Cloning and  Sequence Analysis of Lipase Gene of Halophilic Bacteria Isolated from Mud Crater of Bledug Kuwu, Central Java, Indonesia

Mukhammad Asyari¹,  Pingkan Aditiawati², Akhmaloka¹, Rukman Hertadi^1*

¹Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Ganesha no. 10 Bandung 40132, Indonesia. ²Microbial Biotechnology Research Group, School of Life Sciences and Technology, Institut Teknologi Bandung, Jl. Ganesha  no. 10 Bandung 40132, Indonesia.

DOI : http://dx.doi.org/10.13005/bbra/1856

ABSTRACT: Five lipase genes (lipab4, lipab8, lipab15, lipag18, and lipab18) were isolated from Halomonas and Chromohalobacter local strains of Bledug Kuwu isolates. Based on amino acid sequence analysis, the genes showed some unique motif of amino acid sequences. All of lipases were classified as member of family IV (HSL=hormone-sensitive lipase). These lipases show high similarities of conserved regions with lipolytic of Halomonas and formed a distinct cluster with other types of HSL, such as esterase/lipase and carboxylesterase. All of lipases contain more negative charged of amino acid residues compared to the mesophilic and thermophilic ones, and tend to have similarity to lipases of moderate halophilics. The result of homology and phylogenetic analysis showed that these lipases were clustered into three groups. Group I (lipab8, lipab18 and lipag18) are close to lipolytic gene of Halomonas elongata DSM 2581, while groups II (lipab4) and III (lipab15) create new branches in the phylogenetic tree. In addition, analysis of GC, GC-AT and GC-AT content on the codon usage of the genes revealed the unique profile compared to that the other lipase genes.

KEYWORDS: Halomonas; Chromohalobacter; Halostable lipase; HSL Family; GC-AT content; Bledug Kuwu

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