title: Cloning, expression and characterization of a novel cold-active and organic solvent-tolerant esterase from Monascus ruber M7.
abstract: Cold active esterases are a class of important biocatalysts that exhibit high activity at low temperatures. In this study, a search for putative cold-active esterase encoding genes from Monascus ruber M7 was performed. A cold-active esterase, named Lip10, was isolated, cloned, purified, and characterized. Amino acid sequence analysis reveals that Lip10 contained a conserved sequence motif Gly(173)-Xaa-Ser(175)-Xaa-Gly(177) that is also present in the majority of esterases and lipases. Phylogenetic analysis indicated that Lip10 was a novel microbial esterase. The lip10 gene was cloned and heterologously expressed in Escherichia coli BL21(DE3), resulting in the expression of an active and soluble protein that constituted 40 % of the total cell protein content. Lip10 maintained almost 50 % of its maximal activity at 4-10 °C, with optimal activity at 40 °C. Furthermore, Lip10 retained 184-216 % of its original activity, after incubation in 50 % (v/v) hydrophobic organic solvents for 24 h. The enzyme also exhibited high activity under alkaline conditions and good tolerance to metal ions in the reaction mixture. These results indicate that Lip10 may have potential uses in chemical synthesis and food processing industrial applications as an esterase.
authors.affiliation: Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, Ministry of Education, Wuhan, 430070, Hubei, People's Republic of China.|National Key Laboratory of Agro-Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China.|College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
authors.forename: Hailun
authors.initials: H
authors.lastname: Guo
authors.affiliation: Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, Ministry of Education, Wuhan, 430070, Hubei, People's Republic of China.|National Key Laboratory of Agro-Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China.|College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, Ministry of Education, Wuhan, 430070, Hubei, People's Republic of China.|National Key Laboratory of Agro-Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China.|College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, Ministry of Education, Wuhan, 430070, Hubei, People's Republic of China.|National Key Laboratory of Agro-Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China.|College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
authors.forename: Wanping
authors.initials: W
authors.lastname: Chen
authors.affiliation: Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, Ministry of Education, Wuhan, 430070, Hubei, People's Republic of China.|National Key Laboratory of Agro-Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China.|College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
authors.forename: Fusheng
authors.initials: F
authors.lastname: Chen
authors.affiliation: Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, Ministry of Education, Wuhan, 430070, Hubei, People's Republic of China. limu@mail.hzau.edu.cn.|National Key Laboratory of Agro-Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China. limu@mail.hzau.edu.cn.|College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China. limu@mail.hzau.edu.cn.
authors.forename: Mu
authors.initials: M
authors.lastname: Li
mesh_terms: D020816:Amino Acid Motifs; D001565:Benzoates / Q000494:pharmacology; D003080:Cold Temperature; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004795:Enzyme Stability / Q000187:drug effects; D004950:Esterases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D032741:Monascus / Q000201:enzymology* / Q000235:genetics
chemical_list: D001565:Benzoates; D005656:Fungal Proteins; D004950:Esterases; D004121:Dimethyl Sulfoxide
keywords: Cold active; Esterase; Gene cloning and expression; Industrial applications; Monascus ruber