Oleic acid based experimental evolution of Bacillus megaterium yielding an enhanced P450 BM3 variant

BMC Biotechnology

Table 2 Sequence alignment of new evolved DE mutant compared to members of the CYP102 family

Organism	p450 Cytochrome	Mutations towards consensus									Mutations leaving consensus
Organism	p450 Cytochrome	A28T	V470E	T664A	T715A	A741G	E887G	M967V	A1008D	D1019E	Q546E	K639A	D893G	E947K
Bacillus megaterium	CYP102A1 WT	A	V	T	T	A	E	M	A	D	Q	K	D	E
	CYP102A1 DE	T	E	A	A	G	G	V	D	E	E	A	G	K
Bacillus subtilis	CYP102A2	S	Q	A	L	G	R	K	D	A	Q	K	D	D
	CYP102A3	S	A	G	L	D	G	V	D	A	G		D	E
Bacillus anthracis	CYP102A4	S	E	A	L	D	N	L	D	E	Q	K	N	T
Bacillus cereus	CYP102A5	S	E	A	L	D	N	L	D	E	Q	K	D	T
Bradyrhizobium diazoefficiens	CYP102A6	H	R	M	A	G	G	A	D	E	Q	A	D	D
Bacillus licheniformis	CYP102A7	S	G	A	F	E	G	L	A	I	E	T	D	D
Bacillus thuringiensis	CYP102A8	S	E	A	L	D	N	L	D	E	Q	K	N	T
Bacillus weihenstephanensis	CYP102A9	S	E	A	L	D	N	L	D	E	Q	K	N	T
Streptomyces avermitilis	CYP102D1	Y	Q	L	L	G	T	V	A	K	R	P	D	Q
Actinosynnema pretiosum	CYP102F1	F	V	A	V	G	S	H	A		H	R	D	E
Consensus	PUsc, S/T	E	A	Hsc, A/L	G	G	Hsc	D	E	Q	K	D	NCsc
Secondary consensus					NCsc	Hsc

Pusc Polar uncharged side chain, Hsc Hydrophobic side chain, NCsc Negatively charged side chain

ISSN: 1472-6750