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Method for producing alginate lyase through fermentation by using marine bacteria

A technology of alginate lyase and marine bacteria, which is applied in the field of bioengineering, can solve the problems of lack of high-value processing of brown algae and alginic acid, limit the popularization and application of alginate lyase, and poor enzyme stability, and achieve remarkable fermentation effect and increase Bacterial biomass and the effect of prolonging the enzyme production period

Inactive Publication Date: 2019-09-03
张学花
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the existing alginate lyases generally have problems such as low enzyme activity, poor enzyme stability, tight combination of enzyme and product, and difficult separation. These shortcomings limit the popularization and application of alginate lyase.
At present, the research on alginic acid lyase in my country is relatively basic, and the research at home and abroad is mainly focused on the isolation of strains, the construction of genetically engineered bacteria, the improvement of enzymatic properties, and the structure and performance of enzymatic hydrolysis products. There are few research reports on the fermentation process of lyase, and the lack of industrial enzymes for alginate lyase has become a bottleneck for the high-value processing of brown algae and alginic acid

Method used

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  • Method for producing alginate lyase through fermentation by using marine bacteria
  • Method for producing alginate lyase through fermentation by using marine bacteria
  • Method for producing alginate lyase through fermentation by using marine bacteria

Examples

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Embodiment 1

[0034] A method for producing alginic acid lyase by fermenting marine bacteria, comprising the steps of:

[0035] S1, preparation of seed solution: Flavobacterium aquaticus ( Flavobacterium mizutaii ) comes from the Marine Microorganism Culture Collection and Management Center MCCC, with the preservation number 1A08665. After thawing the frozen and preserved strains, they were streaked and inoculated on 2216E solid medium (recipe: peptone 5g / L, yeast extract powder 1g / L, lemon Ferric acid 0.01g / L, agar 20g / L, aged sea water 1000mL, pH 7.4), cultured at 28°C for 24 hours; put the activated strain into the seed medium, and the liquid volume in the 500mL Erlenmeyer flask was 100mL. The temperature is 28°C, the rotation speed is 180r / min, and the shaker is shaken for 12 hours; the culture is taken for microscopic examination, and it is confirmed that the bacteria grow well without any bacterial contamination, and then they can be inoculated into the seed tank; the formula of the s...

Embodiment 2

[0041] Example 2 Optimization of fermentation conditions

[0042] 1. Optimization of the initial pH value of the fermentation medium

[0043] According to the method provided in Example 1, during fermentation and enzyme production, the initial pH value of the fermentation medium was adjusted to 5.0, 6.0, 7.0, 8.0, and 9.0 respectively. After the fermentation, the crude enzyme liquid was prepared and the alginate lyase activity was measured. Determine the optimal initial pH.

[0044] The initial pH value of the fermentation medium causes the difference in the oxidation-reduction potential of the bacterial cell membrane, which directly affects the progress of the oxidation-reduction reaction and the generation of free energy, thereby affecting the activity of the cell membrane, so the absorption of nutrients in the medium and the secretion of enzymes make an impact. Depend on figure 1 It can be seen that when the pH value is 5.0, the enzyme production ability of Flavobacteri...

Embodiment 3

[0051] Example 3 Optimization of fermentation medium

[0052] 1. Optimization of the amount of bean cake powder added

[0053] According to the method provided in Example 1, when fermenting and producing enzyme culture, the addition amount of bean cake powder in the fermentation medium is respectively set to 5, 10, 15, 20, 25, 30 g / L, and other components remain unchanged. The crude enzyme solution was prepared and the activity of alginate lyase was determined to determine the amount of bean cake powder added.

[0054] Nitrogen source is one of the main components of fermentation medium, which has an important influence on the growth and metabolism of microorganisms. Studies have shown that organic nitrogen sources are easier to be decomposed and utilized by bacteria than inorganic nitrogen sources, and organic nitrogen sources can also provide certain growth factors to enable bacteria to grow efficiently and rapidly. Compared with peptone and yeast extract powder, bean cake...

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Abstract

The invention discloses a method for producing alginate lyase through fermentation by using marine bacteria, which relates to the technical field of sea microbial fermentation. The method adopts the marine bacterium Flavobacterium mizutaii as a starting strain, and obtains a crude enzyme solution of alginate lyase by the steps of preparing the seed liquid, expanding the seed liquid for culture, fermenting for enzyme production culture, and extracting the fermentation liquid. The method aims at enzyme production rule of Flavobacterium mizutaii for optimizing the fermentation conditions and a fermentation medium, and improves the enzyme production ability of the aquatic Flavobacterium mizutaii by a carbon source feeding strategy and a method of adding various enzyme-inducing substances, andprovides important technical parameter guidance for the large-scale production of the alginate lyase in the future.

Description

technical field [0001] The invention belongs to the field of bioengineering and relates to a microbial fermentation process, in particular to a method for producing alginic acid lyase by fermentation of marine bacteria. Background technique [0002] The ocean contains abundant biomass resources and is an important source of human food and medicine. my country is rich in marine resources and is the country with the largest scale of marine algae cultivation, processing and utilization in the world. Seaweed production accounts for about 60% of the world's total output, exceeding 13 million tons. Brown algae is an economical seaweed that is abundant in the ocean. Representative plants include kelp, carrageenan, macroalgae, and sargasso. Alginic acid, also known as alginic acid, alginic acid, seaweed, etc., is a water-soluble acidic polysaccharide in the cell wall and interstitium of brown algae. It is composed of two monomers that are C5 epimers (β-D-mannose uronic acid, α-L-g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/88C12N1/20C12R1/20
CPCC12N9/88C12Y402/02003C12Y402/02011C12N1/20
Inventor 张学花
Owner 张学花
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