Fig. 2
Identification of steroid 11α–hydroxylase gene CYP68J5. a TLC analysis of induction of steroid 11α–hydroxylase activities in TCCC41060. 16,17α-epoxyprogesterone (EP) was used for induction and also as transformation substrate, and petroleum and ethyl acetate (3:2) was used for product separation. 1, EP standard; 2, induction for 0 h; 3, induction for 3 h; and 4, 11α-OH EP standard. b, c Analysis of bioconversion products of the recombinant S. cerevisiae cells expressing either CYP68L8 or CYP68J5. b EP as substrate, petroleum and ethyl acetate (3:2) as the solvent system for TLC; s1, EP standard; s2, S. cerevisiae wild-type; s3, recombinant INVSc1-CYP68L8; s4, INVSc1-CYP68J5; and s5, 11α-OH EP standard. c DE as substrate and petroleum and ethyl acetate (1:1) as the solvent system; s6, DE standard; s7, INVSc1-CYP68J5; s8, S. cerevisiae wild-type; s9, recombinant INVSc1-CYP68L8; and s10, 11α-OH DE standard. d HPLC assays of biotransformation product of recombinant S. cerevisiae cells, acetonitrile: H2O 80:20 (v/v) as the mobile phase. (a) substrate standards (1) EP, (3) DE; (b) 11α-hydroxy-products standards (2) 11α-OH EP, (4) 11α-OH DE; (c) recombinant S. cerevisiae INVSc1-CYP68J5, (d) S. cerevisiae wild-type, and (e) recombinant S. cerevisiae INVSc1-CYP68L8