Vibrio maritimus capable of producing alginate lyase with good thermal stability and application

A technology of alginate lyase and thermal stability, applied in the field of microorganisms, can solve problems such as patent publications and the like that have not yet been found, and achieve the effects of good thermal stability and short growth cycle

Active Publication Date: 2020-11-20
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Through the search, no patent publications related to t

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  • Vibrio maritimus capable of producing alginate lyase with good thermal stability and application
  • Vibrio maritimus capable of producing alginate lyase with good thermal stability and application
  • Vibrio maritimus capable of producing alginate lyase with good thermal stability and application

Examples

Experimental program
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Example Embodiment

[0041] Example 1 Isolation and identification of strains

[0042] The following experiments were performed under sterile conditions.

[0043] 1. Enrichment of strains

[0044] Under aseptic conditions, use a small hammer to break the shell, take out the snails, pick out the intestinal part with scissors, briefly immerse in 75% ethanol to disinfect the surface, and add it to a triangular flask containing 100 mL of 2216E medium. Incubate at ℃ for 48h, and dilute the enriched sample to 10 -7 , Take 100 μL of samples under each gradient and evenly spread them on the 2216E culture plate with sodium alginate as the sole carbon source, incubate at 28°C for 48 hours, and observe the colony growth status.

[0045] The preparation method of the 2216E medium is: peptone 5g, yeast extract 1g, ferric citrate 0.1g, sodium chloride 19.45g, magnesium chloride 5.9g, magnesium sulfate 3.24g, calcium chloride 1.8g, potassium chloride 0.55 g, sodium bicarbonate 0.16g, potassium bromide 0.08g, strontium ...

Example Embodiment

[0060] Example 2 Determination of the optimal growth conditions of Vibrio marine bacteria of the present invention

[0061] The strains were respectively inoculated into 2216E medium with sodium alginate, glucose, sucrose, starch, and xylose as the sole carbon source, cultured at 30°C for 12 hours, and the relative biomass was determined to be 100%, 54.15%, 73.65%, 92.5%, respectively , 15.6%, the relative enzyme activity is 100%, 0%, 37.6%, 55.45%, 2.25%. The result is Figure 4 Shown. It is proved that the most suitable carbon source for the growth and enzyme production of this strain is sodium alginate.

[0062] The strains were inoculated into 2216E medium with yeast extract, tryptone, ammonium sulfate, and ammonium chloride as the sole nitrogen source, and incubated at 30°C for 12 hours. The relative biomass was determined to be 78.85%, 100%, 49.45%, and 40.85, respectively. %, the relative enzyme activities were 81.55%, 100%, 37.6%, 60.55%, 67.7%. The result is Figure 5 S...

Example Embodiment

[0067] Example 3 Determination of Enzymatic Properties of Vibrio marine in the present invention

[0068] 1. Determination of optimum temperature

[0069] The strain was cultured at 30°C for 12h, and its supernatant was determined by DNS method at 25, 30, 35, 40, 45, 50, 55, and 60°C. The relative activity of algin lyase was 60%, respectively. 83.7%, 91.8%, 100%, 92.6%, 72.5%, 39.4%, 6.6%. The result is Picture 10 Shown. It proved that the algin lyase produced by this strain reached the highest enzyme activity at 40°C.

[0070] 2. Determination of heat resistance

[0071] The strain was cultured at 30°C for 12 hours, and the supernatant was heat-treated at 40, 45, 50, 55°C for 5, 15, 30, 60, 90, and 120 minutes, and the relative activity of alginate lyase was determined by DNS method After heat treatment at 40℃, they were 97.3%, 95.8%, 91.1%, 87.4%, 79.1%, 65.9%, and after heat treatment at 45℃, they were 91.1%, 88.6%, 78.4%, 66.4%, 44.3%, 32.7%, 50 After heat treatment at ℃, the...

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Abstract

The invention relates to vibrio maritimus capable of producing alginate lyase with good thermal stability. The name of the vibrio maritimus is ML17, the classification name is Vibrio sp., the preservation number is CGMCC No.20377, the preservation date is July 16, 2020, the preservation unit is China General Microbiological Culture Collection Center, and the preservation site is No.3, yard 1, Beichen West Road, Chaoyang District, Beijing. According to the present invention, the vibrio maritimus is separated from the intestinal tract content of Monodonta Labio, the vibrio maritimus does not have the significant hemolysis phenomenon through the verification of Wagstsuma Blood Agar Base, and the surface of the vibrio maritimus does not have the potential pathogenicity. The vibrio maritimus disclosed by the invention is short in growth cycle and is a dominant strain in intestinal tracts of the Monodonta Labio, and the produced alginate lyase has good thermal stability.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a marine Vibrio strain capable of producing alginate lyase with good thermal stability and its application. Background technique [0002] Alginate is a macromolecular polysaccharide substance that is second only to cellulose in nature. It is mainly extracted from the cell wall and intercellular substance of algal plants such as kelp, macroalgae, and sargassum, which are most widely present in the ocean. It exists widely in the Phaeaceae. Alginate is mainly composed of β-D-mannuronic acid and its C5 epimer α-L-guluronic acid arranged randomly through α / β-1,4 glycosidic bonds, and the two sugar units interact with each other Polymerization forms three different structural units: polymannuronic acid (PolyM), polyguluronic acid (PolyG) and a heteropolymer formed by random combination of M and G (PolyMG). [0003] Alginate lyase is a polysaccharide-degrading enzyme that speci...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/88C12P19/00C12R1/63
CPCC12N9/88C12Y402/02011C12Y402/02003C12P19/00C12R2001/63C12N1/205
Inventor 罗学刚刘耀天李月婵赵阳席茂盛
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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