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

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0021] Example 1. A method for producing α-galactosidase, the following steps are carried out in sequence:

[0022] 1). Preparation of spore suspension:

[0023] The Aspergillus foetidus with the preservation number of CGMCC No.1628 was inoculated on the slant medium according to the streaking method, and cultured at a constant temperature of 28°C for 7 days. At this time, the strain was mature; washed with sterile water and shaken the spores to make the concentration For 10 6 ~10 7 The spore suspension per ml is to be used; the slant medium is a PDA slant medium, namely: 200 g of fresh potatoes, 20 g of glucose, 20 g of agar, 1000 ml of distilled water, and natural pH.

[0024] 2) Preparation of primary seeds:

[0025] First, sterilize the liquid seed culture medium at 121°C for 20 minutes, and then cool it; then add the spore suspension to the cooled liquid seed culture medium at a volume ratio (V / V) of 5%. Cultivate for 18 hours on a shaker at a temperature of 200 rpm at a temp...

Example Embodiment

[0032] Example 2: A method for producing α-galactosidase, which includes the following steps in sequence:

[0033] 1). Preparation of spore suspension:

[0034] The Aspergillus foetidus with the preservation number of CGMCC No.1628 was inoculated on the slant medium according to the streaking method, and cultured at a constant temperature of 28°C for 7 days. At this time, the strain was mature; washed with sterile water and shaken the spores to make the concentration For 10 6 ~10 7 A / ml spore suspension, set aside. The slant medium is a PDA slant medium made of fresh potato-glucose-agar, the same as in Example 1.

[0035] 2) Preparation of primary seeds:

[0036]Sterilize the liquid seed medium at 121°C for 20 minutes, and then cool it; then add the spore suspension to the cooled liquid seed medium at a volume ratio of 5%, at 28°C and rotate at 200 rpm Incubate on a shaker for 18 hours to obtain liquid seeds. The liquid seed medium is a PDA liquid medium made of fresh potato-gluco...

Example Embodiment

[0046] Example 3:

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

[0048] 1). Preparation of spore suspension:

[0049] The same as in Example 2.

[0050] 2) Preparation of primary seeds:

[0051] The same as in Example 2.

[0052] 3) Secondary seed preparation:

[0053] A 500ml Erlenmeyer flask is filled with 40g dry base, which is composed of 32.855g bran, 7.137g soybean meal, 0.004g manganese sulfate and 0.004g copper sulfate. Add 60g of water to the bottle, and then adjust the pH to 5.5-6.0 with 0.5M disodium hydrogen phosphate to obtain a solid seed culture medium.

[0054] Change the culture days to 6 days, and the rest is the same as in Example 2.

[0055] 4) Fermentation of α-galactosidase:

[0056] The same as in Example 2.

[0057] 5), post-processing

[0058] The same as in Example 2.

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