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Alpha-galactosidase solid fermenting producing method

A technology of galactosidase and production method, which is applied in the field of solid-state fermentation to produce α-galactosidase for feed, and can solve the problems of fermentation characteristics of enzyme-producing bacteria, lack of systematic research on nutritional characteristics of fermentation technology, and low enzyme production level of bacteria and other problems, to achieve the effect of low production cost, simple process and high enzyme activity

Inactive Publication Date: 2007-01-24
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are also reports on the production of α-galactosidase by microorganisms, but the enzyme production level of the existing α-galactosidase-producing bacteria is low, and the fermentation characteristics, nutritional characteristics and fermentation technology of the enzyme-producing bacteria have not been systematically studied. Therefore, it is necessary to explore the production method of α-galactosidase with high enzyme activity, which can be inoculated with liquid seeds or solid seeds.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, a kind of α-galactosidase production method, carries out following steps successively:

[0022] 1), the preparation of spore suspension:

[0023] Inoculate Aspergillus foetidus with the preservation number CGMCC No.1628 on the slant medium according to the streaking method, and cultivate it at a constant temperature of 28°C for 7 days. for 10 6 ~10 7 The spore suspension of each / ml is ready for use; the slant medium is a PDA slant medium, namely: 200 grams of fresh potatoes, 20 grams of glucose, 20 grams of agar, 1000 milliliters of distilled water, and a natural pH.

[0024] 2), primary seed preparation:

[0025] Firstly, the liquid seed culture medium was sterilized at 121° C. for 20 min, and then cooled; then the spore suspension was inserted into the above-mentioned cooled liquid seed culture medium at an inoculum size of 5% according to the volume ratio (V / V), and then cooled at 28 Cultivate on a shaker at 200 rpm for 18 hours to obtain liquid see...

Embodiment 2

[0032] Embodiment 2: a kind of α-galactosidase production method, carries out following steps successively:

[0033] 1), the preparation of spore suspension:

[0034] Inoculate Aspergillus foetidus with the preservation number CGMCC No.1628 on the slant medium according to the streaking method, and cultivate it at a constant temperature of 28°C for 7 days. for 10 6 ~10 7 spore suspension per ml, ready for use. The slant medium is the PDA slant medium made from fresh potato-glucose-agar, the same as in Example 1.

[0035] 2), primary seed preparation:

[0036]First, the liquid seed medium was sterilized at 121°C for 20 minutes, and then cooled; then the spore suspension was added to the above-mentioned cooled liquid seed medium according to the volume ratio of 5% inoculum, at 28°C and the rotation speed was 200rpm Cultivate on a shaker for 18 hours to obtain liquid seeds. Described liquid seed medium is the PDA liquid medium that fresh potato-glucose makes; Same as embodi...

Embodiment 3

[0047] A method for producing α-galactosidase, comprising the following steps in sequence:

[0048] 1), the preparation of spore suspension:

[0049] With embodiment 2.

[0050] 2), primary seed preparation:

[0051] With embodiment 2.

[0052] 3) Secondary seed preparation:

[0053] In a 500ml Erlenmeyer flask, 40g dry basis is loaded, and this dry basis is made up of 32.855g bran, 7.137g soybean meal, 0.004g manganese sulfate and 0.004g copper sulfate. Add 60 g of water to the bottle, and adjust the pH to 5.5-6.0 with 0.5 M disodium hydrogen phosphate to obtain a solid seed medium.

[0054] Change the number of days of cultivation into 6 days, and the rest are the same as in Example 2.

[0055] 4), the fermentation of α-galactosidase:

[0056] With embodiment 2.

[0057] 5), post-processing

[0058] With embodiment 2.

[0059] It was determined that the enzyme activity of α-galactosidase was 1928.24U / g dry koji.

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Abstract

The solid fermentation process of producing alpha-galactosidase includes the following steps: 1. preparing spore suspension; 2. preparing the first order seed; and 3. fermentation of alpha-galactosidase. Or, the solid fermentation process of producing alpha-galactosidase includes the following steps: 1. preparing spore suspension; 2. preparing the first order seed; 3. preparing the second order seed; and 4. fermentation of alpha-galactosidase. The alpha-galactosidase produced through the said process has high enzyme generating activity, simple preparation process and easy production application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for producing alpha-galactosidase for feed by solid-state fermentation. Background technique [0002] α-galactosidase is an oligosaccharide characterized by the specific hydrolysis of non-reducing terminal α-1,6-galactosidic linkages of galacto-oligosaccharides and polygalacto-(gluco)mannan Hydrolase has great economic value and application value. Since α-galactosidase can hydrolyze the polysaccharides of oligosaccharides and galactomannans such as melibiose, raffinose, stachyose and verbascose in beans, and can improve sugar metabolism, it is mainly used In feed and food industries. [0003] At present, the feed industry is developing rapidly with the regionalization and scale of animal husbandry production and the increase in the proportion of feed, and the contradiction of shortage of feed resources in my country is becoming increasingly prominent; according to experts'...

Claims

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Application Information

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IPC IPC(8): C12N9/40C12N1/14C12R1/66
Inventor 刘彩琴何国庆陈启和阮晖
Owner ZHEJIANG UNIV
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