Lin_2010_Polar.Res_29_421

Reference

Title : Cloning and heterologous expression of two cold-active lipases from the Antarctic bacterium Psychrobacter sp. G. - Lin_2010_Polar.Res_29_421
Author(s) : Lin X , Cui S , Xu G , Wang S , Du N
Ref : Polar Research , 29 :421 , 2010
Abstract :

Antarctic bacteria producing extracellular lipolytic enzymes with activity at low temperature were isolated, and the most promising strain, named G, was identified as a Psychrobacter species based on 16S rDNA sequence alignment. The genomic DNA of this bacterium was used to construct its plasmid genomic library into pUC118 plasmid vectors, and to screen the cold-active lipolytic enzyme genes. Two genes encoding for cold-active lipolytic enzymes, Lip-1452 (with an open reading frame of 1452 bp in length) and Lip-948 (with an open reading frame of 948 bp in length), were screened. The primary structure of the two lipases deduced from the nucleotide sequence showed a consensus pentapeptide containing the active serine (Lip-1452, GDSAG, and Lip-948, GNSMG) and a conserved His-Gly dipeptide in the N-terminal part of the enzyme. Protein sequence alignment and conserved regions analysis indicated that the two lipases probably belonged to family IV and family V of the bacterial lipolytic enzymes, respectively. The upstream and downstream sequences of the two lipolytic lipases were also obtained. The two lipase genes were cloned into the expression vector pCold III and integrated into Escherichia coli BL21 (DE3). The functional expression of both lipase genes by E. coli BL21 (DE3) cells was observed as the formation of clear haloes around colonies on a 1% (vol/vol) tributyrin plate upon induction with isopropyl-b-Dthiogalactopyranoside at 5C. A lipase activity assay showed that the specific activity of the pCold III+Lip-948 expression system was up to 3.7 U ml-1, whereas that of pCold III+Lip-1452 was very low.

PubMedSearch : Lin_2010_Polar.Res_29_421
PubMedID:
Gene_locus related to this paper: psyck-q1q7w8 , psyck-q1qeu6

Related information

Gene_locus psyck-q1q7w8    psyck-q1qeu6

Citations formats

Lin X, Cui S, Xu G, Wang S, Du N (2010)
Cloning and heterologous expression of two cold-active lipases from the Antarctic bacterium Psychrobacter sp. G.
Polar Research 29 :421

Lin X, Cui S, Xu G, Wang S, Du N (2010)
Polar Research 29 :421