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LIPASE FROM PSEUDOMONAS SP.: REACTIONS, CLONING, AND AMINO ACID SEQUENCE ANALYSIS
Nishioka, Takaaki Chihara-Shiomi, Mikiko Yoshikawa, Kazuhiro Inagaki, Minoru Yamamoto, Yukio Hiratake, Jun Baba, Naomichi Oda, Jun'ichi
Nishioka, Takaaki
Chihara-Shiomi, Mikiko
Yoshikawa, Kazuhiro
Inagaki, Minoru
Yamamoto, Yukio
Hiratake, Jun
Baba, Naomichi
Oda, Jun'ichi
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Issue Date
1991
Submitted date
2024-03-27
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Abstract
Lipases catalyze reactions not only in water but also in organic solvents. We have developed several asymmetric organic syntheses using lipase from Pseudomonas fluorescens in organic solvents. However, all attempts to acylate [1,1’-binaphthyl]-2,2’-diol with the lipase using enol esters failed. Among the lipases we screened for this substrate, lipase from Pseudomonas sp. is the only enzyme that catalyzes both stereoselective acylation of the diol and deacylation of its esters in organic solvents. The lipase shows higher activity in the hydrolysis of triglyceride into glycerol when compared with other lipases. Because of these characteristic reactivities of the lipase from Pseudomonas sp., we were interested in the cloning and sequencing of the gene of the enzyme and amino acid sequence analysis. The gene cloned from Pseudomonas sp. genome DNAwas 933 base pairs and inserted in plasmid pKK233-2. E. ‘coli JM109 transformed with the recombinant plasmid showed lipolysis asa zone of clearing around a colony on agar plate containing tributyrin. Amino acid sequence deduced from the DNA sequence is similar to that of lipase from Pseudomonas fragi (47 % identical). Content of serine residue is unusually high as 12 %. Sequence alignment with other 19 amino acid sequences of lipases suggested that Ser82, His251, and Asp209 are the catalytic residues.
Citation
Lipases : structure, mechanism and genetic engineering, 253 - 262
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Book chapter
conference paper
conference paper
Language
en
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Series/Report no.
GBF monographs ; Volume 16
ISSN
0930-4320
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ISBN
156081165X
3527283323
3527283323
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Attribution-NonCommercial-ShareAlike 4.0 International