. Pasta (products) can also have a filling which mostly consists of meat, fish, cheese, vegetables,... or mixtures thereof.

Pasta products can have many shapes and dimensions and are sold under different type names: (the following list is not limitative) canellonni, capellini, conchiglione, egg noodles, farfalle, fettuccine, gnocchi, lasagne, linguine, macararoni, penne, radiatori, ravioli, spaghetti, spätzle, spirelli, tagliatelle, tortellini, vermicelli. Important pasta product classes are long-cut pasta, short-cut pasta and nest shaped pasta. After forming the pasta structure, the products can be used directly as fresh pasta, stored under conditioned atmosphere, stored cooled or deep-frozen,.... Additional drying steps can be included after the pasta forming process to obtain dry pasta products, which can be stored for longer periods. Drying processes used can be low temperature drying (LT, 20-60oC), high temperature drying (HT, 60-84OC) and ultra high temperature drying (UHT, > 84 C) or combinations or variants of the above. Steaming treatments can be included during the production process of pasta products to fully or partially gelatinise starch. This results in stronger products with eventually a lower cooking time. Other treatments can also be

included in the process to decrease cooking times of the final products. In what follows, the terms pasta (products) are meant for any product that can be produced in the ways described above. With the terms pasta (products) is also meant those products mentioned in the International Patent Classification (Class A23L 1/16) and those products mentioned in the standard pasta literature: Durum Wheat Chemistry and Technology (Fabriani and Lintas 1988), Pasta Technology Today (Milatovic and Mondelli 1991) and Pasta and Noodle Technology (Kruger et al 1996). The term pasta is therefore also including related products as noodles,....

With the term process for producing pasta is meant all processes to produce pasta products, including those mentioned in this text and in the standard pasta literature.

Standard pasta production processes involve one of the following steps or combinations thereof: mixing the pasta ingredients; moistening the pasta ingredients; mixing, kneading or stretching until a desired consistency is reached; shaping the pasta by extruding, molding, sheeting or stretching; pressing or cutting until a desired form is reached; drying the pasta ; and cooking the pasta.

Definitions of other terms used in this text Dietary fiber hydrolysing enzymes: all enzymes able to hydrolyze dietary fiber.

Whole meal pasta products: those pasta products characterised in that they contain a total AX content exceeding 3.00% (expressed on dry basis, d. b.).

Non whole meal pasta products: those pasta products characterised in that they contain a total AX

content lower than or equal 3. 00% (expressed on dry basis, d. b.).

Increased soluble dietary fiber: (a) in the case of AX as source of dietary fiber: (a. 1) for whole meal pasta products: in the most preferred embodiment, a WE-AX concentration higher than 0.80% (on d. b. ) in the whole meal pasta products or 25% of their TOT-AX population is WE-AX or both. in the second most preferred embodiment, a WE-AX concentration higher than 0.85% (on d. b. ) in the whole meal pasta products or 30% of their TOT-AX population is WE-AX or both. in the third most preferred embodiment, a WE-AX concentration higher than 0.90% (on d. b. ) in the whole meal pasta products or 35% of their TOT-AX population is WE-AX or both. in the fourth most preferred embodiment, a WE-AX concentration higher than 0.95% (on d. b. ) in the whole meal pasta products or 40% of their TOT-AX population is WE-AX or both. in the fifth most preferred embodiment, a WE-AX concentration higher than 1.00% (on d. b. ) in the whole meal pasta products or 45% of their TOT-AX population is WE-AX or both. in the sixth most preferred embodiment, a WE-AX concentration higher than 1.05% (on d. b. ) in the whole meal pasta products or 50% of their TOT-AX population are WE-AX or both.

(a. 2) for non whole meal pasta products: in the most preferred embodiment, a WE-AX concentration higher than 0.80% (on d. b. ) in the pasta products or 42% of their TOT-AX population is WE-AX or both.

in the second most preferred embodiment, a WE-AX concentrations higher than 0. 85% (on d. b.) of the pasta products or 45% of their TOT-AX population is WE-AX or both. in the third most preferred embodiment, a WE-AX concentration higher than 0. 90% (on d. b. ) of the pasta products or 50% of their TOT-AX population is WE-AX or both. in the fourth most preferred embodiment, a WE-AX concentration higher than 0.95% (on d. b. ) of the pasta products or 55% of their TOT-AX population is WE-AX or both. in the fifth most preferred embodiment, a WE-AX concentration higher than 1.00% (on d. b. ) of the pasta products or 60% of their TOT-AX population is WE-AX or both. in the sixth most preferred embodiment, a WE-AX concentration higher than 1.05% (on d. b. ) of the pasta products or 65% of their TOT-AX population is WE-AX or both.

(b) for any other source of dietary fiber: contents of soluble dietary fiber resulting from enzymic treatment before or during or after the pasta production process or combinations thereof and consequently higher than those present in pasta products obtained in the corresponding process without use of fiber hydrolysing enzymes.

Well retained or hardly leached or non-leachable or not leached easily: (a) in the case of AX as source of dietary fiber in whole or non whole meal (or combinations thereof) pasta products:

in the most preferred embodiment, not more than 20% of the TOT-AX or WE-AX or both are leached when pasta products are cooked to optimal cooking time (T). in the second most preferred embodiment, not more than 25% of the TOT-AX or WE-AX or both are leached when pasta products are cooked to optimal cooking time (T). in the third most preferred embodiment, not more than 30% of the TOT-AX or WE-AX or both are leached when pasta products are cooked to optimal cooking time (T).

(b) for any other source of dietary fiber in whole or non whole meal (or combinations thereof) pasta products: in the most preferred embodiment, not more than 20% of the specific source of dietary fiber is leached when pasta products are cooked to optimal cooking time (T). in the second most preferred embodiment, not more than 25% of the specific source of dietary fiber is leached when pasta products are cooked to optimal cooking time (T). in the third most preferred embodiment, not more than 30% of the specific source of dietary fiber is leached when pasta products are cooked to optimal cooking time (T).

State of the art AX are an important source of dietary fiber in refined cereal-based products (Theander et al 1993).

Recently, De Vries et al (1999) defined dietary fiber as the remnants of edible plant cell polysaccharides, lignin and associated substances resistant to hydrolysis by human alimentary enzymes.

The importance of dietary fiber has been welldocumented (Trowell 1972, Burkitt 1973, Leveille 1976, Weber and Chaudharry 1987, Kahlon and Chow 1997, Kritchevsky 1997, Meister and Raso 1997). It is certain that not all positive health effects of fiber are understood at present however of the two classes of dietary fiber: insoluble and soluble fiber, the latter have the most prevalent health effects. A diet rich in insoluble fibers is associated with smaller reductions of blood lipids than one rich in soluble dietary fibers (Jenkins et al 1993). Soluble dietary fiber can lower cholesterol levels (Ink and Hurt 1987, Ripsin et al 1992, Haskell et al 1992, Glore et al 1994) and influence human glycemic response positively (Ink and Hurt, 1987, Nutall 1993, Yokoyama et al 1997).

In the past, attempts have been made to increase the levels of (soluble) dietary fiber in pasta by adding high fiber containing material, from sources other than durum wheat. Low glycemic responses in diabetics have been obtained with pastas formulated with guar gum (Gatti et al 1984, Briani et al 1987, Carra et al 1990).

Undesirable side effects of foods formulated with guar gums are reported: negative sensory attributes, vomiting, increased flatulence, diarrhea (Simons et al 1982, Smith et al 1982). Addition of fiber rich durum bran to semolina for pasta production gave a tasteful product, but resulted in increasing cooking losses and reduced firmness of the cooked pasta (Kordonowy and Youngs 1985).

Another disadvantage of the bran incorporation is that it results in decreased mineral availability (Rendleman 1982, Rendleman and Grobe 1982). More generally, indications were found that soluble dietary fibers do not reduce the bioavailibility of minerals to the extent that insoluble fibers do (Drews et al 1979, Behall et al

1983). Dougherty et al (1988) added oat fibers in the pasta dough recipe. However, even with an extra addition of vital wheat gluten, a product of lower quality was obtained. Pasta quality improvements were obtained with xanthan gum, whereas durum wheat WE-AX incorporation gave no difference in quality and pea fiber incorporation and whole wheat pasta yielded lower quality products (Edwards et al 1995). Incorporation of barley -glucan in pasta products yields products with a higher fiber content and modest quality (Knuckles et al 1997, Marconi et al 2000), but which result in lower glycemic responses (Yokoyama et al 1997).

Addition of endoxylanases during pasta making results in products with an increased soluble dietary fiber content. In their effort to understand the role of AX in pasta making, the inventors used endoxylanases as an elegant tool to study the role of AX. While the man aware of the state of the art would have expected that the solubilised AX are leached out during the pasta cooking process, and hence are lost as (soluble) dietary fiber source for the consumer, the inventors surprisingly found that the soluble fiber components obtainable by this process are well retained into the pasta structure after cooking or overcooking, that they are therefore not lost in the cooking water, and hence available to the individual consuming the pasta product.

It was shown previously that pasta made from germinated durum wheat semolina, containing significant levels of a-amylase, results in increased levels of solids in the cooking water. For both pastas made from sound and germinated wheat semolinas, such losses comprise simple sugars and high MW dextrins ( > 250.000) (Kruger and Matsuo 1982). Ingelbrecht et al (1999), as expected by a man knowing the field, showed that the

smaller MW (ca. 22000) NSP component : arabinogalactan peptide (AGP) leached more easily from pasta (produced without addition of enzymes/endoxylanases) during the cooking process, than the higher MW ( > 200000) non enzymically degraded NSP component WE-AX.

In Table I the TOT-AX and WE-AX contents (analytical procedure see infra) and endoxylanase activities of some internationally available commercial pasta samples are shown. Out of these values the percentage of WE-AX in TOT-AX is calculated. The endoxylanase activity is determined with the method as depicted in the Megazyme product sheet 9/95 (Xylazyme AX Test Tablet Procedure) with modifications. Ground pasta (0.5g) was extracted with 5.0 ml buffer (25mM sodium acetate, pH 4.7) during 15 min. After filtration and

preincubation (5 min, 40 C), 1. 0 ml extract was incubated (40oC, 24 hrs) with the tablet. Addition of 10. 0 ml of Trizma base solution (2% w/v) stopped the reaction. After filtration, absorbancy was measured at 590 nm. Incubating 1.0 ml of buffer solution with the tablet provided a substrate blank.

Pastas produced with whole meal semolina (samples nos 29 and 30, Table I) contain the highest percentage of AX. Despite these high contents they contain a relative small portion of WE-AX. In these samples only 7% and 12%, respectively of TOT-AX are WE-AX. Because the inventors see no reason why endoxylanase addition during the production with whole meal samples would not result in pastas with significantly higher levels of WE-AX and thus higher levels of soluble dietary fiber. Hence this is included in the concept of the invention as well. In this text, we use the term whole meal pasta for those products characterised in that they contain a total AX content exceeding 3.00% (expressed on dry basis, d. b.).

For the other pasta products (Table I, all except nos 29 and 30) the contents of TOT-AX varied between 1.75% (no 10) and 2.32% (no 23) (on d. b. ), of WE-AX between 0.19% (no 24) and 0.70% (no 12) (on d. b. ) and the percentage of WE-AX (in TOT-AX) between 12% and 30%.

With low levels [up to 7. 5*10-4 Somogyi units per g semolina (further referred as units)] of endoxylanases only very low increases in the percentage of WE-AX can be obtained (Table II). Higher levels result in substantial increases. The procedure described here (addition of endoxylanases during pasta production) is considered to yield pasta samples with WE-AX contents higher than 0.80% or with a percentage of WE-AX in total AX higher than 40% (25% for whole meal products), measured with the techniques described in the methods of the example.

Since assessment of endoxylanase activities with the colorimetric method with a semisynthetic substrate does not necessarily correlate with their activity in the pasta making process, the values obtained for the percentage of WE-AX (in TOT-AX) (Table I compared to Table II) indicate for the samples available, that no endoxylanases are added during pasta production. Additional evidence is provided by the low endoxylanase activities measured in these samples (Table I). Only for sample no 30 (produced with whole meal) relatively high activities (Table I) are found, as whole meal comprises the outer layer of the grain kernel, which contains higher enzyme levels than the endosperm. Although some samples show higher residual endoxylanase activities (Table I nos 2,10, 12,19, 29,30) than others, their analysed WE-AX contents do not follow the same trend. The lack of this correlation again indicates that endoxylanases are not used in the pasta industry. Residual activities of the samples produced with

endoxylanases are given in Table III. At dosages exceeding 3. 75*10-3 units per g semolina, residual activity is easily detected. In the case of the experimental conditions used in the example, this was also the minimal dosage to obtain a significant increase in WE-AX contents. Strong correlations exist between the measured residual activities and the WE-AX contents of these samples (Table I and III).

Today, endoxylanases are not used during pasta production and certainly not to increase their soluble fiber content. One of the presumed disadvantages is that high enzyme (e. g. due to germination) levels would result in higher cooking losses or desintegration of the pasta structure or both (Kruger and Matsuo, 1982). While enzyme producers (Novo Nordisk (Qi Si et al 1999), Genencore,...) have been looking for applications of endoxylanases in the pasta industry, they were today unable to show clear advantages, such as the unexpected increase of soluble fiber content in the cooked products observed by the inventors.

Not all endoxylanases have the same action pattern. Recent research e. g. to investigate the role of AX in the breadmaking process, by enzymically modifying them by two different endoxylanases, showed clear differences between the two endoxylanases used, i. e. one of Aspergillus aculeatus and one of Bacillus subtilis.

While the former resulted in low molecular weight (LMW) WE-AX, Bacillus subtilis enzyme resulted in higher molecular weight WE-AX (Courtin 2000). Different kinds of endoxylanases or fiber hydrolysing enzymes or combinations thereof offer a valuable tool to regulate the MW distribution of AX.

The present invention aims to provide a method for the preparation of pasta products with increased levels of soluble fiber.

A main aim of the invention is to provide a method to produce pasta products with increased soluble fiber levels and where this soluble fiber is well retained in the pasta structure after cooking.

Another aim of the invention is to obtain with this new pasta production process, a product of comparable or better quality, than pasta products currently available on the market today.

A further aim of the process is to obtain, by choosing different kinds of endoxylanases or fiber hydrolysing enzymes or combinations thereof or processing conditions (temperature, pH, pressure,...) or both, pasta products with varying MW distribution of AX can be obtained.

Another important aim is to obtain cooked pasta products which have positive short or long term health impacts or both when consumed.

A last aim is to obtain an apparatus to dose endoxylanases or fiber hyrolysing enzymes or combinations thereof during pasta production.

Summary of the invention The present invention is related to a process for the preparation of pasta products with increased levels of soluble fiber. This process comprises the addition of endoxylanases or any other fiber hydrolysing enzyme [cellulases (E. C. 3.2. 1.4), glucanases (E. C. 3.2. 1. 6),...] or combinations thereof during pasta production production.

Whole meal pasta products also offer a great potential for the conversion of WU-AX to WE-AX, as source

of soluble dietary fiber, by the addition of endoxylanases during their production.

By such addition of enzymes the quality of the pasta products must largely remain intact or improve.

In spite of what the inventors expected, the solubilised fiber components formed by this process are well retained into the pasta structure after cooking or overcooking.

By choosing different kinds of endoxylanases or other fiber hydrolysing enzymes or combinations thereof or processing conditions (temperature, pH, pressure,...) or both, pasta products with varying MW can be obtained.

It is in general an object of the present invention to provide a new and improved pasta with increased water soluble fiber contents and to provide a process of manufacturing the same. Another object of the invention is to provide a new and improved pasta with increased water soluble fiber contents wherein the water soluble fiber is well retained in said pasta during a normal cooking process of said pasta.

These and other objects are achieved in accordance with the invention by addition of a fiber hydrolysing enzyme at any stage of a normal pasta production process or in combination with adding water insoluble fiber at any stage of the pasta production process. The fiber hydrolysing enzyme or the water insoluble fiber may be added before formation of the pasta structure,-more specifically prior to or during mixing stages of the raw materials.

In addition, the present invention provides a process for manufacturing said pasta products with an increased non-leachable water soluble fiber by addition of fiber hydrolysing enzyme by a raw material enriched with said fiber hydrolysing enzyme, a microbial organism

producing said fiber hydrolysing enzyme, a germinating material producing said fiber hydrolysing enzyme, a material expressing said fiber hydrolysing enzyme, a genetically modified material expressing or overexpressing said fiber hydrolysing enzyme or a combination thereof. The fiber hydrolysing enzyme can be an endoxylanase and it can be an endoxylanase is from Aspergillus niger or Bacillus subtilis.

In its product aspect, the present invention resides in pasta products with an increased non-leachable water-soluble fiber. In another embodiment the invention resides in a pasta products with a water-extractable arabinoxylan concentration of at least 0.80% by dry weight of said pasta material. A further preferred embodiment is non whole meal pasta products having a water-extractable arabinoxylan of at least 42% of the total arabinoxylan population and comprising molecules of a variable molecular weight distribution and preferably a non whole meal pasta products having a water-extractable arabinoxylan of at least 55% of the total arabinoxylan population and comprising molecules of a variable molecular weight distribution. A further preferred embodiment is whole meal pasta products, wherein at least 25% of their total arabinoxylan population is waterextractable and comprising molecules of a variable molecular weight distribution and more preferably whole meal pasta products, wherein at least 30% of their total arabinoxylan population is water-extractable and comprising molecules of a variable molecular weight distribution. The water-extractable arabinoxylan population of these pasta products may be non-leachable. The pasta of present invention can be used for controlling cholesterol and preventing cholesterol damage in a human in need thereto. More specifically it can be

used for reducing blood serum cholesterol levels, reducing absorption of cholesterol from the intestines into the bloodstream or for inducing a more positive glycemic response. It thus is expected to have long term or short term positive health effects or both to consumers.

In its apparatus aspect, the present invention resides in a pasta machine specifically designed for carrying out the process for producing improved pasta with increasing water soluble fiber contents. Such pasta machine comprises structural element to feed or dose the said fiber hydrolysing enzymes to the raw materials. Such pasta machine may further comprise structural elements for mixing the pasta materials, structural elements for moistening the pasta materials and structural elements for mixing, kneading or stretching the pasta materials until a desired consistency is reached, structural elements for shaping the pasta by extruding, molding, stretching, pressing, cutting or any combination thereof until a desired form is reached or a combination thereof.

In yet another aspect, the present invention provides a use of fiber hydrolysing enzyme preparations for enriching pasta with increased water soluble fiber contents which are well retained in the pasta structure during a normal cooking process. The fiber hydrolysing enzyme preparations may be raw material enriched with said fiber hydrolysing enzyme, a microbial organism producing said fiber hydrolysing enzyme, a germinating material producing said fiber hydrolysing enzyme, a material expressing said fiber hydrolysing enzyme, a genetically modified material expressing or overexpressing said fiber hydrolysing enzyme or a combination thereof. In yet another preferred embodiment

the fiber hydrolysing enzyme of said enzyme preparation is an endoxylanase (E. C. 3. 2. 1. 8).

Description of the illustrative enbodiment The present invention is related to a process for the preparation of pasta products with increased levels of soluble (dietary) fiber. This process comprises the addition of endoxylanases (E. C. 3.2. 1.8) or any other fiber hydrolysing enzymes such as cellulases (such as E. C. 3.2. 1.4), glucanases (such as E. C. 3.2. 1.6),... or combinations thereof during pasta production.

By addition of endoxylanases or other fiber hydrolysing enzymes or combinations thereof, the quality of the pasta products largely remains intact or improves.

However, in some cases (e. g. pasta production conditions with low relative humidities), the addition of endoxylanases can result in checking phenomena after drying the pasta samples (unpublished results). Suggested solutions for this problem are changing the drying program or processing the dough at lower water absorptions or both.

In spite of what they expected, the inventors unexpectedly found that the fiber components obtainable by this process are well retained into the pasta structure after cooking or overcooking, that they are therefore hardly leached out in the cooking water, and hence available to the individual consuming the pasta product (cfr. above for definitions) Not only durum wheat (Triticum durum Desf. ) or common wheat (Triticum aestivum L. ) but also every other cereal or pseudocereal such as barley, buckwheat, corn, millet, oat, rice, rye, sorghum, spelt, triticale,... and product thereof such as milling products or any other source of carbohydrates (cassava, pea, soya,...) that is or

can be used for pasta production and has a source of dietary fiber, which can be hydrolyzed by enzymes, is included in the invention. Examples of target dietary fibers are AX, cellulose, ss-glucans, glucomannans, lignin and pectins. Most of them are classified as NSP.

Additions of fiber rich fractions during production such as cereal bran or fiber components as e. g. guar gum, xanthan gum, locust bean gum and pea fiber, which are hydrolysed during the production process by the addition of enzymes are also included in the invention.

It is also logical that, apart from an addition as a microbial preparation, other means to obtain high levels of endoxylanases or other fiber hydrolysing enzymes or combinations thereof in the raw materials for the pasta production are possible, such as adding materials rich in fiber hydrolysing enzymes, obtainable by germination of cereals or any other raw material of the pasta production or by overexpression of endogenous fiber hydrolysing enzymes in cereals or any other raw material of the pasta production, or by expression of fiber hydrolysing enzymes in raw materials of the pasta production or by combinations of the techniques mentioned here above. These methods are also included in the invention.

The terminolgy used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appending claims. This invention is not limited to the particular methodology, protocols and raw materials as these may vary in a pasta production process. Example

Pasta was produced at 800. 0 g scale from two semolinas (semA and semB) using dosages of Bacillus subtilis (XBS) and Aspergillus niger (XAN) endoxylanases varying between 0 and 0.225 Somogyi units (cfr infra) per g semolina (further referred to as units).

High dosages of XAN and XBS resulted in LMW AX, which were expected to easily leach out during the cooking process of pasta and hence to result in a loss of dietary fiber components. Surprisingly, only low levels of AX were found in the cooking water, even with extremely high dosages of endoxylanases used and cooking beyond optimum time. In contrast, AGP were leached more easily, even when the MWs of the enzymically modified AX were lower than those of AGP. The AX, even when they were of low MW, were therefore presumably tightly bound in the gluten network, which is the major component giving strength to pasta strands.

The endoxylanase treatments had no considerable effects on the resulting pasta quality (color of the dry products and surface condition, viscoelastic index and resistance to longitudinal deformations of the cooked products).

I. Materials Chemicals. All reagents were of at least analytical grade. Specialty chemicals were heat-stable aamylase (Termamyl 120 LS, Novo Nordisk, Bagsvaerd, Denmark) and amyloglucosidase (Boehringer Mannheim, Mannheim, Germany). For both enzymes, units were as defined by the respective suppliers. P-D-Allose was obtained from Sigma (Sigma Chemical Co. , St. Louis, MO, USA).

Semolinas. Durum wheat semolinas were from an industrial blend of durum wheats, harvests 1999 (semA)

and 1998 (semB) (Soubry, Roeselare, Belgium). Samples were stored at 4 C until used. Protein contents (N x 5. 7) were determined by a Kjeldahl procedure (AACC method 4611A) as 15. 8% and 14. 3% (d. b.), respectively. Ash (d. b.) contents (AACC method 08-01) were 0.79% for semA and 0.93% for semB. Moisture contents (AACC method 44-15A) were 10.0% and 11. 8% for semA and semB, respectively.

IT. Methods Activity of the Endoxylanase Preparations. Enzyme solutions in deionized water were prepared just before addition with the endoxylanases XBS (from Bacillus subtilis) and XAN (mixture of endoxylanases from Aspergillus niger). The endoxylanases can be isolated from Aspergillus niger and Bacillus subtilis cultures according to the methods or variants thereof described in John et al (1978) and Cleemput et al (1997). Their activities were determined according to a method by Somogyi (1952), with modifications as outlined in Megazyme (Bray, Ireland) product sheet 9/95. One Somogyi unit is the amount of enzyme that releases one micromole of xylose reducing sugar equivalents per minute at 40 C from wheat AX (Megazyme) (1.0% w/v) in sodium phosphate (0.1 M, pH 6.0).

Pasta Production. Pasta (spaghetti) was produced by mixing 800.0 g semolina using a Mini Press (Sercom, Montpellier, France) and slowly adding (during 30 seconds) deionized water to give a total moisture content of 33.80%. Endoxylanase additions (between 0 and 1.30 units/g semolina) replaced part of the water needed. After 20 min of further mixing (120 rpm, direction of the mixing screw is reversed every 10 sec), the resulting dough was extruded (44 C, under partial vacuum: 150 mm Hg) to give pasta strands with a diameter of 1.45 mm.

Pasta was dried to about 12. 5% moisture using a cycle at 70 C in a Secasi-Eurotherm dryer (Chessell, France) (Fig. 1). Samples were stored at 20 C for at least 4 days prior to analysis. All pastas were produced at least in duplicate.

Enzyme Inactivation in Enzyme-Treated or Control Pastas. Dried pastas (100.0 g) were boiled in ethanol (95%, under reflux) for 2 hr. After cooling, the ethanol was removed by vacuum rotary evaporation (45OC), and the material was air-dried. The material was crushed with

mortar and pestle, until it passed a 250 urn sieve. It is hereafter referred to as inactivated pasta.

Determination of Carbohydrate Contents. For the determination of the water-extractable carbohydrates in inactivated pasta, 2.0 g of inactivated material was extracted with 20 mL deionized water (15 min, 4OC). After centrifugation (3,000 x g, 15 min, 4 oC), supernatant (2.5

mL) was hydrolyzed (60 min, 110oC) with 2. 5 mL 4. 0 M trifluoroacetic acid (TFA). Both the extraction and the hydrolysis and derivatization were in duplicate. For the determination of total carbohydrate content of inactivated pastas, 50 mg was hydrolyzed (120 min, 110oC) with 5.0 mL (2.0 M) TFA. After cooling, the hydrolysate was centrifuged (3,000 x g, 15 min). All analyses were at least in duplicate.

In both cases, alditol acetates were prepared according to the method of Englyst and Cummings (1984) and were separated on a Supelco SP-2380 (Bellefonte, PA, USA) column (30 m, 0.32 mm i. d. , 0.2 urn film thickness) in a Chrompack 9011 chromatograph (Middelburg, The Netherlands) equipped with a flame-ionization detector.

The carrier gas was He. Separation was at 225OC, with injection and detection temperatures of 275OC and ss-D

allose as internal standard (1. 0 mL added, with a concentration of 1. 0 mg/mL).

The Xylose (Xyl), Arabinose (Ara) and Galactose (Gal) data led to calculation of the AX and arabinogalactan (AG) contents and the A/X ratio (the arabinose to xylose ratio or substitution degree of AX), using the formulas as in Ingelbrecht et al (2000): AX=

[% Xyl + (% Ara- (0. 7*% Gal)] *0. 88 ; AG= [% Gal*0. 90 + (% Gal*0.7) *0. 88]) and A/X= [% Ara- (% Gal*0. 7)]/ % Xyl. The Ara/Gal ratio was assumed to 0.7 (Ingelbrecht et al 2000), which made calculation of Ara to be assigned to AGP possible. Conversion factors (0.88 and 0.90) were used for calculation of polymeric material contents, consisting of pentose and/or hexose monomers. AG contents are a good estimation of the content of AGP since it can reasonably (e. g. based on the gel permeation behaviour) assumed that, much as T. aestivum L. , the peptide component is only a minor proportion of the structure (Fincher et al 1974).

Purification of Non-Starch Polysaccharides.

Inactivated pasta (80.0 g) was extracted with deionized water (w/v 1/5,15 min, 4 C). The suspension was centrifuged (8,000 x g, 15 min, 4OC), and the supernatant boiled for 10 min. Following a Termamyl (3000 units, 30

min, 90 C) treatment and a centrifugation step (3, 000 x g, 15 min, is-C), samples were treated with amyloglucosidase at pH 4. 5 (50 units, 12 hr, 60oC), centrifuged (8, 000 x g, 40 min, 15OC), and the supernatant was boiled (10 min). After a last centrifugation step (8,000 x g, 40 min, 15 C) to remove the denatured proteins, the supernatant was dialyzed (48 hr, 4 C) and freeze-dried to obtain the NSP material.

Since enzymically degraded AX and AGP have a similar precipitation behaviour in ethanol solutions

(Courtin and Delcour 1998), no further separation between AX and AGP was performed.

Gel Permeation Chromatography. NSP material (6.0 mg) was solubilised in 0.3% NaCl (3.0 mL) and centrifuged (10,000 x g, 10 min). The solution was filtered (0.45 pm) and separated on a Shodex B-804 HQ (Showa Denko K. K., Tokyo, Japan) GPC column (300 x 8 mm) by elution with 0.3% NaCl (0.5 mL/min). The eluate was monitored using a refractive index detector (VDS Optilab, Berlin, Germany).

Molecular weight markers [Shodex P-82 pullulan standards (Showa Denko K. K. ) with MWs of 78.8 x 104, 40.4 X 104,

21. 2 X 104, 11. 2 x 104, 4. 73 x 104, 2. 28 x 104, 1. 18x 104 and 0. 59 x 104] made approximate calculations of MWs possible.

Cooking Losses. Pasta (25.0 g) was cooked to optimal cooking time (T) and (T+11) (min) in 1.0 L of deionized water (containing 0.50 g NaCl). After draining and cooling, the cooking water was freeze-dried. Carbohydrate composition, ash, moisture contents and protein content were determined as outlined above. From these data and comparison with the corresponding analytical data on the uncooked dry pasta, the relative losses in the cooking water of dry matter, AX, AG, glucose, proteins and ash were calculated.

Color of the Dried Pasta. Pasta color results from a desirable yellow component (measured by the index b*) and the undesirable brown (100-L*) and red (a*) components (Laignelet et al 1972). International colorimetric indices L*, a* and b* were determined on the dry, uncooked pasta with a Minolta CR310 (Minolta, Osaka, Japan) colorimeter as in Abécassis et al. (1994). Brown index was expressed as (100-L*), red index as a*, and the

yellow index as b*. Indices were determined at least in triplicate.

Cooked Pasta Quality Assessment.

Surface Condition and Firmness of the Cooked Pasta Pasta strands (50.0 g), broken to a length of ca.

15 cm, were cooked in 1.5 L of salted (7.0 g/L) mineral water (as recommended by AFNOR Standard NF-V 03-714). The optimal cooking time (T) was defined as that needed to gelatinise starch at the center of the pasta strands (Abécassis et al 1994).

As outlined by Abécassis et al (1994), scores between 1 and 9 (1: very bad, 9: excellent) were assigned by a trained panel (4 participants) with photographs as reference, taking into account the general appearance, degree of swelling and stickiness of the cooked pasta samples at T+6 and T+11 (min). For both the determinations of cooking time and surface condition, analyses were performed at least in duplicate for each pasta production.

The viscoelastic index (VI) was assessed with the Viscoelastograph (Chopin, Paris, France) at T+l, T+6 and T+11 (min) as in Abécassis et al (1994) and is a measure of pasta firmness. Analysis was performed at least in five-fold.

Resistance to Longitudinal Deformations of the Cooked Pasta Pasta (15 strands of 15 cm) was cooked to optimal cooking time (T) and (T+11) in 2.0 L deionized water (containing 1.0 g NaCl). After 5 min of cooling, elastic moduli (g/mm) and maximal breaking strengths (g) and distances (mm) were determined with a Texture analyser (TA-XT2I, Stable Micro Systems, Surrey, UK) using the spaghetti/noodle tensile rig A/SPR. The initial distance

between the probes (0. 50 mm) was gradually increased to 100 mm with a speed of 3. 0 mm/sec. The elastic modulus (Hookean modulus) was recorded as the gradient (g/mm) in the linear region between trigger force 5.0 g and 10.0 g.

Analysis of the 15 strands was performed in duplicate.

Statistical Analysis. Statistical analysis was performed using linear mixed models using SAS software (SAS system for Windows v6.12). Productions were considered as random. The dosage of endoxylanases was considered as a continuous variable, statistics were performed using the logarithm in order to avoid heavy influence on the regression parameters of the observations with the high dosages.

Simple linear regression equations were obtained with correlated errors. The correlation between the errors was modelled as a compound symmetric model for which the correlation between errors of the same production was larger than zero and zero otherwise.

Fitting models with random terms involves maximum likelihood techniques. The most frequently used statistic for comparing these kind of models is the Akaike's information criterion. An R2-analog was also calculated by the following formula: 1- (SSE/SSTO) (with SSE: error sum of squares and SSTO: total sum of squares), where SSE was calculated by multiplying the covariance parameter estimate for the residual with its number of degrees of freedom. SSTO was calculated by multiplying the variance of the response by the number of observations minus one.

III. Results Non-Starch Polysaccharide Composition of Enzyme Treated Pastas. Percentages of AX solubilised [ (% WE-AX in endoxylanase-treated samples-% WE-AX in control) /% TOT- AX, calculated from the values in Table II] for semA

pastas produced with 0. 225 units of XAN and XBS, were 40. 9% and 46. 1%, respectively, compared to an initial WE AX percentage of 40.1% for the untreated semA pasta.

Adding 1.30 units XAN resulted in an almost complete solubilization of the WU-AX. The percentage of WE-AX in non-treated semB pasta was 35.3%. With 0.225 units of endoxylanase XAN, 32.0% AX were solubilised.

The A/X ratios of the WE-AX population in control and endoxylanase treated pastas were quite comparable (Table II).

Molecular Weight Profiles of the Non-Starch Polysaccharides. Purification yielded NSP material, which typically contained AGP and WE-AX in a ratio of 1. 0/ 4. 5.

Again, no change in the A/X ratio was noted for WE-AX from pastas produced with endoxylanases (results not shown). The MWs of WE-AX of pasta samples from semA were drastically reduced by the endoxylanase treatment with XBS (Fig. 2). With the highest dosages, MWs of WE-AX were reduced to values lower than that of AGP, which is ca.

22000 (Ingelbrecht et al 2000). The AGP peak could be clearly recognized in the MW profiles of the control and the pastas produced with the lower dosages of XBS.

Similar profiles were obtained using XBS and XAN (the latter with pasta produced from semB) (results not shown).

Cooking Losses. Because of the drastic reduction in MW of the WE-AX by the endoxylanases used and the fact that, for the highest enzyme dosages, almost all AX were solubilised, it was expected that a lot of the soluble fiber would be released in the cooking water. Rather unexpectedly, it turned out that AX were well retained in the cooked pasta, even when large levels of endoxylanases were used (Tables IV and V). Although consistently significant positive relations were found between % AX

leached out and the endoxylanase dosage used, the regression equations had very low slopes (Tables IV and V). At optimal cooking time (Table IV), a maximum percentage of 19.76% AX of TOT-AX and with 11 min overcooking (Table V) maximally 33.7% were leached out from the semA pasta which was produced with 1.30 units XAN. Here, the MW profile of WE-AX, which under such conditions accounted for 97.6% of TOT-AX, was shifted to that below that of AGP (results not shown). In spite of this firm MW reduction, AGP material leached out more easily than AX material (for both cooking times T and T+11). From these data, it could be hypothesized that AX were bound firmer (by physical entrapment or chemical interactions) into the gluten network than AGP. In contrast, for a batter based wheat gluten isolation process Roels et al (1998) found high AGP/AX ratios in gluten fractions isolated from common wheat samples, indicating that AGP are relatively better retained by the gluten network than AX.

For pastas produced with different dosages of endoxylanases and control pasta, no great differences were noted for AGP, protein and glucose losses. A small increase in dry matter loss was observed for the pasta produced with increasing dosages (Tables IV and V).

Further reduction of cooking losses and the concomitant (soluble) dietary fiber can be obtained by optimizing the following parameters, which are all known to have a great influence on cooking losses: cooking time (Binnington et al 1939), product shape and protein content (Holliger 1963), gliadin, globulin, albumin and glutenin contents (Walsh and Gilles 1971), gluten strength (Grzybowski and Donnelly 1979), particle size of the flour used (Breen et al 1977, Alary et al 1979), hardness of the cooking water (Alary et al 1979, Dexter

et al 1983, Seibel and Menger 1985), drying temperature (Dexter et al 1981) and dough temperature at the die (Abécassis et al 1994).

Pressure during Extrusion. Increasing levels of endoxylanase, added before pasta dough mixing, resulted in lower extrusion pressures [Fig. 3 (A) and (B)]. The linear regression equations obtained are as follows: for XAN and semA : P=-6.6 logds + 101.1 with p < 0. 001 and R2 = 0. 89 (eq. 1) for XAN and semB : P=-6.8 logds + 100.5 with p < 0. 01 and R2 = 0. 87 (eq. 2) for XBS and semA : P=-6.4 logds + 98.5 with p < 0. 001 and R2 = 0.91 (eq. 3) With ds: dosage in units + 1 P: Pressure (bar) Logds: log (units + 1) p: p-value, indicating the chance that the slope is not different from zero R2 : correlation coefficient For the highest dosages (0.225 units), almost an halvation of pressure was seen control dough: 105 bar, 64 bar and 67 bar for the XAN and XBS treated doughs (semA), respectively and 107 bar control dough and 62 bar XAN treated dough (semB) (Fig. 3: results not shown).

Further addition of endoxylanase did not result in a substantial reduction [Fig. 3 (B): result not shown: 1.30 units using semA : 67 bar]. These observations confirm earlier data that endoxylanases decrease the maximal consistency of durum semolina doughs prepared in the farinograph significantly and that the simultaneous omission of a certain level of water and addition of a certain level of endoxylanase, restores the maximal dough consistency (Ingelbrecht et al 2000).

Color of the Dried Pasta. The values of the three colorimetric indices (100-L*, a* and b*) for the different dry pastas are given in Table V. Very small but significant trends (regression equations, Table VI) that endoxylanases negatively influence pasta color, were seen. It seems that a less dense structure was obtained with endoxylanase treated samples, probably as a result of lowered extrusion pressures (Fig. 3). If so, the more porous structure may have facilitated the occurrence of oxidation phenomena.

Cooked Quality Assessment.

Surface Condition and Firmness of the Cooked Pasta Increasing dosages of endoxylanases caused a slight but significant decrease in pasta cooking time (Table VII). Surface condition of cooked pastas was generally not negatively influenced by an endoxylanase treatment during production (Table VII). Although not understood at present, an even better product was

obtained with the highest dosages of XAN for semA pastas produced with 0. 225 units (cooked to T+6 and T+ll) and 1. 30 units (cooked to T+11). This was also reflected in the small positive relationships found between surface condition (at T+6 and T+11) and endoxylanase dosage of XAN (Table VII). No significant relationships were found between the dosages of XBS added and surface condition.

The viscoelastic index (VI) of the cooked pastas at T+1, treated with increasing dosages during production, was significantly lowered (regression equations, Table VII) for both endoxylanases XAN and XBS and both semolinas (semA and semB). This negative relationship was also observed for cooking time T+6 when using XAN for both semA and semB. No significant quality

loss for this parameter was seen when drastically overcooking (T+11).

Resistance to Longitudinal Deformations of the Cooked Pasta.

Small significant changes in maximal force before breaking strands (F in g) were observed for semA pastas cooked to time T and containing endoxylanases (regression equations, Table VIII). Even smaller changes (Table VIII, smaller slopes of regression curves for T+ll than for T) were noted for the same pastas, cooked at T+ll. No significant relationships were found between the elastic moduli of cooked strands (E in mm/g) or maximal distances of extension (D in mm) and endoxylanase dosages for pastas cooked to T and T+11. For semB pastas, no negative influence at optimal cooking time of the endoxylanase addition was seen. However, a clear decrease for the measured parameters (E, F and D) was seen at overcooking (Table VIII).

Impact of Endoxylanases on Extrusion Pressure. An important parameter during the pasta production process is extrusion pressure. It has to be sufficiently high to ensure a compact pasta structure, which better resists to cooking (Pagani et al 1989). Recommended pressures for extrusion of long pasta goods are between 90 and 125 bar (Dalbon et al 1996). Forcing the dough through the die holes does not damage the protein network (Dalbon et al 1996). The small quality losses (as mentioned above) seen with the pasta containing endoxylanases, can probably be explained by the lower extrusion pressures during their production [ (Fig. 3 (A) and (B)]. However, as noted earlier (Ingelbrecht et al 2000), lowering the water content of the dough may restore the extrusion pressure and consequently reduce drying times needed for these pasta goods.

It is probable that the lower cooking times found for the endoxylanase containing pasta samples can be explained by the less dense structure (caused by the lower extrusion pressures), which facilitates water penetration during cooking and therefore gelatinization of starch.

IV. Conclusions The use of endoxylanases during pasta production resulted in a high turnover of WU-AX to WE-AX, while the MW of the WE-AX were also drastically reduced. For the highest-osages used, the MW of the WE-AX was even lower than that of AGP. In contrast to expectations, WE-AX from pastas containing very high dosages endoxylanases, were not leached easily during cooking, even at excessive cooking times. AGP were consistently leached more easily, even when the WE-AX had attained a lower MW due to the endoxylanase treatment.

The most obvious effect of adding endoxylanases during pasta production was that the extrusion pressures are significantly lowered. Quality (in terms of color of the dried product and surface condition, viscoelastic index, resistance to longitudinal deformations of the cooked product) of pastas containing endoxylanases, remained fairly constant. Further improvements of the quality are to be expected, when using appropriate (lower) dough hydration levels, yielding normal (90-125 bar) extrusion pressure levels. These lower dough hydration levels could lead to shorter drying times of the extruded pasta.

V. Abbreviations used NSP, non-starch polysaccharides ; AX, arabinoxylans ; TOT-AX, total level of arabinoxylans; WE AX, water-extractable arabinoxylans; WU-AX, water

unextractable arabinoxylans ; AGP, arabinogalactanpeptides ; ara, arabinose ; gal, galactose ; xyl, xylose ; ara/gal, arabinose to galactose ratio ; A/X, arabinose to xylose ratio; FU, farinograph units; GPC, gel permeation chromatography; MM, molecular weight; TFA, trifluoroacetic acid ; d. b. , dry basis; SSE, eeror sum of squares; SSTO, total sum of squares; VI, viscoelastic

2 index ; T, optimal cooking time ; R2, correlation coefficient, P, pressure; p, p-value; XAN, endoxylanase of Aspergillus niger; XBS, endoxylanase of Bacillus subtilis; ds, dosage in units + 1. vi Figure legends Fig. 1. Pasta drying profile: relative humidity (%) () t) and temperature (OC) (+) versus time.

Fig. 2. Gel permeation chromatography profiles of AX-AGP material, purified from pasta samples of semA treated with XBS (control D, 7. 50*10-5 units #, 7. 50*10-4

units +, 3. 75*10-3 units 0, 1. 50*10-2 units , 4. 00*10 units A, 0. 225 units n).

Fig 3. Influence of dosage of XAN (+) and XBS endoxylanases on extrusion pressure (bar) during pasta production, when using semA (A) and semB (B). Units are expressed as Somogyi units per g semolina.

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Table 1. Total arabinoxylan (% TOT-AX) and water-extractable arabinoxylan (% WE-AX) contents, percentage of WE-AX in TOT-AX (% WE AX/% TOT-AX) and endoxylanase activity (Endox-activity, the activity is measured as difference in absorbancy (590 nm) between sample and the blank, and expressed on dry matter) measured in some international commercial pasta and noodle samples. Codes for product origin are as follows 1 : Delhaize, Heverlee, Belgium, 10/02/2000 ; 2 : GB, Herent, Belgium, 15/0612000 ; 3 : Aldi, Herent, Belgium, 15/06/2000 ; 4 : Champion, Leuven, Belgium, 17/06/2000 ; 5 : Colruyt, Heverlee, Belgium, 29/06/2000.

Nr Type Brand Address Cooking Product % TOT-% WE-AX % WEAX/ Endoxtime (min) origin AX % TOTAX activity 1 4 Spaghetti Esselunga Esselunga, Via Gianbologna 1, 10 min 1 2. 03 0. 51 25% 0. 091 1000g Limito, Milan, Italy 2 Gnocchi 85 Barilla Barilla Alimentare S. p. A., Via 14 min 2 1. 96 0. 38 19% 0. 104 500g Mantova, 166, Parma, Italy 3 Spaghetti Wit CABBAC Avenue des Loisirs 10 min 2 2. 28 0. 61 27% 0. 475 500g Product 3C, 1140 Brussels 4 Spaghetti Panzani PAB Benelux Vesten 55, 9120 10 min 2 2. 21 0. 35 16% 0. 108 500g Beveren, Belgium 5 Sport Soubry N. V. Etabl. Joseph Soubry 7 min 2 1. 68 0. 29 17% 0. 118 Spaghetti 250g S. A., 8800 Roeselare, Belgium 6 Spaghetti Soubry N. V. Etabl. Joseph Soubry 8 min 2 2. 19 0. 40 18% 0. 103 Rustica S. A., 8800 Roeselare, Belgium 250g 7 Capellini Soubry N. V. Etabl. Joseph Soubry 4 min 2 2. 00 0. 27 14% 0. 104 250g S. A., 8800 Roeselare, Belgium 8 Tagliatelles Le Goût Le Goût de la Vie, 72301 Sablé 8min 2 2. 09 0. 37 18% 0. 063 Semoule du de la Vie sur Sarthe Cedex France blé dur biologique 500g 9 Spaghetti Buitoni Buitoni Milano, Viale G. 8 min 2 2. 01 0. 57 28% 0. 158 500g Richard 5, Stab. Di Sanse Polero (AR), Italy 10 Pasta met GB GB nv, Nieuwstraat 111, 1000 5 min 2 1. 75 0. 27 15% 0. 409 eieren Brussel Elleboogjes 250g

11 Penne Rigata Panzani PAB Benelux Vesten 55, 9120 3min 2 2. 25 0. 27 12% 0. 118 500g Beveren, Belgium 12 Spaghetti extra Anco NV Anco Guldensporenlei 88, 9 min 2 2. 30 0. 70 30% 0. 313 fine 2300 Turnhout 250g 13 Spaghetti with Panzani PAB Benelux Vesten 55, 9120 7min 2 2. 17 0. 36 17% 0. 126 spinach and Beveren, Belgium tomatoes 500g 14 Radiator Soubry N. V. Etabl. Joseph Soubry 9min 2 2. 22 0. 39 18% 0. 173 250g S. A., 8800 Roeselare, Belgium 15 Macaroni Anco NVAncoGuldensporenlei88, 6min 2 2. 20 0. 45 20% 0. 142 express 2300 Turnhout 250g 16 Capellini 1 Barilla Barilla Alimentare S. p. A., Via 3 min 2 1. 81 0. 42 23% 0. 219 500g Mantova, 166, Parma, Italy 17 Spaghetti fine Soubry N. V. Etabl. Joseph Soubry 7min 2 2. 24 0. 61 27% 0. 200 250g S. A., 8800 Roeselare, Belgium 18 Quick Soubry N. V. Etabl. Joseph Soubry 4 min 2 1. 95 0. 35 18% 0. 145 Spaghetti S. A., 8800 Roeselare, Belgium 250g 19 Tagliatelles Filadelfia Filadelfia S. A., 6700 Arlon, 7-9 min 2 2. 09 0. 55 26% 0. 367 500g Belgium 20 Spiralen Pasta Pasta Zara S. p. A., Rieso Pio X, 6 min 3 2. 17 0. 53 24% 0. 219 500g Mare Italy 21 Spaghetti Pasta Pasta Zara S. p. A., Rieso Pio X, 7 min 3 1. 95 0. 57 29% 0. 258 500g Mare Italy 22 Spaghetti Winny S. C. R. L. Bloc C. V. B. A., rue 11 min 4 2. 31 0. 52 22% 0. 259 500g d'Artoisstraat 4, 1000 Brussel 23 Spaghetti 6 Melissa Melissa-Kikizas S. A. Food 12-13 min 4 2. 32 0. 50 22% 0. 204 500g Manufacturers, Vinos 1, 10443 Athens, Greece

24 Aiki Super Hartog Hartog Union S. A., Unilever 4min 4 1. 91 0. 19 10% 0. 051 Noodles Union Belgium N. V., 80g S. A. Humaniteitslaan 292, 1190 Brussels, Belgium 25 Spirali 29 Delverde Delverde Fara San Martino, 10 min 4 2. 30 0. 40 17% 0. 144 500g 66015 Chieti, Italy 26 45 Ditali La La Molisana, Industrie 4-6 min 4 2. 08 0. 49 24% 0. 094 500g Molisana Alimentari S. p. A., C. da Colle delle Api 100/A, 86100 Campobasso, Italy 27 Pasta Sapore I. A. P. S. p. A. Via-4 2. 12 0. 39 18% 0. 086 lOOOg Mediterra Circonvallazione90, 912026 neo Mazaro del Vallo (TP), Italy Aurora 28 Macaronelli Maxim MaxCrescentini sarl & Cie 12-14 min 4 1. 94 0. 43 22% 0. 357 250g Sees, 4050 Esch-sur-Alzette, Luxembourg 29 Gnocchini Soubry N. V. Etabl. Joseph Soubry 11 min 2 5. 61 0. 40 7% 0. 442 500g S. A., 8800 Roeselare, Belgium 30 Spaghetti Soubry N. V. Etabl. Joseph Soubry 9 min 2 5. 81 0. 69 12% 0. 719 wholemeal S. A., 8800 Roeselare, Belgium 500g

Table II. Non-starch polysaccharide composition [on d. b., arabinoxylans (% AX), arabinogalactans (% AU), arablnose to xylose suostituuo degree (A/X)] of total and water extracts of hydrolysates of pastas produced with different levels of endoxylanase XAN or XBS. n. d. : not determined.

SemA SemB .. ....- . . -G A/X % AG AIX % AG AIX % WE-AX/ % TOT-AX % TOT-AX Total hydrolysate 1. 94 0. 29 0. 83 2. 08 0. 34 0. 80 Water extract hydrolysate Control 0. 78 0. 27 0. 57 40 0. 73 0. 34 0. 56 35 Control+XAN 7. 50*10-5 units 0. 83 0. 29 0. 58 43 n. d. n. d. n. d. n. d.

7. 50*10-4 units 0, 91 0. 28 0. 58 47 0. 82 0. 34 0. 55 39 3. 75*10. 3 units 1. 16 0. 30 0. 58 60 n. d. n. d. n. d. n. d.

1. 50*10-2 units 1. 33 0. 31 0. 58 69 0. 94 0. 35 0. 55 45 4. 00-10-2 units 1. 44 0. 29 0. 58 74 1. 35 0. 36 0. 57 65 0. 225 units 1. 58 0. 29 0. 59 81 1. 40 0. 33 0. 58 67 1. 30 units 1. 90 0. 33 0. 60 98 n. d. n. d. n. d. n. d Control+ XBS 7. 50*10-5 units 0. 87 0. 29 0. 60 45 7. 50*10. 4 units 0, 88 0. 29 0. 58 45 3. 75*10-3 units 0. 89 0. 25 0. 58 46 1. 50*10~2units 1. 34 0. 30 0. 60 69 4. 00-10-2 units 1. 55 0. 26 0. 58 80 0. 225 units 1. 68 0. 33 0. 59 87 Max C. V. a 6% 5% 6% 3% 3% 1%

a : Max C. V. : maximal coefficient of variation

Table III. Residual endoxylanase activities of dried pasta made of semA and stored for 1 year at room temperature. Residual activity is measured as the difference in absorbancy (590 nm) between the sample and the blank and expressed on g dry matter.

Dosage added during pasta production Residual activity (difference in absorbancy/g dry matter) Control 0. 522 Control+XAN 7. 5*10. 5 units 0. 697 3. 75*10"* units 0. 828 0. 225 units 4. 903 Control+ XBS 7. 5*10-5 units 0. 576 3. 75*104 units 0. 907 0. 225 units 4. 898

t aDle I v. lvlaterlal Lary matter t^/oAlil), araDlilogalactal/oAbJ, araD llOxylalls t/oAS), protelns (U/Oprot) glucose (u/oglUC) and ash (% ash)] leached out during cooking (T), expressed as a percentage of their respective contents in the dried pasta, produced with different levels of endoxylanase XAN or XBS, using semolinas semA or semB. Where significant, relationships between % dm or % AX and endoxylanase dosage [logds : log (units + 1)] are given with a : p < 0. 05, b : p < 0. 01 and' : p < 0. 001. n. d. : not determined.

SemA SemB % dm % AG % AX % prot % gluc % dm % AG % AX % prot % gluc Control 6. 25 32. 28 2. 21 3. 70 4. 81 6. 26 25. 46 1. 53 3. 87 4. 46 Control+XAN 7. 50*10-5 units 6. 11 33. 02 2. 30 3. 77 4. 63 n. d. n. d. n. d. n. d. n. d. 7. 50*104 units 6. 13 33. 88 2. 99 3. 67 4. 74 6. 03 24. 10 1. 60 3. 76 4. 21 3. 75*10-3 units 6. 05 31. 57 3. 29 3. 76 4. 24 6. 77 25. 67 2. 42 4. 14 4. 90 1. 50*10-2 units 6. 28 27. 42 3. 84 3. 68 4. 35 n. d. n. d. n. d. n. d. n. d 4. 00-10-2 units 6. 29 30. 92 6. 06 3. 72 4. 18 6. 87 27. 35 5. 12 4. 22 5. 43 0. 225 units 6. 70 31. 02 10. 84 4. 00 4. 50 8. 51 31. 95 10. 91 5. 28 6. 43 1. 30 units 7. 77 32. 51 19. 76 4. 90 4. 70 n. d. n. d. n. d. n. d. n. d.

% dm=0 44blogds+5 96 % AX=5 13'logds+O 81 % dm=0. 94'logds+6 02 % AX=4 00"ogds+0 65 R075 R087 R084 R2=092 Control+XBS 7. 50*10-5 units 6. 12 32. 49 2. 38 3. 66 4. 59 7. 50*104 units 5. 48 27. 72 2. 11 3. 32 3. 98 3. 75*10-3 units 5. 78 28. 79 2. 75 3. 55 4. 27 1. 50*10-2 units 6. 45 33. 67 4. 69 3. 85 4. 74 4. 00-10-2 units 6. 56 32. 22 9. 49 3. 89 4. 77 0. 225 units 6. 72 29. 49 13. 15 3. 85 4. 45 % dm-0 29'logds+6 05 % AX=4 87blogds+l 30 R2= 0 56 R2= 0. 90 Max C. V. 10% 8% 6% 7% 10% 9% 8% 7% 6% 9%

Table 4bis. Non-starch polysaccharide composition (%, dry basis) of purified material of the pastas produced with different levels of endoxylanase, XAN or XBS. n. d. : not determined.

SernA SernB % AX AfX % AG AfX Control 43. 4 10. 6 0. 64 48. 2 15. 1 0. 66 Control+XAN 7. 50*10-5 units 58. 8 12. 9 0. 63 n. d. n. d. n. d. 7. 50*1 04 units 53. 6 11. 0 0. 62 55. 2 15. 7 0. 65 3. 75*10-3 units 72. 5 12. 0 0. 62 n. d. n. d. n. d. 1. 50*10. 2 units 64. 4 9. 3 0. 62 n. d. n. d. n. d.

4. 00-10-2 units 81. 4 10. 9 0. 62 69. 6 12. 7 0. 65 0. 225 units 68. 0 8. 8 0. 65 59. 8 10. 6 0. 67 1. 30 units 56. 5 9. 2 0. 71 n. d. n. d. i. d.

Control+XBS 7. 50*10-5 units 55. 8 12. 0 0. 64 7. 50*104 units 49. 8 11. 0 0. 63 3. 75*10-3 units 47. 6 9. 6 0. 62 1. 50*10-2 units 68. 9 9. 8 0. 63 4. 00-10-2 units 45. 1 9. 1 0. 62 0. 225 units 75. 5 10. 8 0. 65 Max C. V. 9% 10% 2% 9% 10% 2%

Table V. Material [dry matter (% dm), arabinogalactan (% AG), arabinoxylans (% AX), proteins (%oprot), glucose (%ogluc) ana asn (, /oasn) j leached out during overcooking (T+ll), expressed as a percentage of their respective contents in the dried pasta, produced with different levels of endoxylanase XAN or XBS, using semolinas semA or semB. Where significant, relationships between % dm or % AX and endoxylanase dosage [logds : log (units + 1)] are given with a : p < 0. 05, b : p < 0. 01 and : p < 0. 001. n. d. : not determined.

SemA SemB % dm % AG % AX % prot % gluc % dm % AG % AX % prot % gluc Control 7. 92 37. 97 3. 95 4. 60 6. 20 8. 15 36. 18 3. 24 5. 14 6. 96 Control+XAN 7. 50*10-5 units 8. 24 41. 46 4. 83 4. 89 6. 23 n. d. n. d. n. d. n. d. n. d. 7. 50*10-4 units 8. 95 45. 46 5. 67 5. 34 6. 73 8. 01 36. 59 3. 54 4. 91 7. 18 3. 75*10-3 units 7. 64 42. 56 7. 06 4. 50 6. 47 9. 58 42. 34 6. 45 5. 57 8. 71 1. 50*10-2 units 8. 14 39. 27 7. 62 4. 85 5. 98 n. d. n. d. n. d. n. d. n. d.

4. 00-10-2 units 8. 32 40. 67 12. 01 4. 72 6. 25 7. 37 30. 38 7. 73 4. 47 6. 13 0. 225 units 9. 18 41. 42 19. 39 5. 39 6. 76 9. 17 38. 49 15. 84 5. 68 7. 75 1. 30 units 10. 63 45. 75 33. 68 6. 74 7. 46 n. d. n. d. n. d. n. d. n. d.

% dru=0 67'logds+7 90 % AX=8 68'logds+2 29 % AX=5 ogds+269 R'=O 60 R2= 0 90 R= 0 90 Control+XBS 7. 50*10-5 units 7. 83 43. 06 4. 63 4. 63 6. 46 7. 50*104 units 8. 08 39. 78 4. 77 4. 83 6. 25 3. 75*10-3 units 8. 18 43. 80 6. 48 4. 87 7. 02 1. 50*10-2 units 8. 56 43. 78 8. 49 4. 91 6. 51 4. 00-10-2 units 8. 83 41. 55 15. 32 5. 16 6. 66 0. 225 units 9. 69 42. 87 22. 70 5. 17 7. 26 % dam=0 77logds+7 79 % AX=7 98'logds+2 93 R'= 0 97 R2= 0 93 Max C. V. 10% 9% 5% 10% 10% 11% 9% 7% 8%

Table VI. Colorimetric indices (100-L* : brown index, a* : red index, b* : yellow index) of the dried pastas produced with different levels of XAN and XBS endoxylanases, using semolinas semA or semB. Standard deviations are shown between brackets. Relationships between b* and endoxylanase dosage [logds : log (units + 1)] were significant and are given with a : p < 0. 05, b : p < 0. 01 and p < 0. 001. n. d. : not determined.

SernA SemB 100-L* a* b* 100-L* a* b* Control 40. 57 (0. 48) 0. 27 (0. 58) 37. 46 (0. 92) 40. 93 (0. 61) 1. 23 (0. 10) 33. 29 (0. 22) Control+XAN 7. 50*10~5units 40. 93 (0. 51) 0. 17 (0. 35) 36. 46 (0. 28) n. d. n. d. n. d.

7. 50*10-4 units 40. 41 (0. 91) 0. 43 (0. 09) 35. 83 (0. 68) 41. 09 (0. 27) 1. 38 (0. 17) 32. 81 (0. 44) 3. 75*10~3units 40. 31 (0. 23) 0. 65 (0. 30) 36. 36 (1. 24) 41. 14 (0. 08) 1. 55 (0. 11) 32. 85 (0. 02) 1. 50*10-2 units 41. 23 (0. 33) 0. 09 (0. 11) 36. 12 (0. 49) n. d. n. d. n. d.

4. 00-10-2 units 40. 70 (0. 33) 0. 42 (0. 14) 36. 00 (0. 61) 41. 50 (1. 09) 1. 22 (0. 61) 32. 15 (0. 99) 0. 225 units 41. 13 (0. 64) 0. 39 (0. 29) 35. 66 (0. 15) 42. 36 (0. 33) 0. 50 (0. 07) 30. 57 (0. 20) 1. 30 units 41. 19 (0. 24) 1. 41 (0. 07) 35. 96 (0. 21) n. d. n. d. n. d. b*=-0. 17'*togds+36 67 b*=-0. 44blogds+33 26 R2= 0 96 R'= 0. 99 Control 40. 57 (0. 48) 0. 27 (0. 58) 37. 46 (0. 92) Control+XBS 7. 50*10-5 units 40. 06 (0. 30) 0. 87 (0. 38) 37. 19 (0. 71) 7. 50*104units 41. 07 (0. 60) 0. 72 (0. 41) 36. 32 (0. 47) 3. 75*10-3 units 40. 57 (0. 42) 0. 31 (0. 39) 36. 91 (0. 23) 1. 50*10-2 units 39. 92 (0. 40) 0. 72 (0. 20) 37. 87 (0. 14) 4. 00-10-2 units 41. 40 (0. 42) 0. 62 (0. 13) 36. 65 (0. 79) 0. 225 units 42. 04 (0. 86) 0. 77 (0. 42) 35. 96 (0. 65) b*=-O 23'logds+37 27 R'= 0 94

Table VII. Optimal cooking time (T, in min) determined tor pastas produced wnn omerem leveis 01 enuuxynimses -. ui . , uam semolinas semA or semB. Surface condition and viscoelastic index measured at various cooking times (T+1 or 6 or 11 : indicating the number of extra minutes cooked after T). Standard deviations are shown between brackets. Where significant, relationships between T, SC or VI and endoxylanase dosage [logds : log (units +1)] were given with a : p < 0. 05, : p < 0. 01 and : p < 0. 001.

Cooking time (T) Surface condition (SC) Viscoelastic index (VI) (min) T+6 T+l l T+1 T+6 T+l l SemA CM4 Control 7. 6 (0. 4) 5. 5 (0. 6) 4. 6 (0. 8) 5. 8 (0. 7) 4. 7 (0. 9) 2. 8 (1. 0) Control+XAN 7. 50*10-5 units 7, 3 (0. 4) 5. 3 (0. 2) 4. 3 (0. 2) 6. 2 (0. 8) 5. 2 (0. 4) 3. 2 (1. 0) 7. 50*10~4units 7. 6 (0. 4) 5. 1 (0. 4) 4. 1 (0. 1) 5. 5 (0. 5) 4. 5 (0. 4) 2. 1 (1. 3) 3. 75*10-3 units 7. 3 (0. 4) 5. 1 (0. 2) 4. 1 (0. 4) 5. 1 (0. 6) 4. 3 (1. 1) 2. 3 (1. 1) 1. 50*10-2 units 7. 2 (0. 1) 5. 3 (0. 1) 4. 3 (0. 4) 5. 0 (0. 7) 4. 4 (0. 7) 2. 7 (1. 2) 4. 00-10-2 units 7. 2 (0. 1) 5. 3 (0. 4) 4. 1 (0. 5) 4. 0 (0. 7) 3. 7 (0. 7) 2. 0 (0. 8) 0. 225 units 6. 9 (0. 4) 6. 0 (0. 6) 4. 5 (0. 7) 4. 5 (0. 4) 4. 2 (0. 8) 2. 2 (1. 0) 1. 30 units 6. 3 (0. 1) 6. 9 (0. 6) 6. 6 (0. 8) 4. 3 (0. 9) 3. 0 (1. 0) 2. 7 (1. 5) T=-0 2blogds + 7 6 SC (T+6) = 0 2blogds + 5 2 SC (T+t !) = 0 2'logds +4 i VI (T+I) =-0 3'logds + 5 8 VI (T+6) =-0 2'logds + 4 8 R2= 099 R2= 0 98 R2=097 R2=060 R'=049 Control+XBS 7. 50*10-5 units 7, 9 (0. 5) 5. 0 (0. 5) 4. 1 (0. 3) 5. 3 (0. 6) 4. 6 (0. 7) 2. 8 (1. 2) 7. 50*10-4 units 7. 5 (0. 3) 5. 4 (0. 5) 4. 5 (0. 4) 6. 0 (0. 9) 5. 1 (0. 8) 3. 3 (1. 2) 3. 75*10-3 units 7. 7 (0. 3) 5. 3 (0. 4) 4. 3 (0. 5) 5. 7 (0. 8) 4. 8 (0. 7) 2. 9 (1. 1) 1. 50*10. 2 units 7. 7 (0. 3) 5. 2 (0. 4) 4. 5 (0. 4) 5. 4 (0. 7) 4. 3 (0. 8) 2. 9 (1. 0) 4. 00-10-2 units 7. 1 (0. 1) 5. 5 (0. 3) 4. 6 (0. 3) 5. 3 (0. 7) 5. 0 (0. 5) 3. 2 (1. 3) 0. 225 units 7. 0 (0. 1) 5. 8 (0. 3) 4. 5 (0. 3) 5. 1 (0. 6) 4. 5 (0. 7) 2. 4 (1. 2) T=-0 I'logds + 7 7 VI (T+I) =-O I'logds + 5 8 R 0 99R= 0 30 SemB Control 7. 3 (0. 3) 6. 8 (0. 1) 4. 3 (0. 3) 6. 9 (0. 8) 5. 9 (0. 5) 5. 0 (0. 7) Control+XAN 7. 50*10-4 units 7. 2 (0. 2) 5. 8 (0. 3) 4. 2 (0. 3) 6. 1 (1. 0) 5. 6 (0. 6) 4. 2 (0. 4) 3. 75*10. 3 units 7. 3 (0. 3) 6. 0 (0. 4) 4. 4 (0. 4) 5. 6 (0. 3) 5. 4 (0. 5) 3. 8 (0. 5) 4. 00-10-2 units 7. 3 (0. 2) 5. 9 (0. 4) 4. 3 (0. 4) 4. 6 (0. 6) 4. 4 (0. 5) 3. 6 (0. 3) 0. 225 units 6. 3 (0. 3) 6. 5 (0. 4) 4. 4 (0. 4) 5. 2 (0. 5) 5. 2 (0. 4) 4. 2 (0. 3) T=-0 I'logds+74 SC (T+6) =Ot1ogds+63 SC (T+H) =0 !'logds+43 VI (T+1) =-04blogds+64 VI (T+6) =-02"logds+57 R'=0. 71 R R=0 99 R2= 0 99 R2= 0 99R2=0. 71 R2O 81

Table VIII. Elastic moduli (E in g/mm, slope of the initial linear part), maximal forces (F in g) exerted and maximal distances (D in mm) obtained just before breakage of the pasta strands produced with different levels of endoxylanases XAN or XBS, using semolinas semA or semB and cooked to optimal cooking time T ot T+l 1 min. Standard deviations are shown between brackets. Where significant, relationships between E or F or D and endoxylanase dosage [logds : log (units + 1)] are given with p < 0. 05, b : p < 0. 01 and c p < 0. 001.

T T+ll" E (gl mm) F (g) D (mm) E (gl mm) F (g) D (mm) SemA Control 1. 13 (0. 09) 26. 66 (0. 98) 42. 50 (3. 59) 0. 63 (0. 05) 18. 95 (0. 67) 32. 32 (2. 78) Control+XAN 7. 50*10-4 units 1. 37 (0. 08) 25. 83 (1. 51) 30. 93 (1. 47) 0. 69 (0. 04) 18. 47 (0. 61) 27. 14 (2. 22) 1. 50*10-2 units 1. 06 (0. 07) 24. 54 (1. 18) 45. 06 (3. 64) 0. 63 (0. 06) 18. 31 (1. 33) 31. 27 (2. 41) 0. 225 units 1. 07 (0. 15) 24. 35 (1. 60) 42. 48 (5. 08) 0. 69 (0. 09) 17. 44 (1. 48) 25. 86 (4. 97) 1. 30 units 1. 14 (0. 09) 23. 82 (1. 29) 41. 65 (3. 95) 0. 64 (0. 04) 17. 14 (0. 86) 26. 33 (2. 66) F (T) =-0. 3'logds+25 7 F (T+ )) =-0 2'logds+l 8. 8 R'= 0. 54 R2= 0 79 Control+XBS 7. 50*10 units 1. 41 (0. 16) 25. 57 (0. 97) 31. 61 (3. 04) 0. 67 (0. 06) 18. 37 (1. 44) 27. 91 (3. 09) 1. 50*10. 2 units 0. 85 (0. 08) 22. 51 (1. 23) 42. 23 (3. 59) 0. 73 (0. 08) 18. 24 (1. 06) 26. 23 (4. 08) 0. 225 units 1. 12 (0. 12) 24. 21 (1. 17) 41. 51 (5. 03) 0. 67 (0. 07) 17. 56 (1. 08) 28. 02 (4. 12) F (T) =-O. S'logds+25 9 F (T+11) =-0 2'logds+ 18 7 RO. 41R= 0 70 SemB Control 1. 17 (0. 13) 26. 77 (1. 71) 40. 59 (3. 02) 0. 90 (0. 05) 24. 17 (1. 56) 31. 50 (2. 30) Control+XAN 0. 225 units 1. 23 (0. 11) 25. 10 (0. 88) 45. 79 (2. 73) 0. 69 (0. 06) 17. 87 (0. 80) 26. 81 (2. 36)

Claims (24)

1. Claims 1. Process of producing pasta products, comprising increasing water soluble fiber contents by addition of a fiber hydrolysing enzyme at any stage of pasta production.
2. 2. The process according to claim 1, further comprising adding water insoluble fiber at any stage of the pasta production process.
3. 3. The process according to any of the claims 1 or 2, whereby the water soluble fiber is well retained in said pasta during a normal cooking process of said pasta.
4. 4. The process according to any of the claims 1, 2 or 3, wherein a raw material enriched with said fiber hydrolysing enzyme, a microbial organism producing said fiber hydrolysing enzyme, a germinating material producing said fiber hydrolysing enzyme, a material expressing said fiber hydrolysing enzyme, a genetically modified material expressing or overexpressing said fiber hydrolysing enzyme or a combination thereof is added at any stage of said pasta production.
5. 5. The process according to any of the claims 1 to 4, wherein the fiber hydrolysing enzyme is an endoxylanase.
6. 6. The process according to claim 5, wherein the endoxylanase is from Aspergillus niger or Bacillus subtilis.
7. 7. The process according to any of the claims 1 to 6, wherein said fiber hydrolysing enzyme is added

   before formation of the pasta structure, more specifically prior to or during mixing stages of the raw materials.
8. 8. Pasta products with an increased non-leachable water soluble fiber content, which is obtainable of the process according to any of the claims 1 to 7.
9. 9. Pasta products with a water-extractable arabinoxylan concentration of at least 0.80% by dry weight of said pasta material and which is obtainable of a process according to any of the claims 1 to 7.
10. 10. Non whole meal pasta products having a waterextractable arabinoxylan of at least 42% of the total arabinoxylan population and comprising molecules of a variable molecular weight distribution, which is obtainable of the process according to any of the claims 1 to 7.
11. 11. Non whole meal pasta products having a waterextractable arabinoxylan of at least 55% of the total arabinoxylan population and comprising molecules of a variable molecular weight distribution, which is obtainable of the process according to any of the claims 1 to 7.
12. 12. Whole meal pasta products, wherein at least 25% of their total arabinoxylan population is waterextractable and comprising molecules of a variable molecular weight distribution, which is obtainable of a process according to any of the claims 1 to 7.
13. 13. Whole meal pasta products, wherein at least 30% of their total arabinoxylan population is water extractable and comprising molecules of a variable molecular weight distribution, which is obtainable of a process according to any of the claims 1 to 7.
14. 14. The pasta of any of the claims 8 to 13, wherein the water-extractable arabinoxylan population is non-leachable.
15. 15. A pasta machine specifically designed for carrying out the process of any of the claims 1 to 7.
16. 16. The machine of claim 15, comprising structural elements for mixing the pasta materials, structural elements for moistening the pasta materials and structural elements for mixing, kneading or stretching the pasta materials until a desired consistency is reached or structural elements for shaping the pasta by extruding, molding, stretching, pressing or cutting, or any combination thereof until a desired form is reached or a combination thereof.
17. 17. The pasta machine of claim 15 or 16, further comprising structural element to feed or dose the said fiber hydrolyzing enzymes to the raw materials.
18. 18. The use of the pasta products of any of the claims 8 to 14, for controlling cholesterol and preventing cholesterol damage in a human in need thereto.
19. 19. The use of the pasta of any of the claims 8 to 14, for reducing blood serum cholesterol levels, reducing absorption of cholesterol from the intestines

    into the bloodstream or for inducing a more positive glycemic response.
20. 20. The pasta of any of the claims 8 to 14 having long term or short term positive health effects or both to consumers.
21. 21. A pasta for lowering serum cholesterol in humans in the need thereto, obtainable from a process of any of the claims 1 to 7.
22. 22. The use of fiber hydrolysing enzym preparations for enriching pasta with increased water soluble fiber contents which are well retained in the pasta structure during a normal cooking process.
23. 23. Use of an enzyme preparation as in claim 22, wherein said enzyme preparation contains raw material enriched with said fiber hydrolysing enzyme, a microbial organism producing said fiber hydrolysing enzyme, a germinating material producing said fiber hydrolysing enzyme, a material expressing said fiber hydrolysing enzyme, a genetically modified material expressing or overexpressing said fiber hydrolysing enzyme or a combination thereof is added at any stage of said pasta production
24. 24. Use of enzyme preparation as in any of the claims 22 or 23, wherein the fiber hydrolysing enzyme is an endoxylanase.