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Fermentation parameter and partial biochemical characterisation of milk clotting enzyme from Chinese distiller’s yeast

  • Industrial Microbiology
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Abstract

A strain F34 producing milk clotting enzyme with high yield was obtained from Chinese distiller’s yeast, using casein plate prescreening and fermentation reselection. The optimum fermentation medium for milk clotting enzyme production was as follows: 5.0% rice flour and bran digest, 5.0% bean flour digest, 0.5% CaCl2, 0.5% KCl, initial pH 4.5. The optimal fermentation parameters was at 34°C for 60h. The enzyme action was optimal at 50°C and at about pH 5.5. In the absence of substrate the enzyme showed good stability at 40°C and pH 3.5. Ca2+, Zn2+, Fe3+ and Fe2+ showed stimulation on the milk clotting activity; K+ and Mg2+ showed weak stimulation; Na+, Cu2+, Co2+ and Li2+ showed inhibition. The results indicate that this milk clotting enzyme may partially substitute calf rennet in the cheese manufacture.

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References

  • Abdel Fattah A.F., Ismail A.M.S., El-Aasar S.A. (1984). Production of rennin-like enzyme byAbsidia cylindrospora. J. Agric. Waste, 11: 125–131.

    Article  CAS  Google Scholar 

  • Abdel Fattah A.F., Saleh S.A. (1988). Properties of milk-clotting enzyme fromAspergillus versicolor and isolation of renninlike enzyme. J. Biol. Waste., 25: 109–115.

    Article  CAS  Google Scholar 

  • Abdel Malak C.A., Abou El Adab I.F.G., Vukashinovic V., Zalunin I.A., Timokhina E.A., Lavrenovat G.I., Stepanov V.M. (1996). Buffalo (Bos buffali L.) chymosin purification and properties. J. Comp. Biochem. Physiol., 113B (1): 57–62.

    Article  CAS  Google Scholar 

  • Arima K., Iwasake K. (1967). Milk clotting enzyme from microorganisms part I Screening test and the identification of the potent fungus. J. Agric. Biol. Chem., 31: 540–545.

    CAS  Google Scholar 

  • Arima K., Yu J.Y., Iwasaki S. (1968). Milk-clotting Enzyme from Microorganisms V. Purification and Crystallization ofMucor rennin fromMucor pusillus var.Lindt. J. Appl. Microb., 1727–1733.

  • Bergkvist R. (1963). The proteolytic enzyme ofAspergillus oryzae. 1. Methods for the estimation and isolation of the proteolytic enzymes. J. Acta. Chem. Scand., 17: 1521–1540.

    Article  CAS  Google Scholar 

  • Cavalcanti M.T.H., Teixeira M.F.S., Lima F.J.L. (2004). Partial purification of new milk-clotting enzyme produced byNocardiopsis sp. J. Biores. Technol., 93: 29–35.

    Article  CAS  Google Scholar 

  • Chao S. (2004). Studies on the soliding milk characteristic of microbease and calf rennet. J. Ecol. Domest. Anim., 25: 7–9.

    Google Scholar 

  • Chazarra S., Sidrach L., Lopez Molina D., Rodriguez Lopez J.N. (2007). Characterization of the milk-clotting properties of extracts from artichoke (Cynara scolymus L.) flowers. J. Int. Diary J., 17: 1393–1400.

    Article  CAS  Google Scholar 

  • Dunn G.M., Eds (1985). Nutritional requirements of microorganisms. In: MooYoung, M. (Ed.), Comprehensive Biotechnology, Vol. 1, pp. 113–125. Pergamon Press, Oxford, New York

    Google Scholar 

  • Escobar J., Barnett S. (1993). Effect of agitation speed on the synthesis ofMucor miehei acid Protease. J. Enzyme Microb. Technol., 15: 1009–1013.

    Article  CAS  Google Scholar 

  • Hashem A.M. (1999). Optimization of milk-clotting enzyme productivity byPenicillium oxalicum. J. Biores. Technol., 70: 203–207.

    Article  CAS  Google Scholar 

  • Hashem A.M. (2000). Purification and properties of a milk-clotting enzyme produced byPenicillium oxalicum. J. Biores. Technol., 75: 219–222.

    Article  CAS  Google Scholar 

  • Kumar S., Neeru S.S., Saharan M.R. (2004). Extracellular acid protease fromRhizopus oryzae: purification and characterization. J. Process Biochem., 40: 1701–1705.

    Article  Google Scholar 

  • Masanobu K. (1970). Studies on milk clotting enzyme produced bybasidiornycets. J. A Bial. Cheart., 34(2): 164–169.

    Google Scholar 

  • Petit F., Guespin M.J.F. (1992). Production of an extracellular milk-clotting activity during development inMyxococcus xanthus. J. Bacteriol., 174(15): 5136–5140.

    CAS  PubMed  Google Scholar 

  • Porto A.L.F., Campos T.G.M., Lima F.J.L. (1996). Effects of culture conditions on protease production byStreptomyces clavuligerus growing soy bean flour medium. J. Appl. Biochem. Biotechnol., 60: 115–122.

    Article  CAS  Google Scholar 

  • Seker S., Beyenal H., Ayhan F., Tanyolac A. (1998). Production of microbial rennin fromMucor miehei in a continuously fed fermenter. J. Enzyme Microb. Tech., 23: 469–474.

    Article  CAS  Google Scholar 

  • Shieh C.J., Thi L.A.P., Shih I.L. (2008). Milk clotting enzymes produced by culture ofBacillus subtilis natto. Biochem. Eng. J., doi:10.1016/j.bej.2008.09.003.

  • Stoeva S.P., Mesrob B.K. (1977). Proteolytic and esterolytic of a milk-clotting protease (MCP) FromBacillus mesentericus strain 76. J. Febs. Letters, 80 (1): 86–88.

    Article  CAS  Google Scholar 

  • Thakur M. S., Karanth N. G., Nand K. (1990). Production of fungal rennet byMucor miehei using solid state fermentation. J. Appl. Microbiol. Biotechnol., 32: 409–413.

    Article  CAS  Google Scholar 

  • Tubesha Z.A., Al Delaimy K.S. (2003). Rennin-like milk coagulant enzyme produced by a local isolate ofMucor. J. Int. J. Dairy Technol., 56: 237–241.

    Article  CAS  Google Scholar 

  • Venera G.D., Machalinski C., Zumarraga H., Biscoglio de Jiménez Bonino M.J. (1997). Further characterization and kinetic parameter determination of a milk-clotting protease fromMucor bacilliformis. J. Biochem. Biotech., 68: 207–216.

    Article  CAS  Google Scholar 

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Correspondence to Weiping Chen.

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Wu, J., Chen, H. & Chen, W. Fermentation parameter and partial biochemical characterisation of milk clotting enzyme from Chinese distiller’s yeast. Ann. Microbiol. 58, 717–722 (2008). https://doi.org/10.1007/BF03175580

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