Codon Optimization and Cloning of Bovine Chymosin Gene into pTOLT Expression Plasmid of Escherichia coli

Year 2012, Volume: 1 Issue: 1, 33 - 39, 01.01.2012

Yakup Ulusu İsa Gokce

Abstract

Chymosin is a very important industrial enzyme that commonly used in cheese manufacture. Bovine chymosin is an aspartic protease which is extracted from abomasum of suckling calves. In this study, bovine chymosin gene was first optimized and then cloned into pTOLT E.coli expression system for production of chymosin. Protein engineering of chymosin has also been attempted. A number of companies are now producing recombinant chymosin for commercial use in cheese manufacture

Keywords

pTOLT, codon optimization, chymosin

References

• A. Kumar, s. Grover, J. Sharma, V.K. Batish, Chymosin and other milk coagulants: Source and biotechnological interventions. Crit Rev Biotechnol, 2010, Volume:30, pp: 243
• B., Foltman, A review on prorennin and rennin, Compt Rend Trav Lab Carlsberg, 1966, Volume: 35, pp: 143-231.
• X.P. Jang, M.L. Yin, P. Chen, Q. Yang, Constitutive expression, purification and characterization of bovine prochymosin in Pichia pastoris GS115. World J Microbiol Biotechnol, 2012, DOI:10.1007/s11274-012-1012-7.
• F.L. Davis, B.A. Law, Advances in the microbiology and biochemistry of cheese and fermented milk. 1984, Elsevier Applied Science, London.
• A. Nounai, E. Dako, A. Morsli, N. Belhamiche, S. Belbraouet, M.M. Bellal, A. Dadie, Characterization of the purified coagulant extracted derived from artichoke flowers (Cynara scolymus) and from the fig latex (Ficus carica) in light of their use in the manufacture of traditional cheeses in Algeria. 2009, Journal of Food Technol, Volume: 7, pp: 20-29.
• D.F. Steiner, P.S. Quinn, S.J. Chan, J. Marsh, H.S. Tager, Processing mechanisms in the biosynthesis of proteins. Ann NY Acad Sci, 1980, Volume: 343, pp: 1-16.
• B. Foltman, V.B. Pedersen, M. Jacobsen, D. Kauffman, G. Wijbrandt, The complete amino acid sequence of prochymosin. 1977, Volume: 74, pp: 2321-2314.
• A.K., Mohanty, U.K., Mukhopadhyay, S., Grover, V.K., Batish, Bovine chymosin: Production by rRNA technology and application in cheese manufacture. Biotechnology Advances, 1999, Volume: 17, pp: 205-217.
• P.F. Fox, Rennets and their action in cheese manufacture and ripening. Biotechnol Appl Biochem 1988, Volume: 10 pp: 522–35.
• M.P. Mathur, R.D. Dutta, Rennet from living calves. Dairy Guide 1983, Volume: 5(12) pp: 39–47.
• B. Foltman, General and molecular aspects of rennets. In: Fox PF, editor. Cheese: Chemistry, Physics and Microbiology, 1987, Volume: 1 pp: 37–68.
• N.M.C. Kaye, P. Jolles, The involvement of one of the three histidine residues of cow k- casein in the chymosin initiated milk clotting process. 1978, Biochim Biophys Acta, Volume:40, pp: 536:329.
• R.D. Hill, The nature of the rennin sensitive bond in casein and its possible relation to sensitive bonds in other proteins. 1968, Biochem Biophys Res Commun, Volume: 33 pp:659–
• R.D. Hill, Synthetic peptide and ester substrates for rennin. 1969. J Dairy Res, Volume: pp: 409–15.
• S. Jana, J.K. Deb, Strategies for efficient production of heterologous protein in Escherichia coli. Appl Microbiol Biotechnol. 2005, Volume: 67 pp: 289–298.
• T. Makino, G. Skretas, G: Georgiou, Strain engineering for improved expression of recombinant proteins in bacteria. Microb Cell Fact. 2011, 10:32
• Looman AC, Bodlaender J, Comstock LJ, et al. Influence of the codon following the AUG initiation codon on the expression of a modified lacZ gene in Escherichia coli. EMBO Journal. 1987;6(8):2489–2492.
• (http://www.embl.de/pepcore/pepcore_services/protein_expression/ecoli/optimisation _expression_levels/). D. Baev, X.W. Lil, M. Edgerton, Genetically engineered human salivary histatin genes are functional in Candida albicans: development of a new system for studying histatin candidacidal activity, Microbiol-Sgm. 2001, Volume: 147, pp. 3323-3334. http://www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi?species=83333, 13.08.2012. http://blast.ncbi.nlm.nih.gov, 13.08.2012.