Sequence-based approach to finding functional lipases from microbial genome databases.
A sequence-based approach was used to retrieve functional lipases from microbial genome databases. Many novel genes assigned as putative lipases were tested using the criteria of the typical lipase sequence rule, based on a consensus sequence of a catalytic triad (Ser, Asp, His) and oxyanion hole sequence (HG). To obtain the lipase genes satisfying the sequence rule, PCR cloning was performed, while the lipase activities were tested using a tributyrin/tricaprylin plate and p-nitrophenyl caproate. Among nine putative lipases from four strains, five functional lipolytic proteins were obtained from Archaeoglobus fulgidus, Deinococcus radiodurans, and Agrobacterium tumefaciens. All five lipases exhibited a relatively low sequence similarity (less than 26.7%) with known lipases and turned out to belong to different lipase families. Accordingly, the current results indicate that the proposed strategic approach based on the microbial genome is an efficient and rapid method for finding novel and functional lipases.
Kwoun Kim
Hyung
H
Division of Biotechnology, The Catholic University of Korea, 43-1, Yokkok 2-dong, Wonmi-gu, Bucheon, Gyeonggido 420-743, Republic of Korea.
Jung
Yeo-Jin
YJ
Choi
Won-Chan
WC
Ryu
Hyoung Seok
HS
Oh
Tae-Kwang
TK
Lee
Jung-Kee
JK
D016960:Agrobacterium tumefaciens / Q000201:enzymology / Q000235:genetics; D000595:Amino Acid Sequence; D019611:Archaeoglobus fulgidus / Q000201:enzymology / Q000235:genetics; D003001:Cloning, Molecular; D016384:Consensus Sequence; D030541:Databases, Genetic; D034301:Deinococcus / Q000201:enzymology / Q000235:genetics; D020745:Genome, Archaeal; D016680:Genome, Bacterial; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction
D008049:Lipase