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产漆酶真菌血红密孔菌H1对蒽和芘的代谢特性

Peculiarities of metabolism of anthracene and pyrene by laccase-producing fungus Pycnoporus sanguineus H1.

作者信息

Li Xuanzhen, Wang Yan, Wu Shijin, Qiu Lequan, Gu Li, Li Jingjing, Zhang Bao, Zhong Weihong

机构信息

College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou, People's Republic of China.

出版信息

Biotechnol Appl Biochem. 2014 Sep-Oct;61(5):549-54. doi: 10.1002/bab.1197. Epub 2014 Mar 25.

Abstract

The metabolic peculiarities of anthracene and pyrene by Pycnoporus sanguineus H1 were investigated. The fungus H1 could grow on potato dextrose agar plates with anthracene and anthraquinone as carbon sources. In liquid medium, the strain degraded 8.5% of anthracene as the sole carbon source, with no ligninolytic enzymes detected, indicating that intracellular catabolic enzymes might be responsible for the initial oxidation of anthracene. When bran was added to the medium, the degradation rate of anthracene and pyrene increased to 71.3% and 30.2%, respectively, and the laccase activities increased to a maximal value of 501.2 and 587.6 U/L, respectively. By gas chromatography-mass spectrometry analysis, anthraquinone was detected as the unique intermediate product of anthracene oxidation, with a yield molar ratio of 0.3. In vitro experiments showed that the extracellular culture fluid containing laccase transformed anthracene to anthraquinone with a yield molar ratio of 1.0, which was less than that of the in vivo experiment, indicating that anthraquinone could be further metabolized by the strain. Pyrene could not be oxidized by culture fluid. These results showed that both extracellular laccase and intracellular catabolic enzymes might play an important role in the initial oxidation of anthracene, whereas pyrene could be only oxidized by intracellular catabolic enzymes through cometabolism.

摘要

研究了血红密孔菌H1对蒽和芘的代谢特性。该真菌H1能够在以蒽和蒽醌为碳源的马铃薯葡萄糖琼脂平板上生长。在液体培养基中,该菌株以蒽作为唯一碳源时可降解8.5%的蒽,且未检测到木质素分解酶,这表明细胞内分解代谢酶可能负责蒽的初始氧化。当向培养基中添加麸皮时,蒽和芘的降解率分别提高到71.3%和30.2%,漆酶活性分别增加到最大值501.2和587.6 U/L。通过气相色谱-质谱分析,检测到蒽醌是蒽氧化的唯一中间产物,产率摩尔比为0.3。体外实验表明,含有漆酶的细胞外培养液将蒽转化为蒽醌,产率摩尔比为1.0,低于体内实验,这表明蒽醌可被该菌株进一步代谢。芘不能被培养液氧化。这些结果表明,细胞外漆酶和细胞内分解代谢酶可能在蒽的初始氧化中起重要作用,而芘只能通过共代谢被细胞内分解代谢酶氧化。

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