Production of milk-clotting enzyme by Bacillus subtilis B1 from wheat bran
Open accessAbstract
Three strains, Bacillus subtilis B1, B. subtilis B18 and Bacillus thuringiensis B12,were screened from wheat bran to produce milk-clotting enzyme. Among them, B. subtilis B1 exhibited considerable milk-clotting activity with low proteolytic activity. After response surface methodology optimization, milk-clotting activity wasimproved from 782 SU ml-1 to 1129.05 ± 74.55 SU mL-1 with a small inoculum size (0.130%, v/v). The optimized medium contained glucose (16.2 gL-1), wheat bran (30 gL-1), NaCl (5 gL-1), MgSO4·7H2O (5 gL-1), KH2PO4 (2 gL-1) and CaCO3 (3 gL-1). The milk-clotting enzyme from B. subtilis B1 has the optimum pH 5.5 and CaCl2concentration 50 mmol L-1. It is completely inactivated after 5 min at 70°C. The result showed that B. subtilis B1 was a promising strain for industrial milk-clotting enzyme production. Key words: Milk-clotting enzyme, Bacillus subtilis, submerged fermentation, wheat bran.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Reduced MCE production by B. subtilis at high salt concentrations could be attributed to the sensitivity of the bacteria to salt and its limited capacity to synthesize MCE under such conditions.
“Reduced MCE production by B. subtilis at high salt concentrations could be attributed to the sensitivity of the bacteria to salt and its limited capacity to synthesize MCE under such conditions (Zhongyang et al., 2011).”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Zhongyang et al., 2011
Cites
Thompson, J. D., Higgins, D. G., & Gibson, T. J. (1994) · Nucleic Acids Research · doi:10.1093/nar/22.22.4673 · Cited by 64,617 · see citing works
16S ribosomal DNA amplification for phylogenetic study
Weisburg, W. G., Barns, S. M., Pelletier, D. A., & Lane, D. J. (1991) · Journal of Bacteriology · doi:10.1128/jb.173.2.697-703.1991 · Cited by 11,970 · see citing works
Developments in the use of Bacillus species for industrial production
Schallmey, M., Singh, A. V., & Ward, O. P. (2004) · Canadian Journal of Microbiology · doi:10.1139/w03-076 · Cited by 1,215 · see citing works
MEGA: Molecular Evolutionary Genetics Analysis software for microcomputers
Kumar, S., Tamura, K., & Nei, M. (1994) · Computer applications in the biosciences · doi:10.1093/bioinformatics/10.2.189 · Cited by 1,166 · see citing works
Advances in the study of proteolysis during cheese ripening
Sousa-Gallagher, M. J., Ardö, Y., & McSweeney, P. L. H. (2001) · International Dairy Journal · doi:10.1016/s0958-6946(01)00062-0 · Cited by 690 · see citing works
Flavor of Cheddar Cheese: A Chemical and Sensory Perspective
Singh, T. K., Drake, M. A., & Cadwallader, K. R. (2003) · Comprehensive Reviews in Food Science and Food Safety · doi:10.1111/j.1541-4337.2003.tb00021.x · Cited by 414 · see citing works
[30] Milk-clotting enzyme from Mucor pusillus var. Lindt
Arima, Κ., Yu, J., & Iwasaki, S. (1970) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(70)19033-1 · Cited by 170 · see citing works
Production and characterization of a milk-clotting enzyme from Aspergillus oryzae MTCC 5341
Vishwanatha, K. S., Rao, A. G. A., & Singh, S. A. (2009) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-009-2197-z · Cited by 102 · see citing works
Milk-clotting enzymes produced by culture of Bacillus subtilis natto.
Shieh, C.-J., Phan Thi, L.-A., & Shih, I.-L. (2009) · Biochemical Engineering Journal · doi:10.1016/j.bej.2008.09.003 · Cited by 99 · see citing works
Porto, A. L. F., de Campos‐Takaki, G. M., & Filho, J. L. L. (1996) · Applied Biochemistry and Biotechnology · doi:10.1007/bf02788066 · Cited by 64 · see citing works
Effect of pH on the Gelation Properties of Skim Milk Gels Made From Plant Coagulants and Chymosin
Esteves, C. L., Lucey, J. A., Wang, T., & Pires, E. (2003) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(03)73850-8 · Cited by 64 · see citing works
Ageitos, J. M., Vallejo, J. A., Sestelo, A. B. F., Poza, M., & Villa, T. G. (2007) · Journal of Applied Microbiology · doi:10.1111/j.1365-2672.2007.03460.x · Cited by 56 · see citing works
Cheddar cheese flavor. Review of current progress
McGugan, W. A. (1975) · Journal of Agricultural and Food Chemistry · doi:10.1021/jf60202a017 · Cited by 53 · see citing works
Dutt, K., Meghwanshi, G. K., Gupta, P., & Saxena, R. K. (2008) · Letters in Applied Microbiology · doi:10.1111/j.1472-765x.2008.02324.x · Cited by 32 · see citing works
So, S. A. T. O., TOKUDA, H., KOIZUMI, T., & NAKANISHI, K. (2004) · Food Science and Technology Research · doi:10.3136/fstr.10.44 · Cited by 30 · see citing works
Rajagopalan, G., & Krishnan, C. (2008) · Journal of Chemical Technology & Biotechnology · doi:10.1002/jctb.2090 · Cited by 23 · see citing works
Fluorescein Thiocarbamoyl-Kappa-Casein Assay for the Specific Testing of Milk-Clotting Proteases
Ageitos, J. M., Vallejo, J. A., Poza, M., & Villa, T. G. (2006) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(06)72418-3 · Cited by 23 · see citing works
Milk-clotting activity of berries extracts from nine Solanum plants
Guiama, V. D., Libouga, D. G., Ngah, E., & Mbofung, C. M. (2010) · AFRICAN JOURNAL OF BIOTECHNOLOGY · doi:10.5897/ajb2010.000-3266 · Cited by 22 · see citing works
Properties of a Milk-Clotting Microbial Enzyme
Melachouris, N., & Tuckey, S. L. (1968) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(68)87049-3 · Cited by 20 · see citing works
Silva, S. V., & Malcata, F. X. (2005) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(05)72870-8 · Cited by 19 · see citing works
Production and Immobilization of a Bacterial Milk-Clotting Enzyme
D’Souza, T. M., & Pereira, L. H. E. S. (1982) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(82)82464-8 · Cited by 13 · see citing works
Wang, Y., Cheng, Q., Ahmed, Z., Jiang, X., & Bai, X. (2009) · Korean Journal of Chemical Engineering · doi:10.1007/s11814-009-0225-4 · Cited by 13 · see citing works
Purification and properties of milk‐clotting enzyme from Bacillus subtilis K‐26
Rao, L. K., & Mathur, D. K. (1979) · Biotechnology and Bioengineering · doi:10.1002/bit.260210402 · Cited by 9 · see citing works
Clotting of cow (Bos taurus) and goat milk ( Capra hircus ) using calve and kid rennets
Libouga, D. G., Ngah, E., Nono, Y. J., & Bitjoka, L. (2008) · AFRICAN JOURNAL OF BIOTECHNOLOGY · doi:10.4314/ajb.v7i19.59358 · Cited by 8 · see citing works
Flagged first, then by citations
1 of the 25 references couldn’t be matched to a known paper.
Cited by
Sorted by citations
A Sight to Wheat Bran: High Value-Added Products
Katileviciute, A., Plakys, G., Budrevičiūtė, A., Önder, K., Damiati, S., & Kodzius, R. (2019) · Biomolecules · doi:10.3390/biom9120887 · Cited by 116 · see citing works
Novel milk-clotting enzyme from Bacillus stearothermophilus as a coagulant in UF-white soft cheese
Ahmed, S. A., Wehaidy, H. R., Ibrahim, O. A., Abd El Ghani, S., & El-Hofi, M. A. (2016) · Biocatalysis and Agricultural Biotechnology · doi:10.1016/j.bcab.2016.06.011 · Cited by 45 · see citing works
Wu, F., Chang, C.-W., & Shih, I. (2013) · SpringerPlus · doi:10.1186/2193-1801-2-33 · Cited by 38 · see citing works
Meng, F., Chen, R., Zhu, X., Lu, Y., Nie, T., Lu, F., & Lu, Z. (2018) · Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.8b01697 · Cited by 36 · see citing works
Mamo, J., Kangwa, M., Fernandez-Lahore, H. M., & Assefa, F. (2020) · Brazilian Journal of Microbiology · doi:10.1007/s42770-020-00243-y · Cited by 35 · see citing works
Zhang, X., Tong, Y., Wang, J., Lyu, X., & Yang, R. (2021) · Journal of Dairy Science · doi:10.3168/jds.2020-19756 · Cited by 33 · see citing works
High Milk-Clotting Activity Expressed by the Newly Isolated Paenibacillus spp. Strain BD3526
Hang, F., Liu, P., Wang, Q., Han, J., Wu, Z., Gao, C., Liu, Z., Zhang, H., & Chen, W. (2016) · Molecules · doi:10.3390/molecules21010073 · Cited by 34 · see citing works
Hang, F., Wang, Q., Hong, Q., Liu, P., Wu, Z., Liu, Z., Zhang, H., & Chen, W. (2016) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2016.01.028 · Cited by 27 · see citing works
Zhao, X., Cai, M., Yang, Z.-J., Luo, T.-Q., Sarwar, A., Megrous, S., Aziz, T., & Yang, Z.-N. (2019) · European Food Research and Technology · doi:10.1007/s00217-019-03361-6 · Cited by 24 · see citing works
Ding, Z., Wang, W.-F., Wang, B., Ouyang, A., Xiao, S., Wang, Y.-N., Liu, S.-P., Ding, M., Zhang, L., & Shi, G.-Y. (2012) · European Food Research and Technology · doi:10.1007/s00217-011-1650-2 · Cited by 17 · see citing works
Guleria, S., Walia, A., Chauhan, A., & Shirkot, C. K. (2016) · Bioresources and Bioprocessing · doi:10.1186/s40643-016-0108-6 · Cited by 16 · see citing works
Zhang, W., He, X., Liu, H., Guo, H., Ren, F., & Wen, P. (2013) · Indian Journal of Microbiology · doi:10.1007/s12088-013-0391-2 · Cited by 13 · see citing works
Zhang, Y., Xia, Y., Lai, P. F. H., Liu, X., Xiong, Z., Liu, J., & Ai, L. (2019) · Annals of Microbiology · doi:10.1007/s13213-019-01513-3 · Cited by 13 · see citing works
Novel <i>Bacillus</i> Milk-Clotting Enzyme Produces Diverse Functional Peptides in Semihard Cheese
Meng, F., Zhao, H., Lu, F., Bie, X., Lu, Z., & Lu, Y. (2021) · Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.0c08120 · Cited by 11 · see citing works
Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review.
Wiśniewski, P., Małkowska-Kowalczyk, M., Łobacz, A., Zadernowska, A., & Żulewska, J. (2025) · Journal of dairy science · doi:10.3168/jds.2025-26973 · Cited by 10 · see citing works
Wehaidy, H. R., Abdel–Naby, M. A., Shousha, W. G., Mallah, M. Y. E., & Shawky, M. M. (2016) · Egyptian Pharmaceutical Journal · doi:10.4103/1687-4315.197584 · Cited by 9 · see citing works
Optimasi Waktu Inkubasi Lactobacillus rhamnosus SKG 34 Dalam Produksi Enzim Penggumpal Susu
Setiadarma, W., Permana, D. G. M., & Nocianitri, K. A. (2020) · Jurnal Ilmu dan Teknologi Pangan (ITEPA) · doi:10.24843/itepa.2020.v09.i02.p01 · Cited by 5 · see citing works
Zerizer, H., Boughachiche, F., Ait-Kaki, A., Rachedi, K., & Okki, M. E. H. E. (2024) · Biomass Conversion and Biorefinery · doi:10.1007/s13399-024-05612-3 · Cited by 4 · see citing works
Role of Clay Substrate Molecular Interactions in Some Dairy Technology Applications
Azzouz, A., Aruș, V., & Platon, N. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25020808 · Cited by 4 · see citing works
Fileto-Pérez, H. A., Ayón, L. G. M., Soto-García, E., Páez‐Lerma, J. B., Soto-Cruz, N. O., García-Caballero, B. E., & Rutiaga-Quiñones, J. G. (2020) · Journal of Dairy Research · doi:10.1017/s0022029920000655 · Cited by 4 · see citing works
Effect of oxygen supply on milk-clotting activity expressed by Paenibacillus spp. strain BD3526
Hang, F., Wang, Q., & Chen, W. (2017) · LWT - Food Science and Technology · doi:10.1016/j.lwt.2017.04.006 · Cited by 3 · see citing works
Mostafa, E.-S. E., & Saad, M. M. (2013) · American Journal of Biotechnology and Molecular Sciences · doi:10.5251/ajbms.2013.3.1.8.23 · Cited by 2 · see citing works
Talantikite-Kellil, S., Nouani, A., Saunier, M., Gillmann, L., Brahmi, A., & Fazouane, F. (2012) · AFRICAN JOURNAL OF BIOTECHNOLOGY · Cited by 1 · see citing works
Mutagenic breeding of Quambalaria cyanescens strain for the production of milk-clotting enzymes.
Wang, C., Rao, H., Zhou, Z., & Zhang, Y. (2014) · PubMed · Cited by 1 · see citing works
Mamo, J., Kangwa, M., Fernández‐Lahore, M., & Assefa, F. (2021) · Bacterial Empire · doi:10.36547/be.257
Weibing, Z., Qiaoqiao, L., Jiang, M., Min, Y., Pencheng, W., Zhongmin, Z., & Xiaoling, H. (2017) · Journal of Advanced Microbiology · doi:10.5530/jam.3.3.2
Yang, W., Gan, L., Zhang, Y., Wang, Y., Zhang, W., Wang, T., & Qiao, H. (2026) · Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.6c07588
- Authors
- Zhongyang, D., Shuangping, L., Zhenghua, G., Liang, Z., Kechang, Z., & Guiyang, S.
- Venue
- AFRICAN JOURNAL OF BIOTECHNOLOGY
- Publisher
- Academic Journals
- Published
- 2011-08-22
- Volume
- 10
- Issue
- 46
- Pages
- 9370-9378
- ISSN
- 1684-5315 (ISSN-L)
- OpenAlex
- W2116770624
- Cited by
- 28
- References
- 25
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 2 · 2021: 3 · 2020: 3 · 2019: 3 · 2018: 1 · 2017: 2 · 2016: 5 · 2014: 1 · 2013: 4 · 2012: 1
- FWCI
- 1.91
- Type
- journal article
- Language
- en
- Open access
- diamond
- doi.org
- Licence
- cc-by
- Locations
- AFRICAN JOURNAL OF BIOTECHNOLOGY (publishedVersion, cc-by)
- 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, Bacillus subtilis, wheat bran, submerged fermentation, enzyme production optimization, Bacillus thuringiensis, proteolytic activity, response surface methodology, fermentation medium optimization, enzyme characterization, industrial enzyme production
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review