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A kind of preparation method of β-glucanase with good thermostability

A technology of glucanase and thermostability, which is applied in the field of enzyme preparation to achieve the effects of enhancing the body's functionality, wide range of action conditions, and a wide range of pH values

Active Publication Date: 2017-08-04
湖南康捷生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Previously, the high-quality β-glucanase in the market was almost monopolized by foreign enzyme preparation companies, although the enzyme activity of microbial β-glucanase reported in my country in recent years has reached or even exceeded the international level. , but obtaining β-glucanase with high activity and thermostability and its production strain is still a big problem to be solved in industrial application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A preparation method for β-glucanase, comprising the steps of:

[0035] (1) Expanded cultivation of liquid seeds

[0036] ①First-level seed culture: put the activated slant strains into 500 ml shake flasks with 2 rings, the medium capacity is 100 ml, the rotary shaker is 180 rpm, the culture temperature is 40°C, and the culture time is 10 hours;

[0037] ②Cultivation of secondary seeds: Inoculate 10% of primary seeds into 500 ml secondary seed shake flasks, and the cultivation conditions are the same as primary seeds;

[0038] ③Third-level seed cultivation: Inoculate the second-level seeds with 10% inoculum amount into a 5000 ml third-level seed shaker flask, the medium capacity is 1000 ml, the rotary shaker is 100 rpm, the culture temperature is 40°C, and the culture time is 12 hours ;

[0039] ④First-level seed tank cultivation: Put the third-level seeds into the first-level seed tank with a total volume of 150L at 10% inoculum volume, the fermentation medium capacity...

Embodiment 2

[0053] A preparation method for β-glucanase, comprising the steps of:

[0054] (1) Expanded cultivation of liquid seeds

[0055] ①First-level seed culture: put the activated slant strains into 500 ml shake flasks with 2 rings, the medium capacity is 100 ml, the rotary shaker is 180 rpm, the culture temperature is 40°C, and the culture time is 10 hours;

[0056] ②Cultivation of secondary seeds: Inoculate 10% of primary seeds into 500 ml secondary seed shake flasks, and the cultivation conditions are the same as primary seeds;

[0057] ③Third-level seed cultivation: Inoculate the second-level seeds with 10% inoculum amount into a 5000 ml third-level seed shaker flask, the medium capacity is 1000 ml, the rotary shaker is 100 rpm, the culture temperature is 40 ° C, and the culture time is 10 h ;

[0058] ④First-level seed tank cultivation: Put the third-level seeds into the first-level seed tank with a total volume of 150L at 10% inoculum volume, the fermentation medium capacity...

Embodiment 3

[0072] A preparation method for β-glucanase, comprising the steps of:

[0073] (1) Expanded cultivation of liquid seeds

[0074] ①First-level seed culture: put the activated slant strains into 500 ml shake flasks with 2 rings, the medium capacity is 100 ml, the rotary shaker is 180 rpm, the culture temperature is 40°C, and the culture time is 10 hours;

[0075] ②Cultivation of secondary seeds: Inoculate 10% of primary seeds into 500 ml secondary seed shake flasks, and the cultivation conditions are the same as primary seeds;

[0076] ③Third-level seed cultivation: Inoculate the second-level seeds with 10% inoculum amount into a 5000 ml third-level seed shaker flask, the medium capacity is 1000 ml, the rotary shaker is 100 rpm, the culture temperature is 40°C, and the culture time is 15 hours ;

[0077] ④First-level seed tank cultivation: Put the third-level seeds into the first-level seed tank with a total volume of 150L at 10% inoculum volume, the fermentation medium capaci...

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Abstract

The invention discloses a preparation method of beta-glucanase with good thermal stability, and belongs to the technical field of preparation of enzyme preparations. The preparation method comprises the following steps: using bacillus licheniformis beta-10-25 with the preservation number of CCTCC NO: M2013538 as a strain, activating the strain, performing liquid seed culture, and culturing in a fermentation tank. The prepared beta-glucanase with high thermal stability has the enzyme activity of 8,000-10,000 u / ml, which is 2.5 times of that of an original strain; at 50-70 DEG C, the beta-glucanase has relatively high enzymatic activity and the optimal reaction temperature is 65 DEG C; the suitable reaction pH is 5.0; after the beta-glucanaseb is stored for 6 months at room temperature, the loss of the enzymatic activity is less than 25%; after the beta-glucanase is stored for 12 months at 4 DEG C, the loss of the enzymatic activity is less than 20%. Compared with the conventional beta-glucanase, the beta-glucanase has higher enzymatic activity, wider optimal pH range for enzymatic action and higher thermal stability and storage stability, and is especially suitable for industrial demands of feed enzyme.

Description

technical field [0001] The invention belongs to the technical field of preparation of enzyme preparations, in particular to a Bacillus licheniformis capable of producing high-temperature and stable β-glucanase. Background technique [0002] β-glucan is a natural polysaccharide that usually exists in the cell walls of special types of bacteria, yeasts and fungi, or in the coating of higher plant seeds. β-glucan is a structural non-starch polysaccharide that forms the cell wall of grasses and is particularly abundant in the endosperm cell walls of barley, rye, sorghum, rice and millet. Unlike general sugars that are mainly combined with α-1,4-glycosidic bonds to form linear molecules, β-glucan in plant cell walls is a mixed β-(1,3), (1,4) glycoside The D-glucose polymer formed by the bond connection, so β-glucan can be dissolved in water with a large molecular weight, so it has the properties of high viscosity, high hydrophilicity, water swelling force and high water holding ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/24C12R1/10
CPCC12N9/244C12Y302/01006
Inventor 李洪兵张锦杰刘文明刘波封章平王永兰
Owner 湖南康捷生物科技有限公司
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