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A high-tannase-yield strain and a preparing method thereof

A high-yield strain and tannase technology, applied in the field of microorganisms, can solve the problem of low production efficiency of tannase

Inactive Publication Date: 2015-01-07
曹庸
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to address the above existing problems, provide a kind of tannase high-yield bacterial strain and its preparation method, thereby improve the tannase fermentation yield, solve the problem of current tannin The problem of low enzyme production efficiency

Method used

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  • A high-tannase-yield strain and a preparing method thereof
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  • A high-tannase-yield strain and a preparing method thereof

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Embodiment

[0033] 1Materials and methods

[0034] 1.1 Materials and reagents

[0035] 1.1.1 Materials

[0036] Gallnut, collected in Zhangjiajie, Hunan.

[0037] 1.1.2 Reagents

[0038] Gallic acid, propyl gallate, provided by the Key Laboratory of Forest Products and Chemical Engineering of Hunan Province, purity ≥99%; methanol, chromatographic purity, American Fisher Company; glucose, tannic acid, sucrose, sodium nitrate, dipotassium hydrogen phosphate, diphosphate Potassium hydrogen, potassium chloride, iron sulfate, magnesium sulfate, ammonium sulfate, calcium chloride, manganese chloride, sodium molybdate, citric acid, sodium citrate, phenol, lactic acid, glycerin, cotton blue, bromophenol blue, agar are all For domestic analytical products, Guangzhou Chemical Reagent Factory; Fungal Genomic DNA Extraction Kit, Solarbio; D-2000 DNA molecular weight standard Marker, LA Taq DNA polymerase, GCbuffer I enhanced PCR buffer, dNTPs, Takara; Amplification primers, Shanghai Jimei Bi...

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Abstract

The invention provides a tannase high-producing strain and a preparation method thereof. The preparation method comprises the following steps of: taking decayed gallnuts in nature as raw material, screening microorganisms in the decayed gallnuts in a culture medium with gallotannic acid as the only carbon source to obtain a strain T3 which is high in tannase production activity, performing 60 Co gamma ray irradiation induced mutation on the strain, and finally obtaining a strain having a number of T3-5-1 through flat-plate primary screening and shake flask fermentation re-screening, wherein the tannase production activity of the strain is increased by 122.12% in contrast with that of the strain T3. Through strain identification, the strain T3-5-1 is aspergilluseniger and the conservation number of the strain T3-5-1 is CGMCC No. 7423. The strain provided by the invention is capable of producing tanase under optimal cultivation condition; the specific activity of the strain reaches up to 5304.53 U / mg after the strain is separated and purified, which is far higher than the specific activity of 1788.15 U / mg of the tannase produced by American Sigma.

Description

[0001] technical field [0002] The invention relates to the technical field of microorganisms, in particular to the related fields of microorganism screening, strain identification and fermentation enzyme production, and in particular provides a high-yielding strain of tannase and a preparation method thereof. Background technique [0003] Tannase (Tannase, E.C. 3.1.1.20) is an induced enzyme that widely exists in microorganisms and plants, especially in filamentous fungi, it can be produced in large quantities when tannic acid, gallnut, etc. are used as inducers. Cell membrane-bound enzyme that can be secreted extracellularly. Tannase can specifically hydrolyze the ester bond and depsyl carboxyl bond in the hydrolyzed tannin to generate gallic acid, glucose and corresponding alcohols and other substances. [0004] Tannase has been used in many fields such as beverages, brewing, food, medicine, chemical industry, leather and cosmetics, especially in the preparation of g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/18C12R1/685
CPCC12N9/18C12Y301/0102C12N1/145C12R2001/685
Inventor 曹庸张帅林健辉朱华伟农嘉仪
Owner 曹庸
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