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GB2585029A - Xylanase additives for animal feeds - Google Patents

Xylanase additives for animal feeds Download PDF

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Publication number
GB2585029A
GB2585029A GB1909099.2A GB201909099A GB2585029A GB 2585029 A GB2585029 A GB 2585029A GB 201909099 A GB201909099 A GB 201909099A GB 2585029 A GB2585029 A GB 2585029A
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Prior art keywords
animal feed
xylanase
enzyme
feed
additive
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GB1909099.2A
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GB201909099D0 (en
Inventor
Ivan Pletschke Brett
Malgas Samkelo
Mafa Mpho
Nkanyiso Mathibe Brian
Dalitso Chindipha Stones
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Rhodes Univ
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Rhodes Univ
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Priority to GB1909099.2A priority Critical patent/GB2585029A/en
Publication of GB201909099D0 publication Critical patent/GB201909099D0/en
Priority to PCT/IB2020/056003 priority patent/WO2020261164A1/en
Publication of GB2585029A publication Critical patent/GB2585029A/en
Withdrawn legal-status Critical Current

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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/189Enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/14Pretreatment of feeding-stuffs with enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/06Enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135Bacteria or derivatives thereof, e.g. probiotics
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17Amino acids, peptides or proteins
    • A23L33/195Proteins from microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L7/00Cereal-derived products; Malt products; Preparation or treatment thereof
    • A23L7/10Cereal-derived products
    • A23L7/104Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms
    • A23L7/107Addition or treatment with enzymes not combined with fermentation with microorganisms
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y302/00Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01008Endo-1,4-beta-xylanase (3.2.1.8)
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/10Feeding-stuffs specially adapted for particular animals for ruminants
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/70Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75Feeding-stuffs specially adapted for particular animals for birds for poultry

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • Food Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Nutrition Science (AREA)
  • Biotechnology (AREA)
  • Animal Husbandry (AREA)
  • Biochemistry (AREA)
  • Physiology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Mycology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Fodder In General (AREA)

Abstract

An enzyme animal feed or human food additive comprising a GH10 xylanase derived from a bacterium and a GH11 xylanase derived from a fungus. The GH10 xylanase may be derived from Geobacillus stearothermophilus, and may be XT6 encoded by a xt6 gene. The GH11 xylanase may be derived from Trichoderma viride, and may be Xyn2A encoded by a xyn2A gene. The additive may reduce non-starch polysaccharide (NSP) viscosity. A feed composition comprising the enzyme additive, and a method of increasing feed conversion ratio and/or improving nutritional value of a feed by (a) providing cereal-based feed and (b) adding the disclosed enzyme additive to the feed, are also claimed. The feed composition or cereal-based feed may comprise a cereal selected from one or more of barley, maize, oats, rice, rye, sorghum, triticale, wheat. The feed composition or feed may comprise animal feed or human food. Inclusion of the bacterial GH10 and fungal GH11 xylanases in the additive may cause the additive to have a synergistic effect compared to an additive comprising xylanases from a single source. The GH10 and GH11 xylanases may have amino acid sequences as shown in the figures.

Description

XYLANASE ADDITIVES FOR ANIMAL FEEDS
BACKGROUND OF THE INVENTION
This invention relates to an enzyme animal feed additive comprising a GH10 bacterial xylanase, XT6 from Geobacillus stearothermophilus, and a GH11 fungal xylanase, Xyn2A from Trichoderma viride. The present invention also relates to an animal feed composition comprising the animal feed additive described and to the use of the animal feed composition in improving body weight gain and/or feed conversion ratio in an animal. This invention further relates to methods of increasing a feed conversion ratio and/or improving the nutritional value of an animal feed using the enzyme animal feed additive.
Worldwide, agricultural farming systems, such as livestock production, face an increasing challenge of maintaining future global demand for meat and dairy products because of an expected increase in population. The Food and Agriculture Organization (FAO) expects that an increase in purchasing power for food from animal sources will raise the yearly demand to 465 and 1.043 million tonnes for meat and milk products, respectively, by 2050. Besides, the FAO estimates the growth of global population to reach 9.6 billion by the year 2050, with a doubled purchasing power for meat and dairy products.
The price of wheat increased drastically in 2007, and prices on traditional feed ingredients have in general been fluctuating since. This has encouraged meat producers to look for alternative and less expensive ingredients for feed. While most local farmers have resorted to the use of feeds developed on-site using agro-industrial wastes such as corn stover, soybean meal, canola meal, sugarcane bagasse, and wheat bagasse, these feeds contain high quantities of non-starch polysaccharides (NSPs), such as mannan, xylan, pectin and cellulose, which negatively affect the feed's nutrient utilization.
Insoluble NSPs are associated with the encapsulation of nutrients such as starch and protein. This encapsulation causes valuable nutrients to by-pass the digestive tract of the animal, undigested. The soluble NSPs, on the other hand, increase the viscosity of the digesta and as a result, affects the digestion and assimilation of nutrients such as fatty acids by the animal. Moreover, the binding of -2 -NSPs with the intestinal brush border of animals increases the thickness of the unstirred water layer adjacent to the mucosa, leading to impaired nutrient digestion and absorption. NSPs are also associated with stimulating the growth of some pathogenic bacteria species, including Escherichia co/i and Clostridium perfringens. For example, the amount of pectin (about 60 g uronic acids/kg dry matter) and the fibre matrix structure of canola most likely increases the water holding capacity of this raw material, resulting in poor nutrient availability for non-ruminants. Furthermore, the antinutritional factors (ANFs) may reduce the energetic value.
Xylanases (also called 1,4-(3-o-xylan xylanohydrolases, EC 3.2.1.8) are enzymes which randomly cleave the 3-(1,4)-linkages between two D-xylopyranosyl residues in xylan backbones. Xylanases have been classified into glycoside hydrolase (GH) families GH5, GH8, GH10, GH11, GH30, GH43, GH62 and GH98 in the CAZy database of carbohydrate-active enzymes, with GH10 and GH11 being the best characterized families. GH11 xylanases can only hydrolyse xylosidic bonds where the two corresponding xylose moieties in subsites (-1) and (+1) are not branched, while GH10 xylanases attack the glycosidic linkage next to a single or double substituted xylose toward the non-reducing end and require two unsubstituted xylose residues between branched residues. GH11 fungal and bacterial xylanases often contain a catalytic domain linked to one or more non-catalytic modules such as Carbohydrate Binding Molecule Family 22 (CBM22). This appears to favour GH11 xylanases to display high catalytic activity on insoluble xylan substrates, while GH10 xylanases appear to be more catalytically efficient on soluble xylan substrates.
Xylanases are considered one of the industrially important microbial enzymes, which can catabolize xylan residues. Over the years, the use of xylanases at an industrial level has increased significantly. Since applications of xylanases in commercial sectors are widening, an understanding of their nature and properties for efficient and effective usage becomes crucial. Xylanases extracted from microorganisms have been used for pulp bleaching, waste paper treatment and for fabric bio-processing, such as: bio-bleaching, desizing and bio-scouring of fabrics. Xylanases have also been shown to improve rheological properties of dough, bread specific volume and crumb firmness.
Several studies have also demonstrated improvements in nutritive value, feed utilization, body weight gain, composition and activity of intestinal microbiota, and reduction in excreta volume in livestock after supplementation of agro-industrial wastes based diets with NSP-degrading enzymes such as cellulases, pectinases, xylanases -3 -and glucanases. Enzyme supplementation is a well-established method in the animal feed industry to obtain viscosity reduction and fibre degradation in situ in poultry, with xylanases being the enzymes of choice. Recently, the use of exogenous carbohydrolases in aqua feeds has also been getting more attention.
However, others have also reported that exogenous enzymes did not consistently enhance forage quality and utilization by ruminants. This inconsistency may be attributed to several factors such as the source of the enzyme, doses and activities of the enzyme, physical properties of the substrate, treatment duration, enzyme application method, composition of the diet to which enzyme is added and level of animal productivity.
Use of antibiotics in animal feed as growth-promoters appears to promote emergence of antibiotic resistant strains. The Guidelines for Industry issued by the Center for Veterinary Medicines of the Food and Drug Administration, USA recommend use of antibiotics only for the prevention, control and treatment of infections in animals but not for the promotion of growth, increased performance, and improved feed efficiency. Use of fibrolytic enzymes (i.e. xylanases) have been reported to lead to increased populations of probiotics such as Bifidobacteria and Lactobacilli and decreased Gram-negative bacteria overall. This shows that xylanase additives in feeds can potentially replace the use of antibiotics at subtherapeutic levels in feeds as animal growth promoters.
SUMMARY OF THE INVENTION
The present invention relates to an enzyme animal feed additive, an animal feed composition comprising the animal feed additive and to the use of the animal feed composition in improving body weight gain and/or feed conversion ratio in an animal.
According to a first aspect of the present invention there is provided for an enzyme animal feed additive comprising a GH10 xylanase derived from a bacterium and a GH11 xylanase derived from a fungus.
In one embodiment of the invention, the GH10 xylanase derived from a bacterium is derived from Geobacillus stearothermophilus, such as XT6 encoded by a xt6 gene, wherein the GH10 xylanase has an amino acid sequence substantially identical to SEQ ID NO:1.
According to a second embodiment, the GH11 xylanase derived from a fungus is derived from Trichoderma viride, such as Xyn2A encoded by a xyn2A gene, wherein -4 -the GH11 xylanase has an amino acid sequence substantially identical to SEQ ID NO:2.
In a third embodiment of the invention, the enzyme animal feed additive is capable of reducing non-starch polysaccharide viscosity.
According to a fourth embodiment of the present invention, the GH10 xylanase derived from a bacterium and the GH11 xylanase derived from a fungus in the enzyme animal feed additive demonstrate a synergistic effect in hydrolysing non-starch polysaccharides when compared to an enzyme animal feed additive comprising a xylanase from a single source.
Further, the enzyme animal feed additive of the present invention may further comprise a physiologically acceptable carrier.
According to a second aspect of the present invention, there is provided for an animal feed composition comprising the enzyme animal feed additive as described herein. In particular, the animal feed composition may comprise a cereal, such as a cereal selected from the group consisting of barley, maize, oats, rice, rye, sorghum, triticale, wheat, and combinations thereof.
According to a third aspect of the present invention, there is provided for the use of an animal feed composition of the second aspect for improving body weight gain and/or feed conversion ratio in an animal.
According to a fourth aspect of the present invention there is provided for a method of increasing a feed conversion ratio and/or improving the nutritional value of an animal feed comprising: (a) providing a cereal-based animal feed; and (b) adding the enzyme animal feed additive of the invention as described herein to the cereal-based feed.
It will be appreciated by one of skill in the art that the cereal-based animal feed may be selected from the group consisting of barley, maize, oats, rice, rye, sorghum, triticale, wheat, and combinations thereof.
According to a further embodiment of the present invention, the GH10 xylanase derived from a bacterium and the GH11 xylanase derived from a fungus in the enzyme animal feed additive have a synergistic effect in increasing the feed conversion ratio and/or improving the nutritional value of the animal feed compared to an animal feed with an enzyme animal feed additive comprising a xylanase from a single source. -5 -
BRIEF DESCRIPTION OF THE FIGURES
Non-limiting embodiments of the invention will now be described by way of example only and with reference to the following figures: Figure 1: Amino acid sequence of GH10 xylanase XT6 from Geobacillus stearothermophilus (SEQ ID NO:1).
Figure 2: Amino acid sequence of GH11 xylanase Xyn2A from Trichoderma viride (SEQ ID NO:2).
SEQUENCE LISTING
The nucleic acid and amino acid sequences listed in the accompanying sequence listing are shown using standard letter abbreviations for nucleotide bases, and the standard three letter abbreviations for amino acids. It will be understood by those of skill in the art that only one strand of each nucleic acid sequence is shown, but that the complementary strand is included by any reference to the displayed strand. In the accompanying sequence listing: SEQ ID NO:1 -amino acid sequence of XT6 from Geobacillus stearothermophilus SEQ ID NO:2 -amino acid sequence of Xyn2A from Trichoderma viride
DETAILED DESCRIPTION OF THE INVENTION
The present invention will now be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments of the invention are shown.
The invention as described should not be limited to the specific embodiments disclosed and modifications and other embodiments are intended to be included within the scope of the invention. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.
As used throughout this specification and in the claims that follow, the singular forms "a", "an" and "the" include the plural form, unless the context clearly indicates otherwise.
The terminology and phraseology used herein is for the purpose of description and should not be regarded as limiting. The use of the terms "comprising", "containing", "having" and "including" and variations thereof used herein, are meant to encompass the items listed thereafter and equivalents thereof as well as additional items. -6 -
The present invention relates to the use of a synergistic combination of xylanases, specifically a combination of a GH10 bacterial xylanase and a GH11 fungal xylanase, as an additive to animal feed, and to feeds that contains the synergistic combination of xylanases. The invention also relates to a method for improving the feed conversion ratio and/or the apparent metabolizable energy of feedstuffs using the synergistic combination of xylanases.
As used herein a "xylanase" means a protein, or a polypeptide, having xylanase activity. Xylanase activity can be measured using any assay, in which a substrate is employed, that includes 1,4-p-o-xylosidic endo-linkages in xylans. Different types of substrates are available for the determination of xylanase activity e.g. Xylazyme cross-linked arabinoxylan tablets, or insoluble powder dispersions and solutions of azo-dyed arabinoxylan.
The xylanases of the present invention may be formulated as a feed additive by methods known to those skilled in the art. Physiologically acceptable ingredients may be used. The term "physiologically acceptable" refers to properties and/or substances which are acceptable for administration to an animal from a toxicological point of view. Further "physiologically acceptable" refers to factors such as formulation, stability, patient acceptance and bioavailability, which will be known to a person skilled in the art.
As used herein the term "derived from" with reference to a xylanase refers to any wild-type xylanase isolated from the organism in question, synthetic versions thereof, as well as variants or fragments thereof which retain xylanase activity.
The xylanases of the invention may be "purified", in that they may be comprised in a protein-enriched preparation, in which a substantial amount of low molecular components, typical residual nutrients and minerals have been removed. Such purification can e.g. be by conventional chromatographic methods such as ion-exchange chromatography, hydrophobic interaction chromatography and size exclusion chromatography.
The terms polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymer.
As used herein a "substantially identical" amino acid sequence is an amino acid sequence that differs from a reference sequence only by one or more non-conservative substitutions, deletions, or insertions located at positions of the sequence that do not destroy or substantially reduce the activity of the expressed protein or polypeptide. Alignment for purposes of determining percent sequence identity can be achieved in various ways that are within the knowledge of those with skill in the art. These include using, for instance, computer software such as CLUSTALW or BLAST software. Those skilled in the art can readily determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. In one embodiment of the invention there is provided for a polypeptide sequence that has at least about 80% sequence identity, at least about 90% sequence identity, or even greater sequence identity, such as about 95%, about 96%, about 97%, about 98% or about 99% sequence identity to the sequences described herein. This definition also refers to, or may be applied to, the compliment of a given sequence.
The term "animal" refers to all animals, including human beings. Examples of animals are non-ruminants, and ruminants. Ruminant animals include, for example, animals such as sheep, goat, and cattle, e.g. cow such as beef cattle and dairy cows. In a particular embodiment, the animal is a non-ruminant animal. Non-ruminant animals include mono-gastric animals, e.g. pig or swine (including, but not limited to, piglets, growing pigs, and sows); poultry such as turkeys, ducks and chickens (including but not limited to broiler chicks, layers); fish (including but not limited to salmon, trout, tilapia, catfish and carp); and crustaceans (including but not limited to shrimp and prawn). The xylanase compositions of the invention can be fed to the animal before, after, or simultaneously with the diet.
The term "feed", "feed composition", or "diet' means any compound, preparation, mixture, or composition suitable for, or intended for intake by an animal.
The "suitable forms" of the feed additive may be combined with "physiologically acceptable carriers" and other elements known in the art to produce additives for feeds. The feed additive may further be combined with other ingredients which promote absorption by the digestive tract. By "physiologically acceptable carrier" is meant a solid or liquid filler, diluent or encapsulating substance which may be safely used for the administration of the xylanases and/or feed additives to an animal. The physiologically acceptable carrier may be a cereal or derived from a cereal. Such cereals include milled wheat, maize, soya, sugars, starches or a by-product of any of these. Such carriers are conventional in this technical art, and so are not described in any further detail. -8 -
The feed additive of present invention may be combined with other feed components to produce a cereal-based feed. Such other feed components include one or more other enzyme supplements, vitamin feed additives, mineral feed additives and amino acid feed additives. The resulting feed additive may then be mixed in an appropriate amount with the other feed components such as cereal and protein supplements to form an animal feed. Although the cereal component of a cereal-based feed constitutes a source of protein, it may be necessary to include sources of supplementary protein in the feed such as those derived from fish-meal, meat-meal or vegetables. Sources of vegetable proteins include at least one of full fat soybeans, rapeseeds, canola, soybean-meal, rapeseed-meal and canola-meal.
The feed provided by the present invention may also include other enzyme supplements such as one or more of P-glucanase, glucoamylase, mannanase, agalactosidase, phytase, lipase, a-arabinofuranosidase, protease, a-amylase and pectinase.
Processing of these components into an animal feed can be performed using any of the currently used processing methods and apparatuses such as a doublepelleting machine, a steam pelleter, an expander or an extruder.
The use of the xylanases, or feed additives or feeds containing the xylanases, entails administration of an effective amount of the xylanases, or feed additives or feeds containing the xylanases, to an animal in order to promote digestion and improve the nutritional value of an animal feed. "Improving the nutritional value of a feed" means improving the availability of nutrients, whereby the growth rate, weight gain, and/or feed conversion (i.e. the weight of ingested feed relative to weight gain) of the animal is/are improved.
Although some indications have been given as to suitable dosage amounts of the xylanases, or feed additives or feeds containing the xylanases, the exact amount and frequency of administration will be dependent on several factors and can be optimised. These factors include the individual components used, the formulation of the feed additives or feeds containing the xylanases, the age, weight and general physical condition of the animal, and other factors as are known to those skilled in the art.
The following examples are offered by way of illustration and not by way of limitation. -9 -
EXAMPLE 1
Formulation of an Enzyme Cocktail for Use as an Animal Feed Additive Using commercial xylanases supplied by Megazyme (Ireland, Wicklow) and Sigma Aldrich (USA, St. Louis), namely the GH11 xylanase Xyn2A from the fungus Trichoderma viride and the GH10 xylanase XT6 from the bacterium Geobacillus stearothermophilus, an enzyme cocktail relevant for use as an animal feed additive was formulated. The enzyme formulation comprises 75:25 % of Xyn2A to XT6 at protein mass dosage/loading.
EXAMPLE 2
Comparison of the Enzyme Formulation Activity to Other Xylanase Animal Feed Products The activity of the enzyme formulation from Example 1 was compared to that of commercially available xylanases; XynA from Thermomyces lanuginosus, XT6 from Geobacillus stearothermophilus, Xyn2A from Trichoderma viride, and Xyn100 from Cellvibrio japonicus, during the degradation of wheat-flour arabinoxylan, a predominant NSP in agricultural grains and feedstocks. The enzyme loading was kept at 0.25 mg protein/g biomass in all the reactions. Activity of the xylanases was determined by incubating them with 1 % (w/v) wheat-flour arabinoxylan (in 50 mM sodium citrate buffer, pH 5.0) at 50 °C for 12 hours and estimating the reducing sugar release using the 3,5-dinitrosalicylic acid (DNS) method with absorbance readings at 540 nm. Table 1 below shows the sugar release in mg/ml of the xylanases.
Table 1. Evaluation of wheat-flour arabinoxylan hydrolysis by the enzyme formulation and other xylanases from different sources and/or GH families. ANOVA analysis for differences in hydrolysis with respect to reducing sugar release by the different enzymes (p value < 0.05). Different superscript letters denote statistical significant difference between values. Values are represented as mean values ± SD (n=4).
Enzyme Formulation XynA Xyn2A XT6 Xyn1OD Sugar release (mg/ml) 2.89±0.39a 2.07±0.46° 2.47±0.47b 2.51±0.486 2.61±0.516 As can be seen from Table 1 above, the enzyme formulation was shown to have higher hydrolytic efficiency against arabinoxylan compared to some of commercially available enzyme formulations used as feed additives.
EXAMPLE 3
Arabinoxylan Viscosity Reduction by the Enzyme Formulation The NSP (wheat-flour arabinoxylan) viscosity reduction ability of the enzyme formulation of Example 1 was determined using a Cannon-Manning Semi-Micro viscometer (size 50) at room temperature upon hydrolysis of 0.5 % (w/v) wheat flour arabinoxylan by 0.25 mg protein/g biomass of the enzyme formulation at pH 5.0 and 50 C after 12 hours and compared to an undigested/un-hydrolysed sample (negative control). Table 2 below shows the viscosity reduction of the enzyme formulation relative to the negative control.
Table 2. Viscosity reduction of 0.5 % wheat flour arabinoxylan by the enzyme formulation. ANOVA analysis for differences in hydrolysis with respect to reducing sugar release by the different enzymes (p value < 0.01). Different superscript letters denote statistical significant difference between values. Values are represented as mean values ± SD (n=4).
Sample Negative control Formulation treated Viscosity (cSt) 3.202a 1.551b As can be seen from Table 2 above, significant reduction in NSP viscosity was proven using the enzyme formulation.

Claims (16)

  1. CLAIMS1. An enzyme animal feed additive comprising a GH10 xylanase derived from a bacterium and a GH11 xylanase derived from a fungus.
  2. 2. The enzyme animal feed additive of claim 1, wherein the GH10 xylanase derived from a bacterium is derived from Geobacillus stearothermophilus.
  3. 3. The enzyme animal feed additive of claim 1 or 2, wherein the GH10 xylanase is XT6 encoded by a xt6 gene.
  4. 4. The enzyme animal feed additive of any one of claims 1 to 3, wherein the GH11 xylanase derived from a fungus is derived from Trichoderma viride.
  5. 5. The enzyme animal feed additive of any one of claims 1 to 4, wherein the GH11 xylanase is Xyn2A encoded by a xyn2A gene.
  6. 6. The enzyme animal feed additive of any one of claims 1 to 5, wherein the GH10 xylanase has an amino acid sequence substantially identical to SEQ ID NO:1 and the GH11 xylanase has an amino acid sequence substantially identical to SEQ ID NO:2.
  7. 7. The enzyme animal feed additive of any one of claims 1 to 6, wherein the enzyme animal feed additive reduces non-starch polysaccharide viscosity.
  8. 8. The enzyme animal feed additive of any one of claims 1 to 7, wherein the GH10 xylanase derived from a bacterium and the GH11 xylanase derived from a fungus have a synergistic effect in hydrolysing non-starch polysaccharide compared to an enzyme animal feed additive comprising a xylanase from a single source.
  9. 9. The enzyme animal feed additive of any one of claims 1 to 8, further comprising a physiologically acceptable carrier.
  10. 10. An animal feed composition comprising the animal feed additive of any one of claims 1 to 9.
  11. -12 - 11. The animal feed composition of claim 10, wherein the animal feed composition comprises a cereal.
  12. 12. The animal feed composition of claim 11, wherein the cereal is selected from the group consisting of barley, maize, oats, rice, rye, sorghum, triticale, wheat, and combinations thereof.
  13. 13. Use of an animal feed composition of any one of claims 10 to 12 for improving body weight gain and/or feed conversion ratio in an animal.
  14. 14. A method of increasing a feed conversion ratio and/or improving the nutritional value of an animal feed comprising: (a) providing a cereal-based animal feed; and (b) adding the enzyme animal feed additive of any one of claims 1 to 9 to the cereal-based feed.
  15. 15. The method of claim 14, wherein the cereal-based animal feed is selected from the group consisting of barley, maize, oats, rice, rye, sorghum, triticale, wheat, and combinations thereof.
  16. 16. The method of claim 14 or 15, wherein the GH10 xylanase derived from a bacterium and a GH11 xylanase derived from a fungus in the enzyme animal feed additive have a synergistic effect in increasing the feed conversion ratio and/or improving the nutritional value of the animal feed compared to an animal feed with an enzyme animal feed additive comprising a xylanase from a single source.
GB1909099.2A 2019-06-25 2019-06-25 Xylanase additives for animal feeds Withdrawn GB2585029A (en)

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GB1909099.2A GB2585029A (en) 2019-06-25 2019-06-25 Xylanase additives for animal feeds
PCT/IB2020/056003 WO2020261164A1 (en) 2019-06-25 2020-06-25 Xylanase additives for food or feeds

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CN107836572A (en) * 2017-11-09 2018-03-27 上海欧耐施生物技术有限公司 A kind of xylanase enzyme preparation for effectively improving feed grain nutritive value and its application

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