产多酚氧化酶茶树内生真菌的筛选及产酶条件优化

doi:10.13305/j.cnki.jts.2015.03.010

摘要/Abstract

摘要： 以愈创木酚、α-萘酚、没食子酸、L-酪氨酸和单宁酸等5种酚类物质为底物,采用鉴别培养基筛选法从14株茶树内生真菌中初筛获得4株产多酚氧化酶的真菌。根据变色圈的大小、颜色深浅和摇瓶发酵的结果复筛获得产多酚氧化酶能力较强的菌株CSN-13。对菌株CSN-13产酶营养条件进行初步分析,结果表明,在供试的6种碳源物质中,以麸皮对菌株CSN-13产多酚氧化酶的促进作用最为明显;供试的5种氮源物质中,以硝酸铵的促进作用最为明显;在发酵培养基中添加茶水,对产酶有明显的促进作用。采用正交设计对CSN-13产酶发酵培养基进行初步优化,优化后的培养基配方为：麸皮（40βg·L^-1）、硝酸铵（15βg·L^-1）、茶水（4βg·L^-1）、KH₂PO₄（2βg·L^-1）、MgSO₄·7H₂O（0.5βg·L^-1）、无水CaCl₂（0.075βg·L^-1）、CuSO₄·5H₂O（0.01βg·L^-1）。采用优化后的培养基,菌株CSN-13在28℃下培养5βd,酶活力达到241βU·mL^-1·min^-1,比优化前提高8.5倍。茶树内生真菌菌株CSN-13及其发酵产酶培养基的研究为多酚氧化酶的进一步开发打下了基础。

关键词: 茶树, 内生真菌, 多酚氧化酶, 菌株筛选, 培养基优化

Abstract: Five kinds of phenolic materials, including guaiacol, alpha-naphthol, gallic acid, L-tyrosine and tannic acid, were used as the substrates to screen polyphenol oxidase-producing fungal strains. Four out of 14 endophytic fungal strains of tea plant (Camellia sinensis) were selected by agar plate screening methods. Strain CSN-13 was obtained for its color ring size and color depth, which owned a powerful production capacity of polyphenol oxidase. A preliminary analysis for nutritional components of strain CNS-13 was conducted. The results showed that among 6 kinds of carbon sources, wheat bran was the best for its promoting effects upon polyphenol oxidase production. Ammonium nitrate showed the most significant promotion among 5 kinds of nitrogen sources. Enzyme production was significantly promoted when tea infusion was added into the fermentation medium. With the adoption of orthogonal design, the fermentation medium for enzyme production was preliminarily optimized. The optimized medium contains wheat bran (40βg·L^-1), ammonium nitrate (15βg·L^-1), tea (4βg·L^-1), KH₂PO₄ (2βg·L^-1), MgSO₄·7H₂O (0.5βg·L^-1), CaCl₂ (0.075βg·L^-1), and CuSO₄·5H₂O (0.01βg·L^-1). By using the optimized medium, enzyme activity could reach 241 U·mL^-1·min^-1 under culture conditions with 28℃ temperature for 5 days, which was 8.5 times higher than that of before optimization. Strain CSN-13 and the fermentation medium in present study offered an alternative for the further development of polyphenol oxidase.

Key words: Camellia sinensis, endophyte, polyphenol oxidase, fungal strain screening, medium optimization

中图分类号:

张婉蓉, 巫婷玉, 杨民和. 产多酚氧化酶茶树内生真菌的筛选及产酶条件优化[J]. 茶叶科学, 2015, 35(3): 271-280. doi: 10.13305/j.cnki.jts.2015.03.010.

ZHANG Wanrong, WU Tingyu, YANG Minhe. Screening and Culture Medium Optimization of Polyphenol Oxidase Producing-Fungi from Endophytes of Tea Plant (Camellia sinensis)[J]. Journal of Tea Science, 2015, 35(3): 271-280. doi: 10.13305/j.cnki.jts.2015.03.010.

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