Fig.1 HPLC profiles of standards（17α-hydroxyprogest-erone and 11α，17α-dihydroxy progesterone）（a）and biotransformation sample（b）

Fig. 2 TLC results of 17α-hydroxyprogesterone at 48 h by 10 molds 1：Standard of 17α-hydroxyprogesterone. 2：Standard of 11α，17α-dihydroxy progesterone. 3：Aspergillus ochraceus. 4：Cunningpamycetes elegans. 5：Absidia coerulea. 6：Beauveria bassiana. 7：Metarhizium anisopliae. 8：Aspergillus niger. 9：Penicillium citreoviridin. 10：Aspergillus flavus. 11：Penicillium citrinum. 12：Penicillium raistrickii

Fig. 3 Molar production rate（a）and production intensity（b）of 11α，17α-dihydroxy progesterone from 17α-hydroxyprogesterone by Aspergillus ochraceus and Cunningpamycetes elegans，respectively

Fig. 4 Verification of the recombinant P. pastoris（a），S. cerevisiae（b）and R. rhodochrous（c）expressing CYP68J5 M：DL 2000 DNA marker；1：PCR product of pYES2-11αAo；2：PCR product of pNV18-11αAo

Fig. 5 TLC biotransformation results of 17α-hydroxypro-gesterone at 48 h by the recombinant S. cerevisiae，P. pastoris and R. rhodochrous expressing CYP68J5，respectively 1：Standard of 17α-hydroxyprogesterone. 2. Standard of 11α，17α-dihydroxy progesterone. 3：Saccharomyces cerevisiae. 4：Pichia pastoris. 5：Rhodococcus rhodochrous

Fig. 6 Molar production rate（a）and production inten-sity（b）of 11α, 17α-dihydroxy progesterone from 17α-hydroxyprogesterone by the recombinant S. cer-evisiae and P. pastoris expressing CYP68J5, respec-tively

Fig. 7 Biotransformation performance of recombinant S. cerevisiae strains expressing CYP68J5

Fig. 8 Predicted overall structure of CYP68J5 and the molecular docking with 17α-hydroxyprogesterone α helix，β folding and random curl are colored in bright blue，bright purple and pink，respectively. The active site residues and the substrate are colored in bright yellow and dark red，respectively

Fig. 9 Relative molar production rate of 11α, 17α-dihydr-oxy progesterone from 17α-hydroxyprogesterone by the mutants compared to that of the wild-type CYP68J5 The data represent the mean value of three independent experiments. The relative molar production rate is calculated by using the molar production rate of wild-type CYP68J5 as 100%

Fig. 10 Local model of the key amino acid site residues D118（a），F216（b）and M488（c）of CYP68J5 and the alanine mutation site residues A118（d），A216（e）and A488（f）