PsoP1, a Milk-Clotting Aspartic Peptidase from the Basidiomycete Fungus Piptoporus soloniensis
Abstract
The first enzyme of the basidiomycete Piptoporus soloniensis, a peptidase (PsoP1), was characterized after isolation from submerged cultures, purification by fractional precipitation, and preparative native-polyarylamide gel electrophoresis (PAGE). The native molecular mass of PsoP1 was 38 kDa with an isoelectric point of 3.9. Similar to chymosin from milk calves, PsoP1 showed a maximum milk-clotting activity (MCA) at 35-40 °C and was most stable at pH 6 and below 40 °C. The complete inhibition by pepstatin A identified this enzyme as an aspartic peptidase. Electrospray ionization-tandem MS showed an amino acid partial sequence that was more homologous to mammalian milk clotting peptidases than to the chymosin substitute from a fungal species, such as the Zygomycete Mucor miehei. According to sodium dodecyl sulfate-PAGE patterns, the peptidase cleaved κ-casein in a way similar to chymosin and hydrolyzed β-casein slowly, as it would be expected from an efficient chymosin substitute.
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?
A broader range of stability has been observed for proteases derived from Rhizopus oryzae (pH 4.0 to 9.0), Thermoascus aurantiacus (pH 3.0 to 6.0), and Piptoporus soloniensis (pH 3.0 to 9.0).
“A broader range of stability has been observed for proteases derived from R. oryzae (pH 4.0 to 9.0; Kumar et al., 2005), Thermoascus aurantiacus (pH 3.0 to 6.0; Merheb et al., 2007), and Piptoporus soloniensis (pH 3.0 to 9.0; El-Baky et al., 2011).”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as El-Baky et al., 2011
Most aspartyl proteases synthesized by fungi exhibit the highest activity under acidic conditions.
“as most aspartyl proteases synthesized by fungi exhibit the highest activity under acidic conditions.”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as El-Baky et al., 2011
Cites
PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT
Lowry, O. H., Rosebrough, N., Farr, A. L., & Randall, R. (1951) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)52451-6 · Cited by 315,771 · see citing works
Kappeler, S. R., van den Brink, H. M., Rahbek-Nielsen, H., Farah, Z., Puhan, Z., Hansen, E. B., & Johansen, E. (2006) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2006.02.014 · Cited by 204 · 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
Chymosin and Other Milk Coagulants: Sources and Biotechnological Interventions
Kumar, A., Grover, S., Sharma, J., & Batish, V. K. (2010) · Critical Reviews in Biotechnology · doi:10.3109/07388551.2010.483459 · Cited by 160 · see citing works
Egito, A. S., Girardet, J.-M., Laguna, L. E., Poirson, C., Mollé, D., Miclo, L., Humbert, G., & Gaillard, J.-L. (2007) · International Dairy Journal · doi:10.1016/j.idairyj.2006.09.012 · Cited by 138 · see citing works
488. An improved method of observing the clotting of milk containing rennin
Berridge, N. J. (1952) · Journal of Dairy Research · doi:10.1017/s0022029900006567 · Cited by 104 · 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
Merheb‐Dini, C., Gomes, E., Boscolo, M., & da Silva, R. (2009) · Food Chemistry · doi:10.1016/j.foodchem.2009.09.075 · Cited by 98 · see citing works
Aspartic proteinases from Mucor spp. in cheese manufacturing
Yeğin, S., Fernández‐Lahore, M., Salgado, A. J. G., Güvenç, U., Göksungur, Y., & Tarı, C. (2010) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-010-3020-6 · Cited by 93 · see citing works
Chymosin and aspartic proteinases
Chitpinityol, S., & Crabbe, M. J. C. (1998) · Food Chemistry · doi:10.1016/s0308-8146(97)00090-3 · Cited by 92 · see citing works
Azocasein assay for alkaline protease in complex fermentation broth
Iversen, S. L., & J�rgensen, M. H. (1995) · Biotechnology Techniques · doi:10.1007/bf00152446 · Cited by 69 · see citing works
Recombinant Lamb Chymosin as an Alternative Coagulating Enzyme in Cheese Production
Rogelj, I., Perko, B., Francky, A., Penca, V., & Pungerčar, J. (2001) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(01)74561-4 · Cited by 59 · see citing works
Cloning and structural analysis of the calf prochymosin gene
Hidaka, M., Katsutoshi, S., Uozumi, T., & Beppu, T. (1986) · Gene · doi:10.1016/0378-1119(86)90207-6 · Cited by 54 · see citing works
The first characterized asparaginase from a basidiomycete, Flammulina velutipes
Eisele, N., Linke, D., Bitzer, K., Na’amnieh, S., Nimtz, M., & Berger, R. G. (2010) · Bioresource Technology · doi:10.1016/j.biortech.2010.10.098 · Cited by 54 · see citing works
Vairo-Cavalli, S., Claver, S., Priolo, N., & Natalucci, C. (2005) · Journal of Dairy Research · doi:10.1017/s0022029905000749 · Cited by 54 · see citing works
Yang, J. P., Teplyakov, A., & Quail, J. W. (1997) · Journal of Molecular Biology · doi:10.1006/jmbi.1997.0968 · Cited by 42 · see citing works
Proteolytic Activity of Some Milk-Clotting Enzymes on κ-Casein
Shammet, K. M., Brown, R. J., & McMahon, D. J. (1992) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(92)77889-8 · Cited by 32 · see citing works
Crystal Structure of Aspartic Proteinase from Irpex lacteus in Complex with Inhibitor Pepstatin
Fujimoto, Z., Fujii, Y., Kaneko, S., Kobayashi, H., & Mizuno, H. (2004) · Journal of Molecular Biology · doi:10.1016/j.jmb.2004.06.049 · Cited by 32 · see citing works
Thermal stability of acid proteinases
Walsh, M. K., & LI, X. (2000) · Journal of Dairy Research · doi:10.1017/s0022029900004532 · Cited by 22 · see citing works
Complete primary structure of lamb preprochymosin deduced from cDNA
Pungerčar, J., Štrukelj, B., Gubenšek, F., Türk, V., & Kregar, I. (1990) · Nucleic Acids Research · doi:10.1093/nar/18.15.4602 · Cited by 18 · see citing works
Milk-clotting enzyme from Irpex lacteus as a calf rennet substitute for cheddar cheese manufacture.
Kobayashi, H., Kusakabe, I., & Murakami, K. (1985) · Agricultural and Biological Chemistry · doi:10.1271/bbb1961.49.1605 · Cited by 13 · see citing works
Characterization of Aspartic Proteinase from Basidiomycete, Laetiporus sulphureus
Kobayashi, H., & Hyonok, K. I. M. (2003) · Food Science and Technology Research · doi:10.3136/fstr.9.30 · Cited by 13 · see citing works
Isolation of Milk-Clotting Enzyme from Transgenic Sheep Milk and Its Comparison with Calf Chymosin
Мезина, М. Н., Lavrenova, G. I., Прокофьев, М. И., Starovoitova, V. V., Ермолаев, В. И., Chernyh, V. Y., Balandina, G. N., & Demidovich, S. S. (2001) · Biochemistry (Moscow) · doi:10.1023/a:1010289010462 · Cited by 12 · see citing works
Substrate Specificity of the Milk-clotting Enzyme from Irpex lacteus on κ- and β-Casein
Kobayashi, H., Kusakabe, I., Yokoyama, S., & Murakami, K. (1985) · Agricultural and Biological Chemistry · doi:10.1080/00021369.1985.10866962 · Cited by 10 · see citing works
Studies on Milk Clotting Enzymes Produced by Basidiomycetes
KAWAI, M. (1970) · Agricultural and Biological Chemistry · doi:10.1271/bbb1961.34.164 · Cited by 9 · see citing works
Kawai, M. (1970) · Agricultural and Biological Chemistry · Cited by 7 · see citing works
Screening for Milk-clotting Enzyme from Basidiomycetes
Kobayashi, H., Kim, H., Itoh, T., Kasamo, K., Kusakabe, I., & Murakami, K. (1994) · Bioscience Biotechnology and Biochemistry · doi:10.1271/bbb.58.440 · Cited by 6 · see citing works
Production of milk-clotting enzymes by basidiomycetes
Nerud, F., Mišurcová, Z., & Musilek, V. (1989) · Folia Microbiologica · doi:10.1007/bf02814473 · Cited by 5 · see citing works
Kleinert, T., Lange, I., Rösicke, B., Honig, A. B., & Schleusener, R. (1988) · Acta Biotechnologica · doi:10.1002/abio.370080415 · Cited by 3 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Secreted fungal aspartic proteases: A review
Mandujano-González, V., Villa‐Tanaca, L., Anducho-Reyes, M. Á., & Mercado-Flores, Y. (2016) · Revista Iberoamericana de Micología · doi:10.1016/j.riam.2015.10.003 · Cited by 102 · see citing works
Sun, Q., Chen, F., Geng, F., Luo, Y., Gong, S., & Jiang, Z. (2017) · Food Chemistry · doi:10.1016/j.foodchem.2017.10.113 · Cited by 90 · see citing works
Yeğin, S., & Dekker, P. (2013) · Dairy Science and Technology · doi:10.1007/s13594-013-0137-2 · Cited by 68 · see citing works
Fungal Enzymes: Sources and Biotechnological Applications
Kango, N., Jana, U. K., & Choukade, R. (2019) · Springer eBooks · doi:10.1007/978-981-13-9349-5_21 · Cited by 69 · see citing works
Rahmad, N. B., Al‐Obaidi, J. R., Rashid, N. M. N., Ng, B. Z., Yuswan, M. H., Shaharuddin, N. S., Jamil, N. A. M., & Saleh, N. M. (2014) · Biological Research · doi:10.1186/0717-6287-47-30 · Cited by 51 · see citing works
da Silva, R. R., de Oliveira, L. C. G., Juliano, M. A., Juliano, L., de Oliveira, A. H. C., Rosa, J. C., & Cabral, H. (2017) · Food Chemistry · doi:10.1016/j.foodchem.2017.01.009 · Cited by 47 · see citing works
Silva, B. L., Geraldes, F. M., Murari, C. S., Gomes, E., & Da-Silva, R. (2014) · Applied Biochemistry and Biotechnology · doi:10.1007/s12010-013-0655-7 · Cited by 35 · see citing works
Yin, C., Zheng, L., Chen, L., Tan, Q., Shang, X., & Ma, A. (2013) · Applied Biochemistry and Biotechnology · doi:10.1007/s12010-013-0674-4 · Cited by 27 · see citing works
Kangwa, M., Salgado, J. A. G., & Fernández‐Lahore, M. (2018) · AMB Express · doi:10.1186/s13568-018-0691-3 · Cited by 26 · see citing works
Purification and characterization of a chymosin from Rhizopus microsporus var. rhizopodiformis.
Sun, Q., Wang, X.-P., Yan, Q.-J., Chen, W., & Jiang, Z.-Q. (2014) · Applied Biochemistry and Biotechnology · doi:10.1007/s12010-014-1044-6 · Cited by 25 · see citing works
Mandujano-González, V., Arana-Cuenca, A., Anducho-Reyes, M. Á., Téllez‐Jurado, A., González-Becerra, A. E., & Mercado-Flores, Y. (2013) · Protein Expression and Purification · doi:10.1016/j.pep.2013.10.003 · Cited by 21 · see citing works
Escherichia coli as a production host for novel enzymes from basidiomycota
Zelena, K., Eisele, N., & Berger, R. G. (2014) · Biotechnology Advances · doi:10.1016/j.biotechadv.2014.08.006 · Cited by 19 · see citing works
Hernández-Domínguez, E. M., Rios-Latorre, R. A., Álvarez-Cervantes, J., Loera‐Corral, O., Román‐Gutiérrez, A. D., Dı́az-Godı́nez, G., & Mercado-Flores, Y. (2014) · BioResources · doi:10.15376/biores.9.2.2341-2358 · Cited by 16 · see citing works
Evaluation of the milk clotting properties of an aspartic peptidase secreted by Rhizopus microsporus
da Silva, R. R., Souto, T. B., da Rosa-Garzon, N. G., de Oliveira, L. C. G., Juliano, M. A., Juliano, L., Rosa, J. C., & Cabral, H. (2019) · Preparative Biochemistry & Biotechnology · doi:10.1080/10826068.2019.1683861 · Cited by 16 · see citing works
Potential to develop functional food products from mushroom bioactive compounds.
Shamtsyan, M. (2016) · Journal of Hygienic Engineering and Design · Cited by 15 · see citing works
Pleurotus albidus: A new source of milk-clotting proteases
Martim, S. R., Larissa, S. C. S., Leilane, B. D. S., Edson, J. D. C., Alecrim, M. M., de Vasconcellos, M. C., Ila, M. D. A. O., & Maria, F. S. T. (2017) · African Journal of Microbiology Research · doi:10.5897/ajmr2017.8520 · Cited by 15 · see citing works
Martim, S. R., Silva, L. S. C., Alecrim, M. M., Teixeira, L. S., & Teixeira, M. F. S. (2021) · Acta Scientiarum Biological Sciences · doi:10.4025/actascibiolsci.v43i1.57275 · Cited by 8 · 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
Purification and Characterization of Milk Clotting Enzyme from Edible Mushroom (Pleurotus florida)
(2021) · Letters in Applied NanoBioScience · doi:10.33263/lianbs112.33623373 · Cited by 7 · see citing works
Proteases ácidas de cogumelo comestível da Amazônia para aplicabilidade industrial
Martim, S. R., Silva, L. S. C., Alecrim, M. M., Souza, B. C. D., Oliveira, I. M. D. A., & Teixeira, M. F. S. (2017) · Boletim do Museu Paraense Emílio Goeldi - Ciências Naturais · doi:10.46357/bcnaturais.v12i3.86 · Cited by 6 · see citing works
Brito, É. C. D. M., Braga, R. D. S., Teixeira, M. F. S., & Martim, S. R. (2019) · Boletim do Museu Paraense Emílio Goeldi - Ciências Naturais · doi:10.46357/bcnaturais.v14i3.232 · Cited by 6 · see citing works
Kishimoto, M., Nakamura, K. T., Kanemaru, K., Tasaki, T., Nakamura, T., Sato, K., & Tanimoto, M. (2018) · Food Science and Technology Research · doi:10.3136/fstr.24.139 · Cited by 5 · see citing works
Braga, R. D. S., Brito, É. C. D. M., de Souza, R. Á. T., Teixeira, M. F. S., & Martim, S. R. (2020) · Brazilian Journal of Development · doi:10.34117/bjdv6n11-127 · Cited by 4 · see citing works
Kishimoto, M., Nakamura, K. T., Tasaki, T., MATSUMOTO, K., Nakano, R., & Tanimoto, M. (2020) · Food Science and Technology Research · doi:10.3136/fstr.26.93 · Cited by 4 · see citing works
Guo, Y., Zhou, J., Jia, W., Gao, H., Zhang, H., & Zhang, C. (2023) · Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.2c07303 · Cited by 2 · see citing works
Yao, L., Sun, Y., Ji, R.-Y., Tian, Z., Zhang, S., Zhang, G.-W., & Lv, J. (2025) · Natural Product Research · doi:10.1080/14786419.2025.2565702 · Cited by 1 · see citing works
Minakov, D. V., Уразова, Я. В., & Минакова, А. А. (2022) · Ползуновский вестник · doi:10.25712/astu.2072-8921.2022.03.024 · Cited by 1 · see citing works
Guo, L.-C., Li, Z., Zhang, J. B., Yao, L., & Lv, J.-H. (2026) · Natural Product Research · doi:10.1080/14786419.2026.2689484
da Silva, B. L. (2013) · Aleph UCLA Undergraduate Research Journal for the Humanities and Social Sciences
- Authors
- El-Baky, H. A., Linke, D., Nimtz, M., & Berger, R. G.
- Affiliations
- Leibniz University Hannover (DE)
- Helmholtz Centre for Infection Research (DE)
- Corresponding author
- Berger, Ralf Günter
- Venue
- Journal of Agricultural and Food Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2011-09-02
- Volume
- 59
- Issue
- 18
- Pages
- 10311-10316
- ISSN
- 0021-8561 (ISSN-L)
- 1520-5118
- OpenAlex
- W2326260557
- Cited by
- 30
- References
- 29
- Citations by year
- 2026: 1 · 2025: 2 · 2023: 1 · 2022: 1 · 2021: 2 · 2020: 2 · 2019: 4 · 2018: 2 · 2017: 4 · 2016: 2 · 2014: 4 · 2013: 5
- FWCI
- 2.45
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Agricultural and Food Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Max Planck Institute for Plasma Physics (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
- Life Sciences › Agricultural and Biological Sciences › Food Science › Proteins in Food Systems
- Keywords
- aspartic protease, milk-clotting enzyme, basidiomycete fungi, enzyme characterization, pepstatin inhibition, κ-casein hydrolysis, β-casein hydrolysis, enzyme stability, amino acid sequence homology
- MeSH terms
- Animals, Basidiomycota, Caseins, Milk, Peptide Hydrolases, Substrate Specificity, Aspartic Acid Proteases
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
