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Vibiro splendidus highly yielding alginate lyase and application thereof

A technology of alginate lyase and alginate, which is applied in the field of marine vibrio with high production of alginate lyase, can solve problems such as poor product uniformity, and achieve the effects of short fermentation period and high enzymatic activity

Active Publication Date: 2017-05-24
上海海糖生物医药科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Alginate lyase can also be applied to algin fermentation to produce bioethanol, but the currently used alginate lyase has the problem of poor product uniformity

Method used

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  • Vibiro splendidus highly yielding alginate lyase and application thereof
  • Vibiro splendidus highly yielding alginate lyase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: Screening and identification of microorganisms

[0017] 1. Screening of microorganisms

[0018] The fresh live scallop and abalone were dissected under aseptic conditions, and the digestive glands of the two were separated. Sterile physiological saline is added to the extracted digestive gland tissue, and the suspension of scallop and abalone digestive gland tissue is obtained after leaching. Dilute the suspension with sterile saline, 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 The gradient was diluted. Take 100-200 μL of the diluted liquid and spread it on the 2216E alginate solid selection medium. Place the culture plate in a 25°C incubator for cultivation. Pick colonies with good growth status from the plate and inoculate them in 2216E alginate liquid selection medium, and culture them in a shaking table with shaking. After 24 hours, the alginate-producing enzyme activity of each strain was measured, and the strain with high activity was sel...

Embodiment 2

[0030] Embodiment 2: Fermentation research of brilliant vibrio (Vibiro splendidus) OU02 strain

[0031] 1. Strain fermentation culture

[0032] The highly active bacterial strain obtained through the screening is fermented and cultured to obtain the growth curve of the microorganism in the alginate medium.

[0033] According to the inoculum amount of 1-3%, put the seed liquid into the alginate liquid medium, cultivate it in a shaker at 25°C and 150rpm, collect the fermentation liquid at different times, use the uninoculated medium as a blank control, and measure the bacteria The absorbance value at 600nm of the solution was used to draw the growth curve of the bacteria ( figure 1 ). As a result, it was found that the bacteria reached the plateau stage of growth in about 6-10 hours in the alginate medium, and at this time the OD 600 = about 2.5. This is the best time to collect bacteria in the future.

[0034] 2. Extraction and activity determination of enzymes in differen...

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PUM

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Abstract

The invention provides a vibiro splendidus OU02 strain highly yielding alginate lyase, the preservation number of which is CGMCC No.13478. The vibiro splendidus OU02 strain screened by the invention is used for producing and preparing alginate lyase. The alginate lyase produced by the vibiro splendidus OU02 strain screened can degrade alginate to generate alginate oligomers with potential biological activity. The microorganism is short in fermenting period and high in enzymatic activity, and almost activities intracellular activities, so that a sample is favorably collected and stored, thereby laying a foundation for follow-up industrial application.

Description

technical field [0001] The invention belongs to the technical field of beneficial microorganism screening, and in particular relates to a marine vibrio with high yield of alginate lyase and its application Background technique [0002] Alginate is a non-homopolymeric linear molecule composed of α-L-guluronic acid (G) and β-D-mannuronic acid (M) through 1,4-glycosidic bonds in different combinations. Alginate lyase catalyzes the 1,4-glycosidic bond between monomers through the β-elimination reaction, and forms an unsaturated double bond between C4 and C5 at the non-reducing end. The double bond has a maximum absorbance at 235nm. In the determination of enzyme activity, mainly by OD 235 The generation of unsaturated double bonds was detected by ultraviolet absorption method and the homogeneity of degradation products was determined by thin layer chromatography (TLC). [0003] At present, alginate lyase, especially endo-type alginate lyase, has been widely used in the product...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/88C12P19/00C12R1/63
CPCC12N9/88C12P19/00C12Y402/02003C12N1/205C12R2001/63
Inventor 刘伟治律倩倩张柯柯李智剑
Owner 上海海糖生物医药科技有限公司
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