白腐菌Phanerochaete sordida YK-624产锰过氧化物酶的生产及初步纯化

林产化学与工业 ›› 2003, Vol. 23 ›› Issue (04): 22-26.

白腐菌Phanerochaete sordida YK-624产锰过氧化物酶的生产及初步纯化

谢慧芳¹, 近藤隆一郎², 李忠正¹

1. 南京林业大学化工学院, 江苏南京 210037;2. 九州大学农学部, 日本九州福冈 812-81

收稿日期:2003-01-15 修回日期:1900-01-01 出版日期:2003-12-30 发布日期:2003-12-30

PRODUCTION AND PURIFICATION OF MANGANESE PEROXIDASE BY WHITE ROT FUNGI PHANEROCHAETE SORDIDA YK-624

XIE Hui-fang¹, KONDO Ryuichiro², LI Zhong-zheng¹

1. College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China;2. Graduate School of Bioresource and Bioenvironmental Science, Kyushu University, Fukuoka Kyushu 812-81, Japan

Received:2003-01-15 Revised:1900-01-01 Online:2003-12-30 Published:2003-12-30

摘要/Abstract

摘要： 利用白腐菌Phanerochaete sordida YK 624(AT CC90872),采用Kirk液体培养基在富氧条件下进行振动培养,可得到锰过氧化物酶(MnP)活性较高和漆酶活性微量的酶液。该酶液通过超滤、透析、二乙氨基乙基琼脂糖凝胶(DEAE-sepharose CL-6B,pharmacia)离子交换柱层析的方法进行初步纯化后,得到比活性很高(1941.54IU/mg)的MnP酶液。漆酶活性和MnP活性比值的变化、SDS-PAGE凝胶电泳试验和406nm附近的吸光值变化均表明,经离子交换柱层析后的YK1部分是主要的MnP,电泳试验同时表明纯化后的主要MnP的分子质量约为43kDa。经超滤、透析、离子交换柱层析纯化后得到的酶液YK1可用于进一步地研究及应用。

关键词: 锰过氧化物酶, 液体培养, 离子交换柱层析

Abstract: Crude enzymes liquor, which contained high manganese peroxidase(MnP) activity and trace laccase (activity), was produced by cultivating white rot fungi Phanerochaete sordida YK-624(ATCC 90872) under shaking and high O₂ content conditions in Kirk liquor culture medium.Ultra-filtration, dialysis and DEAE-sepharose CL-6B(pharmacia) anion-exchange chromatography were used to purify the enzymes solutions.Thus, enzymes solution of high specific MnP activity marked as YK1(1941.54 IU/mg) was obtained.The ratio of MnP to laccase, the results of SDS-polyacrylamide gel electrophoresis and the absorption spectra around 406 nm proved that YK1, was the main part of MnP activity.Similarly, SDS-polyacrylamide gel electrophoresis showed molecular weight of YK1 to be about (43 kDa).Thus, YK1, obtained from ultrafiltration, dialysis and anion-exchange chromatography, could be chosen as the enzyme solution for more application research, such as biobleaching and improvement of fiber quality.

Key words: manganese peroxidase(MnP), liquid cultivation, anion-exchange

中图分类号:

谢慧芳;近藤隆一郎;李忠正. 白腐菌Phanerochaete sordida YK-624产锰过氧化物酶的生产及初步纯化[J]. 林产化学与工业, 2003, 23(04): 22-26.

XIE Hui-fang;KONDO Ryuichiro;LI Zhong-zheng. PRODUCTION AND PURIFICATION OF MANGANESE PEROXIDASE BY WHITE ROT FUNGI PHANEROCHAETE SORDIDA YK-624[J]. , 2003, 23(04): 22-26.

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