title: Genome shuffling of Colletotrichum lini for improving 3β,7α,15α-trihydroxy-5-androsten-17-one production from dehydroepiandrosterone.
abstract: 3β,7α,15α-Trihydroxy-5-androsten-17-one (7α,15α-diOH-DHEA) is a key intermediate of the novel oral contraceptive Yasmin. It can be catalyzed from dehydroepiandrosterone (DHEA) through Colletotrichum lini. Improvement of 7α,15α-diOH-DHEA production was performed through recursive protoplast fusion of C. lini ST in a genome shuffling format. 7α,15α-diOH-DHEA yield of the best performing recombinant C. lini ST-F307 reached 6.08 g/L from 10 g/L DHEA, and this was 94.9% higher than that of the initial C. lini ST strain. Through optimized conditions, the 7α,15α-diOH-DHEA yield was increased to 9.32 g/L from 12 g/L DHEA, with 1.5% ethanol as cosolvent. This is the highest reported substrate concentration and 7α,15α-diOH-DHEA production with one-step substrate addition. Moreover, C. lini ST-F307 showed high P450 enzyme activity and gene transcript levels of several cytochrome P450s, and this might contribute to the enhancement of 7α,15α-diOH-DHEA production. Genome shuffling was an efficient approach to breed high-yield strains.
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Jin
authors.initials: J
authors.lastname: Sun
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, People's Republic of China. lihui@jiangnan.edu.cn.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ni
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Zhang
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Jinsong
authors.initials: J
authors.lastname: Shi
authors.affiliation: School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, People's Republic of China.|National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, 214122, Jiangsu, People's Republic of China.|Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Zhenghong
authors.initials: Z
authors.lastname: Xu
mesh_terms: D000737:Androstenols / Q000378:metabolism*; D020231:Colletotrichum / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D032621:DNA Shuffling; D003687:Dehydroepiandrosterone / Q000378:metabolism*; D016681:Genome, Fungal
chemical_list: C000593271:3beta,7alpha,15alpha-trihydroxy-5-androsten-17-one; D000737:Androstenols; D003687:Dehydroepiandrosterone; D003577:Cytochrome P-450 Enzyme System
keywords: Bioconversion; Colletotrichum lini; DHEA; Ethanol; Genome shuffling