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Method for fermentative production of laccase using fungoid co-cultivated

A production method and co-cultivation technology, which are applied in the field of preparation of industrial enzyme preparations, can solve the problems of low yield of laccase, difficulty in meeting industrial production requirements, and no patent protection in the production process, and achieve simple processing and production technology. Safety, environmental protection and low cost

Inactive Publication Date: 2006-07-26
ANHUI UNIVERSITY
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the production of laccase in these studies is not high, it is difficult to meet the requirements of industrial production
On the other hand, there is no patent protection for the production process of fungal laccase produced by co-cultivation of microorganisms

Method used

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Examples

Experimental program
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Effect test

Embodiment Construction

[0017] 1. Preparation of Trametes AH28-2 and Trichoderma ZH1 bacterial suspension (seed solution)

[0018] In potato medium (1L medium: 20% potato juice, 20g glucose, 3g KH 2 PO 4 , 1.5g MgSO 4 ·7H 2 0, 0.4g vitamin B, add 1.5% agar to make solid medium) Trametes AH28-2 and Trichoderma ZH1 preserved at 4 ℃ on the slant are respectively inoculated in high-temperature sterilized seed liquid medium (1L medium: 15g Glucose, 2.5g Asparagine, 1.0g KH 2 PO 4 , 0.5g MgSO 4 ·7H 2 O, 0.1g Na 2 HPO 4 ·5H 2 O, 0.01g CaCl 2 , 0.002g CuSO 4 ·5H 2 O, 0.001gFeSO 4 ·7H 2 O, 0.0275g adenine and 0.05g vitamin B, natural pH), 28°C, 100-200rpm culture for 1-3 days, 3,000rpm homogenization for about 10s to make bacterial suspension.

[0019] 2. Preparation of co-culture fermentation medium

[0020] Fermentation enzyme production medium (1L): 10~30g carbon source, 1.0~4.0g nitrogen source, 1.0g KH 2 PO 4 , 0.5g MgSO 4 ·7H 2 O, 0.1g Na 2 HPO 4 ·5H 2 O, 0.01g CaCl 2 , 0.002g Cu...

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Abstract

This invention relates to a epiphyte co-culture ferment Laccase production method, which is to inoculate the bacterium detergent of the Trametes AH28-2(Trametes sp. AH28-2) in the ferment culture medium under 22-37deg C for 1-4 days, then adding the bacterium detergent of Trichoderma ZH1(Trichoderma sp. ZH1) to culture for 5-10 days, finally separating to obtain the Laccase preparation, wherein the enzyme activity is up to 6, 000U / L(guaiacol method). The culture medium is the liquid culture medium, which mainly contains a certain quantity of carbon, azotes, phosphate of kalium and natrium, sulfate of copper and ferrum, adenine and vitamin B and so on. This invention doesn't' use the aromatic compound and heavy metal iron induce, which is safe and clean, and the enzyme activity is high and stable. '

Description

1. Technical field [0001] The invention relates to a method for preparing an industrial enzyme preparation, in particular to a method for preparing an industrial laccase, specifically a method for producing laccase by fungal co-cultivation and fermentation. 2. Background technology [0002] Laccase (Laccase, EC 1.10.3.2) is a class of copper-containing polyphenol oxidases that catalyze the oxidation of a variety of phenolic and nonphenolic compounds with concomitant reduction of oxygen to water. Laccase widely exists in higher plants and higher fungi (especially basidiomycetes). Laccase has a wide range of substrates and high catalytic efficiency. It is widely used in biopulping and bleaching, improvement of food flavor, improvement of feed nutrition, degradation and conversion of textile dyes, softening and refinement of textile fibers, development of new drugs, and application of new biosensors. It has potential and important application value in research and new energy d...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N1/14C12R1/885
Inventor 肖亚中洪宇植张赫袁璟房伟
Owner ANHUI UNIVERSITY
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