Table 1 Strains and plasmids used in this study

Strains

  Escherichia coli DH 5α

E. coli cloning host

Transgen Biotech

  Mycolicibacterium neoaurum

Source of G6PDH and NADK genes

This lab

  M3

Ksdd and KshA deletion mutant of MNR ATCC 25,795

This lab

  M3-261

M3 containing pMV261 as control

This study

  M3-BM3

M3 expressing original P450-BM3 gene

This study

  M3-mBM3

M3 expressing mP450-BM3 gene (F87A/T260G)

This study

  M3-mBM3-0

M3 expressing mP450-BM3-0 gene (S72W/V78L/A82L

This study

/T88S/A328G/A330W)

  M3-mBM3-0-NADK2

M3 expressing mP450-BM3-0 and NADK2 genes

This study

  M3-mBM3-0-G6PDH2

M3 expressing mP450-BM3-0 and G6PDH2 genes

This study

  M3-mBM3-0-NADK2-G6PDH2

M3 expressing mP450-BM3-0, NADK2 and G6PDH2 genes

This study

Plasmids

  pUC57-BM3

The codon-optimized original P450-BM3 gene delivered

Shanghai Generay

by pUC57, Amp^R

Biotech Co. Ltd

  pUC57-mBM3

The codon-optimized mP450-BM3 gene delivered by

Shanghai Generay

pUC57, Amp^R

Biotech Co. Ltd

  pMV261

Shuttle vector of Mycobacterium and E. coli, P_hsp60, Kan^R

Dr. W. R. Jacobs Jr. for

providing pMV261

  pMV261-BM3

pMV261 containing original P450-BM3 gene, Kan^R

This study

  pMV261-mBM3

pMV261 containing mP450-BM3 gene, Kan^R

This study

  pMV261-mBM3-0

pMV261 containing mP450-BM3-0 gene, Kan^R

This study

  pMV261-NADK

pMV261 containing NADK gene from M. neoaurum, Kan^R

This study

  pMV261-G6PDH

pMV261 containing G6PDH gene from M. neoaurum, Kan^R

This study

  pMV261-mBM3-0-NADK2

pMV261 containing mP450-BM3-0 gene and NADK2

This study

Gene from M. neoaurum, Kan^R

  pMV261-mBM3-0-G6PDH2

pMV261 containing mP450-BM3-0 gene and G6PDH2

This study

Gene from M. neoaurum, Kan^R

  pMV261-mBM3-0-NADK2-

pMV261 containing mP450-BM3-0 gene, NADK2 and

This study

  G6PDH2

G6PDH2 genes from M. neoaurum, Kan^R