African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12196

Full Length Research Paper

Purification and characterization of thermostable and alcohol tolerant lipase from Pseudoxanthomonas sp.

Syifa F. Syihab
  • Syifa F. Syihab
  • Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Jl. Ganesha 10, Bandung Indonesia.
  • Google Scholar
Fida Madayanti
  • Fida Madayanti
  • Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Jl. Ganesha 10, Bandung Indonesia.
  • Google Scholar
Akhmaloka Akhmaloka
  • Akhmaloka Akhmaloka
  • Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Jl. Ganesha 10, Bandung Indonesia.
  • Google Scholar
Made Puspasari Widhiastuty
  • Made Puspasari Widhiastuty
  • Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Jl. Ganesha 10, Bandung Indonesia.
  • Google Scholar


  •  Received: 23 April 2017
  •  Accepted: 25 July 2017
  •  Published: 02 August 2017

Abstract

An extracellular lipase from Psedoxanthomonas sp. was purified 47.3 folds with an overall yield of 27% through purification procedure of acetone precipitation, ion exchange and gel filtration chromatography. Protein precipitation using acetone fractionation showed that the enzyme was precipitated at the fraction of 0 to 40%. Further purification of the enzyme by ion exchange followed by gel filtration chromatography showed that there were two types of lipases with similar molecular size at around 50 kDa. Characterization of optimum pH, temperature, and substrate specificity were carried out against the isolated lipase. Lip1 showed specific substrate preferences towards p-nitro phenyl myristate meanwhile Lip2 exhibited hydrolytic activity towards short and medium acyl chain of the substrate. Further analysis of both enzyme activities on variation of pH and temperature showed that the optimum pH and temperature for Lip1 were at pH 10.0 and 70°C, respectively while pH 8.0 and 50°C were the optimum pH and temperature for Lip2 respectively. Lyophilized lipase isolated from acetone fraction showed lipolytic activity under variation of methanol concentration up to 30%.

Key words: Pseudoxanthomonas lipase, ion exchange chromatography, gel filtration chromatography.