JPH0591851A - Production of sustained release dairy product flavor - Google Patents

Abstract

(57) [Summary] [Objective] To provide a novel lipase obtained from a microorganism belonging to Penicillium chrysogenum and a method for producing a flavor of dairy products such as milk, cheese and butter using the lipase. [Composition] Penicillium C
Dairy flavor with excellent palatability, flavor intensity and longevity by enzymatically treating food materials containing milk fat with a novel lipase that has a high ability to produce low molecular weight fatty acids, separated and collected from microorganisms belonging to hrysogenum To get

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to Penicillium chrysogenum.
The present invention relates to a method for producing a dairy product flavor using a lipase produced by a microorganism belonging to m). More specifically, the present invention uses a milk fat-containing food material such as milk, whole milk powder, butter, cheese and cream as a substrate in the presence of a lipase produced by a microorganism belonging to Penicillium chrysogenum. By subjecting the material to an enzymatic reaction, the flavor of the material is, for example, tens of times the flavor intensity, and the heat resistance and durability of butter, cheese, cream and milk are also excellent. And other methods for providing dairy flavors industrially advantageously.

[0002]

2. Description of the Related Art Milk raw materials such as milk fat, whole-fat milk powder, cheese, cheese whey, and milk are treated with a microorganism or a lipolytic enzyme produced by the microorganism to produce a cheese or butter-like flare. There are many reports on how to make bars. For example, fats and oils such as butter oil are decomposed by adding microorganisms such as Aspergillus spp, Mucor spp, Rhizopus spp, and lipolytic enzymes obtained from swine pancreas or oral secretory glands of small livestock. In addition, a method for producing a butter flavour, which further comprises decomposing lipoxygenase (Japanese Patent Publication No. 57-59743), and a mixture of fats and oils, non-fat milk solids and water, as exemplified above. A method for producing a butter flavour, which comprises decomposing by adding the same lipolytic enzyme and protein degrading enzyme / lactose degrading enzyme (JP-B-57
41898), and after decomposing the milk protein and milk fat to some extent by treating the animal milk with a protein degrading enzyme and pancreatin, and a lipolytic enzyme produced by Candida syrindraase, which is a type of yeast. , A method for producing a cheese flavoring substance by inoculating lactic acid bacteria to carry out lactic acid fermentation (Japanese Patent Publication No. 46-23578), and a cheese or a cheese-related material containing lipolytic enzyme, Method for producing a tease flavor which acts on a white degrading enzyme and microorganisms of the genus Streptococcus, Lactobacillus, Propionibacterium, Penicillium and Saccharomyces (Japanese Patent Publication No. 5325024), skim milk, whole Lipase / protease sources (enzymes or Candida spp.) Can be added to flavor-generating medium such as milk, butter, cream, dried whey. Chi cultured by adding biological) and lactic acid-producing microorganisms - method of manufacturing's flavor substances (JP 60-7
No. 8582), or a method for producing a dairy flavor by adding sugar to a milk fat-containing material and further simultaneously allowing yeast and lipase to act (JP-A-62-960).
39), many proposals have been made regarding a method for producing a flavor of dairy products using microorganisms or enzymes.

[0003]

However, the lipases of microbial origin that have been conventionally proposed such as the above examples generally produce a small amount of lower fatty acids, and all of them are natural cheeses produced by a traditional method. Compared with the delicate balance and flavor with high taste produced by dairy products such as butter and butter, they are not always satisfactory, and in particular, they have the drawbacks of poor flavor intensity and persistence. Further, when a forestomach lipase of an animal system such as a calf or a goat is used, a lower fatty acid is produced to some extent, but the decomposition rate of fat is low, so that it is still unsatisfactory in terms of flavor intensity.

[0004]

Means for Solving the Problems As a result of intensive studies to solve the above-mentioned drawbacks, the present inventors have found that a milk-fat-containing material is treated with an enzyme in the presence of a lipase produced by a microorganism belonging to Penicillium chrysogenum. The reaction produces a significantly higher amount of lower fatty acids as compared with the conventional proposals, an excellent and good flavor can be obtained, and a flavor having excellent palatability of natural milk, cheese and butter. -A dairy flavor which can faithfully reproduce the balance, whose flavor intensity is remarkably strengthened as compared to the original milk fat-containing food material, and which also has excellent heat resistance and durability. The present invention has been completed by finding that − can be obtained.

It is well known that, for example, Penicillium rockforti P. roqueforti and Penicillium camembertii belonging to the genus Penicillium greatly contribute to the production of flavors during cheese production. Also, lipase produced by P. cyclopium has been reported [Science and Industry, 52 ,
48, 93 (1978)]. However, the microorganisms belonging to Penicillium chrysogenum have been utilized for the production of the drug penicillin, but the lipases produced by them have not been known at all in the past, and have not been suggested, and were first discovered by the present inventors. It was the one.

Therefore, an object of the present invention is to provide a method for producing a dairy flavor utilizing a lipase produced by a microorganism belonging to Penicillium chrysogenum. Less than,
The aspects of the present invention will be described in more detail.

The lipase used in the present invention can be isolated and collected from a culture of a microorganism belonging to Penicillium chrysogenum. Examples of such microorganisms belonging to Penicillium chrysogenum include Penicillium chrysogenum IFO 6143 strain,
Same IFO 6144, Same IFO 8644, Same IFO 8645, Same I
FO 9646, IFO 8647, IFO 864 and IFO
Known freely-distributed strains such as 8649 strain can be mentioned.

In order to obtain lipase from these microorganisms belonging to Penicillium chrysogenum, the cells are cultured in a medium in which a strain of the genus Penicillium known per se is grown,
The enzyme lipase may be collected from the obtained culture. Hereinafter, the composition of the medium that can be used to grow the above strain will be described, but this is merely for the purpose of explanation and the present invention is not limited to the medium having this composition.

Examples of the carbon source include carbohydrates such as glucose and fructose; organic acids such as palmitic acid and stearic acid; oils and fats such as palm oil, soybean oil, olive oil and butter oil, and one or more of these. A mixture of two or more can be mentioned. The nitrogen source is not particularly limited, but inorganic nitrogen compounds such as ammonium sulfate and ammonium nitrate, and organic nitrogen sources such as peptone can be used. As the inorganic salts, various phosphates and sulfates can be used. Further, a trace amount of metal (iron salt, calcium salt, etc.) may be contained in the medium.

As the culture method, a shaking culture method, a deep aeration stirring culture method and the like can be used. The culture temperature is preferably in the range of, for example, about 20 to about 40 ° C. and pH of about 4 to about 8. Further, the number of days for culturing is not particularly limited, but is usually in the range of about 1 to about 7 days.

In this way, the bacterial cells are separated and collected by centrifugation from the culture in which the enzyme is produced and accumulated, and the obtained bacterial cells are dispersed in water.
The cell-bound enzyme is extracted by adjusting to ~ 9. After extraction, the bacterial cells are removed by centrifugation to obtain an enzyme extract. This enzyme solution can be used as it is as the crude enzyme solution of the present invention, but usually, acetone, alcohol, ammonium sulfate or the like is added to collect the enzyme precipitate, which is appropriately dried by a drying means to be used as a dried crude enzyme. .. The enzyme itself thus obtained or the enzyme solution thereof has a strong lipase activity and can be suitably used for the decomposition of the above-mentioned milk fat-containing food material.

The lipase derived from a microorganism belonging to Penicillium chrysogenum of the present invention can be obtained as described above, and its enzymatic action is an αβ type lipase that randomly releases fatty acids from triglyceride, which has been known so far. It is completely different from other lipases derived from other microorganisms and was first discovered by the present inventors. Hereinafter, various properties of the lipase of the present invention and differences from known lipases will be specifically described.

(1) Enzymatic action It has a lipase activity which hydrolyzes triglyceride in the vicinity of neutrality to separate fatty acid.

(2) Substrate Specificity The substrate specificity of the lipase of the present invention for various monoacidic triglycerides is shown in Table 1 below.

[0015]

TABLE 1 group cytoplasm relative activity (%) ※ Triacetin C2 98 Tripropionin C3 448 Tributyrin C4 486 Tricaproin C6 62 Tricaprylin C8 261 Tricaprin C10 152 Trilaurin C12 148 Trimyristin C14 19 Tripalmitin C16 20 Tristearin C18 3 Triolein C18-1 100 ※ substrate Triolein Relative activity (%) when the activity against is defined as 100%.

As is clear from the results shown in Table 1, the lipase of the present invention specifically acts on a lower fatty acid triglyceride substrate having about 3 to 8 carbon atoms, while on the other hand, it acts on a fatty acid triglyceride having a relatively large molecular weight such as olive oil. It was found that some fractions worked well, but at the present time, it has not been possible to isolate each fraction.

The substrate specificity when the lipase of the present invention and the lipases of other microbial origins known in the prior art are allowed to act on the same monoacidic triglyceride as shown in Table 1 above is shown in “Oil Chemistry, 29, (8) 587”.
■ 591 (1980) ”is as shown in Table 2 below.
It can be seen that it is clearly different from the lipase obtained from Penicillium chrysogenum of the present invention.

[0018]

[Table 2]Penicillium Aspergillus Rhizopus Geotrichum  Substrate cyclopium niger delmar candidum  C2 22 54 40 4 C3 164 123 41 4 C4 375 77 84 4 C6 123 37 52 4 C8 220 85 165 60 C10 162 170 138 47 C12 63 146 88 41 C14 19 70 85 19 C16 9 42 58 8 C18 8 45 32 8 C18-1 100 100 100 100

(3) Optimum pH FIG. 1 shows the optimum pH when the lipase of the present invention is allowed to act on olive oil. The optimum pH when acting on olive oil was found to be 6-8. Further, the optimum pH when acting on tributyrin is shown in FIG. The optimum pH when acting on tributyrin is 8-10. Therefore, the lipase of the present invention is presumed to be a mixture of two types.

(4) Stable pH range The stable pH range when acting on olive oil is pH 6 to 10 as shown in FIG. 3, and when acting on tributyrin is pH 4 to 10 as shown in FIG.

(5) Optimum temperature range of action The temperature-activity curve of this enzyme is about 30 to 50 ° C. when olive oil (shown in FIG. 5) and tributyrin (shown in FIG. 6) are used as substrates. ..

(6) Deactivation condition by temperature The enzyme was heat-treated at each temperature for 10 minutes or 30 minutes, immediately cooled in ice water, and then reacted with olive oil and tributyrin. The results are shown in FIGS. 7 and 8, respectively. Shown in.
As is clear from this result, the enzyme starts to be inactivated at about 40 ° C and is completely inactivated by heating at about 60 to 70 ° C for 10 minutes.

(7) Regiospecificity In accordance with the "index for quantitatively expressing the regiospecificity of lipase" described in the Journal of Fermentation Engineering, 66, 5, 406-408 (1988), the lipase of the present invention has different carbon numbers. Positional specificity index [hereinafter referred to as PS
I (%)] was calculated. The results are shown in Table 3 below.

[0024]

[Table 3]Peak area PSI (%)*1,2 [2,3] DG 1,3 DG  2.5913 1.4238-4.7 2.6886 1.4283-3.0 3.0743 1.5454-0.3 3.0864 1.6296-2.7 ave. -2.7

* PSI (%) = {[1,2 (2,3) DG- (1,3DG) × 2] / [1,2 [2,3] DG + (1,3DG) × 2} × However, DG is a diglyceride, and 1,2,1,3,2,3 are fatty acid binding positions. α-type lipase → + 100% αβ-type lipase → 0% As is clear from the results in Table 3, this enzyme has αβ-type lipase activity.

In order to clarify the difference between the regiospecificity of the lipase produced by P. chrisogenum of the present invention and the regiospecificity of the conventionally known microbial lipase, the PSI (%) of the commercially available microbial lipase is described above. It is shown in Table 4 below, quoted from the Journal of Fermentation Engineering.

[0027]

[Table 4]Trade name Origin PSI (%)  Lipase AP (Amano) Aspergillus niger 100 Lipase LP (Toyo jozo) Chromobacterium viscosum 100 Lipase M (Amano) Mucor japonicum 100 Palatase M (Novo) Mucor miehei 100 Lipase M (Amano) Penicillium cyclopium 100 Lipase (Sigma) Porcine pancreas 100 Lipase ( BMY) Porcine pancreas 100 Lipase (Sigma) Rhizopus arrhizus 100 Lipase (Seikagaku) Rhizopus delemar 100 Lipase F (Amano) Rhizopus japonicus 100 (their laboratory) Saccharomycopsis liplytica 89.8 Lipase PN (Takeda) Phycomyces nites 75.3zo Lipase P (Amano) Pseudomonas fluorescens 74.5 Lipoprotein lipase (Meito) Alcaligenes sp. 73.2 Lipase B (Sapporo) Pseudomonas fragi 72.0 Lipoprotein lipase (toyobo) Pseudomonas sp. 59.6 Lipase AY (Amano) Candida cylindrasea (29.6 Alkaline lipase) 16.0 Cholesterol esterase (Meito) Candida cylindrasea 11.0 Lipase (Meito) Candida cylindrasea 10.0 Lipase MY (Meito) Candida cylindrasea 6.1 Lipase OF (Meito) Candida cylindrasea -11.2 (their Laboratory) Geotricum sp. -24.0 Palatase A (Novo) Aspergillus sp impossible Lipase This invention Penicillium chrysogenum -2.7

Next, the method for producing the dairy flavor of the present invention will be specifically described. Examples of the milk-fat-containing food material that can be used in the present invention include milk such as raw milk, normal milk and processed milk; cream such as cream and fat-replacement cream; whole milk powder; condensed milk such as sugar-free condensed milk and sweetened condensed milk. Examples thereof include cheeses, butter and buttermilk. In addition to these milk-fat-containing food materials, milk materials such as skimmed milk, skim milk powder, and sweetened skimmed condensed milk can be appropriately blended, if desired. Such milk fat-containing food material, it is desirable to select a material suitable for the intended flavor, for example, when using the cheese flavor as the intended product, for example, emmental cheese, It is advantageous to use natural cheese such as cheddar cheese, gold cheese, and parmesan cheese as a main raw material. Similarly, when the target substance is butter flavor, it is preferable to add natural butter such as unsalted butter or salted butter in addition to milk components such as whole milk powder. Further, when a milk type flavor is used as a target product, it is preferable to use milk raw materials such as milk, whole milk powder, and sweetened whole milk condensed milk. However, it is needless to say that the raw materials are not limited to the exemplified raw materials, and all the above-mentioned dairy raw materials can be appropriately selected and used in an arbitrary combination as desired.

To illustrate the preferred embodiments of these dairy flavor preparations, first a milk fat containing food material consisting of whole milk powder, buttermilk, butter, cheese and any mixture thereof. For example, about 1 to about 10 parts by weight of soft water or a buffer is added to 1 part by weight and dissolved. At this time, a cheese melting agent may be added, if desired. Furthermore, if desired, an emulsifier such as lecithin,
Polyglycerin fatty acid ester, glycerin fatty acid ester, sucrose fatty acid ester and the like can also be added. The obtained solution or paste is sterilized at about 110 to about 130 ° C. for about 10 to about 30 minutes. If desired, emulsification treatment can be performed before and after the sterilization treatment.

The sterilized mixture was immediately cooled to a temperature suitable for enzymatic reaction, for example, about 25 to about 55 ° C., and separated and collected from microorganisms belonging to Penicillium chrysogenum.
Add ze. The amount of the lipase used can be appropriately selected depending on the degree of purification. For example about 0.0 based on the weight of milk fat
The amount used is, for example, about 5 to about 5% by weight, preferably about 0.2 to about 2% by weight. The addition method of the lipase can be appropriately selected, and for example, it may be directly added to the sterilized product of the milk fat-containing food material and dispersed, but is preferably dissolved or dispersed in a small amount of water in advance and added. Is good.

In this case, in addition to the lipase produced by Penicillium chrysogenum, a lipase derived from other microorganisms
Ze can also be added. As such a lipase,
For example, a commercially available lipase produced from microorganisms of the genus Aspergillus, genus Mucor, genus Rhizopus, genus Candida and the like can be mentioned. The amount of these lipases added is, for example, about 0.1 based on the weight of the milk fat-containing food material.
A range of about 1 to about 1% by weight is exemplified.

The conditions for the enzymatic reaction treatment are, for example, about 3
0 to about 60 ° C., about 1 to about 120 hours, preferably about 4
Conditions such as enzyme treatment at 0 to about 55 ° C. for about 8 to about 24 hours can be exemplified. After completion of the enzyme reaction, the mixture is heat-treated at about 80 ° C. to about 90 ° C. for about 10 to about 20 minutes to inactivate the enzyme and sterilize it.

The obtained product can be used as it is as the dairy flavor of the present invention, but if desired, it can be further added to the enzyme-inactivated product such as glucose, fructose, sucrose, isomerized sugar and Saccharides such as sugar reduction products, emulsifiers such as those exemplified above and untreated milk-fat-containing food materials are uniformly mixed and dissolved or emulsified, and further sterilized to give rich milk with excellent flavor. Product flavors are provided.

Further, a powdering aid such as gum arabic, starch, starch derivative, dextrin, xanthan gum and cyclodextrin is blended with the above-mentioned enzymatic reaction-treated product, and spray drying, vacuum drying, freeze drying and the like are appropriately used in a usual manner. It can also be made into a powder form by drying using the above-mentioned drying means.

The dairy flavor obtained as described above can be blended and utilized in various foods in a wide range of fields including foods and drinks and favorite products. For example, butter, margarine,
Oils and fats such as shortings; milk drinks such as lactose coffee; creams such as creams and fat-substituted creams; yo
Fermented dairy products such as grout and lactic acid bacteria beverages; ice cream
Frozen desserts such as wheat, ice milk, lacto ice, and soft cream; cheeses; confectionery such as caramel, candy, crackers, biscuits; soft drinks; other dairy processed foods and various instant foods When added to the food or drink of the present invention, a dairy product-like flavor having a natural flavor and having a long-lasting and enhanced flavor is provided. The present invention will be described in detail below with reference to Examples, Comparative Examples and Reference Examples.

[0036]

[Reference Example 1] Preparation of lipase (1) A basal medium having the following composition was prepared and placed in a 50 liter jar fermenter. Peptone 40g yeast extract _{40g K 2 HPO 4 10g MgSO 4} · 7H 2 O 5g olive 400g soft water 20000g

After adjusting the pH of the above medium to 7.0, 120 ° C.
Sterilize for 15 minutes with Penicillium chrysogenum Penic
50 g of a culture of the illium chrysogenum IFO6143 strain was added, and the mixture was agitated and aerated at 30 ° C. for 50 hours. After the culture was completed, 2.0 kg of cells were separated and collected by centrifugation. Next, 14 kg of soft water was added to the separated cells, and the pH was adjusted to 8.0 with 1N-NaOH, and the cell-bound enzyme was extracted with stirring. After extraction, the bacterial cells were removed by centrifugation to obtain 14 kg of enzyme extract. 35 kg of acetone was added to the obtained enzyme extract to precipitate the enzyme, and 250 g of the precipitate was recovered by centrifugation, and this was vacuum dried to obtain 85 g of dried crude enzyme. The lipase activity of this crude enzyme was 10,000 u / g.

[0038]

Reference Example 2 Preparation of Lipase (2) In Reference Example 1, Penicillium chrysogenum Penicill
The ium chrysogenum SK-10 strain was replaced with the Penicillium chrysogenum IFO 6144 strain and cultured under the same conditions to obtain 1.8 kg of bacterial cells. Then, 72 g of dried crude enzyme was obtained by the same operation as in Reference Example 1.
The lipase activity of this crude enzyme was 8000 u / g.

[0039]

[Reference Example 3] Preparation of lipase (3) In the same manner as in Reference Example 1, Penicillium chrysogenum Peni
Using the cillium chrysogenum IFO8644 strain, 53 g of dried crude enzyme was obtained. The enzyme activity of this crude enzyme is 3500u / g
Met.

[0040]

Example 1 Commercially available raw cream (fat content 20%) 50
Sterilize 0g by heating at 85 ℃ for 15 minutes, cool to 40 ℃,
0.6 g of the crude enzyme lipase obtained in Reference Example 1 was added to 200 g of phosphate buffer (1.0 g of sodium hydrogen phosphate,
A solution dissolved in sodium hydrogenphosphate (4.4 g) and soft water (194.6 g) was added, and an enzymatic reaction was carried out at 35 ° C. for 18 hours under stirring conditions. The reaction-treated product is heated at 85 ° C. for 15 to deactivate the enzyme, and the acid value (K required to neutralize 1 g of the treated product)
Mg of OH. Hereinafter, 700 g of an excellent buttery flavor having a value of 35 (AV) was obtained (invention product 1).

[0041]

Comparative Example 1 In Example 1, the same amount of commercially available Aspergillus microorganism-derived lipase AP (manufactured by Amano Pharmaceutical Co., Ltd.) was added in place of 0.6 g of the crude enzyme lipase obtained in Reference Example 1, and the enzyme reaction was performed. Example 1 except that the time was changed to 24 hours
Enzyme treatment was carried out under the same conditions as above to obtain 495 g of butter flavor. The AV of this butterflavor was 33 (Comparative Product 1).

[0042]

Comparative Example 2 In Example 1, in place of 0.6 g of the crude enzyme lipase obtained in Reference Example 1, the same amount of commercially available Candida cylindrace origin lipase MY (manufactured by Meito Sangyo Co., Ltd.) was added. The same operation as in Example 1 was carried out except that the enzyme reaction was carried out at 20 ° C. for 20 hours to obtain 500 g of butter flavor. The AV of this butterflavor was 35 (Comparative Product 2).

[0043]

[Reference Example 4] The butter flavors obtained in Example 1, Comparative Example 1 and Comparative Example 2 were added to a cookie dough having the following formulation, and baked at 220 ° C for 7 minutes to prepare a cookie.

[0044]Mix recipe Wheat flour (soft flour) 1000 g Sugar 220 g Shortening (M.P 37 ° C) 350 g Full fat sugar condensed condensed milk 110 g Whole fat milk powder 15 g Salt 8 g Sodium bicarbonate 3 g Ammonium bicarbonate 4 g Yocrecitin 1 g Water 120 g Butter flavor6 g  1837g in total

Cookers in which the product 1 of the present invention, the comparative product 1 and the comparative product 2 were added to the above butter flavour, were designated as 1, 2 and 3, respectively. For these cookies,
Sensory evaluation was performed by 20 well-trained sensory inspectors. Regarding heat resistance, evaluation items 1 and 3 immediately after firing
Butter flavor and palatability were evaluated together. Further, regarding the sustainability, the product stored at 30 ° C. for 2 weeks after firing was evaluated. The results are shown in Table 5.

[0046]

[Table 5] Sensory evaluation Cookie-1 Cookie-2 Cookie-3 Butter-excellent in flavor 20 people 0 0 0 Excellent in persistence of scent-flavor 18 18 1 1 Highly palatable 18 people 2 people 0 people

As is clear from the results of Table 5, the product 1 of the present invention
In comparison with Cookies-2 and 3 to which Comparative Products 1 and 2 were added, respectively, the Cookies to which was added were superior in all of the flavor, sustainability and palatability at a significant level of 0.1%.

[0048]

Example 2 500 g of unsalted butter and phosphate buffer
200 g (1.0 g of potassium hydrogen phosphate, 4.4 g of sodium hydrogen phosphate, 194.6 g of soft water) were added to give 8
After heat-dissolving at 5 ° C for 15 minutes, sterilization, and cooling to 50 ° C, 2.0 g of the crude enzyme lipase obtained in Reference Example 2 was added, and the enzyme reaction was performed at 30 ° C for 60 hours under stirring conditions. It was
Then, the mixture was heated at 90 ° C. for 15 minutes to deactivate the enzyme to obtain 690 g of cheese flavor. A of this butter flavor
V was 85 (invention product 2).

[0049]

Comparative Example 3 2.0 g of the lipase obtained in Reference Example 1 in Example 2 was used as a commercially available lipase derived from a microorganism of the genus Rhizopus.
ZE (Saiken) (manufactured by Osaka Bacterial Research Institute) except that the same amount was used, and the enzymatic reaction was carried out under the same conditions as in Example 2, and AV
682 g of cheese flavor of 82 was obtained (Comparative Product 3).

[0050]

Reference Example 5 Using the cheese flavors of the product 2 of the present invention and the comparative product 3 obtained in Example 2 and Comparative Example 3, a cracker was prepared by a conventional method according to the following formulation, and the same functionalities as in Reference Example 4 were prepared. An evaluation was made. The results are shown in Table 6.

[Cracker Formulation]Cracker 1 (invention product) Cracker 2 (comparative product) Wheat medium strength flour 1000 (g) Same as left (g) Sugar 15 〃 Fructose fructose liquid sugar (75%) 40 〃 Shortening (mp37 °) 150 〃 Salt 14 〃 Sodium glutamate 1 〃 Non-fat dry milk 16 〃 Ammonium bicarbonate 12 〃 Sodium bicarbonate 10 〃 Monocalcium phosphate 12 〃 Takafud B (Sankyo) 1.5 〃 Cheese flavor (Invention product 2) 17.5 0 Cheese flavor (Comparative product 3) 0 17.5 Water300 300  Total 1589g 1589g

[0052]

[Table 6] Sensory Evaluation Results Cracker-1 Cracker-2 Cheese Excellent in flavor 20 0 0 Excellent in sustaining cheese flavor 20 0 Highly palatable 19 1 Name

As is clear from the results in Table 6, the cracker-1 using the product 2 of the present invention is much closer to the flavor of natural cheese than the cracker-2 using the comparative product 3, and has a good palatability and longevity. It was excellent at a significance level of 0.1%.

[0054]

Example 3 500 g of commercially available milk was sterilized by heating at 85 ° C. for 15 minutes, cooled to 45 ° C., 0.1 g of the crude enzyme lipase obtained in Reference Example 3 was added, and the mixture was stirred at 37 ° C. for 24 hours. The enzyme reaction was performed below. After heating at 90 ° C for 15 minutes to deactivate the enzyme, 495 g of milk flavor containing AV8 was obtained (invention product 3).

[0055]

[Comparative Example 4] In Example 3, 0.1 g of the crude enzyme lipase obtained in Reference Example 3 was replaced with 0.1 g of commercially available Mucor microbial origin lipase M-AP (manufactured by Amano Pharmaceutical Co., Ltd.). Other than that, the enzymatic reaction was performed under the same conditions as in Example 3 to obtain 490 g of a milk flavor containing AV9 (Comparative product 4).

The milk flavors obtained in Example 3 and Comparative Example 4 (invention product 3 and comparative product 4) were heated to 1
It diluted to weight% and sensory evaluation was performed by 20 sensory inspectors. The results are shown in Table 7.

[0057]

[Table 7] Sensory evaluation results Inventive product 3 Comparative product 4 People with excellent milk-like flavor 20 0 People with excellent sustainability of milk-like flavor 20 0 0 People with high palatability 19 1 1

As is clear from the results in Table 7, the milk flavor of Inventive Product 3 is much closer to the natural rich milk-like flavor than the milk flavor of Comparative Product 4 and has palatability and sustainability. It was excellent at a significance level of 0.1%.

[0059]

According to the present invention, there is provided a lipase found from a microorganism belonging to Penicillium chrysogenum. Also, by enzymatically treating a milk fat-containing material using this lipase, it is possible to reproduce a flavor excellent in freshness, flavor balance and palatability that could not be obtained by a conventionally known lipase, Moreover, the flavor intensity thereof is increased to several tens to 100 times as high as that of the raw material containing milk fat, and in addition, a dairy flavor having excellent heat resistance and durability is provided. It

[Brief description of drawings]

FIG. 1 is a diagram showing an optimum pH range when the enzyme of the present invention is allowed to act on olive oil.

FIG. 2 is a diagram showing an optimum pH range when the enzyme of the present invention is allowed to act on tributyrin.

FIG. 3 is a view showing an optimum temperature range when the enzyme of the present invention is allowed to act on olive oil.

FIG. 4 is a diagram showing an optimum temperature range when the enzyme of the present invention is allowed to act on tributyrin.

FIG. 5 is a graph showing optimum action temperature when the enzyme of the present invention is allowed to act on olive oil.

[Fig. 6] Fig. 6 is a view showing a suitable action temperature when the enzyme of the present invention is allowed to act on tributyrin.

FIG. 7 is a diagram in which the inactivation condition of the enzyme of the present invention is measured using olive oil as a substrate.

FIG. 8 is a diagram in which the inactivating condition of the enzyme of the present invention is measured using tributyrin as a substrate.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Takuhiro Kawabata 335 Hasegawa Kaori Co., Ltd., Nakahara-ku, Kawasaki-shi, Kanagawa Kawasaki Research Institute Co., Ltd. Company Kawasaki Research Center

Claims (1)

[Claims] 1. A milk-fat-containing food material is provided as Penicillium chrysogenum.
num), the enzymatic reaction in the presence of a lipase produced by a microorganism belonging to num).