Table 1 Summary of CYPs variants-catalyzed regio- and stereoselective steroid hydroxylation by directed evolution

CYP102A1 (Bacillus megaterium)

KSA-1

97% 2β-hydroxytestosterone

79

Directed evolution based on CAST, ISM, MLs, MDs, etc. (Kille et al. 2011)

KSA-1

91% 16β-hydroxyprogesterone

n.d.*

KSA-9

100% 2β-hydroxyprogesterone

n.d.*

KSA-14

94% 15β-hydroxytestosterone

85

M01-A82W

85% 16β-hydroxytestosterone

n.d.

Rational design based on site-directed mutagenesis. (Rea et al. 2012; Venkataraman et al. 2012)

M01-A82W

88% 16β-hydroxynorethisterone

n.d.*

M11-A82W

75% 16β-hydroxytestosterone

n.d.*

M11-A82W

78% 16β-hydroxynorethisterone

n.d.*

M01-A82W/S72I

81% 16α-hydroxytestosterone

n.d.*

WIFI-WC

96% 16β-hydroxyprogesterone

95%

Directed evolution based on CAST, ISM, MLs, MDs, etc. (Acevedo-Rocha et al. 2018)

WIFI-WC

95% 16α-hydroxyandrostenedione

85%

WWV-QRS

92% 16β- hydroxyprogesterone

92%

WWV-HMQ

100% 16β-hydroxyandrostenedione

93%

WWV-Q

90% 16β-hydroxynandrolone

91%

WWV-Q

72% 16β-hydroxyboldenone

71%

WWI

91% 16β-hydroxynorethindrone

59%

LIFI-CW

98% 16β-hydroxynandrolone

71%

LIFI-CW

97% 16β-hydroxyboldenone

70%

139-3

100% 1α-hydroxyandrostenedione

37%

Random mutagenesis. (Liu and Kong 2017)

CYP106A2 (Bacillus megaterium ATCC 13368)

T89N/A395I

81% 11α- hydroxyprogesterone

24.4- fold than WT

Directed evolution based on CAST, ISM, MLs, MDs, etc. (Nguyen et al. 2012; Nikolaus et al. 2017)

A243S

88.8% 6β-hydroxyprogesterone

93.1%

CYP260A1 (myxobacterium Sorangium cellulosum Soce56)

S276N

57% 1α- hydroxyprogesterone

n.d.*

Directed evolution based on CAST, ISM, MLs, MDs, etc. (Khatri et al. 2018)

S276C

48% 1α- hydroxyprogesterone

n.d.*

S276I

64% 17α-hydroxyprogesterone

n.d.*

S276L

62% 17α-hydroxyprogesterone

n.d.*

S276V

58% 17α-hydroxyprogesterone

n.d.*

CYP260B1 (Sorangium cellulosum)

T224A

75% 9α-hydroxy-11-deoxycorticosterone

100%

Rational design based on site-directed mutagenesis (Litzenburger and Bernhardt 2017)