期刊
MOLECULAR BIOLOGY REPORTS
卷 47, 期 5, 页码 3919-3928出版社
SPRINGER
DOI: 10.1007/s11033-020-05484-6
关键词
Metagenomic library; 1; 4-Butanediol diacrylate esterase; Family VIII; Porticoccus; Heterologous expression
资金
- National Council for Scientific and Technological Development (CNPq)
- Foundation for Scientific and Technological Development of Ceara (FUNCAP) [PRONEX-PR2-0101-00012.01.00/15]
A functional screening of 1152 clones from a plasmid library constructed with DNA extracted from Brazilian mangrove sediments revealed 3 positive clones for ester-hydrolyzing enzymes, or about one lipolytic gene per 1.2 Mb DNA, which corroborates the idea that oil-contaminated mangroves are a good source of novel microbial lipases/esterases. The partial sequence of the clone LipG7 (1179 bp) showed 30.2% of predicted structure identity with a known esterase, confirming LipG7 as a new member of family VIII esterases. LigG7 shared 80% sequence identity with 1,4-butanediol diacrylate esterase from the Gammaprotebacteria Porticoccus hydrocarbonoclasticus, suggesting it belongs to the Porticoccaceae family. LipG7 was heterologously expressed in Escherichia coli Rosetta-Gami DE3; the purified recombinant enzyme exhibited a predicted molecular weight of 45.2 kDa and exceptional activity towards 4-nitrophenyl butyrate, compared with other recombinant esterases, highlighting its enormous potential for biological applications.
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