STUDIES ON A NOVEL RECOMBINANT LIPASE A FROM BACILLUS SUBTILIS FS2
PDF

Keywords

Bacillus subtilis FS2
lipase
gelatinase
enzyme activity
recombinant LipA

Abstract

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.  


https://doi.org/10.29037/ajstd.263
PDF

References

Jaeger, K.E., Dijkstra, B.W., and Reetz, M.T. (1999), Bacterial catalysts: molecular biology, three dimension structure and biological application of Lipase, Annual Rev Microbial 53, pp. 315-351.

Jaeger, K.E. and Eggert, T. (2002), Lipases for biotechnology. Curr Opin Biotechnol 13(4), pp. 390-397

Jaeger, K.E. and Reetz, T.M. (1998), Microbial lipases from versatile tools for biotechnology. Trends Biotechnol 16(9), pp. 396-403.

Kourist, R., Bartsch, S., Fransson, L., Hult, K., and Bornscheuer, U.T. (2008), Understanding promiscuous amidase activity of an esterase from Bacillus subtilis, Chembiochem 9(1), pp. 67-9.

Lesuisse, E., Schanck, K., and Colson, C. (1993), Purification and preliminary characterization of the extracellular lipase of Bacillus subtilis 168, an extremely basic pH tolerant enzyme, Eur J Biochem 216, 155-160.

Nguyen Hong Thanh, Phung Thu Nguyet, Truong Nam Hai (2007), Expression of gene encoding lipase (LipA) from Bacillus subtilis FS2, J Biotech 5(1), pp. 41-46.

Phung Thu Nguyet, Tran Minh Tri, Nguyen Hong Thanh, To Kim Anh, Truong Nam Hai (2005) Cloning and expression of gene encoding for collagenase from B. subtilis FS-2. Proceeding of International Vietnam - Korea Symposium:16-21.

Rani, G., Pooja, R., Namita, G., Sapna, B (2003), Lipase assays for conventional and molecular screening: an overview, Biotechnol Appl Biochem. 37, pp. 63-71.

Rhohit, S., Yusuf, C., Uttam. CB (2001), Production, purification, characterization and applications of lipases, Biotechnol Adv 19, pp. 627-662.

Sambrock, J. et al. (1989) Molecular Cloning: A Laboratory Manual. 2d Edition. Cold Spring, Harbor Press, Cold Spring Harbor.

Throsten, E., Gertie, P., Gaelle, P, Isabelle, D., Robert, V., Bauke, W., Dijksra, Karl. E.J. (2002), Biochemical properties and three dimensional structure of two extracellular lipolytic enzymes from Bacillus subtilis, Biointerfaces 26, pp. 37-46.

Uchida, H., Kondo, D., Yamashita, S., Tanakanm T., Tran, L.H., Nagano, H., Uwajima, T. (2004), Purification and properties of protease produced by Bacillus subtilis CN2 from Vietnamese fish sauce, World J Microbiol Biotech 20, pp. 579-582.

Downloads

Download data is not yet available.