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Method for preparing vegetable seed peptide with single bioactivity by mix bacterium solid state fermentation

A solid-state fermentation and biological activity technology, applied in microorganism-based methods, peptide preparation methods, biochemical equipment and methods, etc., can solve the problems of destroying the original configuration of amino acids, limited sources of enzymes, and severe reaction conditions. Achieve the effects of being beneficial to environmental protection, saving biological enzyme preparations, and simple production process

Inactive Publication Date: 2011-06-29
NANJING UNIV OF FINANCE & ECONOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chloropropanol carcinogens will be produced when chemical methods acid or alkali hydrolyze vegetable protein, and the reaction conditions are severe, destroying the original configuration of amino acids, and the discharge of hydrolysis waste liquid will also cause certain environmental pollution
Enzymatic hydrolysis is currently the main method for the preparation of rapeseed peptides, but since the anti-nutritional factors in rapeseed meal or rapeseed protein isolate cannot be completely removed, the activity of protease is affected, the degree of hydrolysis is not high, and the flavor of rapeseed protein hydrolyzate In particular, the problem of bitter taste has not been fundamentally resolved. At the same time, the sources of enzymes used for the hydrolysis of rapeseed protein are limited, and the cost remains high. Enzymatic preparation of rapeseed peptides with specific biological activity is limited.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Screening of strains for the production of biologically active rapeseed peptides by solid-state fermentation of rapeseed meal

[0020] Screening objects: Aspergillus niger, Geotrichum candidum, Aspergillus oryzae, Aspergillus usami, Actinomucor elegans, lactic acid bacteria, Candida utilis, Bacillus licheniformis and Bacillus subtilis.

[0021] Preparation of starter: after the expanded culture of the activated bacterial classification (cultivated by a conventional method), dilute the fermented seeds with sterile water to make the concentration of thalline or spores reach 3×10 7 Each / mL is the starter.

[0022] The double-low rapeseed meal was pulverized to a particle size of about 500 μm, and sterilized by moist heat at 121°C for 30 minutes. In the fermenter, the starter is added according to the solid-liquid ratio of 1:1-1:2 (mass ratio), the fermentation temperature is 28°C-35°C, the ambient humidity is 80%-90%, stirring and ventilating. After 3 days of f...

Embodiment 2

[0024] Example 2 Preparation of Bacillus subtilis suspension with specific biological activity of rapeseed peptide produced by solid-state fermentation of mixed bacteria of Bacillus subtilis and Mucor radiata:

[0025] Incline medium: beef extract 3g, peptone 10g, NaCl 5, agar 20g, tap water 1000mL, pH 7.2-7.4.

[0026] Seed medium: beef extract 3g, peptone 10, NaCl 5g, tap water 1000mL, pH 7.2-7.4.

[0027] Nutrient solution composition: glucose 2.6g, KH 2 PO 4 2.6g, sterile water 1000mL, pH 7.0.

[0028] Pick two rings of Bacillus subtilis from the activated strain preservation slant and inoculate it in the seed medium, seal it with 8 layers of gauze, culture it on a shaker at 35°C, 120r / min for 24 hours, and the concentration of bacteria reaches 10 8 individual / mL.

[0029] Dilute the fermented liquid with sterile nutrient solution to prepare the cell concentration of 3×10 7 Individual / mL suspension of Bacillus subtilis.

[0030] Preparation of Mucor actinosa suspensi...

Embodiment 3

[0043] Example 3 Preparation of Rapeseed Peptide Suspension of Mucor actinosa and Candida utilis Synchronous Mixed Bacteria Solid-State Fermentation Production:

[0044] Slant medium: 300g potato, 20g glucose, 20g agar, 1000mL tap water.

[0045] Plate expansion medium: 300g potato, 20g glucose, 20g agar, 1000mL tap water.

[0046] Sterile nutrient solution: glucose 5g, KH 2 PO 4 2g, 1000mL sterile water, pH 6.5.

[0047] Mucor actinosa was inoculated on a slant medium and cultured at a constant temperature of 28°C for 4 days, and then inoculated on a flat plate for expansion culture. After 4 days, the spores were washed with sterile nutrient solution, and the concentration was adjusted to 3×10 7 individual / mL.

[0048] Preparation of Candida utilis suspension:

[0049] Incline medium: 3g of malt extract, 10g of glucose, 3g of yeast extract, 5g of peptone, 20g of agar, 1000mL of tap water.

[0050] Seed medium: 3g of malt extract, 10g of glucose, 3g of yeast extract, 5g ...

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Abstract

The invention provides a method for preparing rapeseed peptide with uniform biological activity by means of mixed solid-state fermentation. In the method, one of bacillus subtilis, lactobacillus or candida utili is combined with actinomucor elegans or aspergillus usamii to prepare leaven; the leaven is inoculated in smashed rapeseed dregs for carrying out the solid-state fermentation; culture medium obtained after the solid-state fermentation is extracted to obtain crude extract of the rapeseed peptide; the crude extract is separated, purified, sterilized and dried to obtain the rapeseed peptide with uniform biological activity. The method is environment-friendly with mild reaction conditions and can give full play to prolease hydrolytic action, degradation of sulfuric glucoside of microorganism, therefore, the method of the invention is a method suitable for industrialized preparation of rapeseed bioactive peptide in a large scale with low cost.

Description

technical field [0001] The invention relates to a preparation method of rapeseed peptide with specific biological activity, in particular to a method for preparing rapeseed active peptide with specific biological activity through mixed culture of two kinds of microorganisms and solid-state fermentation of rapeseed dregs. Background technique [0002] Rapeseed meal is a by-product of rapeseed oil production, which is rich in protein and is an important plant protein resource. Because rapeseed meal contains anti-nutritional components such as glucosinolates and phytic acid, rapeseed meal is used as a plant protein The application of resources in the food industry is limited. Rapeseed protein is mainly composed of 12S globulin and 2S or 1.7S albumin. The content of lysine, cystine and methionine is relatively high. The amino acid balance is better than that of soybean protein. It is a high-quality protein. Rapeseed protein is hydrolyzed into polypeptides Finally, it has better...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P39/00C12P21/06C07K1/34C07K1/20C07K1/18C12R1/66C12R1/785C12R1/72C12R1/225C12R1/125
Inventor 鞠兴荣何荣袁建王立峰
Owner NANJING UNIV OF FINANCE & ECONOMICS
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