African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 5173

Full Length Research Paper

Production of lipase from genetically improved Streptomyces exfoliates LP10 isolated from oil-contaminated soil

Magda M. Aly1,2*, Sanaa Tork1,3, Saleh M. Al-Garni1 and Lubna Nawar1
1Biology Department, Faculty of Science, King Abdulaziz University, Saudi Arabia. 2Botany Department, Faculty of Science, Kafr El-Sheikh University, Egypt. 3Microbial Genetics Department, National Research Center, Dokki, Cairo, Egypt.
Email: [email protected]

  •  Accepted: 30 November 2011
  •  Published: 16 February 2012

Abstract

Lipases (triacyl glycerol acyl hydrolase) catalyze hydrolysis and syntheses of ester formed from glycerol and long-chain fatty acids. They have many industrial applications, especially in food and detergent industries. Out of 33 bacterial isolates, a group of 20 bacterial isolates produced lipase enzyme on tributyrin agar and Tween 80 agar media. In liquid medium, the lipase activity was ranged from 1.5 to 6.9 IU/ml. Among the evaluated bacteria, the isolate LP10 that was isolated from soil collected from fuel station. It was the most active isolate in lipase production (6.9 IU/ml). Using morphological, physiological and biochemical studies, it was identified as an isolate belonging to the genus Streptomycesand identified as Streptomyces exfoliates LP10. Identification was confirmed using 16S rDNA analysis. Growth of the selected bacterium in medium containing tributyrin and Tween 60 at initial pH 6 in addition to incubation at 37°C for three days yielded the maximum lipase production. The molecular weight of the purified enzyme was 60 kDa, determined using gel electrophoresis. Improvement of lipase production was carried out between Streptomyces exfoliates LP10 and Streptomyces niveus using protoplast fusion. Five fusants were obtained. Fusant LP3 was the best lipase producer (3 times higher) compared to its parents.

 

Key words: Lipase, Streptomyces exfoliates, tributyrin, protoplast fusion, 16S rDNA.