Cited by 42Enzyme Production and CharacterizationProteins in Food SystemsEnzyme Catalysis and Immobilization

Purification, physico-chemico-kinetic characterization and thermal inactivation thermodynamics of milk clotting enzyme from Bacillus subtilis MTCC 10422

Kumari Narwal, Rajesh, Bhushan, Bharat, Pal, Ajay, Panwar, Anil, Malhotra, Sarla·2016·LWT

Abstract

Extracellular milk clotting enzyme from Bacillus subtilis MTCC 10422, which has the capacity of forming milk curds, was purified 32.9 fold with 20.82% recovery using sequential chromatographic techniques. The molecular weight of the enzyme was found to be 27 kDa which is of monomeric nature. The optimum temperature of the enzyme was found to be 45 °C and it was quite stable in the temperature range of 35–65 °C. The enzyme showed the pH optima of 6.0, and quite stable in broad pH range of 5.0–8.0 and showed a typical hyperbolic velocity saturation curve with Km value of 5 mg mL−1 with skim milk as a substrate. Calcium chloride at the concentration of 10 mM was found to be the most effective stimulator, accelerating the enzyme activity by about 2.8 folds. Various kinetic parameters towards thermo-inactivation of milk clotting enzyme were studied. Thermal inactivation behaviour of the purified enzyme suggested its thermostability and also its sensitivity to duration of heat treatment. After purification, both yield and recovery of purified preparation were found appreciable. Since this enzyme has wider pH stability and thermostability, usage of this high activity microbial enzyme for various cheese preparations can be evaluated at commercial scale.

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Crossref · 10.1016/j.lwt.2015.08.065

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36 of 36 references verified0 citation errors · 0 not checkable

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  • The low thermal stability of milk-clotting enzymes enabled the denaturation of MCE residues through temperature control, which can minimize the adverse impact of milk-clotting activity residues on the texture and flavor of cheese throughout the aging process.

    “In addition, the low thermal stability enabled the denaturation of MCE residues through temperature control, which can minimize the adverse impact of milk-clotting activity residues on the texture and flavor of cheese throughout the aging process (Kumari Narwal et al., 2016).”
  • The milk-clotting enzyme from Bacillus subtilis exhibits its highest activity at pH 6.0, with only a modest decline in clotting efficiency at both lower and higher pH values.

    “Kumari Narwal et al. (2016) reported that the milk-clotting enzyme from B. subtilis exhibits its highest activity at pH 6.0, with only a modest decline in clotting efficiency at both lower and higher pH values.”

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Authors
Kumari Narwal, R., Bhushan, B., Pal, A., Panwar, A., & Malhotra, S.
Affiliations
  • Central Institute of Post-Harvest Engineering and Technology (IN)
  • Chaudhary Charan Singh Haryana Agricultural University (IN)
Corresponding author
Bhushan, Bharat
Venue
LWT
Publisher
Elsevier BV
Published
2016-01
Volume
65
Pages
652-660
ISSN
0023-6438 (ISSN-L)
OpenAlex
W2190240927
Cited by
42
References
40
Citations by year
2026: 1 · 2025: 2 · 2024: 1 · 2023: 6 · 2022: 2 · 2021: 9 · 2020: 3 · 2019: 8 · 2017: 6 · 2016: 4
FWCI
2.77
Type
journal article
Language
en
Open access
closed
Locations
LWT (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Proteins in Food Systems
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
Keywords
milk-clotting enzyme, Bacillus subtilis, enzyme purification, thermal stability, thermal inactivation, enzyme kinetics, calcium chloride activation, cheese production, milk curdling, pH stability, molecular weight determination

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