Purification and characterisation of xylitol dehydrogenase from Neurospora crassa NCL communication No. 6347

Abstract The purification and characterisation of one of the key enzymes in the conversion of xylose to ethanol, xylitol dehydrogenase (XD), from Neurospora crassa is reported. This organism has the unique ability to directly convert biomass to ethanol. The enzyme was purified 81-fold by fractional...

Full description

Saved in:
Bibliographic Details
Published in:FEMS microbiology letters 1997-01, Vol.146 (1), p.79-83
Main Authors: Phadtare, S.U, Rawat, U.B, Rao, M.B
Format: Article
Language:English
Subjects:
Ammonium
Ammonium sulfate
Calcium
Calcium ions
Cellulose
Column chromatography
Dehydrogenase
Dehydrogenases
Enzymes
Ethanol
Gel electrophoresis
Gel filtration
Magnesium
Microbiology
NAD
Neurospora
Neurospora crassa
pH effects
Purification
Sodium lauryl sulfate
Sorbitol
Xylitol
Xylitol dehydrogenase
Xylose
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c2889-730f461ea56508b16e91c6865ec04372e5bf02b81b3ab41d67417f1c38fbac8b3
cites cdi_FETCH-LOGICAL-c2889-730f461ea56508b16e91c6865ec04372e5bf02b81b3ab41d67417f1c38fbac8b3
container_end_page 83
container_issue 1
container_start_page 79
container_title FEMS microbiology letters
container_volume 146
creator Phadtare, S.U
Rawat, U.B
Rao, M.B
description Abstract The purification and characterisation of one of the key enzymes in the conversion of xylose to ethanol, xylitol dehydrogenase (XD), from Neurospora crassa is reported. This organism has the unique ability to directly convert biomass to ethanol. The enzyme was purified 81-fold by fractional ammonium sulfate precipitation, column chromatography on DEAE-cellulose, Sephacryl S-200 and β-NAD agarose. The purified enzyme had a native molecular mass of 87 kDa (gel filtration) and was composed of two identical subunits of molecular mass 43.6 kDa (SDS-PAGE). The enzyme was most active at pH 8.4 and at 28°C. It was most stable at pH 7 and at 4°C. It was activated by Mg2+ and stabilised by Ca2+, Mg2+ and Mn2+. It showed activity towards xylitol and sorbitol, the respective Kms being 28.5 and 100 mM and the Km for NAD was 0.7 mM.
doi_str_mv 10.1111/j.1574-6968.1997.tb10174.x
format article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_journals_2331801098</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><oup_id>10.1111/j.1574-6968.1997.tb10174.x</oup_id><sourcerecordid>2331801098</sourcerecordid><originalsourceid>FETCH-LOGICAL-c2889-730f461ea56508b16e91c6865ec04372e5bf02b81b3ab41d67417f1c38fbac8b3</originalsourceid><addsrcrecordid>eNqVkMFOhDAQQBujievqPzR6BlsKbTHxYDaumuDqQc9NKa1CgK4txOXvhbB60oNzmWRm3kzmAXCOUYjHuKxCnLA4oCnlIU5TFnY5RpjF4e4ALH5ah2CBCOMBRik7BifeVwihOEJ0Aern3pWmVLIrbQtlW0D1Lp1UnXaln4vWwN1Ql52tYaHfh8LZN91Kr6FxtoEb3Tvrt9ZJqJz0XsLNKoPKNk3ffq_d2BBSErNTcGRk7fXZPi_B6_r2ZXUfZE93D6ubLFAR52nACDIxxVomNEE8x1SnWFFOE61QTFikk9ygKOc4JzKPcUFZjJnBinCTS8VzsgQX896tsx-99p2obO_a8aSICMEcjSL4OHU1T6nxAe-0EVtXNtINAiMx2RWVmBSKSaGY7Iq9XbEb4esZ_ixrPfyDFOvHjKUjn8y87bd_0MFvd78AWTeSag</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2331801098</pqid></control><display><type>article</type><title>Purification and characterisation of xylitol dehydrogenase from Neurospora crassa NCL communication No. 6347</title><source>Oxford Journals Online</source><creator>Phadtare, S.U ; Rawat, U.B ; Rao, M.B</creator><creatorcontrib>Phadtare, S.U ; Rawat, U.B ; Rao, M.B</creatorcontrib><description>Abstract The purification and characterisation of one of the key enzymes in the conversion of xylose to ethanol, xylitol dehydrogenase (XD), from Neurospora crassa is reported. This organism has the unique ability to directly convert biomass to ethanol. The enzyme was purified 81-fold by fractional ammonium sulfate precipitation, column chromatography on DEAE-cellulose, Sephacryl S-200 and β-NAD agarose. The purified enzyme had a native molecular mass of 87 kDa (gel filtration) and was composed of two identical subunits of molecular mass 43.6 kDa (SDS-PAGE). The enzyme was most active at pH 8.4 and at 28°C. It was most stable at pH 7 and at 4°C. It was activated by Mg2+ and stabilised by Ca2+, Mg2+ and Mn2+. It showed activity towards xylitol and sorbitol, the respective Kms being 28.5 and 100 mM and the Km for NAD was 0.7 mM.</description><identifier>ISSN: 0378-1097</identifier><identifier>EISSN: 1574-6968</identifier><identifier>DOI: 10.1111/j.1574-6968.1997.tb10174.x</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Ammonium ; Ammonium sulfate ; Calcium ; Calcium ions ; Cellulose ; Column chromatography ; Dehydrogenase ; Dehydrogenases ; Enzymes ; Ethanol ; Gel electrophoresis ; Gel filtration ; Magnesium ; Microbiology ; NAD ; Neurospora ; Neurospora crassa ; pH effects ; Purification ; Sodium lauryl sulfate ; Sorbitol ; Xylitol ; Xylitol dehydrogenase ; Xylose</subject><ispartof>FEMS microbiology letters, 1997-01, Vol.146 (1), p.79-83</ispartof><rights>Copyright © 1997 Federation of European Microbiological Societies. Published by Elsevier Science B.V. 1997</rights><rights>Copyright © 1997 Federation of European Microbiological Societies. Published by Elsevier Science B.V.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c2889-730f461ea56508b16e91c6865ec04372e5bf02b81b3ab41d67417f1c38fbac8b3</citedby><cites>FETCH-LOGICAL-c2889-730f461ea56508b16e91c6865ec04372e5bf02b81b3ab41d67417f1c38fbac8b3</cites></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></links><search><creatorcontrib>Phadtare, S.U</creatorcontrib><creatorcontrib>Rawat, U.B</creatorcontrib><creatorcontrib>Rao, M.B</creatorcontrib><title>Purification and characterisation of xylitol dehydrogenase from Neurospora crassa NCL communication No. 6347</title><title>FEMS microbiology letters</title><description>Abstract The purification and characterisation of one of the key enzymes in the conversion of xylose to ethanol, xylitol dehydrogenase (XD), from Neurospora crassa is reported. This organism has the unique ability to directly convert biomass to ethanol. The enzyme was purified 81-fold by fractional ammonium sulfate precipitation, column chromatography on DEAE-cellulose, Sephacryl S-200 and β-NAD agarose. The purified enzyme had a native molecular mass of 87 kDa (gel filtration) and was composed of two identical subunits of molecular mass 43.6 kDa (SDS-PAGE). The enzyme was most active at pH 8.4 and at 28°C. It was most stable at pH 7 and at 4°C. It was activated by Mg2+ and stabilised by Ca2+, Mg2+ and Mn2+. It showed activity towards xylitol and sorbitol, the respective Kms being 28.5 and 100 mM and the Km for NAD was 0.7 mM.</description><subject>Ammonium</subject><subject>Ammonium sulfate</subject><subject>Calcium</subject><subject>Calcium ions</subject><subject>Cellulose</subject><subject>Column chromatography</subject><subject>Dehydrogenase</subject><subject>Dehydrogenases</subject><subject>Enzymes</subject><subject>Ethanol</subject><subject>Gel electrophoresis</subject><subject>Gel filtration</subject><subject>Magnesium</subject><subject>Microbiology</subject><subject>NAD</subject><subject>Neurospora</subject><subject>Neurospora crassa</subject><subject>pH effects</subject><subject>Purification</subject><subject>Sodium lauryl sulfate</subject><subject>Sorbitol</subject><subject>Xylitol</subject><subject>Xylitol dehydrogenase</subject><subject>Xylose</subject><issn>0378-1097</issn><issn>1574-6968</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><recordid>eNqVkMFOhDAQQBujievqPzR6BlsKbTHxYDaumuDqQc9NKa1CgK4txOXvhbB60oNzmWRm3kzmAXCOUYjHuKxCnLA4oCnlIU5TFnY5RpjF4e4ALH5ah2CBCOMBRik7BifeVwihOEJ0Aern3pWmVLIrbQtlW0D1Lp1UnXaln4vWwN1Ql52tYaHfh8LZN91Kr6FxtoEb3Tvrt9ZJqJz0XsLNKoPKNk3ffq_d2BBSErNTcGRk7fXZPi_B6_r2ZXUfZE93D6ubLFAR52nACDIxxVomNEE8x1SnWFFOE61QTFikk9ygKOc4JzKPcUFZjJnBinCTS8VzsgQX896tsx-99p2obO_a8aSICMEcjSL4OHU1T6nxAe-0EVtXNtINAiMx2RWVmBSKSaGY7Iq9XbEb4esZ_ixrPfyDFOvHjKUjn8y87bd_0MFvd78AWTeSag</recordid><startdate>19970101</startdate><enddate>19970101</enddate><creator>Phadtare, S.U</creator><creator>Rawat, U.B</creator><creator>Rao, M.B</creator><general>Blackwell Publishing Ltd</general><general>Oxford University Press</general><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope></search><sort><creationdate>19970101</creationdate><title>Purification and characterisation of xylitol dehydrogenase from Neurospora crassa NCL communication No. 6347</title><author>Phadtare, S.U ; Rawat, U.B ; Rao, M.B</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2889-730f461ea56508b16e91c6865ec04372e5bf02b81b3ab41d67417f1c38fbac8b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Ammonium</topic><topic>Ammonium sulfate</topic><topic>Calcium</topic><topic>Calcium ions</topic><topic>Cellulose</topic><topic>Column chromatography</topic><topic>Dehydrogenase</topic><topic>Dehydrogenases</topic><topic>Enzymes</topic><topic>Ethanol</topic><topic>Gel electrophoresis</topic><topic>Gel filtration</topic><topic>Magnesium</topic><topic>Microbiology</topic><topic>NAD</topic><topic>Neurospora</topic><topic>Neurospora crassa</topic><topic>pH effects</topic><topic>Purification</topic><topic>Sodium lauryl sulfate</topic><topic>Sorbitol</topic><topic>Xylitol</topic><topic>Xylitol dehydrogenase</topic><topic>Xylose</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Phadtare, S.U</creatorcontrib><creatorcontrib>Rawat, U.B</creatorcontrib><creatorcontrib>Rao, M.B</creatorcontrib><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health &amp; Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health &amp; Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><jtitle>FEMS microbiology letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Phadtare, S.U</au><au>Rawat, U.B</au><au>Rao, M.B</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification and characterisation of xylitol dehydrogenase from Neurospora crassa NCL communication No. 6347</atitle><jtitle>FEMS microbiology letters</jtitle><date>1997-01-01</date><risdate>1997</risdate><volume>146</volume><issue>1</issue><spage>79</spage><epage>83</epage><pages>79-83</pages><issn>0378-1097</issn><eissn>1574-6968</eissn><abstract>Abstract The purification and characterisation of one of the key enzymes in the conversion of xylose to ethanol, xylitol dehydrogenase (XD), from Neurospora crassa is reported. This organism has the unique ability to directly convert biomass to ethanol. The enzyme was purified 81-fold by fractional ammonium sulfate precipitation, column chromatography on DEAE-cellulose, Sephacryl S-200 and β-NAD agarose. The purified enzyme had a native molecular mass of 87 kDa (gel filtration) and was composed of two identical subunits of molecular mass 43.6 kDa (SDS-PAGE). The enzyme was most active at pH 8.4 and at 28°C. It was most stable at pH 7 and at 4°C. It was activated by Mg2+ and stabilised by Ca2+, Mg2+ and Mn2+. It showed activity towards xylitol and sorbitol, the respective Kms being 28.5 and 100 mM and the Km for NAD was 0.7 mM.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><doi>10.1111/j.1574-6968.1997.tb10174.x</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0378-1097
ispartof FEMS microbiology letters, 1997-01, Vol.146 (1), p.79-83
issn 0378-1097
1574-6968
language eng
recordid cdi_proquest_journals_2331801098
source Oxford Journals Online
subjects Ammonium
Ammonium sulfate
Calcium
Calcium ions
Cellulose
Column chromatography
Dehydrogenase
Dehydrogenases
Enzymes
Ethanol
Gel electrophoresis
Gel filtration
Magnesium
Microbiology
NAD
Neurospora
Neurospora crassa
pH effects
Purification
Sodium lauryl sulfate
Sorbitol
Xylitol
Xylitol dehydrogenase
Xylose
title Purification and characterisation of xylitol dehydrogenase from Neurospora crassa NCL communication No. 6347
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-04T16%3A11%3A14IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Purification%20and%20characterisation%20of%20xylitol%20dehydrogenase%20from%20Neurospora%20crassa%20NCL%20communication%20No.%206347&rft.jtitle=FEMS%20microbiology%20letters&rft.au=Phadtare,%20S.U&rft.date=1997-01-01&rft.volume=146&rft.issue=1&rft.spage=79&rft.epage=83&rft.pages=79-83&rft.issn=0378-1097&rft.eissn=1574-6968&rft_id=info:doi/10.1111/j.1574-6968.1997.tb10174.x&rft_dat=%3Cproquest_cross%3E2331801098%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c2889-730f461ea56508b16e91c6865ec04372e5bf02b81b3ab41d67417f1c38fbac8b3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2331801098&rft_id=info:pmid/&rft_oup_id=10.1111/j.1574-6968.1997.tb10174.x&rfr_iscdi=true