Purification and characterization of a milk clotting protease from Mucor bacilliformis

An acid protease having milk clotting activity has been isolated from Mucor bacilliformis cultures. The enzyme was basically purified by ionic exchange chromatography. An average yield of 29 mg purified product was obtained from 100 mL crude extract. As purity criteria, SDS-PAGE, reverse-phase HPLC,...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Applied biochemistry and biotechnology 1992-12, Vol.37 (3), p.283-294
Hauptverfasser: ARECES, L. B, DE JIMENEZ BONINO, M. B, PARRY, M. A. A, FRAILE, E. R, FERNANDEZ, H. M, CASCONE, O
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 294
container_issue 3
container_start_page 283
container_title Applied biochemistry and biotechnology
container_volume 37
creator ARECES, L. B
DE JIMENEZ BONINO, M. B
PARRY, M. A. A
FRAILE, E. R
FERNANDEZ, H. M
CASCONE, O
description An acid protease having milk clotting activity has been isolated from Mucor bacilliformis cultures. The enzyme was basically purified by ionic exchange chromatography. An average yield of 29 mg purified product was obtained from 100 mL crude extract. As purity criteria, SDS-PAGE, reverse-phase HPLC, and N-terminal analysis were performed. The protease is a protein composed of a single polypeptide chain with glycine at the N-terminus. The mol wt is approx 32,000, and its amino acid composition is very similar to those of other fungal proteases. As expected, its clotting activity was drastically inhibited by pepstatin A action. On the other hand, its instability against heat treatment and its clotting/proteolytic activity ratio indicate that it may be considered as a potential substitute for bovine chymosin.
doi_str_mv 10.1007/BF02788880
format Article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_73521367</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>73521367</sourcerecordid><originalsourceid>FETCH-LOGICAL-f294t-9609a4e8dd20f032a6d8db6a87083df1c8e31c0c983f4f4b9b1a793b20f2dae93</originalsourceid><addsrcrecordid>eNo9kM1LxDAQxYMouq5evCs5iLfqJOlHctTFL1AU_LiWaZqs0bZZk_agf72BXXyXgfd-zDyGkCMG5wyguri6AV7JJNgiM1YUKgOu2DaZJVtknEu1R_Zj_ARgXBbVLtllAgSUxYy8P0_BWadxdH6gOLRUf2BAPZrgftemtxRp77ovqjs_jm5Y0lXwo8FoqA2-p4-T9oE2qF3XOetD7-IB2bHYRXO4mXPydnP9urjLHp5u7xeXD5nlKh8zVYLC3Mi25WBBcCxb2TYlygqkaC3T0gimQSspbG7zRjUMKyWaRPMWjRJzcrbemxp9TyaOdTquTdfhYPwU60oUnImySuDxBpya3rT1Krgew0-9eUTKTzc5Ro2dDThoF_-xvMqTIGEna8yir3EZEvL2wpTKAVJaKvEHb991Qw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>73521367</pqid></control><display><type>article</type><title>Purification and characterization of a milk clotting protease from Mucor bacilliformis</title><source>MEDLINE</source><source>SpringerNature Journals</source><creator>ARECES, L. B ; DE JIMENEZ BONINO, M. B ; PARRY, M. A. A ; FRAILE, E. R ; FERNANDEZ, H. M ; CASCONE, O</creator><creatorcontrib>ARECES, L. B ; DE JIMENEZ BONINO, M. B ; PARRY, M. A. A ; FRAILE, E. R ; FERNANDEZ, H. M ; CASCONE, O ; UBA, CONICET, Buenos Aires, Argentina</creatorcontrib><description>An acid protease having milk clotting activity has been isolated from Mucor bacilliformis cultures. The enzyme was basically purified by ionic exchange chromatography. An average yield of 29 mg purified product was obtained from 100 mL crude extract. As purity criteria, SDS-PAGE, reverse-phase HPLC, and N-terminal analysis were performed. The protease is a protein composed of a single polypeptide chain with glycine at the N-terminus. The mol wt is approx 32,000, and its amino acid composition is very similar to those of other fungal proteases. As expected, its clotting activity was drastically inhibited by pepstatin A action. On the other hand, its instability against heat treatment and its clotting/proteolytic activity ratio indicate that it may be considered as a potential substitute for bovine chymosin.</description><identifier>ISSN: 0273-2289</identifier><identifier>EISSN: 1559-0291</identifier><identifier>DOI: 10.1007/BF02788880</identifier><identifier>PMID: 1303065</identifier><identifier>CODEN: ABIBDL</identifier><language>eng</language><publisher>Heidelberg: Springer</publisher><subject>Amino Acids - analysis ; Animals ; Aspartic Acid Endopeptidases - isolation &amp; purification ; Aspartic Acid Endopeptidases - metabolism ; Biological and medical sciences ; Biotechnology ; Caseins - metabolism ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; cuajo ; Culture Media ; Electrophoresis, Polyacrylamide Gel ; Enzyme engineering ; Fundamental and applied biological sciences. Psychology ; Hydrogen-Ion Concentration ; Improved methods for extraction and purification of enzymes ; Kinetics ; Methods. Procedures. Technologies ; microorganisme ; microorganismos ; microorganisms ; Milk - metabolism ; Molecular Weight ; mucor ; Mucor - enzymology ; presure ; proteolisis ; proteolyse ; proteolysis ; purificacion ; purification ; rennet</subject><ispartof>Applied biochemistry and biotechnology, 1992-12, Vol.37 (3), p.283-294</ispartof><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=4744440$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1303065$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>ARECES, L. B</creatorcontrib><creatorcontrib>DE JIMENEZ BONINO, M. B</creatorcontrib><creatorcontrib>PARRY, M. A. A</creatorcontrib><creatorcontrib>FRAILE, E. R</creatorcontrib><creatorcontrib>FERNANDEZ, H. M</creatorcontrib><creatorcontrib>CASCONE, O</creatorcontrib><creatorcontrib>UBA, CONICET, Buenos Aires, Argentina</creatorcontrib><title>Purification and characterization of a milk clotting protease from Mucor bacilliformis</title><title>Applied biochemistry and biotechnology</title><addtitle>Appl Biochem Biotechnol</addtitle><description>An acid protease having milk clotting activity has been isolated from Mucor bacilliformis cultures. The enzyme was basically purified by ionic exchange chromatography. An average yield of 29 mg purified product was obtained from 100 mL crude extract. As purity criteria, SDS-PAGE, reverse-phase HPLC, and N-terminal analysis were performed. The protease is a protein composed of a single polypeptide chain with glycine at the N-terminus. The mol wt is approx 32,000, and its amino acid composition is very similar to those of other fungal proteases. As expected, its clotting activity was drastically inhibited by pepstatin A action. On the other hand, its instability against heat treatment and its clotting/proteolytic activity ratio indicate that it may be considered as a potential substitute for bovine chymosin.</description><subject>Amino Acids - analysis</subject><subject>Animals</subject><subject>Aspartic Acid Endopeptidases - isolation &amp; purification</subject><subject>Aspartic Acid Endopeptidases - metabolism</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Caseins - metabolism</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Chromatography, Ion Exchange</subject><subject>cuajo</subject><subject>Culture Media</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzyme engineering</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hydrogen-Ion Concentration</subject><subject>Improved methods for extraction and purification of enzymes</subject><subject>Kinetics</subject><subject>Methods. Procedures. Technologies</subject><subject>microorganisme</subject><subject>microorganismos</subject><subject>microorganisms</subject><subject>Milk - metabolism</subject><subject>Molecular Weight</subject><subject>mucor</subject><subject>Mucor - enzymology</subject><subject>presure</subject><subject>proteolisis</subject><subject>proteolyse</subject><subject>proteolysis</subject><subject>purificacion</subject><subject>purification</subject><subject>rennet</subject><issn>0273-2289</issn><issn>1559-0291</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kM1LxDAQxYMouq5evCs5iLfqJOlHctTFL1AU_LiWaZqs0bZZk_agf72BXXyXgfd-zDyGkCMG5wyguri6AV7JJNgiM1YUKgOu2DaZJVtknEu1R_Zj_ARgXBbVLtllAgSUxYy8P0_BWadxdH6gOLRUf2BAPZrgftemtxRp77ovqjs_jm5Y0lXwo8FoqA2-p4-T9oE2qF3XOetD7-IB2bHYRXO4mXPydnP9urjLHp5u7xeXD5nlKh8zVYLC3Mi25WBBcCxb2TYlygqkaC3T0gimQSspbG7zRjUMKyWaRPMWjRJzcrbemxp9TyaOdTquTdfhYPwU60oUnImySuDxBpya3rT1Krgew0-9eUTKTzc5Ro2dDThoF_-xvMqTIGEna8yir3EZEvL2wpTKAVJaKvEHb991Qw</recordid><startdate>19921201</startdate><enddate>19921201</enddate><creator>ARECES, L. B</creator><creator>DE JIMENEZ BONINO, M. B</creator><creator>PARRY, M. A. A</creator><creator>FRAILE, E. R</creator><creator>FERNANDEZ, H. M</creator><creator>CASCONE, O</creator><general>Springer</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>19921201</creationdate><title>Purification and characterization of a milk clotting protease from Mucor bacilliformis</title><author>ARECES, L. B ; DE JIMENEZ BONINO, M. B ; PARRY, M. A. A ; FRAILE, E. R ; FERNANDEZ, H. M ; CASCONE, O</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-f294t-9609a4e8dd20f032a6d8db6a87083df1c8e31c0c983f4f4b9b1a793b20f2dae93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Amino Acids - analysis</topic><topic>Animals</topic><topic>Aspartic Acid Endopeptidases - isolation &amp; purification</topic><topic>Aspartic Acid Endopeptidases - metabolism</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Caseins - metabolism</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Chromatography, Ion Exchange</topic><topic>cuajo</topic><topic>Culture Media</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzyme engineering</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hydrogen-Ion Concentration</topic><topic>Improved methods for extraction and purification of enzymes</topic><topic>Kinetics</topic><topic>Methods. Procedures. Technologies</topic><topic>microorganisme</topic><topic>microorganismos</topic><topic>microorganisms</topic><topic>Milk - metabolism</topic><topic>Molecular Weight</topic><topic>mucor</topic><topic>Mucor - enzymology</topic><topic>presure</topic><topic>proteolisis</topic><topic>proteolyse</topic><topic>proteolysis</topic><topic>purificacion</topic><topic>purification</topic><topic>rennet</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>ARECES, L. B</creatorcontrib><creatorcontrib>DE JIMENEZ BONINO, M. B</creatorcontrib><creatorcontrib>PARRY, M. A. A</creatorcontrib><creatorcontrib>FRAILE, E. R</creatorcontrib><creatorcontrib>FERNANDEZ, H. M</creatorcontrib><creatorcontrib>CASCONE, O</creatorcontrib><creatorcontrib>UBA, CONICET, Buenos Aires, Argentina</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Applied biochemistry and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>ARECES, L. B</au><au>DE JIMENEZ BONINO, M. B</au><au>PARRY, M. A. A</au><au>FRAILE, E. R</au><au>FERNANDEZ, H. M</au><au>CASCONE, O</au><aucorp>UBA, CONICET, Buenos Aires, Argentina</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification and characterization of a milk clotting protease from Mucor bacilliformis</atitle><jtitle>Applied biochemistry and biotechnology</jtitle><addtitle>Appl Biochem Biotechnol</addtitle><date>1992-12-01</date><risdate>1992</risdate><volume>37</volume><issue>3</issue><spage>283</spage><epage>294</epage><pages>283-294</pages><issn>0273-2289</issn><eissn>1559-0291</eissn><coden>ABIBDL</coden><abstract>An acid protease having milk clotting activity has been isolated from Mucor bacilliformis cultures. The enzyme was basically purified by ionic exchange chromatography. An average yield of 29 mg purified product was obtained from 100 mL crude extract. As purity criteria, SDS-PAGE, reverse-phase HPLC, and N-terminal analysis were performed. The protease is a protein composed of a single polypeptide chain with glycine at the N-terminus. The mol wt is approx 32,000, and its amino acid composition is very similar to those of other fungal proteases. As expected, its clotting activity was drastically inhibited by pepstatin A action. On the other hand, its instability against heat treatment and its clotting/proteolytic activity ratio indicate that it may be considered as a potential substitute for bovine chymosin.</abstract><cop>Heidelberg</cop><pub>Springer</pub><pmid>1303065</pmid><doi>10.1007/BF02788880</doi><tpages>12</tpages></addata></record>
fulltext fulltext
identifier ISSN: 0273-2289
ispartof Applied biochemistry and biotechnology, 1992-12, Vol.37 (3), p.283-294
issn 0273-2289
1559-0291
language eng
recordid cdi_proquest_miscellaneous_73521367
source MEDLINE; SpringerNature Journals
subjects Amino Acids - analysis
Animals
Aspartic Acid Endopeptidases - isolation & purification
Aspartic Acid Endopeptidases - metabolism
Biological and medical sciences
Biotechnology
Caseins - metabolism
Chromatography, High Pressure Liquid
Chromatography, Ion Exchange
cuajo
Culture Media
Electrophoresis, Polyacrylamide Gel
Enzyme engineering
Fundamental and applied biological sciences. Psychology
Hydrogen-Ion Concentration
Improved methods for extraction and purification of enzymes
Kinetics
Methods. Procedures. Technologies
microorganisme
microorganismos
microorganisms
Milk - metabolism
Molecular Weight
mucor
Mucor - enzymology
presure
proteolisis
proteolyse
proteolysis
purificacion
purification
rennet
title Purification and characterization of a milk clotting protease from Mucor bacilliformis
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T13%3A41%3A36IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Purification%20and%20characterization%20of%20a%20milk%20clotting%20protease%20from%20Mucor%20bacilliformis&rft.jtitle=Applied%20biochemistry%20and%20biotechnology&rft.au=ARECES,%20L.%20B&rft.aucorp=UBA,%20CONICET,%20Buenos%20Aires,%20Argentina&rft.date=1992-12-01&rft.volume=37&rft.issue=3&rft.spage=283&rft.epage=294&rft.pages=283-294&rft.issn=0273-2289&rft.eissn=1559-0291&rft.coden=ABIBDL&rft_id=info:doi/10.1007/BF02788880&rft_dat=%3Cproquest_pubme%3E73521367%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=73521367&rft_id=info:pmid/1303065&rfr_iscdi=true