Engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.

Hexokinase 2 (Hxk2p) from Saccharomyces cerevisiae is a bi-functional enzyme being both a catalyst and an important regulator in the glucose repression signal. In the presence of xylose Hxk2p is irreversibly inactivated through an autophosphorylation mechanism, affecting all functions. Consequently,...

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Autores principales:	Basti Bergdahl, Anders G Sandström, Celina Borgström, Tarinee Boonyawan, Ed W J van Niel, Marie F Gorwa-Grauslund
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d866290bca7a4d60963f074c389649e6
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spelling	oai:doaj.org-article:d866290bca7a4d60963f074c389649e62021-11-18T08:56:26ZEngineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.1932-620310.1371/journal.pone.0075055https://doaj.org/article/d866290bca7a4d60963f074c389649e62013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24040384/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Hexokinase 2 (Hxk2p) from Saccharomyces cerevisiae is a bi-functional enzyme being both a catalyst and an important regulator in the glucose repression signal. In the presence of xylose Hxk2p is irreversibly inactivated through an autophosphorylation mechanism, affecting all functions. Consequently, the regulation of genes involved in sugar transport and fermentative metabolism is impaired. The aim of the study was to obtain new Hxk2p-variants, immune to the autophosphorylation, which potentially can restore the repressive capability closer to its nominal level. In this study we constructed the first condensed, rationally designed combinatorial library targeting the active-site in Hxk2p. We combined protein engineering and genetic engineering for efficient screening and identified a variant with Phe159 changed to tyrosine. This variant had 64% higher catalytic activity in the presence of xylose compared to the wild-type and is expected to be a key component for increasing the productivity of recombinant xylose-fermenting strains for bioethanol production from lignocellulosic feedstocks.Basti BergdahlAnders G SandströmCelina BorgströmTarinee BoonyawanEd W J van NielMarie F Gorwa-GrauslundPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 9, p e75055 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Basti Bergdahl Anders G Sandström Celina Borgström Tarinee Boonyawan Ed W J van Niel Marie F Gorwa-Grauslund Engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.
description	Hexokinase 2 (Hxk2p) from Saccharomyces cerevisiae is a bi-functional enzyme being both a catalyst and an important regulator in the glucose repression signal. In the presence of xylose Hxk2p is irreversibly inactivated through an autophosphorylation mechanism, affecting all functions. Consequently, the regulation of genes involved in sugar transport and fermentative metabolism is impaired. The aim of the study was to obtain new Hxk2p-variants, immune to the autophosphorylation, which potentially can restore the repressive capability closer to its nominal level. In this study we constructed the first condensed, rationally designed combinatorial library targeting the active-site in Hxk2p. We combined protein engineering and genetic engineering for efficient screening and identified a variant with Phe159 changed to tyrosine. This variant had 64% higher catalytic activity in the presence of xylose compared to the wild-type and is expected to be a key component for increasing the productivity of recombinant xylose-fermenting strains for bioethanol production from lignocellulosic feedstocks.
format	article
author	Basti Bergdahl Anders G Sandström Celina Borgström Tarinee Boonyawan Ed W J van Niel Marie F Gorwa-Grauslund
author_facet	Basti Bergdahl Anders G Sandström Celina Borgström Tarinee Boonyawan Ed W J van Niel Marie F Gorwa-Grauslund
author_sort	Basti Bergdahl
title	Engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.
title_short	Engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.
title_full	Engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.
title_fullStr	Engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.
title_full_unstemmed	Engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.
title_sort	engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/d866290bca7a4d60963f074c389649e6
work_keys_str_mv	AT bastibergdahl engineeringyeasthexokinase2forimprovedtolerancetowardxyloseinducedinactivation AT andersgsandstrom engineeringyeasthexokinase2forimprovedtolerancetowardxyloseinducedinactivation AT celinaborgstrom engineeringyeasthexokinase2forimprovedtolerancetowardxyloseinducedinactivation AT tarineeboonyawan engineeringyeasthexokinase2forimprovedtolerancetowardxyloseinducedinactivation AT edwjvanniel engineeringyeasthexokinase2forimprovedtolerancetowardxyloseinducedinactivation AT mariefgorwagrauslund engineeringyeasthexokinase2forimprovedtolerancetowardxyloseinducedinactivation
_version_	1718421170445877248

Engineering yeast hexokinase 2 for improved tolerance toward xylose-induced inactivation.

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