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KMID : 1160220190470030301
Mycobiology
2019 Volume.47 No. 3 p.301 ~ p.307
Heterologous Expression of Rhizopus Oryzae CYP509C12 Gene in Rhizopus Nigricans Enhances Reactive Oxygen Species Production and 11¥á-Hydroxylation Rate of 16¥á, 17-Epoxyprogesterone
Shen Chaohui

Gao Xiyang
Li Tao
Zhang Jun
Gao Yuqian
Qiu Liyou
Zhang Guang
Abstract
The 11¥á-hydroxylation of 16¥á, 17-epoxyprogesterone (EP) catalyzed by Rhizopus nigricans is crucial for the steroid industry. However, lower conversion rate of the biohydroxylation restricts its potential industrial application. The 11¥á-steroid hydroxylase CYP509C12 from R. oryzae were reported to play a crucial role in the 11¥á-hydroxylation in recombinant fission yeast. In the present study, the CYP509C12 of R. oryzae (RoCYP) was introduced into R. nigricans using the liposome-mediated mycelial transformation. Heterologous expression of RoCYP resulted in increased fungal growth and improved intracellular reactive oxygen species content in R. nigricans. The H2O2 levels in RoCYP transformants were approximately 2-folder that of the R. nigricans wild type (RnWT) strain, with the superoxide dismutase activities increased approximately 45% and catalase activities decreased approximately 68%. Furthermore, the 11¥á-hydroxylation rates of EP in RoCYP transformants (C4, C6 and C9) were 39.7%, 38.3% and 38.7%, which were 12.1%, 8.2% and 9.4% higher than the rate of the RnWT strain, respectively. This paper investigated the effect of heterologous expression of RoCYP in R. nigricans, providing an effective genetic method to construct the engineered strains for steroid industry.
KEYWORD
11¥á-hydroxylation, cytochrome P450, heterologous expression, Rhizopus nigricans
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