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Studies on Ganoderma Lucidum Laccase and Some Extracellular Enzymes

Author: LiuJianPeng
Tutor: LanJin
School: Peking Union Medical College , China
Course: Pharmacognosy
Keywords: Ganoderma lucidum extracellular enzyme laccase inducer electrophoresis
CLC: S567.31
Type: Master's thesis
Year: 2009
Downloads: 80
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Abstract


Ganoderma lucidum is a famous fungi with very high medicinal value in China. The extracellular enzymes involved in the process to get nutrition for G. lucidum when cultured, were related to its output and quality. As one of extracellular enzymes, laccase from G. lucidum is also a multifunction phenol oxidase with wide application. In order to elucidate the relation between extracellular enzymes and output and quality of G. lucidum and promote the application of G. lucidum laccase, we studied the variation of extracellular enzymes and polysaccharides of G. lucidum S3 cultured. After screening inducers of laccase, we also used a procedure which involved ammonium sulfate precipitation, DEAE-sepharose anion exchange chromatography and Source-15PHE hydrophobic chromatography to purify laccase and then characterized it in detail.It was showed that the lignin-modifying enzymes of G. lucidum grown with cottonseed skin as the sole carbon and energy source consisted of laccase and manganese-dependent peroxidase, no lignin peroxidase was observed. Every enzyme had a different variation trend. Culture methods had an effect on the polysaccharide content.The maximum mycelium biomass appeared on the 7th day, while the maximum activity of the laccase enzyme appeared on the 9th day. The majority of the potential tested were able to serve as inducers for laccase to some extent, among which 2,6-dimethylaniline was found to be the best when the concentration was 0.8mmol/L. With the native-PAGE zymogram, it was inferred that G. lucidum secreted only a single laccase in liquid fermentation, not affected by inducers.The purified laccase was a single submit protein with a molecular weight of 59.6KDa by SDS-PAGE. The Km was 9.02×10-6mol/L with ABTS as substrate, while 1.08×10-3mol/L with guaiacol as substrate. This laccase was stable from pH 6.0 to 8.0. The temperature optimum for its activity was 45℃. The research also showed that a lot of compounds tested could affect the laccase activity, some acted as activators while others acted as inhibitors.

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CLC: > Agricultural Sciences > Crop > Economic crops > Medicinal crops > Fungus > Ganoderma lucidum
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