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Ocean catalase production bacteria and directional screening method

A technology of catalase and enzyme bacteria, which is applied in the field of marine bioengineering, can solve the problem of low catalase production and achieve the effects of low cost, wide industrial use value and high enzyme activity

Active Publication Date: 2013-01-09
SECOND INST OF OCEANOGRAPHY MNR
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to the US patent report, the yield of catalase obtained by fermentation is low

Method used

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  • Ocean catalase production bacteria and directional screening method
  • Ocean catalase production bacteria and directional screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Conventional method, liquid medium screening method and semi-solid medium screening method

[0038] Collect seawater samples, take 150 μL of gradient dilution and spread them on natural seawater oligonutrient medium plates, incubate at 28°C for about 7 days to observe the results, and pick single colonies for slant preservation. The strains with catalase activity were initially screened by 3% hydrogen peroxide bubbling method. The natural seawater oligonutrient medium is 0.5g / L yeast extract, 0.1g / L peptone, pH is 7.2, and the rest is natural seawater.

[0039] Take 4.5mL seawater sample and add it to 4.5mL sterile aged seawater, add filter-sterilized 30% hydrogen peroxide 4μL and filter-sterilized carbon and nitrogen sources (final concentration: 0.25% peptone, 0.25 %Casamino acid and 0.01% yeast extract). Shake enrichment culture at 28°C for about 4 days, dilute the enrichment solution to an appropriate multiple aseptically, spread the plate, and select a ...

Embodiment 2

[0043] Example 2 Isolation and screening of P.chinense CGMCC No.6043

[0044] Seawater samples were collected, and 4.5 mL of seawater samples were added to the semi-solid medium under sterile conditions, as well as carbon and nitrogen sources that had been pre-filtered and sterilized, with a final concentration of 0.25% peptone, 0.25% Casamino acid and 0.01% yeast extract. Shake enrichment culture at 28°C for about 4 days, dilute the enrichment solution to an appropriate multiple aseptically, spread the plate, and select a single clone for slant preservation.

[0045] The bacterial strain isolated by the above method is transferred to natural seawater liquid medium for shake flask fermentation, and the catalase activity re-screening is carried out by spectrophotometry. The natural seawater medium is: sterile aged seawater with a final concentration of 0.25% peptone, 0.25% Casamino acid and 0.01% yeast extract, pH 7.2.

[0046] The hydrogen peroxide stimulation method was opti...

Embodiment 3

[0047] Example 3 Morphological identification and biological characteristics of P.chinense CGMCC No.6043

[0048] P.chinense (P.chinense) CGMCC No.6043 was inoculated in natural seawater liquid medium. To prepare solid medium, add 15g / L agar. 121°C damp heat sterilization for 20 minutes. After inoculation and culture, it was identified that the bacterium had the following characteristics: (1) Colony morphology: the colony was round, with a smooth surface, flat edges, slightly raised, and yellow. (2) Cell morphology: Gram-positive bacteria, under the cell microscope (Olympus, BX40), show motility, and the morphology is mostly spherical (0.8×1.0μm). (3) Physiological and biochemical characteristics: aerobic growth; hydrolyzed gelatin, non-hydrolyzed starch, casein, Tween 80, and aescin; can use glucose to produce acid, but cannot use sucrose, raffinose, xylan, lactose, and arabinose , cellobiose, xylose, rhamnose, melibiose, mannose or mannitol to produce acid.

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Abstract

The invention relates to a method for directionally screening catalase high-production bacterial strain by a self-creative semi-solid culture medium and ocean catalase high-production bacteria obtained by the method. The method includes that hydrogen peroxide is filled into a lower semi-solid agar layer and slowly permeates into an upper liquid culture medium layer and stably supplies a certain oxidation pressure to microbes in the upper liquid culture medium layer, and thereby the purpose that directional enrichment of the catalase high-production bacteria is achieved and the ocean catalase production bacteria have the advantages of high selectivity and high efficiency. A mobility bacillus CGMCC (China General Microbiological Culture Collection Center) No.6043 can be screened from ocean water by means of directional enrichment of the semi-solid culture medium, has the advantages of short growth period, high enzymatic activity and low cost of fermenting catalase production, and is widely applicable to aspects of the textile industry, food industry and waste water treatment and the like. Accordingly, the method for directionally screening catalase high-production bacterial strain and the catalase strain have high industrial utilization value and evident economic benefit prospect.

Description

technical field [0001] The invention belongs to the technical field of marine bioengineering, and in particular relates to a marine catalase high-yielding strain and a method for directional screening of a catalase high-yielding strain using a self-created semi-solid medium. Background technique [0002] The textile industry is a traditional industry and a pillar industry in China, and occupies an important position in the gross domestic product and foreign trade export value. The current textile industry is seriously polluted, especially in the process of printing and dyeing. The traditional process consumes a lot of water and chemicals, which not only consumes resources, but also causes environmental pollution and destroys the ecological balance. To change the traditional extensive economic growth mode of high energy consumption and high pollution in the textile industry, it is necessary to replace and transform the textile industry process with green biotechnology industr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/08C12Q1/30C12Q1/04C12R1/01
Inventor 吴月红许学伟王春生吴敏张心齐潘杰
Owner SECOND INST OF OCEANOGRAPHY MNR
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