Partial Purification and Characterization of Exoinulinase from Kluyveromyces marxianus YS-1 for Preparation of High-Fructose Syrup
An extracellular exoinulinase (2,1-β-D fructan fructanohydrolase, EC 3.2.1.7), which catalyzes the hydrolysis of inulin into fructose and glucose, was purified 23.5-fold by ethanol precipitation, followed by Sephadex G-100 gel permeation from a cell-free extract of Kluyveromyces marxianus YS-1. The...
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| Veröffentlicht in: | Journal of microbiology and biotechnology 2007-05, Vol.17 (5), p.733-738 |
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| creator | Singh, Ram Sarup (Punjabi University, Punjab, India), E-mail: rssingh11@lycos.com Dhaliwal, Rajesh (Punjabi University, Punjab, India) Puri, Munish (Punjabi University, Punjab, India) |
| description | An extracellular exoinulinase (2,1-β-D fructan fructanohydrolase, EC 3.2.1.7), which catalyzes the hydrolysis of inulin into fructose and glucose, was purified 23.5-fold by ethanol precipitation, followed by Sephadex G-100 gel permeation from a cell-free extract of Kluyveromyces marxianus YS-1. The partially purified enzyme exhibited considerable activity between pH 5 to 6, with an optimum pH of 5.5, while it remained stable (100%) for 3 h at the optimum temperature of 50℃. Mn²+ and Ca²+ produced a 2.4-fold and 1.2-fold enhancement in enzyme activity, whereas Hg²+ and Ag²+ completely inhibited the inulinase. |
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The partially purified enzyme exhibited considerable activity between pH 5 to 6, with an optimum pH of 5.5, while it remained stable (100%) for 3 h at the optimum temperature of 50℃. Mn²+ and Ca²+ produced a 2.4-fold and 1.2-fold enhancement in enzyme activity, whereas Hg²+ and Ag²+ completely inhibited the inulinase.</description><identifier>ISSN: 1017-7825</identifier><identifier>PMID: 18051293</identifier><language>eng</language><publisher>Seoul: Korean Society for Applied Microbiology</publisher><subject>Asparagus Plant - chemistry ; Asparagus racemosus ; Biological and medical sciences ; Biotechnology ; Cations, Divalent - pharmacology ; Chemical Fractionation ; Chromatography, Gel ; Dextrans ; Enzyme Activators - pharmacology ; Enzyme Inhibitors - pharmacology ; Enzyme Stability ; exoinulinase ; Fructose - metabolism ; Fundamental and applied biological sciences. Psychology ; Glycoside Hydrolases - isolation & purification ; Glycoside Hydrolases - metabolism ; high-fructose syrup ; Hydrogen-Ion Concentration ; Inulin - isolation & purification ; Inulin - metabolism ; Kluyveromyces - enzymology ; KLUYVEROMYCES MARXIANUS ; Liliaceae ; Metals - pharmacology ; Orchidaceae ; partial purification ; Plant Extracts - metabolism ; Raffinose - metabolism ; Substrate Specificity ; Sucrose - metabolism ; Temperature ; Time Factors</subject><ispartof>Journal of microbiology and biotechnology, 2007-05, Vol.17 (5), p.733-738</ispartof><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18781587$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18051293$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Singh, Ram Sarup (Punjabi University, Punjab, India), E-mail: rssingh11@lycos.com</creatorcontrib><creatorcontrib>Dhaliwal, Rajesh (Punjabi University, Punjab, India)</creatorcontrib><creatorcontrib>Puri, Munish (Punjabi University, Punjab, India)</creatorcontrib><title>Partial Purification and Characterization of Exoinulinase from Kluyveromyces marxianus YS-1 for Preparation of High-Fructose Syrup</title><title>Journal of microbiology and biotechnology</title><addtitle>J Microbiol Biotechnol</addtitle><description>An extracellular exoinulinase (2,1-β-D fructan fructanohydrolase, EC 3.2.1.7), which catalyzes the hydrolysis of inulin into fructose and glucose, was purified 23.5-fold by ethanol precipitation, followed by Sephadex G-100 gel permeation from a cell-free extract of Kluyveromyces marxianus YS-1. The partially purified enzyme exhibited considerable activity between pH 5 to 6, with an optimum pH of 5.5, while it remained stable (100%) for 3 h at the optimum temperature of 50℃. Mn²+ and Ca²+ produced a 2.4-fold and 1.2-fold enhancement in enzyme activity, whereas Hg²+ and Ag²+ completely inhibited the inulinase.</description><subject>Asparagus Plant - chemistry</subject><subject>Asparagus racemosus</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cations, Divalent - pharmacology</subject><subject>Chemical Fractionation</subject><subject>Chromatography, Gel</subject><subject>Dextrans</subject><subject>Enzyme Activators - pharmacology</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Enzyme Stability</subject><subject>exoinulinase</subject><subject>Fructose - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glycoside Hydrolases - isolation & purification</subject><subject>Glycoside Hydrolases - metabolism</subject><subject>high-fructose syrup</subject><subject>Hydrogen-Ion Concentration</subject><subject>Inulin - isolation & purification</subject><subject>Inulin - metabolism</subject><subject>Kluyveromyces - enzymology</subject><subject>KLUYVEROMYCES MARXIANUS</subject><subject>Liliaceae</subject><subject>Metals - pharmacology</subject><subject>Orchidaceae</subject><subject>partial purification</subject><subject>Plant Extracts - metabolism</subject><subject>Raffinose - metabolism</subject><subject>Substrate Specificity</subject><subject>Sucrose - metabolism</subject><subject>Temperature</subject><subject>Time Factors</subject><issn>1017-7825</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpN0M9LwzAUwPEeFDenf4KSi94KSfoj6VHG5mQDh9ODp_KaJlukbWrSyOrRv9zKNvD0wuPDF_LOgjHBhIWM02QUXDr3gXFKKE8vghHhOCE0i8bBzxpsp6FCa2-10gI6bRoETYmmO7AgOmn192FpFJrtjW58pRtwEilrarSsfP8lh1cvpEM12L2Gxjv0vgkJUsaitZXtEDoVFnq7C-fWi84MiU1vfXsVnCuonLw-zknwNp-9Thfh6vnxafqwChVNSRcWvISCZAKXksg0oQAxFVmUsULKImYqKjhkMsKSxTimwIDGHMqyFJAxLOIymgT3h25rzaeXrstr7YSsKmik8S5PM0yGW7EB3h6hL2pZ5q3Vw8f6_HS1AdwdATgBlbLQCO3-OcZJwv9CNwenwOSwtYNZvlCMGcZRlKXRLzk0gPE</recordid><startdate>20070501</startdate><enddate>20070501</enddate><creator>Singh, Ram Sarup (Punjabi University, Punjab, India), E-mail: rssingh11@lycos.com</creator><creator>Dhaliwal, Rajesh (Punjabi University, Punjab, India)</creator><creator>Puri, Munish (Punjabi University, Punjab, India)</creator><general>Korean Society for Applied Microbiology</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>20070501</creationdate><title>Partial Purification and Characterization of Exoinulinase from Kluyveromyces marxianus YS-1 for Preparation of High-Fructose Syrup</title><author>Singh, Ram Sarup (Punjabi University, Punjab, India), E-mail: rssingh11@lycos.com ; Dhaliwal, Rajesh (Punjabi University, Punjab, India) ; Puri, Munish (Punjabi University, Punjab, India)</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-f261t-b8dab19c0de1e652aa42c9397beeb47f3b8a9e30e74042a7a248adddca970c4d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Asparagus Plant - chemistry</topic><topic>Asparagus racemosus</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cations, Divalent - pharmacology</topic><topic>Chemical Fractionation</topic><topic>Chromatography, Gel</topic><topic>Dextrans</topic><topic>Enzyme Activators - pharmacology</topic><topic>Enzyme Inhibitors - pharmacology</topic><topic>Enzyme Stability</topic><topic>exoinulinase</topic><topic>Fructose - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glycoside Hydrolases - isolation & purification</topic><topic>Glycoside Hydrolases - metabolism</topic><topic>high-fructose syrup</topic><topic>Hydrogen-Ion Concentration</topic><topic>Inulin - isolation & purification</topic><topic>Inulin - metabolism</topic><topic>Kluyveromyces - enzymology</topic><topic>KLUYVEROMYCES MARXIANUS</topic><topic>Liliaceae</topic><topic>Metals - pharmacology</topic><topic>Orchidaceae</topic><topic>partial purification</topic><topic>Plant Extracts - metabolism</topic><topic>Raffinose - metabolism</topic><topic>Substrate Specificity</topic><topic>Sucrose - metabolism</topic><topic>Temperature</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Singh, Ram Sarup (Punjabi University, Punjab, India), E-mail: rssingh11@lycos.com</creatorcontrib><creatorcontrib>Dhaliwal, Rajesh (Punjabi University, Punjab, India)</creatorcontrib><creatorcontrib>Puri, Munish (Punjabi University, Punjab, India)</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>Journal of microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Singh, Ram Sarup (Punjabi University, Punjab, India), E-mail: rssingh11@lycos.com</au><au>Dhaliwal, Rajesh (Punjabi University, Punjab, India)</au><au>Puri, Munish (Punjabi University, Punjab, India)</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Partial Purification and Characterization of Exoinulinase from Kluyveromyces marxianus YS-1 for Preparation of High-Fructose Syrup</atitle><jtitle>Journal of microbiology and biotechnology</jtitle><addtitle>J Microbiol Biotechnol</addtitle><date>2007-05-01</date><risdate>2007</risdate><volume>17</volume><issue>5</issue><spage>733</spage><epage>738</epage><pages>733-738</pages><issn>1017-7825</issn><abstract>An extracellular exoinulinase (2,1-β-D fructan fructanohydrolase, EC 3.2.1.7), which catalyzes the hydrolysis of inulin into fructose and glucose, was purified 23.5-fold by ethanol precipitation, followed by Sephadex G-100 gel permeation from a cell-free extract of Kluyveromyces marxianus YS-1. The partially purified enzyme exhibited considerable activity between pH 5 to 6, with an optimum pH of 5.5, while it remained stable (100%) for 3 h at the optimum temperature of 50℃. Mn²+ and Ca²+ produced a 2.4-fold and 1.2-fold enhancement in enzyme activity, whereas Hg²+ and Ag²+ completely inhibited the inulinase.</abstract><cop>Seoul</cop><pub>Korean Society for Applied Microbiology</pub><pmid>18051293</pmid><tpages>6</tpages></addata></record> |
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| subjects | Asparagus Plant - chemistry Asparagus racemosus Biological and medical sciences Biotechnology Cations, Divalent - pharmacology Chemical Fractionation Chromatography, Gel Dextrans Enzyme Activators - pharmacology Enzyme Inhibitors - pharmacology Enzyme Stability exoinulinase Fructose - metabolism Fundamental and applied biological sciences. Psychology Glycoside Hydrolases - isolation & purification Glycoside Hydrolases - metabolism high-fructose syrup Hydrogen-Ion Concentration Inulin - isolation & purification Inulin - metabolism Kluyveromyces - enzymology KLUYVEROMYCES MARXIANUS Liliaceae Metals - pharmacology Orchidaceae partial purification Plant Extracts - metabolism Raffinose - metabolism Substrate Specificity Sucrose - metabolism Temperature Time Factors |
| title | Partial Purification and Characterization of Exoinulinase from Kluyveromyces marxianus YS-1 for Preparation of High-Fructose Syrup |
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