Production of a Milk-Clotting Enzyme by Glutinous Rice Fermentation and Partial Characterization of the Enzyme: Production and Characterization of a Milk-Clotting Enzyme
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Fermentation parameters affecting a milk-clotting enzyme (MCE) production by glutinous rice fermentation were evaluated using selected rice starters called Jiuqu. The MCE was produced under the following conditions: rice inoculation starter at 3%, fermentation time of 5 days, temperature at 30C, initial rice medium pH of 5.0, rice concentration of 70% and rotation speed of 120 rpm. The enzyme preparation obtained by 70% ethanol precipitation of the culture supernatant had milk-clotting activity (MCA) of 6,160 Su/g and a C/P ratio (the ratio between MCA and proteolytic activity) of 10.81 at a recovery rate of 44.82%. The molecular mass of the MCE was 39.7 kDa. MCE production was optimal at 35C and a pH of 5.4. The presence of Na+, K+, Mg2+, Ca2+ and Al3+ stimulated enzyme activity. The Km and Vmax values were 22.90 g/L and 21.11 mL/s, respectively, with skim milk as the substrate. Practical Applications In the last two decades, different sources of milk-clotting enzymes (MCEs) have been explored to meet the increasing demand for cheese production worldwide. This study describes a MCE isolated by ethanol fractionation from glutinous rice wine fermented by Jiuqu. The MCE, with a relatively high C/P ratio, is potentially applicable as a coagulant in cheese making. The property of thermal instability of the enzyme is advantageous as it would be inactivated during curd scalding (at about 50C), thus formation of bitter-taste peptides caused by residual proteolytic activity of the enzyme during cheese ripening could be prevented. In addition, the optimal temperature (35C) of the MCE is within the range of temperature (30–35C) generally used during milk coagulation processes. Therefore, the results suggest that a MCE that could have potential application in cheese production may be obtained by glutinous rice fermentation under the conditions of this study.
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The microbial rennet from glutinous rice-fermented wine was characterized as having strong milk-clotting activity and moderate proteolytic activity.
“The microbial rennet was characterized as having strong milk‐clotting activity and moderate proteolytic activity.”
From Change of proteolysis and sensory profile during ripening of Cheddar-style cheese as influenced by a microbial rennet from rice wine (2019) · cites this paper as Zhao, Wang, Zheng, Zhao, & Yang, 2015
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References 39 · Cited by 11
Research works printed but not linked: 2
- CHEESEMAN, G.C. 1981. Rennet and cheesemaking. In Enzymes and Food Processing (G.G. Birch, N. Blakebrough and K.J. Parker, eds) pp. 195-211, Springer Netherlands, U.K. CHENG, Q., BAI, X. and WANG, Y. 2008. Isolation and fermentation condition of milk-clotting enzyme producing strain from glutinous rice wine. Chinese J. Biotech. 24, 999-1003.
- HORNE, D.S. and BANKS, J.M. 2004. Rennet-induced coagulation of milk. In Cheese: Chemistry, Physics and Microbiology (D.S. Horne and J.M. Banks, eds) pp. 47-71, Elsevier Applied Science, U.K. FIG. 7. (A) THE MILK-CLOTTING ACTIVITY (MCA) CURVE WITH INCREASING CONCENTRATION OF SUBSTRATE; (B) PLOT OF 1/MCA AGAINST 1/[S]. VALUES PRESENTED ARE MEANS ± STANDARD DEVIATION OF SAMPLES ANALYZED IN TRIPLICATE. VERTICAL LINES REPRESENT STANDARD DEVIATIONS
- Authors
- Zhao, X., Wang, J., Zheng, Z., Zhao, A., & Yang, Z.
- Affiliations
- Beijing Technology and Business University (CN)
- School of Food Science; Beijing Laboratory of Food Quality and Safety; Beijing Technology and Business University; No. 11 Fu-Cheng Road Hai-Dian District Beijing 100048 China
- Corresponding author
- Yang, Zhennai
- Venue
- Journal of Food Biochemistry
- Publisher
- Wiley
- Published
- 2015-01-15
- Volume
- 39
- Issue
- 1
- Pages
- 70-79
- ISSN
- 0145-8884 (ISSN-L)
- OpenAlex
- W1574354885
- Cited by
- 11
- References
- 39
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 2 · 2022: 1 · 2020: 1 · 2019: 4 · 2017: 1
- FWCI
- 0.28
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Journal of Food Biochemistry (publishedVersion)
- Funders
- The Key Project of the Educational Committee of Beijing City (KZ201310011011)
- National Public Benefit Research (Agriculture) Foundation (201303085)
- Natural Science Foundation of China (31371804)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Agricultural and Biological Sciences › Food Science › Advanced Scientific Research Methods
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
- Keywords
- milk-clotting enzyme, rice wine, cheese coagulants, enzyme production optimization, ethanol precipitation, C:P ratio, proteolytic activity, thermal stability, enzyme kinetics
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