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 se...

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Bibliographic Details
Published in:FEMS microbiology letters 2004-06, Vol.235 (2), p.349-355
Main Authors: Kwoun Kim, Hyung, Jung, Yeo-Jin, Choi, Won-Chan, Ryu, Hyoung Seok, Oh, Tae-Kwang, Lee, Jung-Kee
Format: Article
Language:English
Subjects:
Agrobacterium tumefaciens
Agrobacterium tumefaciens - enzymology
Agrobacterium tumefaciens - genetics
Amino Acid Sequence
Archaeoglobus fulgidus
Archaeoglobus fulgidus - enzymology
Archaeoglobus fulgidus - genetics
Bacteriology
Biological and medical sciences
Biological Sciences
Cloning, Molecular
computer science
Consensus Sequence
Databases, Genetic
Deinococcus - enzymology
Deinococcus - genetics
Deinococcus radiodurans
Fundamental and applied biological sciences. Psychology
Genome, Archaeal
Genome, Bacterial
information science
libraries
Lipase
Lipase - chemistry
Lipase - genetics
Lipase - metabolism
Microbial genome database
Microbiology
Miscellaneous
Polymerase Chain Reaction
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Summary: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.
ISSN:0378-1097
1574-6968
DOI:10.1016/j.femsle.2004.05.004