Preparation method of laccase from sparassis crispa

Abstract

The invention discloses a preparation method of laccase from sparassis crispa. The preparation method comprises the following steps (1) taking mixed fruit peel as a fermentation substrate, mixing themixed fruit peel with activated carbon, performing homogenizing, performing filtration, and performing sterilization so as to obtain a fruit peel solid-state culture medium; (2) inoculating the fruitpeel solid-state culture medium with activated sparassis crispa strains, and performing primary fermentation culture; (3) performing laccase enzyme activity monitoring on primary fermentation products, after enzyme activity reaches the peak value, selecting sparassis crispa mycelium balls obtained in the step (2), inoculating liquid nutrient mediums containing Cu2+ and inducers with the selected sparassis crispa mycelium balls, performing shaking flask culture to produce enzymes, and then conveying the enzymes into a fermenter; and (4) adding amino-oligosaccharin and biological fermentation antifoaming agents to the fermenter, and performing secondary deep fermentation so as to obtain fermentation products namely the laccase produced from sparassis crispa. The laccase is high in enzyme activity, the sparassis crispa is appropriate in fermentation bacterium age, and fermentation strains are high in purity and high in yield.

Description

technical field [0001] The invention relates to the field of hydrangea, in particular to a method for preparing laccase produced by hydrangea. Background technique [0002] Lignin and pollutants similar in structure to lignin precursors are one of the important sources of environmental pollution. In recent years, the use of fungal laccase to degrade such pollutants has become a hot spot in environmental protection research, especially in pulp bleaching, dye decolorization, Laccase has shown great research value and application potential in detoxification and degradation of environmental pollutants. [0003] Laccase is a copper-containing polyphenol oxidase, which belongs to the blue multi-copper oxidase family with ascorbate oxidase, mammalian ceruloplasmin and bilirubin oxidase, and Hydrangea can secrete laccase, manganese peroxidase Several common extracellular enzymes such as oxidase or lignin peroxidase, these lignin degrading enzymes are distributed outside the cell, b...

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Problems solved by technology

[0004] In the prior art, there are methods for producing laccase by fungal fermentation, such as liquid submerged fermentation or solid fermentation, and submerged liquid fermentation is likely to cause poor oxygen circulation and is unfavorable for the growth and development of aerobic hydrangea. At the same time, due to embroidered Coccus is a giant fungus, the mycelium is easy to wrap around the power equipment, block the pipeline, and the disordered expansion of the mycelium will also increase the viscosity of the fermentation broth, seriously affecting the uniform transfer and diffusion of materials, oxygen and heat; and Solid fermentation The solid fermentation cycle is long and the enzyme production is not high. When the strain is transferred to reproductive growth, the mycelium is easy to kink with each other, thereby inhibiting the extracellular enzyme production of the strain. And because the solid fermentation space is relatively open to liquid fermentation, it is easy to mix in Other miscellaneous bacteria reduce the efficiency of enzyme production

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