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Production and characterization of a milk-clotting enzyme from Aspergillus oryzae MTCC 5341

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Abstract

Microbial milk-clotting enzymes are valued as calf rennet substitutes in the cheese industry. Aspergillus oryzae MTCC 5341 was identified to produce the highest milk-clotting activity during screening of 16 fungal strains. Solid state fermentation using wheat bran along with 4% defatted soy flour and 2% skim milk powder as substrate was optimal for growth of A. oryzae and production of the enzyme. Nearly 40,000 U/g bran of milk-clotting activity was present at the end of 120 h. The enzyme could be recovered by percolating the bran with 0.1 M sodium chloride for 60 min at 4°C. The decolorized enzyme preparation had high ratio of milk clotting to proteolytic activity. Affinity precipitation with alginate and subsequent elution with 0.5 M sodium chloride containing 0.2 M CaCl2 resulted in an enzyme preparation with specific activity of 3,500 U/mg and 72% yield. Optimum pH and temperature for activity of the enzyme were characterized as 6.3 and 55°C, respectively. Milk-clotting enzyme showed differential degree of hydrolysis on casein components. High ratio of milk clotting to proteolytic activity coupled with low thermal stability strengthens the potential usefulness of milk-clotting enzyme of A. oryzae MTCC 5341 as a substitute for calf rennet in cheese manufacturing.

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Acknowledgements

The authors wish to thank Dr. V. Prakash, Director, Central Food Technological Research Institute, Mysore, for his valuable support and suggestions. Department of Biotechnology, Govt. of India is thanked for providing the funds for this research. KSV thanks the Council for Scientific and Industrial Research, Govt. of India, for a research fellowship.

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Authors and Affiliations

  1. Department of Protein Chemistry & Technology, Central Food Technological Research Institute, Mysore, 570020, India

    Kurutahalli S. Vishwanatha, A. G. Appu Rao & Sridevi Annapurna Singh

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  1. Kurutahalli S. Vishwanatha
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  2. A. G. Appu Rao
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  3. Sridevi Annapurna Singh
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Correspondence to Sridevi Annapurna Singh.

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The Central Food Technological Research Institute is a constituent laboratory of Council for Scientific and Industrial Research

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Vishwanatha, K.S., Appu Rao, A.G. & Singh, S.A. Production and characterization of a milk-clotting enzyme from Aspergillus oryzae MTCC 5341. Appl Microbiol Biotechnol 85, 1849–1859 (2010). https://doi.org/10.1007/s00253-009-2197-z

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  • DOI: https://doi.org/10.1007/s00253-009-2197-z

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