Production of rhamnolipids-producing enzymes of Pseudomonas in E. coli and structural characterization
- 45 Downloads
Rhamnolipids are a class of biosurfactants that have a great potential to be used in industries. Five proteins/enzymes, namely RhlA, RhlB, RhlC, RhlG and RhlI, are critical for the production of rhamnolipids in Pseudomonas aeruginosa. Four of the 5 proteins except RhlC were successfully over-expressed in E. coli and three of them (RhlA, RhlB and RhlI) were purified and obtained in milligram quantities. The purified proteins were shown to be folded in solution. Homology models were built for RhlA, RhlB and RhlI. These results lay a basis for further structural and functional characterization of these proteins in vitro to favor the construction of super strains for rhamnolipids production.
Keywordsrhamnolipids Pseudomonas RhlA RhlB RhlI protein folding
Unable to display preview. Download preview PDF.
This research is supported by the Science and Engineering Research Council (SERC) of the Agency for Science, Technology and Research (A*STAR) of Singapore (SERC grant number: 1526004161).
- 2.Desai J D, Banat I M. Microbial production of surfactants and their commercial potential. Microbiology and Molecular Biology Reviews, 1997, 61(1): 47–64Google Scholar
- 9.Wittgens A, Tiso T, Arndt T T, Wenk P, Hemmerich J, Muller C, Wichmann R, Kupper B, Zwick M, Wilhelm S, et al. Growth independent rhamnolipid production from glucose using the nonpathogenic Pseudomonas putida KT2440. Microbial Cell Factories, 2011, 10(80), doi: 10.1186/1475-2859-10-80Google Scholar
- 11.Ochsner U A, Fiechter A, Reiser J. Isolation, characterization, and expression in Escherichia coli of the Pseudomonas aeruginosa rhlAB genes encoding a rhamnosyltransferase involved in rhamnolipid biosurfactant synthesis. Journal of Biological Chemistry, 1994, 269(31): 19787–19795Google Scholar
- 12.Rahim R, Ochsner U A, Olvera C, Graninger M, Messner P, Lam J S, Soberon-Chavez G. Cloning and functional characterization of the Pseudomonas aeruginosa rhlC gene that encodes rhamnosyltransferase 2, an enzyme responsible for di-rhamnolipid biosynthesis. Molecular Microbiology, 2001, 40(3): 708–718CrossRefGoogle Scholar
- 19.Hoang T T, Ma Y, Stern R J, McNeil M R, Schweizer H P. Construction and use of low-copy number T7 expression vectors for purification of problem proteins: Purification of mycobacterium tuberculosis RmlD and Pseudomonas aeruginosa LasI and RhlI proteins, and functional analysis of purified RhlI. Gene, 1999, 237(2): 361–371CrossRefGoogle Scholar
- 20.Biasini M, Bienert S, Waterhouse A, Arnold K, Studer G, Schmidt T, Kiefer F, Cassarino T G, Bertoni M, Bordoli L, et al. SWISSMODEL: Modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Research, 2014, 42(Web Server issue): W252–258Google Scholar
© Higher Education Press and Springer-Verlag Berlin Heidelberg 2017