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Method for producing alpha-galactosidase by three-stage solid fermentation

A solid-state fermentation and galactose technology, applied in the biological field, can solve the problems of affecting fermentation, affecting economic benefits, and low vitality, and achieve the effects of stable fermentation process, shortened fermentation time, and improved vitality

Inactive Publication Date: 2012-11-21
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current α-galactosidase solid-state fermentation production process has unstable fermentation conditions, which often affect the smooth progress of fermentation, resulting in unstable product quality and low vitality. The monitoring of fermentation conditions requires a lot of manpower and material resources, which affects the economy. benefit

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 3

[0020] Example 1 Three-stage solid-state fermentation to produce α-galactosidase

[0021] (1) Preparation of spore suspension: Aspergillus niger was inoculated on the PDA slant medium, and cultivated for 144h at 28° C. in a 60% constant temperature and humidity incubator until the spores covered the slant; 6 -10 7 Individual / mL spore suspension is ready for use;

[0022] (2) Preparation of liquid seeds: Sterilize the PDB liquid medium at 121°C for 20 minutes, and cool to room temperature; insert the spore suspension into the above-mentioned cooled PDB liquid medium according to the volume ratio of 5% inoculum, and incubate at 28°C 1. Cultivate for 18 hours on a shaker with a rotating speed of 150rpm, and harvest liquid seeds;

[0023] (3) Preparation of solid-state fermentation medium: the solid-state fermentation medium is sterilized at 121° C. for 20 minutes, and then used; the solid-state fermentation medium is made by mixing and stirring dry base and distilled water with...

Embodiment 2 3

[0028] Example 2 Three-stage solid-state fermentation to produce α-galactosidase

[0029] (1) Preparation of spore suspension: Aspergillus niger was inoculated on the PDA slant medium, and cultivated for 160 h at 28° C. in a 60% constant temperature and humidity incubator until the spores covered the slant; made 100 h with normal saline 6 -10 7 Individual / mL spore suspension is ready for use;

[0030] (2) Preparation of liquid seeds: Sterilize the PDB liquid medium at 121°C for 20 minutes, and cool to room temperature; insert the spore suspension into the above-mentioned cooled PDB liquid medium according to the volume ratio of 5% inoculum, and incubate at 28°C 1. Cultivate for 18 hours on a shaker with a rotating speed of 150rpm, and harvest liquid seeds;

[0031] (3) Preparation of solid-state fermentation medium: After the solid-state fermentation medium is sterilized at 121°C for 20 minutes, it is ready for use; after the solid-state fermentation medium is mixed with dry...

Embodiment 3

[0036] Embodiment 3 common solid-state fermentation produces α-galactosidase

[0037] (1) Preparation of spore suspension: Aspergillus niger was inoculated on the PDA slant medium, and cultivated for 144h at 28° C. in a 60% constant temperature and humidity incubator until the spores covered the slant; 6 -10 7 Individual / mL spore suspension for use;

[0038] (2) Liquid seed preparation: sterilize the PDB liquid medium at 121°C for 20 minutes, and cool to room temperature; add the spore suspension into the above-mentioned cooled PDB liquid medium according to the volume ratio of 5% inoculum, and incubate at 28°C, Cultivate for 18 hours on a shaker with a rotating speed of 150rpm, and harvest liquid seeds;

[0039] (3) Preparation of solid-state fermentation medium: After the solid-state fermentation medium is sterilized at 121° C. for 20 minutes, it is ready for use; after the solid-state fermentation medium is mixed with dry basis and distilled water with a weight ratio of 2...

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Abstract

The invention discloses a method for producing alpha-galactosidase by three-stage solid fermentation, which comprises: inoculating aspergillus niger onto a sterilized solid fermentation culture medium; and placing the culture medium in a sterile environment for staged fermented, wherein in a first stage, the fermentation time is 22 to 26 hours, the environment temperature is 26 to 30 DEG C and the environment humidity is 60 to 70 percent; in a second stage, the fermentation time is 32 to 40 hours, the environment temperature is 23 to 25 DEG C, and the environment humidity is 85 to 95 percent;and in a third stage, the fermentation time is 28 to 62 hours, the environment temperature is 26 to 30 DEG C and the environment humidity is 90 to 95 percent. The process for produces alpha-galactosidase by solid fermentation is divided into three stages with different conditions; and compared with the conventional solid method, the activity of the alpha-galactosidase product produced by the production method is improved by about 40 percent, the fermentation time is reduced by more than 20 hours, the operation is simple and convenient, the cost is low, and the fermentation process is stable.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for producing α-galactosidase by three-stage solid-state fermentation. Background technique [0002] α-galactosidase is a glycoside hydrolase characterized by the specific hydrolysis of non-reducing terminal α-1,6-galactosidic bonds, so it can hydrolyze the Sugar and other α-galactomannoligosaccharides and heteropolysaccharides such as guar gum and locust bean gum. It has been widely used in food, feed and other industries. [0003] At present, most livestock and poultry feed types in my country are mainly corn-soybean meal, and soybean meal contains about 7% of α-galactosides such as closeose, raffinose, stachyose and verbascose. Nutritional factors are metabolized by anaerobic bacteria in the back of the digestive tract, producing carbon dioxide, hydrogen and a small amount of methane gas, causing indigestion in animals and reducing the digestion and absorption of nutrien...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/40C12R1/685
Inventor 何国庆顾丰颖阮晖高洁
Owner ZHEJIANG UNIV