A novel bacterial carboxylesterase identified in a metagenome derived-clone from Brazilian mangrove sediments.
Francisco J AraujoDenise C HissaGabrielly O SilvaAndré S L M AntunesVanessa L R NogueiraLuciana R B GonçalvesVânia Maria Maciel MeloPublished in: Molecular biology reports (2020)
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.
Keyphrases
- heavy metals
- cell free
- escherichia coli
- circulating tumor
- risk assessment
- single molecule
- polycyclic aromatic hydrocarbons
- microbial community
- wastewater treatment
- drinking water
- crispr cas
- copy number
- human health
- single cell
- genome wide
- amino acid
- organic matter
- klebsiella pneumoniae
- biofilm formation
- fatty acid
- heat shock protein
- gene expression
- cystic fibrosis
- pseudomonas aeruginosa
- circulating tumor cells