Enhancing the biotransformation efficiency of human CYP17A1 in Pichia pastoris by co-expressing CPR and glucose-6-phosphate dehydrogenase simultaneously
نویسندگان
چکیده
Steroids are a widely used class of drugs, and their hydroxylation modification has important pharmacological significance. Cytochrome P450 is the core enzyme for steroid molecules, but its use significantly restricted because low activity poor catalytic efficiency. In present study, we optimized codons human CYP17A1 (hCYP17A1) gene performed functional expression in Pichia pastoris GS115. The GS115-hCYP17A1 cells exhibited activities dual function, following two products were detected: 17α-hydroxyprogesterone (17-HP, titer: 40.12 ± 3.16 mg/L) androstenedione (AD, 4.70 0.31 mg/L). Subsequently, compared hCYP17A1 co-expression with NADPH-cytochrome oxidoreductases (CPRs) from five different strains P. pastoris. Moreover, to strengthen NADPH regeneration system, glucose-6-phosphate dehydrogenase (G6PDH) three species was introduced into pathway progesterone 17-HP AD by cells. After optimizing CPRs G6PDHs, foreign proteins co-expressed host cell, namely CYP17A1, CPRYP Yorkshire pig, G6PDHc Candida tropicalis. Finally, substrate conversion efficiency found be increased 2.21-fold (46.88%) that starting strains. We obtained heterologous system highest biotransformation achieved date. This work provided an essential reference study introduction crucial systems model organisms construct efficient system.
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ژورنال
عنوان ژورنال: Systems microbiology and biomanufacturing
سال: 2021
ISSN: ['2662-7663', '2662-7655']
DOI: https://doi.org/10.1007/s43393-021-00063-7