NATURAL PRODUCT RESEARCH AND DEVELOPMENT ›› 2020, Vol. 32 ›› Issue (2): 296-304. doi: 10.16333/j.1001-6880.2020.2.014

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Fermentation techonology of Fomes fomentarius SO3 producing laccase and partial enzymatic properties

XIE Yu-qing1,2,CHEN Jin1,2 ,DAI Jin-ping1,2,YANG Xin-ping1,2,GuliAhmat1,2,WANG Zhi-fang1,2,WANG Xiao-wu1,2,FENG Lei1,2*   

  1.  1Institute of Microbiology,Xinjiang Academy of Agricultural Sciences;2Xinjiang Key Laboratory of Special Enviromental Microbiology,Urumqi 830091,China
  • Online:2020-02-29 Published:2020-04-23

Abstract: Strain Fomes fomentarius SO3 with high-yield laccase was screened from the Alter Mountains in Xinjiang.We optimized the fermentation technology and studied the partial enzymatic properties.The experimental result showed that the optimal carbon and nitrogen sources were wheat bran and yeast extract powder.Adding Mn2+,Ca22+,Zn2+ and Mg2+ could promote the yield of laccase ,the effect of Cu2+ was adverse.Gallic acid and tannic acid made enzyme production peak two days ahead and ABTS may increase enzyme activity from 6 736 U/L to 8 470 U/L,orthogonal experiment L9(34) proved that optimum medium consists of 0.5% yeast extract powder,2.5 % wheat bran,0.5 % glucose and 0.5 %(NH4)H2PO4,in this case,enzyme activity could reach to 10 863 U/L.The study of enzymatic properties showed optimal reaction temperature of laccase is 50 ℃ and there was still activity at 80 ℃,optimal reaction pH was 3.0 and enzyme activity was stable when pH was 4.0-5.0 and temperature was under 40 ℃.At the same time,Co2+,Cd2+,Cr2+nd Pb2+ could damage enzyme stability.Bidirectional electrophoresis was performed on the bacteria and fementation liquid in which isoelectric point of laccase was determined to be 4.1.A pure enzyme protein sequcence was obtained by mass spectrometry that showed the protein to be LaccaseE (Trames sp. 420).

Key words: Fomes fomentarius SO3, laccase, optimizing fermentation technology, enzymatic properties, mass spectrometry sequencing

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