Cloning and sequencing of a poly(DL-lactic acid) depolymerase gene from Paenibacillus amylolyticus strain TB-13 and its functional expression in Escherichia coli.
The gene encoding a poly(DL-lactic acid) (PLA) depolymerase from Paenibacillus amylolyticus strain TB-13 was cloned and overexpressed in Escherichia coli. The purified recombinant PLA depolymerase, PlaA, exhibited degradation activities toward various biodegradable polyesters, such as poly(butylene succinate), poly(butylene succinate-co-adipate), poly(ethylene succinate), and poly(epsilon-caprolactone), as well as PLA. The monomeric lactic acid was detected as the degradation product of PLA. The substrate specificity toward triglycerides and p-nitrophenyl esters indicated that PlaA is a type of lipase. The gene encoded 201 amino acid residues, including the conserved pentapeptide Ala-His-Ser-Met-Gly, present in the lipases of mesophilic Bacillus species. The identity of the amino acid sequence of PlaA with Bacillus lipases was no more than 45 to 50%, and some of its properties were different from those of these lipases.
PRESTO, Japan Science and Technology Corporation, Tsukuba, Ibaraki 305-8572, Japan.
Yukie
Y
Akutsu-Shigeno
Teerawat
T
Teeraphatpornchai
Kamonluck
K
Teamtisong
Nobuhiko
N
Nomura
Hiroo
H
Uchiyama
Tadaatsu
T
Nakahara
Toshiaki
T
Nakajima-Kambe
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D001419:Bacteria / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D001673:Biodegradation, Environmental; D002265:Carboxylic Ester Hydrolases / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D007700:Kinetics; D008049:Lipase / Q000235:genetics; D008969:Molecular Sequence Data; D011091:Polyesters / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D004269:DNA, Bacterial; D011091:Polyesters; D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C425772:poly(lactic acid) depolymerase; D008049:Lipase