Molecular cloning and characterization of fengycin synthetase gene fenB from Bacillus subtilis.
A fengycin synthetase gene, fenB, has been cloned and sequenced. The protein (FenB) encoded by this gene has a predicted molecular mass of 143.6 kDa. This protein was overexpressed in Escherichia coli and was purified to near homogeneity by affinity chromatography. Experimental results indicated that the recombinant FenB has a substrate specificity toward isoleucine with an optimum temperature of 25 degrees C, an optimum pH of 4.5, a Km value of 922 microM, and a turnover number of 236 s(-1). FenB also consists of a thioesterase domain, suggesting that this protein may be involved in the activation of the last amino acid of fengycin.
Lin
G H
GH
Graduate Institute of Botany, National Taiwan University, Taipei.
Chen
C L
CL
Tschen
J S
JS
Tsay
S S
SS
Chang
Y S
YS
Liu
S T
ST
D000595:Amino Acid Sequence; D049148:Aminoacylation; D000935:Antifungal Agents / Q000096:biosynthesis*; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D007532:Isoleucine / Q000378:metabolism; D007700:Kinetics; D055666:Lipopeptides; D008074:Lipoproteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010453:Peptide Synthases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D000935:Antifungal Agents; D055666:Lipopeptides; D008074:Lipoproteins; D011994:Recombinant Proteins; D007532:Isoleucine; C049972:fengycin; D010453:Peptide Synthases; C110972:fengycin synthetase