Chaperone-dependent gene expression of organic solvent-tolerant lipase from Pseudomonas aeruginosa strain S5

Syarul Nataqain Baharum, Raja Noor Zaliha Raja Abdul Rahman, Mahiran Basri, Abu Bakar Salleh

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The gene coding for the intracellular organic solvent-tolerant lipase of Pseudomonas aeruginosa strain S5 was isolated from a genomic DNA library and cloned into pRSET. The cloned sequence included two open reading frames (ORF) of 1575 bp for the first ORF (ORF1), and 582 bp for the second ORF (ORF2). The ORF2, known as chaperone, plays an important role in the expression of the S5 gene. The ORF2 is located downstream of lipase gene, and functions as the act gene for ORF1. The conserved pentapeptide, Gly-X-Ser-X-Gly, is located in the ORF1. A sequence coding for a catalytic triad that resembles that of a serine protease, consisting of serine, histidine, and aspartic acid or glutamic acid residues, was present in the lipase gene. Expression of the S5 lipase gene in E. coli resulted in a 100-fold increase in enzyme activity 9 h after induction with 0.75 mM IPTG. The recombinant protein revealed a size of 60 kDa on SDS-PAGE. The Lip S5 gene was stable in the presence of 25% (v/v) n-dodecane and n-tetradecane after 2 h incubation at 37 °C.

Original languageEnglish
Pages (from-to)346-354
Number of pages9
JournalProcess Biochemistry
Volume45
Issue number3
DOIs
Publication statusPublished - Mar 2010
Externally publishedYes

Fingerprint

Lipases
Lipase
Gene expression
Organic solvents
Pseudomonas aeruginosa
Genes
Gene Expression
Open Reading Frames
Isopropyl Thiogalactoside
Genomic Library
Recombinant proteins
Serine Proteases
Lip
Acids
Gene Library
Recombinant Proteins
Histidine
Aspartic Acid
Enzyme activity
Serine

Keywords

  • Chaperone
  • Expression
  • Organic solvent-tolerant lipase
  • Phylogenetic tree analysis
  • Pseudomonas aeruginosa
  • Shot-gun cloning

ASJC Scopus subject areas

  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Chaperone-dependent gene expression of organic solvent-tolerant lipase from Pseudomonas aeruginosa strain S5. / Baharum, Syarul Nataqain; Rahman, Raja Noor Zaliha Raja Abdul; Basri, Mahiran; Salleh, Abu Bakar.

In: Process Biochemistry, Vol. 45, No. 3, 03.2010, p. 346-354.

Research output: Contribution to journalArticle

Baharum, Syarul Nataqain ; Rahman, Raja Noor Zaliha Raja Abdul ; Basri, Mahiran ; Salleh, Abu Bakar. / Chaperone-dependent gene expression of organic solvent-tolerant lipase from Pseudomonas aeruginosa strain S5. In: Process Biochemistry. 2010 ; Vol. 45, No. 3. pp. 346-354.
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