CN106987613B

CN106987613B - Industrial preparation method and application of wheat peptone

Info

Publication number: CN106987613B
Application number: CN201710239828.6A
Authority: CN
Inventors: 郭文杰; 崔凤杰; 方颂平; 孙文敬; 辛晓亚; 刘飞翔; 田启梅; 周鹏磊
Original assignee: Anhui Ruifuxiang Food Co ltd
Current assignee: ANHUI RUN'AN XINKE DETECTION TECHNOLOGY Co.,Ltd.
Priority date: 2017-04-13
Filing date: 2017-04-13
Publication date: 2021-06-01
Anticipated expiration: 2037-04-13
Also published as: CN106987613A

Abstract

The invention relates to an industrial preparation method of wheat peptone and application thereof, wherein the method comprises the following steps: firstly, taking wet-based wheat protein (wet gluten) as a raw material, efficiently crushing and homogenizing the raw material, hydrolyzing the raw material by using a multi-enzyme system, filtering the hydrolyzed wheat protease liquid, and performing cooking, injection treatment enzyme deactivation and spray drying to obtain wheat peptone; the protein content is 60-90%; contains 18 amino acids, wherein the content of glutamine/glutamic acid is more than 25%; the content of peptide fragment with molecular mass less than 1000Da is more than 80 percent; the wheat peptone can completely or partially replace a nitrogen source in a microorganism growth/fermentation culture medium, is used for producing fermentation products such as probiotics, antibiotics, enzyme preparations, amino acids, organic acids, vitamins, vaccines, antibodies and the like, provides a technical basis for industrially producing and preparing wheat peptone products with high dispersibility and high solubility, and simultaneously prolongs the fine and deep processing industrial chain of wheat resources and improves the economic benefits of the wheat resources.

Description

Industrial preparation method and application of wheat peptone

Technical Field

The invention relates to the field of agricultural and sideline product deep processing and fermentation engineering, in particular to an industrial preparation method of wheat peptone and application thereof.

Background

Peptone is a water-soluble mixture obtained by utilizing proteolysis, and mainly comprises peptone, peptide and amino acid. Peptone can be classified from a source into animal-derived peptone, plant-derived peptone and microbial peptone. The conventional fermentation industry, antibiotic enterprises and part of biological pharmaceutical enterprises mainly adopt peptone from which casein is taken as a source to carry out daily production. However, animal-derived peptones have significant disadvantages, such as microbial contamination due to Rana virus, etc., large batch-to-batch variation, undefined components, and difficulty in separation and purification of target products such as vaccines and monoclonal antibodies. In the field of microbial fermentation, in particular, the use safety concerns of the biopharmaceutical industry (biological products such as vaccines, antibodies, interferon and the like) on meat products and animal-derived peptone are increasing, and related production enterprises are required to provide raw material safety certificates during use. And the non-animal-derived culture medium has no risk of polluting a target product, has higher safety guarantee, and is one of the priorities in the fields of biological pharmacy and the like.

Wheat protein is a wheat starch processing byproduct, has a protein content of 72-85%, and is widely applied to bread baking, noodle making, meat product processing, cosmetics, feed industry and the like. The wheat protein resource in China is rich, the price of food-grade wheat protein in China is 9000-10000 yuan/ton generally at present, is lower than that of casein, whey protein and soybean protein isolate, and is a food protein resource with moderate and lower added value. Meanwhile, wheat protein has poor solubility, which limits the application of wheat protein in food and fermentation industries.

At present, the research on wheat protein at home and abroad mainly focuses on the aspect of active polypeptide. For example, CN105331663A discloses a method for preparing low-bitter peptide powder by enzymatic hydrolysis of wheat gluten, wherein wheat gluten (wheat gluten) is hydrolyzed and dried by protease to obtain peptide powder with low bitter value, good palatability and good flavor, which is expected to be applied in the fields of food and feed; CN104263794A discloses that wheat gluten (wheat gluten) peptide is prepared by adopting an ultrasonic-assisted enzymolysis and aerobic-anaerobic two-step fermentation process, and the obtained wheat gluten peptide product has white color, powder shape, no impurity, no bitter taste and certain antioxidant activity; CN103484246A discloses that after wheat gluten protein (vital gluten) is subjected to enzymolysis, amino acid and oligopeptide with higher proportion content are obtained, and the wheat gluten protein has strong delicate flavor and can be applied to improving the delicate flavor intensity of essence; CN102911995A discloses a process for preparing wheat peptone from pulverized whole wheat flour as a raw material by enzymolysis at a relatively low temperature, but a large amount of starch in wheat increases the difficulty of filtration at the later stage and high-temperature cooking cannot be used.

Based on the current research situation, the invention provides a method for efficiently producing high-quality wheat peptone by fusion processing of wheat protein by multiple technologies. The method maximally converts the wheat protein resource into a high-quality peptone product which has excellent solubility and good dispersibility and is suitable for microbial growth and fermentation, expands the industrial chain of deep processing of wheat, promotes the additional value of wheat protein, meets the market demand, and provides a new option for processing and producing high-value products by utilizing low-valued plant protein resources.

Disclosure of Invention

The invention provides an industrial preparation method of wheat peptone, which is characterized in that the content of protein in the wheat peptone prepared by the method is 60-90%; contains 18 amino acids, wherein the content of glutamine/glutamic acid is more than 25%; the content of peptide fragment with molecular mass less than 1000Da is more than 80 percent; the method can completely or partially replace a nitrogen source in a microorganism growth/fermentation culture medium, is used for producing fermentation products such as probiotics, antibiotics, enzyme preparations, amino acids, organic acids, vitamins, vaccines, antibodies and the like, provides a technical basis for industrially producing and preparing the wheat peptone product with high dispersibility and high solubility, and simultaneously prolongs the fine and further processing industrial chain of wheat resources and improves the economic benefit of the wheat resources.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the present invention provides an industrial preparation method of wheat peptone, which comprises the following steps:

(1) taking wet-based wheat protein as a raw material, crushing and homogenizing the wet-based wheat protein, and then performing enzymolysis on the wet-based wheat protein by adding protease and amylase;

(2) and filtering the hydrolyzed wheat protease liquid, and carrying out steaming, spraying, enzyme deactivation and spray drying to obtain the wheat peptone.

The wet base wheat protein in the present invention means "wet gluten"; the crushing in the step (1) adopts high-efficiency crushing treatment, and the treatment is favorable for full enzymolysis of wet-based wheat protein.

The preparation method can realize the maximized transformation of wheat protein resources into peptone products which have excellent solubility and good dispersibility and are suitable for microbial growth and fermentation, expand the industrial chain of wheat deep processing and improve the additional value of wheat protein.

Preferably, the wheat protein material in step (1) is derived mainly from: the wheat starch processing byproduct, or the wet-based wheat protein obtained by processing the wheat into special wheat flour and separating the starch and the protein in the wheat flour through a cyclone process.

The invention uses the wet wheat protein (wet gluten) as the raw material, reduces the drying link, reduces the production cost, reduces the use of water, fully maintains the inherent activity of the wheat protein, and obviously improves the comprehensive utilization level and the application value of the wheat protein; meanwhile, the wheat peptone prepared by using the wheat protein as a raw material has high quality, and realizes the maximized transformation of wheat protein resources into peptone products which have excellent solubility and good dispersibility and are suitable for microbial growth and fermentation.

In the invention, before the wheat protein raw material is hydrolyzed by an enzyme system, the wheat protein raw material needs to be added with water to prepare a solution with the concentration of 10-300 g/L, such as 10g/L, 20g/L, 50g/L, 100g/L, 120g/L, 150g/L, 200g/L, 220g/L, 250g/L or 300 g/L; adding 0.1-1.0 mol/L (such as 0.1mol/L, 0.5mol/L, 1.0mol/L) HCl solution or 0.5-2 mol/L (such as 0.5mol/L, 1.0mol/L, 2.0mol/L) NaOH solution to adjust the pH value of the wheat protein solution to 3.5-6.5, 6.5-7.5 or 7.5-11.0, so as to facilitate enzymolysis.

Preferably, the protease is any one of acid protease, neutral protease or alkaline protease; wherein the pH value of the acid protease is 3.5-6.5, such as 3.5, 4, 4.5, 5, 6 or 6.5, and the enzymolysis temperature is 40-60 ℃, such as 40 ℃, 42 ℃, 45 ℃, 50 ℃, 55 ℃ or 60 ℃; the neutral protease has an enzymolysis pH value of 6.5-7.5, such as 6.5, 7, 7.1, 7.2 or 7.5, and an enzymolysis temperature of 40-60 ℃, such as 40 ℃, 42 ℃, 45 ℃, 50 ℃, 55 ℃ or 60 ℃; the alkaline protease has an enzymolysis pH of 7.5-11.0, such as 7.5, 8, 8.5, 9, 10.5 or 11, and an enzymolysis temperature of 40-60 deg.C, such as 40 deg.C, 42 deg.C, 45 deg.C, 50 deg.C, 55 deg.C or 60 deg.C.

Preferably, the amylase is a moderate temperature amylase or a high temperature amylase; wherein the enzymolysis temperature of the medium-temperature amylase is 50-90 ℃, such as 50 ℃, 52 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃, 80 ℃ or 90 ℃, and the enzymolysis pH value is 5.0-8.0, such as 5.0, 5.5, 6, 7, 7.5 or 8; the enzymolysis temperature of the high-temperature amylase is 90-110 ℃, such as 90 ℃, 92 ℃, 95 ℃, 100 ℃, 105 ℃ or 110 ℃, and the enzymolysis pH value is 4.0-8.0, such as 4.0, 4.5, 5, 7, 7.5 or 8.

In the invention, the enzymolysis is carried out by adding protease and then adding amylase, which is beneficial to the full enzymolysis of wheat protein; meanwhile, aiming at different proteases or amylases, the pH value, the enzymolysis temperature and the enzymolysis time of a reaction system need to be adaptively adjusted during enzymolysis, for example, when acid protease is added, the pH value of the reaction system needs to be adjusted to 3.5-6.5, the enzymolysis temperature is 40-60 ℃, and the enzymolysis time is 0.5-3.0 h; when neutral protease is added, the pH value of the reaction system needs to be adjusted to 6.5-7.5, the enzymolysis temperature is 40-60 ℃, and the enzymolysis time is 0.5-3.0 h; when the medium-temperature amylase is added, the enzymolysis temperature needs to be adjusted to be 50-90 ℃, the pH value needs to be 5.0-8.0, and the enzymolysis time needs to be 0.5-5.0 h.

Preferably, the filtration in the step (2) is to remove solid matters from the wheat protease hydrolysate obtained by hydrolysis through a separation system; wherein the separation system comprises a plate and frame filtration system or a membrane module separation system.

Preferably, the plate-and-frame filtration system is a plate-and-frame filtration system containing porous ceramic, plastic or sintered metal media; the membrane component separation system is a membrane component separation system with molecular weight cutoff of 5kDa and 1 kDa.

Preferably, the cooking temperature in step (2) is 80-100 ℃, for example 80 ℃, 90 ℃ or 100 ℃, preferably 90-100 ℃.

Preferably, the temperature of the spraying treatment in the step (2) is 100 to 180 ℃, for example 100 ℃, 110 ℃, 115 ℃, 120 ℃, 135 ℃, 145 ℃, 150 ℃, 162 ℃ or 180 ℃, preferably 120 to 170 ℃; the time is 3 to 50s, for example 3s, 5s, 10s, 15s, 20s, 30s, 40s or 50s, preferably 5 to 15 s.

In the invention, the pH value of the wheat protease hydrolysate is adjusted to 5.0-8.0, and the wheat protease hydrolysate is cooked for 0.5-3 h at 80-100 ℃ in a reaction tank; the injection treatment is carried out by treating the wheat protein enzymatic hydrolysate steamed in the reaction tank for 3-50s by using a high-temperature injection device. By adopting the cooking treatment at 80-100 ℃ and the spraying treatment for 3-50s, the surface property, the solubility and the dispersibility of the composite material can be effectively improved.

Preferably, the conditions of the spray drying in step (2) are: the concentration of a feed sample is 10-40% (w/v); the air inlet temperature is 120-220 ℃; the hot air flow rate is 10-50 m³/h。

The industrial preparation method of the wheat peptone specifically comprises the following steps:

(1) crushing and homogenizing a wet-based wheat protein raw material, adding water to prepare a solution with the concentration of 10-300 g/L, adding 0.1-1.0 mol/L HCl solution or 0.5-2 mol/L NaOH solution to adjust the pH value of the wheat protein solution to 3.5-6.5, 6.5-7.5 or 7.5-11.0;

(2) adding acid protease, neutral protease or alkaline protease, respectively 0.5-500U/g of substrate, adjusting the temperature of a reaction system to 40-60 ℃, and performing enzymolysis for 0.5-3.0 h to obtain protease hydrolysate of wheat protein;

(3) adding medium-temperature amylase or high-temperature amylase which is 0.1-50U/g of substrate respectively, setting the enzymolysis temperature to be 50-100 ℃, and carrying out enzymolysis for 0.5-5.0 h to obtain protease-amylase hydrolysate;

(4) removing solid matters from protease-amylase hydrolysate of wheat protein by a separation system; wherein the separation system comprises a disc centrifuge, a plate-and-frame filtration system containing porous ceramic/plastic/sintered metal medium or a membrane component separation system with molecular weight cutoff of 5kDa and 1 kDa;

(5) the protease-amylase hydrolysate of the filtered wheat protein is subjected to cooking treatment at 80-100 ℃ to inactivate enzyme and sprayedSpraying for 3-50s, and spray drying to obtain the wheat peptone; wherein the spray drying conditions are as follows: the concentration of a feed sample is 10-40% (w/v); the air inlet temperature is 120-220 ℃; the hot air flow rate is 10-50 m³/h。

In a second aspect, the invention also provides wheat peptone prepared by the method of the first aspect.

The wheat peptone prepared by the invention has the following characteristics: the appearance is light yellow powder; the protein content is 60-90%; contains 18 amino acids, wherein the content of glutamine/glutamic acid is more than 25%; the content of peptide fragments with molecular mass of more than 5000Da is less than 5 percent; the content of peptide fragments with the molecular mass of 1000-5000 Da is less than 15 percent; the content of peptide fragment with molecular mass less than 1000Da is more than 80 percent; the content of reducing sugar is 2.0-10%; the dissolution performance is good, and the dissolution rate is less than 30 min; the dispersion degree of 60s is 70-100%, and the dispersion degree of 20s is 50-90%; the appearance of the solution is light yellow, clear and bright.

In a third aspect, the present invention also provides the use of wheat peptone as described in the second aspect, for total or partial replacement of total nitrogen source in laboratory or large-scale production medium of probiotics, antibiotics, enzyme preparations, amino acids, organic acids, vitamins, vaccines or antibodies.

In particular, the wheat peptone can be applied in the following fields:

1) the prepared wheat peptone totally replaces probiotic bacteria agents (such as: the total nitrogen source (two or more of peptone, yeast powder/paste, beef extract and the like are added simultaneously) in the laboratory or large-scale production culture medium of Lactococcus lactis subsp lactis, Lactobacillus reuteri, Lactobacillus plantarum (Lactobacillus reuteri), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) and Streptococcus lactis (Streptococcus lactis) and the like, and has the same or higher viable count and/or yield of probiotics under the condition of the original nitrogen source.

Or replace animal, microorganism or other plant source peptones in laboratory or large-scale production culture medium of probiotic bacteria (such as Lactococcus lactis subsp lactis, Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus casei, Streptococcus lactis, etc.), and has the same or higher viable count and/or yield of probiotics under the condition of original nitrogen source.

2) The prepared wheat peptone was used in the same amount as the whole in place of antibiotics (e.g.: natamycin, nisin, abamectin, polymyxin, etc.) has the same or higher potency and/or yield of antibiotics under the condition of the original nitrogen source (two or more peptone, yeast powder/extract, beef extract, etc. are added simultaneously).

Or replacing antibiotics (such as natamycin, nisin, abamectin, polymyxin and the like) in the peptone of animal, microorganism or other plant sources in a laboratory or large-scale production culture medium, and has the same or higher titer and yield of the antibiotics under the condition of a primary nitrogen source.

3) The prepared wheat peptone was used in the same amount as the whole in place of the enzyme preparation (e.g.: cellulase, xylanase, amylase, glucoamylase, protease, and the like) in a laboratory or in a large-scale production medium (two or more of peptone, yeast powder/extract, beef extract, and the like are added simultaneously), and the activity and/or the yield of the enzyme preparation under the condition of the original nitrogen source are equal to or higher than those of the enzyme preparation.

Or animal, microorganism or other plant-derived peptones in laboratory or large-scale production media in place of enzyme preparations (e.g., cellulases, xylanases, amylases, saccharifying enzymes, proteases, etc.) with activity and/or yield equal to or higher than that of enzyme preparations under the conditions of the original nitrogen source.

4) The prepared wheat peptone was used in the same amount as the whole to replace animal or plant vaccines (e.g.: brucella, etc.) or in a large-scale production medium (two or more peptone, yeast powder/extract, beef extract, etc. are added simultaneously), has the same or higher activity and/or yield than the vaccine under the original nitrogen source condition.

Or replace animal, microorganism or other plant source peptone in the laboratory or large-scale production culture medium of animal or plant vaccines such as Brucella, and the like, and has the same or higher activity and/or yield than the vaccine under the original nitrogen source condition.

5) The prepared wheat peptone was used in the same amount as that used in place of all monoclonal antibodies (e.g.: recombinant escherichia coli laboratory or large-scale production of monoclonal antibodies such as beta-galactosidase and the like) laboratory or large-scale production culture medium (two or more than two peptone, yeast powder/extract, beef extract and the like are added simultaneously), and the expression quantity and/or yield of the monoclonal antibodies under the condition of the original nitrogen source are equal to or higher than those of the monoclonal antibodies under the condition of the original nitrogen source.

Or replacing animal, microorganism or other plant source peptone in the laboratory or scale production culture medium of the monoclonal antibody (such as recombinant Escherichia coli laboratory or scale production of the monoclonal antibody such as beta-galactosidase, etc.), and the expression quantity and/or the yield of the monoclonal antibody are equal to or higher than those of the monoclonal antibody under the condition of a raw nitrogen source.

6) The prepared wheat peptone was used in the same amount as used in place of all the organic acids (e.g.: citric acid, lactic acid, alpha-ketoglutaric acid, pyruvic acid, etc.), amino acids (such as: glutamic acid, arginine, lysine, etc.), vitamins (e.g.: vitamin C, vitamin B2, vitamin B12, and the like) and the like, and has the same or higher yield than the fermentation product under the condition of the original nitrogen source.

Or replacing organic acid (such as citric acid, lactic acid, alpha-ketoglutaric acid, pyruvic acid, and the like), amino acid (such as glutamic acid, arginine, lysine, and the like), vitamin (such as vitamin C, vitamin B2, vitamin B12, and the like), and peptone of animal, microorganism or other plant sources in a laboratory or large-scale production culture medium of fermentation products such as the fermentation products has the same or higher yield of the fermentation products under the original nitrogen source condition.

Compared with the prior art, the invention has at least the following beneficial effects:

the preparation method provided by the invention can utilize plant proteins such as wheat protein with low added value as raw materials, convert the plant proteins into biochemical reagent grade peptone with high added value to the maximum extent, and has the same or higher microbial thallus growth efficiency and titer and yield of fermentation products under the condition of original nitrogen source when the nitrogen source is completely or partially replaced by the microbial growth and fermentation nitrogen source, thereby obviously improving the comprehensive utilization level and application field of the wheat protein.

Detailed Description

For the purpose of facilitating an understanding of the present invention, the present invention will now be described by way of examples. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.

Example 1

(1) Taking a wheat starch processing byproduct as a wet-based wheat protein raw material, crushing, homogenizing, adding water to prepare a solution with the concentration of 300g/L, and adding 0.1mol/L HCl solution or 0.5mol/L NaOH solution to adjust the pH value of the wheat protein solution to 4.0; adding acid protease with the addition of 50U/g substrate, adjusting the temperature of a reaction system to 55 ℃, and performing enzymolysis for 2.0 hours to obtain protease hydrolysate; adding high-temperature amylase, adding 15U/g substrate, adjusting enzymolysis pH to 6.0, performing enzymolysis at 90 deg.C for 1.0h to obtain protease-amylase hydrolysate;

(2) removing solid matters by adopting a membrane component separation system with molecular weight cutoff of 5kDa and 1 kDa; the filtered protease-amylase hydrolysate of the wheat protein is subjected to cooking treatment at 100 ℃ and spraying treatment for 5s, and the concentration of feed protein is 30% (w/v), the inlet air temperature is 160 ℃ and the hot air flow is 20m³Spray drying is carried out under the condition of/h, and the wheat peptone with light yellow powder appearance is obtained.

The wheat peptone obtained by the preparation has the protein content of 80 percent; contains 18 kinds of amino acids, wherein the content of glutamine/glutamic acid is 35%; the peptide segment content with the molecular mass of more than 5000Da is 1.0 percent; the peptide segment content with the molecular mass of 1000 Da-5000 Da is 13.0%; the content of peptide with the molecular mass of less than 1000Da is more than 87 percent; the reducing sugar content is 8%; the dissolution rate is 15 min; the dispersion degree of 60s is 100 percent, and the dispersion degree of 20s is 90 percent; the appearance of the solution is light yellow, clear and bright.

Example 2

(1) Processing wheat into special wheat flour, separating starch and protein in the wheat flour by a cyclone process to obtain a wet-based wheat protein raw material, crushing, homogenizing, adding water to prepare a solution with the concentration of 200g/L, and adding 0.1mol/L HCl solution or 0.5mol/L NaOH solution to adjust the pH value of the wheat protein solution to 7.0; adding neutral protease with the addition amount of 100U/g substrate, adjusting the temperature of the reaction system to 60 ℃, and performing enzymolysis for 1.0h to obtain protease hydrolysate; then adding medium temperature amylase, adding 30U/g substrate, adjusting enzymolysis pH to 6.5, hydrolyzing at 70 deg.C for 1.0h to obtain protease-amylase hydrolysate;

(2) removing solid matters by adopting a high-speed disc centrifuge and a plate-frame filtering system of porous ceramic medium; the protease-amylase hydrolysate of the filtered wheat protein is subjected to cooking treatment at 90 ℃ and spraying treatment for 15 s; at a feed protein concentration of 20% (w/v), an inlet air temperature of 120 ℃ and a hot air flow rate of 40m³Spray drying is carried out under the condition of/h, and the wheat peptone with light yellow powder appearance is obtained.

The wheat peptone obtained by the preparation has the protein content of 75 percent; contains 18 kinds of amino acids, wherein the content of glutamine/glutamic acid is 30%; the peptide segment content with the molecular mass of more than 5000Da is 2.5 percent; the peptide segment content with the molecular mass of 1000 Da-5000 Da is 14.0%; the content of peptide fragment with molecular mass less than 1000Da is more than 83.5 percent; the content of reducing sugar is 10 percent; the dissolution rate is 20 min; the dispersion degree of 60s is 90%, and the dispersion degree of 20s is 80%; the appearance of the solution is light yellow, clear and bright.

Example 3

(1) Taking a wheat starch processing byproduct as a wet-based wheat protein raw material, crushing, homogenizing, adding water to prepare a solution with the concentration of 150g/L, and adding 0.1mol/L HCl solution or 0.5mol/L NaOH solution to adjust the pH value of the wheat protein solution to 10.0; adding alkaline protease with the addition amount of 150U/g substrate, adjusting the temperature of the reaction system to 60 ℃, and performing enzymolysis for 3.0h to obtain protease hydrolysate; adding high-temperature amylase, adding 40U/g substrate, adjusting enzymolysis pH to 6.0, performing enzymolysis at 95 deg.C for 3.0h to obtain protease-amylase hydrolysate;

(2) removing solid matters by adopting a membrane component separation system with molecular weight cutoff of 5kDa and 1 kDa; the protease-amylase hydrolysate of the filtered wheat protein is subjected to cooking treatment at 80 ℃ and spraying treatment for 5 s; at a feed protein concentration of 25% (w/v), an inlet air temperature of 200 ℃ and a hot air flow rate of 50m³Spray drying is carried out under the condition of/h, and the wheat peptone with light yellow powder appearance is obtained.

The wheat peptone obtained by the preparation has the protein content of 85 percent; contains 18 amino acids, wherein the content of glutamine/glutamic acid is 27%; the content of peptide fragments with the molecular mass of more than 5000Da is 0.5 percent; the peptide segment content with the molecular mass of 1000 Da-5000 Da is 10.0%; the content of peptide fragment with molecular mass less than 1000Da is more than 89.5 percent; the reducing sugar content is 8%; the dissolution rate is 10 min; the dispersity of 60s is 100%, and the dispersity of 20s is 85%; the appearance of the solution is light yellow, clear and bright.

Example 4

The wheat peptone prepared in the example 1 partially replaces tryptone in the modified MRS culture medium, and the addition amount is 10 g/L; the other components and contents are kept unchanged, and are respectively 10g/L of beef extract, 5g/L of yeast extract and K₂HPO₄·3H₂O₂2g/L, 5g/L sodium acetate, 20g/L glucose, 801 mL/L Tween-801, 2g/L diammonium hydrogen citrate, MgSO₄·7H₂O 0.58g/L，MnSO₄·H₂O0.25 g/L; after adjusting the pH to 6.5, the medium was sterilized at 121 ℃ for 20 min. Inoculating 3% (v/v) activated Lactococcus lactis subsp lactis, Lactobacillus reuteri, Lactobacillus plantarum and Lactobacillus casei seed solutions of lactic acid bacteria, standing and culturing at 37 deg.C for 12h to obtain the final product with OD600nm values of 1.95, 1.87, 1.78 and 2.32, and viable count of 1.64 × 10⁹CFU/m，2.45×10⁹CFU/m，1.76×10⁹CFU/m and 1.88X 10⁹CFU/mL, allThe growth performance of the lactobacillus is more than 8 percent higher than that of the prior modified MRS culture medium.

Example 5

The wheat peptone prepared in example 2 was partially substituted for the enzyme production medium (microcrystalline cellulose 10g/L, tryptone 1g/L, yeast extract 1g/L, KH) of Trichoderma reesei (Trichoderma reesei)₂PO₄ 2g/L、CaCl₂ 0.3g/L、(NH₄)₂SO₄1.4g/L、MgSO₄0.3g/L, tween-801 mL/L and trace elements 1mL/L), the addition amount of the nitrogen sources such as peptone and the like is 1.0g/L, the fermentation is carried out for 5 days at the temperature of 28 ℃, the filter paper enzyme activity produced by the fermentation of trichoderma reesei is 49U/mL, the CMC enzyme activity is 18U/mL, the xylanase activity is 303U/mL, and the enzyme production performance is obviously higher than that of the original trichoderma reesei enzyme production culture medium by more than 10%.

Example 6

The wheat peptone prepared in the example 3 partially replaces a yeast extract nitrogen source in a fermentation medium (30 g/L of glucose, 2g/L of yeast extract, 10g/L of beef extract, 1g/L of NaCl and 7.0 of pH) of streptomyces natalensis, the addition amount is 2g/L, and other components and contents are kept unchanged; inoculating with 6% inoculum size, and culturing at 28 deg.C and 180r/min for 108 h; the yield of the natamycin reaches 612mg/L, which is 37 percent higher than that of a control group under the original culture medium condition.

Example 7

The wheat peptone prepared in example 1 is used for replacing peptone in a growth improved Martin medium (tryptone 5.0g/L, dipotassium hydrogen phosphate 1.0g/L, yeast extract powder 2.0g/L, magnesium sulfate 0.5g/L and glucose 20.0g/L) of a Brucella S2 strain, the addition amount is 5.0g/L, and other components and contents are kept unchanged; inoculating seed liquid of Brucella S2 strain at 1% inoculation amount, culturing at 28 deg.C and 150r/min for 36 hr to obtain viable count of 4.06 × 10⁹CFU/mL is 22% higher than that of the control group under the original culture medium condition, and has the function of promoting the proliferation of the bacteria.

Comparative example 1

The procedure of example 1 was followed, except that the multi-enzyme system of protease and amylase was replaced with a single acid protease, i.e., the high temperature amylase was omitted, as compared with example 1.

The wheat peptone prepared by the comparative example has a protein content of 75%; contains 18 kinds of amino acids, wherein the content of glutamine/glutamic acid is 32%; the peptide fragment content with the molecular mass of more than 5000Da is 6.8 percent; the peptide segment content with the molecular mass of 1000 Da-5000 Da is 17.0%; the content of peptide fragments with the molecular mass of less than 1000Da is less than 75 percent; the reducing sugar content is 8%; the dissolution rate is 25 min; the dispersity for 60s is 85%, and the dispersity for 20s is 72%; the appearance of the solution is dark yellow, and the appearance of the solution is light yellow, clear and bright.

The wheat peptone prepared in example 1 used in example 4 was replaced with the wheat peptone prepared in this comparative example, and the OD600nm values of the 4 lactic acid bacteria obtained were 1.35, 1.12, 0.96 and 1.44, respectively, and the viable count reached 7.17X 10, respectively⁸CFU/m，6.77×10⁸CFU/m，5.59×10⁸CFU/m and 3.14X 10⁸CFU/mL, the growth performance of the lactobacillus is obviously reduced compared with that of the lactobacillus in example 4.

Comparative example 2

The same as example 1, except that the multi-enzyme system of protease and amylase was replaced with a single high temperature amylase, i.e., the acid protease was omitted, as compared to example 1.

The wheat peptone prepared by the comparative example has a protein content of 65%; contains 18 amino acids, wherein the content of glutamine/glutamic acid is 28%; the peptide segment content with the molecular mass of more than 5000Da is 12.4 percent; the peptide segment content with the molecular mass of 1000 Da-5000 Da is 20.5%; the content of peptide with molecular mass less than 1000Da is less than 70 percent; the reducing sugar content is 15%; the dissolution rate is 25 min; the dispersity of 60s is 80%, and the dispersity of 20s is 70%; the solution was pale yellow in appearance, turbid and a small amount of insoluble material precipitated.

The wheat peptone prepared in example 1 used in example 4 was replaced with the wheat peptone prepared in this comparative example, and the OD600nm values of the 4 lactic acid bacteria obtained were 1.04, 0.87, 0.96 and 1.12, respectively, and the viable count reached 5.43X 10, respectively⁸CFU/m，4.69×10⁸CFU/m，6.75×10⁸CFU/m and 4.37X 10⁸CFU/mL, lactic acid bacteria growth performance compared with example 4 allAnd is significantly reduced.

Comparative example 3

The same as example 1 except that the steaming treatment and the spraying treatment were omitted, as compared with example 1.

The dissolution rate of the wheat peptone prepared by the comparative example is 35 min; the dispersity for 60s was 87% and for 20s 74%, which was significantly lower than that of example 1.

The wheat peptone prepared in example 1 used in example 4 was replaced with the wheat peptone prepared in this comparative example, and the OD600nm values of the 4 lactic acid bacteria obtained were 1.63, 1.57, 1.29 and 2.01, respectively, and the viable count reached 1.04X 10, respectively⁹CFU/m，1.85×10⁹CFU/m，1.35×10⁹CFU/m and 1.06X 10⁹CFU/mL, lactic acid bacteria growth performance compared with example 4 uniform degree of decline.

The results show that the wheat peptone which can completely or partially replace a nitrogen source in a microorganism growth/fermentation culture medium is obtained by using plant proteins such as wheat protein with low added value as raw materials and hydrolyzing the plant proteins by using a multienzyme system, then filtering the wheat protease hydrolysate obtained by hydrolysis, and performing cooking, spraying treatment and spray drying.

The applicant states that the present invention is illustrated by the above examples to show the detailed process equipment and process flow of the present invention, but the present invention is not limited to the above detailed process equipment and process flow, i.e. it does not mean that the present invention must rely on the above detailed process equipment and process flow to be implemented. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Claims

1. An industrial preparation method of wheat peptone, which is characterized by comprising the following steps:

(1) taking wet-based wheat protein as a raw material, crushing, homogenizing, and then performing enzymolysis on the wet-based wheat protein by adopting a mode of firstly adding protease and then adding amylase;

(2) filtering the hydrolyzed wheat protease liquid, steaming at 80-100 ℃, performing spray treatment at 100-180 ℃ for 3-50s to inactivate enzyme, and performing spray drying to obtain the wheat peptone;

the wet-based wheat protein raw material in the step (1) is prepared from the following raw materials in parts by weight: processing wheat starch into byproduct or processing wheat into special wheat flour, and separating starch and protein in the wheat flour by cyclone process to obtain wet-based wheat protein;

the enzymolysis mode in the step (1) is as follows: adding acid protease, neutral protease or alkaline protease, namely 50-150U/g of substrate, adjusting the temperature of a reaction system to 55-60 ℃, and performing enzymolysis for 1-3.0 hours to obtain protease hydrolysate of wheat protein; adding medium-temperature amylase or high-temperature amylase which is 15-40U/g of substrate respectively, setting the enzymolysis temperature to be 70-95 ℃, and carrying out enzymolysis for 1-3 hours to obtain protease-amylase hydrolysate;

the characteristics of the wheat peptone are as follows: the appearance is light yellow powder; the protein content is 75-85%; contains 18 amino acids, wherein the content of glutamine/glutamic acid is more than 25%; the content of peptide fragments with molecular mass of more than 5000Da is less than 5 percent; the peptide segment content with the molecular mass of 1000 Da-5000 Da is less than 15 percent; the content of peptide fragment with molecular mass less than 1000Da is more than 80 percent; the content of reducing sugar is 2.0-10%; dissolution rate <30 min; the dispersion degree of 60s is 70-100%, and the dispersion degree of 20s is 50-90%.

2. The method according to claim 1, wherein the filtration in the step (2) is to remove solids from the hydrolyzed wheat protease hydrolysate by a separation system.

3. The method of claim 2, wherein the separation system comprises a disk centrifuge, a plate and frame filtration system, or a membrane module separation system.

4. The method of claim 3, wherein the plate and frame filter system is a plate and frame filter system comprising a porous ceramic, plastic, or sintered metal media.

5. The method of claim 3, wherein the disk centrifuge is an automatic discharge centrifuge.

6. The method of claim 3, wherein the membrane module separation system is a 5kDa and 1kDa molecular weight cut-off membrane module separation system.

7. The method of claim 1, wherein the cooking temperature in step (2) is 90-100 ℃.

8. The method according to claim 1, wherein the temperature of the spraying treatment is 100 to 180 ℃ and the time is 5 to 15 seconds.

9. The method of claim 1, wherein the spray drying conditions are: the concentration of a feed sample is 10-40% (w/v); the air inlet temperature is 120-220 ℃; the hot air flow rate is 10-50 m³/h。

10. Wheat peptone prepared by the process according to any one of claims 1 to 9.

11. Wheat peptone as claimed in claim 10, characterized in that it is: the appearance is light yellow powder; the protein content is 75-85%; contains 18 amino acids, wherein the content of glutamine/glutamic acid is more than 25%; the content of peptide fragments with molecular mass of more than 5000Da is less than 5 percent; the peptide segment content with the molecular mass of 1000 Da-5000 Da is less than 15 percent; the content of peptide fragment with molecular mass less than 1000Da is more than 80 percent; the content of reducing sugar is 2.0-10%; dissolution rate <30 min; the dispersion degree of 60s is 70-100%, and the dispersion degree of 20s is 50-90%.

12. Use of wheat peptone as claimed in claim 10, characterized in that said wheat peptone replaces totally or partially the total nitrogen source in laboratory or scale production media of probiotics, antibiotics, enzyme preparations, amino acids, organic acids, vitamins, vaccines or antibodies.

13. The use according to claim 12, wherein the probiotic bacteria are any of lactococcus lactis subsp.

14. The use of claim 12, wherein the antibiotic is any one of natamycin, nisin, abamectin or polymyxin.

15. The use of claim 12, wherein the enzyme preparation is any one of cellulase, xylanase, amylase, glucoamylase or protease.

16. The use of claim 12, wherein the amino acid is any one of glutamic acid, arginine or lysine.

17. Use according to claim 12, wherein the organic acid is any one of citric acid, lactic acid, α -ketoglutaric acid or pyruvic acid.

18. The use of claim 12, wherein the vitamin is any one of vitamin C, vitamin B2, or vitamin B12.