title: Cloning, Expression, and Characterization of a Cold-Active and Organic Solvent-Tolerant Lipase from Aeromicrobium sp. SCSIO 25071.
abstract: The gene encoding lipase (Lip98) from Aeromicrobium sp. SCSIO 25071 was cloned and functionally expressed in Escherichia coli. Lip98 amino acid sequence shares the highest (49%) identity to Rhodococcus jostii RHA1 lipase and contains a novel motif (GHSEG), which is different from other clusters in the lipase superfamily. The recombinant lipase was purified to homogeneity with Ni-NTA affinity chromatography. Lip98 showed an apparent molecular mass of 30 kDa on SDS gel. The optimal temperature and pH value for enzymatic activity were recorded at 30°C and 7.5, respectively. Lip98 exhibited high activity at low temperatures with 35% maximum activity at 0°C and good stability at temperatures below 35°C. Its calculated activation energy was 4.12 kcal/mol at the low temperature range of 15-30°C. Its activity was slightly affected by some metal ions such as K(+), Ca(2+), and Na(+). The activity of Lip98 was increased by various organic solvents such as DMSO, ethanol, acetone, and hexane with the concentration of 30% (v/v) and retained more than 30% residual activity in neat organic solvent. The unique characteristics of Lip98 imply that it is a promising candidate for industrial application as a nonaqueous biocatalyst and food additive.
authors.affiliation: Key Laboratory of Tropical Marine Bio-Resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, P.R. China.|University of Chinese Academy of Sciences, Beijing 100049, P.R. China.
authors.forename: Hongfei
authors.initials: H
authors.lastname: Su
authors.affiliation: Key Laboratory of Tropical Marine Bio-Resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, P.R. China.
authors.forename: Zhimao
authors.initials: Z
authors.lastname: Mai
authors.affiliation: Key Laboratory of Tropical Marine Bio-Resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, P.R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Yang
authors.affiliation: Key Laboratory of Tropical Marine Bio-Resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, P.R. China.|University of Chinese Academy of Sciences, Beijing 100049, P.R. China.
authors.forename: Yunzhu
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Key Laboratory of Tropical Marine Bio-Resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, P.R. China.
authors.forename: Xinpeng
authors.initials: X
authors.lastname: Tian
authors.affiliation: Key Laboratory of Tropical Marine Bio-Resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, P.R. China.
authors.forename: Si
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000096:Acetone / Q000494:pharmacology; D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D003001:Cloning, Molecular / Q000379:methods; D003080:Cold Temperature; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D000431:Ethanol / Q000494:pharmacology; D006586:Hexanes / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008970:Molecular Weight; D016415:Sequence Alignment; D012997:Solvents / Q000378:metabolism* / Q000494:pharmacology*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D006586:Hexanes; D012997:Solvents; D000096:Acetone; D000431:Ethanol; D008049:Lipase; D004121:Dimethyl Sulfoxide
keywords: Aeromicrobium sp. SCSIO 25071; Cold-active; Lipase; Solvent-tolerant