Pseudomonas sp. 1401의 5-chloroindole 산화 유전자 클로닝 및 산화 효소를 이용한 물질 전환
Cloning of a gene encoding 5-chloroindole oxygenase from Pseudomonas sp. 1401 and biotransformation application of the oxygenase
Pseudomonas sp. 1401 5-chloroindole 산화 유전자 클로닝 산화효소 염기서열;
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We have cloned a gene from the genomic library of Pseudomonas sp. 1401. This gene was responsible for catalyzing the oxidation of 5-chloroindole to blue pigment, indigo. The open reading frame was 1,242 bp nucleotide sequence, which encodes a protein of 413 amino acids. Oxygenase of 413 amino acid residues has at the N-terminal FAD binding domain and middle and C-end oxygenase domain of p-hydroxybenzoate hydroxylase. The molecular mass of the protein was estimated from amino acid sequence to be 45 kDa with calculated isoelectric point of 5.65. The 5-chloroindole oxygenase showed high amino acid sequence identity to StyA from other Pseudomonas strains and also showed high sequence identities to CDSs with unknown function from genomic sequences of Ralstonia, Streptomyces, Bacillus and Leptospira. StyA from Pseudomonas sp. is large subunit of a two-component styrene monooxygenase, which catalyzes the oxidation of styrene to styrene oxide. But In Pseudomonas sp. 1401, it has no neighboring ORFs similar to the monooxygenase small subunit. In addition, the identified oxygenase catalyzed the oxidation of styrene to various products such as benzaldehyde, benzyl alcohol, phenetyl alcohol, styrene oxide, hydroquinone, phenylethanediol, and 4-hydroxy benzaldehyde. In order to understand the biochemical properties such as substrate specificities, 5-chloroindole oxygenase was expressed in the recombinant E. coli CGSC#7692 strain. The recombinant E. coli CGSC#7692(pJH692) expressing the 5-chloroindole oxygenase was capable of producing dyestuffs from 19 different indole derivatives of 30 test compounds. Thus, 5-chloroindole oxygenase can be applied to generate new dyestuffs from indole derivatives, which may have bioactivity. This results indicate that the 5-chloroindole oxygenase could be a useful biocatalyst to carry out unusual hydroxylation or oxidation.