High-yield alginate lyase strain and application thereof

A technology of alginate lyase and strain, applied in the field of microorganisms, can solve the problems of restricting wide application, low enzyme-producing ability of strains, unsuitable for industrial production and the like

Pending Publication Date: 2022-06-24
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the low enzyme production ability of the strain, it is not suitable for industrial production, which restricts its wide application.

Method used

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  • High-yield alginate lyase strain and application thereof
  • High-yield alginate lyase strain and application thereof
  • High-yield alginate lyase strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Isolation and screening of Vibrio HB161653GDMCC No.61929

[0035] Sargassum samples were collected from Tanmen Port, Qionghai, Hainan Province. Take 10g of Sargassum algae sample, cut it into pieces and dissolve it in a sterile state, put it in 90ml of sterile seawater, shake it in a constant temperature shaker at 28°C and 200r / min for 30min, and after standing for 5min, take the supernatant to prepare 10 -2 ~10 -4 series of suspensions. Pipette 0.1 ml of the suspension, spread it on 2116E medium supplemented with 0.05% sodium alginate, and place it in a 30°C incubator for upside-down culture. When the colony grows, according to the phenotypic characteristics of the colony shape, color, edge state, transparency, surface dry and wet state, etc., a single colony is picked and streaked and purified to obtain a pure culture. After preliminary sorting, number and save.

[0036] The isolated bacterial strains were tested for alginate lyase activity, and the stra...

Embodiment 2

[0043] Example 2 Species identification of Vibrio HB161653

[0044] A single colony of strain HB161653 was picked by plate streak method, inoculated on 2116E solid medium, and incubated at 37°C at a constant temperature. When cultured for 2 days, the colony was 2-4mm in diameter, milky yellow, round, slightly convex, translucent, moist and smooth on the surface, and neat in the edges; Vibrio HB161653 can grow normally under the conditions of 25~37℃, pH 6~9, 1.5~4.0%(w / v) NaCl.

[0045] The physiological and biochemical characteristics of the strain HB161653 were identified by referring to the Manual for the Identification of Common Bacterial Systems. The results showed that the strain HB161653 could utilize glucose, mannitol and inositol, etc. Gram stain negative, alkaline phosphatase, esterase (C4), esterase lipase (C8), acid phosphatase, glucosaminidase, trypsin, lipase (C14) test positive, can reduce nitrate.

[0046] The genomic DNA of strain HB161653 was extracted with...

Embodiment 3

[0050] Embodiment 3 Alginate lyase and preparation method thereof

[0051] The alginate lyase in the present invention is prepared by fermentation of Vibriosp.HB161653 whose deposit number is GDMCC No.61929 in Example 1, and the specific method includes the following steps:

[0052] (1) Strain activation: inoculate the Vibrio HB161653 in Example 1 on a solid medium, and cultivate at 30°C for 36 hours to obtain an activated strain;

[0053] (2) Liquid culture: insert the activated strain into the liquid seed medium, and shake at 30°C and 180r / min for 12-18h to prepare seed liquid, OD 600 Controlled between 0.8 and 1.2;

[0054] (3) Fermentation culture: inoculate the seed liquid into the liquid fermentation medium by volume percentage of 2%, shake and culture at 30°C, 180r / min for 32-36h, centrifuge, and collect the supernatant to obtain the crude algin lyase enzyme solution;

[0055] The solid medium in step (1) is composed of: sodium alginate 5g / L, peptone 5g / L, yeast powd...

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Abstract

The invention relates to the technical field of microorganisms, in particular to a high-yield alginate lyase strain and application thereof. The invention provides a bacterial strain HB161653 for producing alginate lyase. The bacterial strain is classified and named as Vibrio sp., and the preservation number of the bacterial strain is GDMCC No.61929. The invention further provides a preparation method of the bacterial strain HB161653 for producing alginate lyase. The alginate lyase produced by the strain has the characteristics of high enzyme yield and wide pH stability. The invention further provides alginate lyase and a preparation method thereof, and application of the strain HB161653 and the alginate lyase in degradation of alginate to produce alginate oligosaccharide and degradation of alginate bodies, and the alginate lyase has a good application prospect.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to an alginate lyase-producing Vibrio strain HB161653 and applications thereof. Background technique [0002] Alginate, also known as sodium alginate and sodium alginate, is a water-soluble acidic polysaccharide derived from brown algae cell wall and intercellular substance. It is widely distributed in brown algae such as kelp, sargassum and macroalgae. Pseudomonas) is an important component of biofilms. Alginate can be degraded to generate oligosaccharide fragments composed of 2 to 20 monosaccharides, showing a variety of biological activities, such as promoting growth, enhancing immunity, neuroprotection, anti-inflammatory, anti-virus, etc. It has a wide range of application value in agriculture, health care products and other fields. The degradation methods of alginate are divided into chemical method, physical method and enzymatic hydrolysis method. Among them, the enz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/88C12P19/00C12P19/12C12P19/04C12R1/63
CPCC12N9/88C12P19/00C12P19/12C12P19/04
Inventor 鲍时翔朱军黄惠琴邹潇潇
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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