2种野外采集白腐真菌分离及其产漆酶液体培养基优化

李环明 杨洪升 薛春梅

摘要[目的]优化2种野外采集白腐真菌菌丝产漆酶液体培养基。[方法]对2种采集的白腐真菌轮纹韧革菌（Stereum ostrea）和朱红密孔菌（Pycnoporus cinnabarinus）通過分离纯化得到白腐真菌菌丝，应用ABTS法测定其产漆酶活性，研究碳源、氮源、金属阳离子、pH、温度、转速对白腐真菌产漆酶的影响。[结果]轮纹韧革菌最佳培养基是葡萄糖20.00 g、硝酸钾5.00 g、NaH2PO4 5.00 g、CuSO4 1.00 g、初始pH 6、温度25 ℃、转速170 r/min，在第11天其产漆酶活性达到最大，为1.557 U/mL，是PDA液体培养基产漆酶活性的2.7倍。朱红密孔菌最佳培养基是麦芽糖25.00 g、酵母膏5.00 g、NaH2PO4 1.00 g、KCl 2.00 g、初始pH 7、溫度30 ℃、转速200 r/min，在第9天其产漆酶活性达到最大，为1.478 U/mL，是PDA液体培养基产漆酶活性的2.3倍。[结论]筛选出2种野外白腐真菌产漆酶最佳培养基，为漆酶的进一步研究提供了参考。

关键词 白腐真菌；漆酶；培养基；筛选

中图分类号 S182 文献标识码 A 文章编号 0517-6611（2017）03-0001-03

Abstract[Objective] The aim was to optimize the liquid culture medium of two species of field acquisition white rot fungi.[Method] By isolating and purifying two species of white rot fungi，Stereum ostrea and Pycnoporus cinnabarinus，white rot fungus hyphae were collected，and the liquid culture medium of laccase was screened.ABTS method was applied to measure the activity of laccase.Effects of carbon sources，nitrogen sources，metal cations，pH，temperature and rotational speed on laccase produced by white rot fungi were studied.[Result] The optimum medium of Stereum ostrea was glucose 20.00 g，potassium nitrate 5.00 g，NaH2PO4 5.00 g，CuSO4 1.00 g，the initial pH 6，temperature 25 ℃， rotational speed 170 r/min，and the laccase activity reached the maximum on the 11th day the maximum value reached 1.557 U/mL which was 2.7 times of the laccase activity of PDA liquid culture.The optimum medium of Pycnoporus cinnabarinus was maltose 25.00 g，yeast extract 5.00 g，NaH2PO4 1.00 g，KCl 2.00 g，the initial pH 7，temperature 30 ℃，rotational speed 200 r/min，and the laccase activity reached the maximum in the 9 d，the maximum value 1.478 U/mL which was 2.3 times of the laccase activity of PDA liquid culture.[Conclusion] The opitimal liquid culture medium of two species of field acquisition white rot fungi is obtained so as to provide reference for further study of laccase.

Key words White rot fungus；Laccase；Culture medium；Screen

白腐真菌是一类使木材呈白色腐朽的真菌，可在特定条件下产各种酶系[1]，包括漆酶、木质素过氧化氢酶和锰过氧化氢酶等[2-3]。在白腐真菌产的酶系中，漆酶的应用非常广泛。在染料脱色方面，马利[4]、傅恺[5]、张丽[6]、王天女等[7]对重组血红密孔菌产漆酶对活性蓝等染料进行脱色结果分析，其脱色率达93%。在环境污染的处理方面，彭丹对黄孢原毛平革菌在固态发酵体系产真菌漆酶对五氯酚（PCP）降解进行研究，结果表明在没有氧化还原介体时粗漆酶液能降解PCP，反应6 h降解率为37.8%，粗酶液中加入5 mmol/L氧化还原介体（ABTS）能获得更高的降解率，达97.0%；将粗漆酶液用（NH4）2SO4盐析纯化，用提纯后漆酶降解PCP，6 h后降解率为81.8%[8-9]。……