2017-M2 bacterial strain containing mannanase gene

A 2017-M2, mannanase technology, which is applied in glycosylase, genetic engineering, plant genetic improvement, etc., can solve the problem that the type and quantity of mannanase strains cannot meet the needs of natural red pigment producing bacteria resources and other problems to achieve the effect of low requirements for culture conditions and fast growth

Active Publication Date: 2019-12-17
武汉设计工程学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The object of the present invention is to overcome the defective of prior art, and the kind and quantity of existing mannanase bacterial classification can not satisfy industry to the demand of natural red pigment producing bacterium resource

Method used

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  • 2017-M2 bacterial strain containing mannanase gene
  • 2017-M2 bacterial strain containing mannanase gene
  • 2017-M2 bacterial strain containing mannanase gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Isolation, screening and identification of Talaromyces atroroseus 2017-M2 strain

[0042] 1. Isolation of Talaromyces atroroseus 2017-M2 strain

[0043] (1) Talaromyces atroroseus 2017-M2 strain was isolated from the soil 5 cm below the ground in Tangxun Lake area, Jiangxia District, Wuhan City, Hubei Province, where Wuhan Institute of Design and Engineering is located in Wuhan City, Hubei Province. The specific steps are as follows: take 10 g of soil, dilute it 10 times with 90 mL of sterile water, shake well, clarify, and then continue to dilute to 1000 times; take 0.1 mL of the 1000-fold dilution and spread it on a Martin Bengal red solid plate, cultivate for 3 days, The culture was continued at room temperature until the 7th day, and the growth of the colonies in the petri dish and the production of water-soluble red pigment in the medium were observed. Pick Penicillium-like colonies that produce red pigment. Pure cultures were streaked in Gaw's No. 1 so...

Embodiment 2

[0053] Example 2 Application of Talaromyces atroroseus2017-M2 strain fermentation production and extraction of red pigment

[0054] In this example, the Talaromyces atroroseus 2017-M2 strain was inoculated into a fermentation medium (a 500mL conical flask was filled with 100mL fermentation medium), and the fermentation medium was a PDA liquid medium: peeled potatoes 200g to 30min, take 1000mL of potato juice, add Glucose 20g, packaged and sterilized, pH 7.0, cultured at 30°C, 200rpm shaker for 5d-7d. Bacterial liquid was centrifuged at 8000rpm and 10min to obtain fermentation broth, and sterilized by bacterial filter filtration, and the obtained filtrate was the crude extract of red pigment. The crude extract of red pigment was placed in a rotary evaporator at 70°C for vacuum concentration until it became a paste, and the obtained product was the crude extract of red pigment.

[0055] The obtained red pigment crude extract was dissolved in the same amount of distilled water a...

Embodiment 3

[0059] Example 3 Identification of cellulase produced by Talaromyces atroroseus 2017-M2 strain

[0060] Pick the strains and spot them on the cellulose identification medium with a toothpick, incubate at 28°C for 5-7 days, cover the medium with 1 mg / ml Congo red solution for 15 minutes, pour off the Congo red solution, and wash with 1 mol / L NaCl solution The Congo red solution was removed for 15 minutes, the NaCl solution was poured out, and a transparent circle was observed, indicating that the Talaromyces atroroseus 2017-M2 strain had the ability to produce cellulase.

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Abstract

The invention belongs to the application field of agricultural microorganisms, and relates to a 2017-M2 bacterial strain containing a mannanase gene. The 2017-M2 bacterial strain can produce edible red pigment, mannanase and cellulase and can be obtained from soil through separation. The 2017-M2 bacterial strain containing the mannanase gene is Talaromyces atroroseus, and the sequence of the mannanase gene is shown in SEQ ID NO: 2. The similarity with known mannan beta-1,4-endonuclease gene is only 71%, and the 2017-M2 bacterial strain is a novel functional gene and has wide application value.The 2017-M2 bacterial strain grows well at 22DEG C-35 DEG C, produces the cellulase, the mannanase and amylase, does not produce protease, and has no bacteriostatic effect. The red pigment with a maximum light absorption value being 600 nm can be obtained in a PDA medium. The active gene can be expressed in Escherichia coli. The best induction conditions is 1<m>M IPTG 1.5 [mu]L, and induction iscarried out at 37 DEG C for 3 h.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganism application, and in particular relates to a Talaromyces atroroseus 2017-M2 strain containing a mannanase gene and capable of producing edible red pigment, cellulase and mannanase. Isolated from soil, it is Talaromyces.atroroseus, which is evolved from Penicillium branch, belongs to a new strain, contains mannanase gene, the mannan expressed by this gene The enzyme is β-1,4-endomannanase. Background technique [0002] Talaromyce atroroseus is a new species of fungus discovered in 2013, which is classified as Talaromyces.atroroseus. The fungus is present in soil and fruit and was originally identified from house dust collected in South Africa. It produces large amounts of red pigment and does not contain any known mycotoxins. The genome of the strain contains a gene encoding β-1,4-mannanase, and β-mannanase (endo-1,4-β-mannanase) is an endohydrolase, which can act on the mannos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N15/56C12N9/42C12P1/02C12R1/645
CPCC12N9/2491C12Y302/01025C12N9/2437C12Y302/01004C12P1/02C12R2001/645C12N1/145
Inventor 石慧翟淑红张娟莫晨阳欧炜明
Owner 武汉设计工程学院
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