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Novel cytochrome P450s for various hydroxylation of steroids from filamentous fungi.
Yi, Guojuan; Zou, Hanlu; Long, Tao; Osire, Tolbert; Wang, Lin; Wei, Xiaoyun; Long, Mengfei; Rao, Zhiming; Liao, Guojian.
Afiliación
  • Yi G; College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
  • Zou H; College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
  • Long T; College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
  • Osire T; Faculty of Biology, Shenzhen MSU-BIT University, 1 University Park Road, Shenzhen, Guangdong 518172, China.
  • Wang L; College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
  • Wei X; College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
  • Long M; College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China. Electronic address: mengfeilong@swu.edu.cn.
  • Rao Z; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
  • Liao G; College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China. Electronic address: gjliao@swu.edu.cn.
Bioresour Technol ; 394: 130244, 2024 Feb.
Article en En | MEDLINE | ID: mdl-38145763
ABSTRACT
Hydroxylated steroids are value-added products with diverse biological activities mediated by cytochrome P450 enzymes, however, few has been thoroughly characterized in fungi. This study introduces a rapid identification strategy for filamentous fungi P450 enzymes through transcriptome and bioinformatics analysis. Five novel enzymes (CYP68J5, CYP68L10, CYP68J3, CYP68N1 and CYP68N3) were identified and characterized in Saccharomyces cerevisiae or Aspergillus oryzae. Molecular docking and dynamics simulations were employed to elucidate hydroxylation preferences of CYP68J5 (11α, 7α bihydroxylase) and CYP68N1 (11α hydroxylase). Additionally, redox partners (cytochrome P450 reductase and cytochrome b5) and ABC transporter were co-expressed with CYP68N1 to enhance 11α-OH-androstenedione (11α-OH-4AD) production. The engineered cell factory, co-expressing CPR1 and CYP68N1, achieved a significant increase of 11α-OH-4AD production, reaching 0.845 g·L-1, which increased by 14 times compared to the original strain. This study provides a comprehensive approach for identifying and implementing novel cytochrome P450 enzymes, paving the way for sustainable production of steroidal products.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Esteroides / Sistema Enzimático del Citocromo P-450 Idioma: En Revista: Bioresour Technol Asunto de la revista: ENGENHARIA BIOMEDICA Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Esteroides / Sistema Enzimático del Citocromo P-450 Idioma: En Revista: Bioresour Technol Asunto de la revista: ENGENHARIA BIOMEDICA Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido