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Yeast cultures expressing the Ffase from Schwanniomyces occidentalis, a simple system to produce the potential prebiotic sugar 6-kestose
The β-fructofuranosidase Ffase from the yeast Schwanniomyces occidentalis produces potential prebiotic fructooligosaccharides with health-promoting properties, making it of biotechnological interest. Ffase is one of the highest and more selective known producers of 6-kestose by transfructosylation o...
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Published in: | Applied microbiology and biotechnology 2019-01, Vol.103 (1), p.279-289 |
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description | The β-fructofuranosidase Ffase from the yeast
Schwanniomyces occidentalis
produces potential prebiotic fructooligosaccharides with health-promoting properties, making it of biotechnological interest. Ffase is one of the highest and more selective known producers of 6-kestose by transfructosylation of sucrose. In this work, production of 6-kestose was simplified by directly using cultures of
S. occidentalis
and
Saccharomyces cerevisiae
expressing both the wild-type enzyme and a mutated Ffase variant including the Ser196Leu substitution (Ffase-Leu196). Best results were obtained using yeast cultures supplemented with sucrose and expressing the Ffase-Leu196, which after only 4 h produced ~ 116 g/L of 6-kestose, twice the amount obtained with the corresponding purified enzyme. 6-Kestose represented ~ 70% of the products synthesized. In addition, a small amount of 1-kestose and the neofructoligosaccharides neokestose and blastose were also produced. The Ser196Leu substitution skewed production of 6-kestose and neofructooligosaccharides resulting in an increase of ~ 2.2- and 1.5-fold, respectively, without affecting production of 1-kestose. Supplementing yeast cultures with glucose clearly showed that blastose originates from direct fructosylation of glucose, a property that has not been described for other similar proteins from yeasts. Modeling neokestose and blastose into the Ffase-active site revealed the molecular basis explaining the peculiar specificity of this enzyme. |
doi_str_mv | 10.1007/s00253-018-9446-y |
format | article |
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Schwanniomyces occidentalis
produces potential prebiotic fructooligosaccharides with health-promoting properties, making it of biotechnological interest. Ffase is one of the highest and more selective known producers of 6-kestose by transfructosylation of sucrose. In this work, production of 6-kestose was simplified by directly using cultures of
S. occidentalis
and
Saccharomyces cerevisiae
expressing both the wild-type enzyme and a mutated Ffase variant including the Ser196Leu substitution (Ffase-Leu196). Best results were obtained using yeast cultures supplemented with sucrose and expressing the Ffase-Leu196, which after only 4 h produced ~ 116 g/L of 6-kestose, twice the amount obtained with the corresponding purified enzyme. 6-Kestose represented ~ 70% of the products synthesized. In addition, a small amount of 1-kestose and the neofructoligosaccharides neokestose and blastose were also produced. The Ser196Leu substitution skewed production of 6-kestose and neofructooligosaccharides resulting in an increase of ~ 2.2- and 1.5-fold, respectively, without affecting production of 1-kestose. Supplementing yeast cultures with glucose clearly showed that blastose originates from direct fructosylation of glucose, a property that has not been described for other similar proteins from yeasts. Modeling neokestose and blastose into the Ffase-active site revealed the molecular basis explaining the peculiar specificity of this enzyme.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-018-9446-y</identifier><identifier>PMID: 30357454</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Baking yeast ; Biomedical and Life Sciences ; Biotechnologically Relevant Enzymes and Proteins ; Biotechnology ; Cultures (Biology) ; Enzymes ; Fructooligosaccharides ; Glucose ; Health promotion ; Invertase ; Life Sciences ; Microbial Genetics and Genomics ; Microbiological research ; Microbiology ; Oligosaccharides ; Prebiotics ; Probiotics ; Proteins ; Saccharomyces cerevisiae ; Substitutes ; Sucrose ; Sugar ; Yeast ; Yeasts ; Yeasts (Fungi)</subject><ispartof>Applied microbiology and biotechnology, 2019-01, Vol.103 (1), p.279-289</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2018</rights><rights>COPYRIGHT 2019 Springer</rights><rights>Applied Microbiology and Biotechnology is a copyright of Springer, (2018). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c619t-3c7c3b60d2c7ce8b3cb55145896883833fb9793a5ac777e0c7182425c97952b43</citedby><cites>FETCH-LOGICAL-c619t-3c7c3b60d2c7ce8b3cb55145896883833fb9793a5ac777e0c7182425c97952b43</cites><orcidid>0000-0003-0778-7443</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2124646303/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$H</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2124646303?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,780,784,11688,27924,27925,36060,36061,44363,74767</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30357454$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rodrigo-Frutos, David</creatorcontrib><creatorcontrib>Piedrabuena, David</creatorcontrib><creatorcontrib>Sanz-Aparicio, Julia</creatorcontrib><creatorcontrib>Fernández-Lobato, María</creatorcontrib><title>Yeast cultures expressing the Ffase from Schwanniomyces occidentalis, a simple system to produce the potential prebiotic sugar 6-kestose</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>The β-fructofuranosidase Ffase from the yeast
Schwanniomyces occidentalis
produces potential prebiotic fructooligosaccharides with health-promoting properties, making it of biotechnological interest. Ffase is one of the highest and more selective known producers of 6-kestose by transfructosylation of sucrose. In this work, production of 6-kestose was simplified by directly using cultures of
S. occidentalis
and
Saccharomyces cerevisiae
expressing both the wild-type enzyme and a mutated Ffase variant including the Ser196Leu substitution (Ffase-Leu196). Best results were obtained using yeast cultures supplemented with sucrose and expressing the Ffase-Leu196, which after only 4 h produced ~ 116 g/L of 6-kestose, twice the amount obtained with the corresponding purified enzyme. 6-Kestose represented ~ 70% of the products synthesized. In addition, a small amount of 1-kestose and the neofructoligosaccharides neokestose and blastose were also produced. The Ser196Leu substitution skewed production of 6-kestose and neofructooligosaccharides resulting in an increase of ~ 2.2- and 1.5-fold, respectively, without affecting production of 1-kestose. Supplementing yeast cultures with glucose clearly showed that blastose originates from direct fructosylation of glucose, a property that has not been described for other similar proteins from yeasts. Modeling neokestose and blastose into the Ffase-active site revealed the molecular basis explaining the peculiar specificity of this enzyme.</description><subject>Baking yeast</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnologically Relevant Enzymes and Proteins</subject><subject>Biotechnology</subject><subject>Cultures (Biology)</subject><subject>Enzymes</subject><subject>Fructooligosaccharides</subject><subject>Glucose</subject><subject>Health promotion</subject><subject>Invertase</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiological research</subject><subject>Microbiology</subject><subject>Oligosaccharides</subject><subject>Prebiotics</subject><subject>Probiotics</subject><subject>Proteins</subject><subject>Saccharomyces cerevisiae</subject><subject>Substitutes</subject><subject>Sucrose</subject><subject>Sugar</subject><subject>Yeast</subject><subject>Yeasts</subject><subject>Yeasts 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biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rodrigo-Frutos, David</au><au>Piedrabuena, David</au><au>Sanz-Aparicio, Julia</au><au>Fernández-Lobato, María</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Yeast cultures expressing the Ffase from Schwanniomyces occidentalis, a simple system to produce the potential prebiotic sugar 6-kestose</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2019-01-01</date><risdate>2019</risdate><volume>103</volume><issue>1</issue><spage>279</spage><epage>289</epage><pages>279-289</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>The β-fructofuranosidase Ffase from the yeast
Schwanniomyces occidentalis
produces potential prebiotic fructooligosaccharides with health-promoting properties, making it of biotechnological interest. Ffase is one of the highest and more selective known producers of 6-kestose by transfructosylation of sucrose. In this work, production of 6-kestose was simplified by directly using cultures of
S. occidentalis
and
Saccharomyces cerevisiae
expressing both the wild-type enzyme and a mutated Ffase variant including the Ser196Leu substitution (Ffase-Leu196). Best results were obtained using yeast cultures supplemented with sucrose and expressing the Ffase-Leu196, which after only 4 h produced ~ 116 g/L of 6-kestose, twice the amount obtained with the corresponding purified enzyme. 6-Kestose represented ~ 70% of the products synthesized. In addition, a small amount of 1-kestose and the neofructoligosaccharides neokestose and blastose were also produced. The Ser196Leu substitution skewed production of 6-kestose and neofructooligosaccharides resulting in an increase of ~ 2.2- and 1.5-fold, respectively, without affecting production of 1-kestose. Supplementing yeast cultures with glucose clearly showed that blastose originates from direct fructosylation of glucose, a property that has not been described for other similar proteins from yeasts. Modeling neokestose and blastose into the Ffase-active site revealed the molecular basis explaining the peculiar specificity of this enzyme.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>30357454</pmid><doi>10.1007/s00253-018-9446-y</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0003-0778-7443</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Baking yeast Biomedical and Life Sciences Biotechnologically Relevant Enzymes and Proteins Biotechnology Cultures (Biology) Enzymes Fructooligosaccharides Glucose Health promotion Invertase Life Sciences Microbial Genetics and Genomics Microbiological research Microbiology Oligosaccharides Prebiotics Probiotics Proteins Saccharomyces cerevisiae Substitutes Sucrose Sugar Yeast Yeasts Yeasts (Fungi) |
title | Yeast cultures expressing the Ffase from Schwanniomyces occidentalis, a simple system to produce the potential prebiotic sugar 6-kestose |
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