STUDIES ON A NOVEL RECOMBINANT LIPASE A FROM BACILLUS SUBTILIS FS2

Phung Thu Nguyet (Institute of Biotechnology, Vietnamese Academy of Science and Technology, 18 Hoang Quoc Viet Rd., Cau Giay Dist., Hanoi)
Nguyen Hong Thanh (Institute of Biotechnology, Vietnamese Academy of Science and Technology, 18 Hoang Quoc Viet Rd., Cau Giay Dist., Hanoi)
Truong Nam Hai (Institute of Biotechnology, Vietnamese Academy of Science and Technology, 18 Hoang Quoc Viet Rd., Cau Giay Dist., Hanoi)
Jan-Christer Janson (Department of Surface Biotechnology, Biomedical Center, Uppsala University, Uppsala)

ASEAN Journal on Science and Technology for Development
Vol. 25 No. 2 (2008): ASEAN Journal on Science and Technology for Development (AJSTD)

A strain of Bacillus subtilis FS2 isolated from a traditional fish source in Vietnam has been shown to produce an enzyme possessing both a gelatinase and a lipase A (LipA) activity when tested on agar plates containing 0.1% tributyrin and 0.3% gelatine. A gene encoding LipA  was isolated and expressed in E. coli BL21 (DE3). The enzyme has got an apparent Mr of 24 kDa, a pI of 9.2 and was purified to electrophoretical homogeneity by affinity chromatography. Both the lipase and the gelatinase showed optimum activity at pH 10 and at 30 0 C, with highest activity in the range 25-37°C. The metal ions Fe++, Co++, and Mn++ at 10 mM had strong inhibitory effect on both activities whereas Ca++ had no effect and Zn++  had a slight enhancing effect. The specific enzymatic activities obtained were 19814 U/mg for the lipase and 1245 U/mg for the gelatinase.