Engineering of yeast pyruvate decarboxylase for enhanced selectivity towards carboligation
•Cys221 mutation increases the affinity of Pdc towards pyruvate.•The increased affinity for pyruvate leads to an increase carboligation capacity.•It increases the R-PAC production.•Over-expression in pdc null strain leads to maximum R-PAC production.•Mutant Pdc is able to accept exogenous acetaldehy...
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| Veröffentlicht in: | Bioresource technology 2015-09, Vol.192, p.90-96 |
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| creator | Agarwal, Praveen Kumar Uppada, Vanita Swaminathan, A.G. Noronha, Santosh B. |
| description | •Cys221 mutation increases the affinity of Pdc towards pyruvate.•The increased affinity for pyruvate leads to an increase carboligation capacity.•It increases the R-PAC production.•Over-expression in pdc null strain leads to maximum R-PAC production.•Mutant Pdc is able to accept exogenous acetaldehyde as substrate.
The aim of the study was to increase production of (R)-PAC by altering carboligation activity of Pdc in Saccharomyces cerevisiae. Pdc1 activity was modified by over-expression as well as changing the rate of decarboxylation and carboligation by site specific mutation in Pdc1. Over-expression of mutant Pdc1 resulted in 50±2.5% increase in levels of (R)-PAC in wild-type and further 30–40% in pdc null background. The combination of mutant Pdc1 in pdc null background was successfully evaluated for production of (R)-PAC at industrial scale. This is the first report of enhancing (R)-PAC product in yeast by recombinant technology with capability of commercial production. |
| doi_str_mv | 10.1016/j.biortech.2015.05.060 |
| format | Article |
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The aim of the study was to increase production of (R)-PAC by altering carboligation activity of Pdc in Saccharomyces cerevisiae. Pdc1 activity was modified by over-expression as well as changing the rate of decarboxylation and carboligation by site specific mutation in Pdc1. Over-expression of mutant Pdc1 resulted in 50±2.5% increase in levels of (R)-PAC in wild-type and further 30–40% in pdc null background. The combination of mutant Pdc1 in pdc null background was successfully evaluated for production of (R)-PAC at industrial scale. This is the first report of enhancing (R)-PAC product in yeast by recombinant technology with capability of commercial production.</description><identifier>ISSN: 0960-8524</identifier><identifier>EISSN: 1873-2976</identifier><identifier>DOI: 10.1016/j.biortech.2015.05.060</identifier><identifier>PMID: 26022970</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>(R)-phenyl acetyl carbinol ; Biotransformation ; Carboligation ; Decarboxylation ; Decarboxylation - genetics ; Methanol - metabolism ; Mutation - genetics ; Pyruvate decarboxylase ; Pyruvate Decarboxylase - genetics ; Pyruvate Decarboxylase - metabolism ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae - metabolism ; Saccharomyces cerevisiae Proteins - genetics ; Saccharomyces cerevisiae Proteins - metabolism</subject><ispartof>Bioresource technology, 2015-09, Vol.192, p.90-96</ispartof><rights>2015 Elsevier Ltd</rights><rights>Copyright © 2015 Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c368t-6bed2fc73773d3564f6d031519dcd59b0541bfbb65d181833682e388e28b346c3</citedby><cites>FETCH-LOGICAL-c368t-6bed2fc73773d3564f6d031519dcd59b0541bfbb65d181833682e388e28b346c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.biortech.2015.05.060$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26022970$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Agarwal, Praveen Kumar</creatorcontrib><creatorcontrib>Uppada, Vanita</creatorcontrib><creatorcontrib>Swaminathan, A.G.</creatorcontrib><creatorcontrib>Noronha, Santosh B.</creatorcontrib><title>Engineering of yeast pyruvate decarboxylase for enhanced selectivity towards carboligation</title><title>Bioresource technology</title><addtitle>Bioresour Technol</addtitle><description>•Cys221 mutation increases the affinity of Pdc towards pyruvate.•The increased affinity for pyruvate leads to an increase carboligation capacity.•It increases the R-PAC production.•Over-expression in pdc null strain leads to maximum R-PAC production.•Mutant Pdc is able to accept exogenous acetaldehyde as substrate.
The aim of the study was to increase production of (R)-PAC by altering carboligation activity of Pdc in Saccharomyces cerevisiae. Pdc1 activity was modified by over-expression as well as changing the rate of decarboxylation and carboligation by site specific mutation in Pdc1. Over-expression of mutant Pdc1 resulted in 50±2.5% increase in levels of (R)-PAC in wild-type and further 30–40% in pdc null background. The combination of mutant Pdc1 in pdc null background was successfully evaluated for production of (R)-PAC at industrial scale. 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The aim of the study was to increase production of (R)-PAC by altering carboligation activity of Pdc in Saccharomyces cerevisiae. Pdc1 activity was modified by over-expression as well as changing the rate of decarboxylation and carboligation by site specific mutation in Pdc1. Over-expression of mutant Pdc1 resulted in 50±2.5% increase in levels of (R)-PAC in wild-type and further 30–40% in pdc null background. The combination of mutant Pdc1 in pdc null background was successfully evaluated for production of (R)-PAC at industrial scale. This is the first report of enhancing (R)-PAC product in yeast by recombinant technology with capability of commercial production.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>26022970</pmid><doi>10.1016/j.biortech.2015.05.060</doi><tpages>7</tpages></addata></record> |
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| subjects | (R)-phenyl acetyl carbinol Biotransformation Carboligation Decarboxylation Decarboxylation - genetics Methanol - metabolism Mutation - genetics Pyruvate decarboxylase Pyruvate Decarboxylase - genetics Pyruvate Decarboxylase - metabolism Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - metabolism |
| title | Engineering of yeast pyruvate decarboxylase for enhanced selectivity towards carboligation |
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