Cloning and expression of the FR901379 acylase gene from Streptomyces sp. no. 6907.
FR901379 acylase, an enzyme that catalyzes the hydrolysis of the palmitoyl moiety of the antifungal lipopeptide FR901379, was purified from the culture broth of Streptomyces sp. no. 6907 (FERM BP-5809), revealing the 80 kDa, two-subunit heterodimeric protein characteristic of the β-lactam acylase family. Using oligodeoxyribonucleotide primers constructed on the basis of the N-terminal amino acid sequence of each purified subunit, the gene was identified from a cosmid library of Streptomyces sp. no. 6907 DNA. The deduced 775 amino acid sequence corresponded to a single polypeptide chain containing two subunits, and it shared 41.7% identity with aculeacin A acylase from Actinoplanes utahensis NRRL12052. FR901379 acylase activity was found to be 250-fold higher in the recombinant Streptomyces lividans 1326 carrying the cloned gene than in the original Streptomyces sp. no. 6907 strain.
Fermentation and Biotechnology Laboratories, Astellas Pharma, Aichi, Japan.
Satoshi
S
Ueda
Takashi
T
Shibata
Katsuhiko
K
Ito
Nobutaka
N
Oohata
Michio
M
Yamashita
Motohiro
M
Hino
Masato
M
Yamada
Yasuhiro
Y
Isogai
Seiji
S
Hashimoto
D000581:Amidohydrolases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010456:Peptides, Cyclic / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D021122:Protein Subunits; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics*
D017931:DNA Primers; D004269:DNA, Bacterial; C110368:FR 901379; D010456:Peptides, Cyclic; D021122:Protein Subunits; D000581:Amidohydrolases; C019665:amidase