Improved Production of Recombinant Myrosinase in Pichia pastoris

The effect of the deletion of a 57 bp native signal sequence, which transports the nascent protein through the endoplasmic reticulum membrane in plants, on improved AtTGG1 plant myrosinase production in Pichia pastoris was studied. Myrosinase was extracellularly...

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Autores principales:	Zuzana Rosenbergová, Zuzana Hegyi, Miroslav Ferko, Natália Andelová, Martin Rebroš
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	signal sequence <i>Pichia pastoris</i> myrosinase <i>Arabidopsis thaliana</i> plant enzymes Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/f9a3e6dad4c4487384b229e48f5920b5
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spelling	oai:doaj.org-article:f9a3e6dad4c4487384b229e48f5920b52021-11-11T17:18:16ZImproved Production of Recombinant Myrosinase in <i>Pichia pastoris</i>10.3390/ijms2221118891422-00671661-6596https://doaj.org/article/f9a3e6dad4c4487384b229e48f5920b52021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11889https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067The effect of the deletion of a 57 bp native signal sequence, which transports the nascent protein through the endoplasmic reticulum membrane in plants, on improved <i>At</i>TGG1 plant myrosinase production in <i>Pichia pastoris</i> was studied. Myrosinase was extracellularly produced in a 3-liter laboratory fermenter using α-mating factor as the secretion signal. After the deletion of the native signal sequence, both the specific productivity (164.8 U/L/h) and volumetric activity (27 U/mL) increased more than 40-fold compared to the expression of myrosinase containing its native signal sequence in combination with α-mating factor. The deletion of the native signal sequence resulted in slight changes in myrosinase properties: the optimum pH shifted from 6.5 to 7.0 and the maximal activating concentration of ascorbic acid increased from 1 mM to 1.5 mM. Kinetic parameters toward sinigrin were determined: 0.249 mM (K<sub>m</sub>) and 435.7 U/mg (V<sub>max</sub>). These results could be applied to the expression of other plant enzymes.Zuzana RosenbergováZuzana HegyiMiroslav FerkoNatália AndelováMartin RebrošMDPI AGarticlesignal sequence<i>Pichia pastoris</i>myrosinase<i>Arabidopsis thaliana</i>plant enzymesBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11889, p 11889 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	signal sequence <i>Pichia pastoris</i> myrosinase <i>Arabidopsis thaliana</i> plant enzymes Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	signal sequence <i>Pichia pastoris</i> myrosinase <i>Arabidopsis thaliana</i> plant enzymes Biology (General) QH301-705.5 Chemistry QD1-999 Zuzana Rosenbergová Zuzana Hegyi Miroslav Ferko Natália Andelová Martin Rebroš Improved Production of Recombinant Myrosinase in <i>Pichia pastoris</i>
description	The effect of the deletion of a 57 bp native signal sequence, which transports the nascent protein through the endoplasmic reticulum membrane in plants, on improved <i>At</i>TGG1 plant myrosinase production in <i>Pichia pastoris</i> was studied. Myrosinase was extracellularly produced in a 3-liter laboratory fermenter using α-mating factor as the secretion signal. After the deletion of the native signal sequence, both the specific productivity (164.8 U/L/h) and volumetric activity (27 U/mL) increased more than 40-fold compared to the expression of myrosinase containing its native signal sequence in combination with α-mating factor. The deletion of the native signal sequence resulted in slight changes in myrosinase properties: the optimum pH shifted from 6.5 to 7.0 and the maximal activating concentration of ascorbic acid increased from 1 mM to 1.5 mM. Kinetic parameters toward sinigrin were determined: 0.249 mM (K<sub>m</sub>) and 435.7 U/mg (V<sub>max</sub>). These results could be applied to the expression of other plant enzymes.
format	article
author	Zuzana Rosenbergová Zuzana Hegyi Miroslav Ferko Natália Andelová Martin Rebroš
author_facet	Zuzana Rosenbergová Zuzana Hegyi Miroslav Ferko Natália Andelová Martin Rebroš
author_sort	Zuzana Rosenbergová
title	Improved Production of Recombinant Myrosinase in <i>Pichia pastoris</i>
title_short	Improved Production of Recombinant Myrosinase in <i>Pichia pastoris</i>
title_full	Improved Production of Recombinant Myrosinase in <i>Pichia pastoris</i>
title_fullStr	Improved Production of Recombinant Myrosinase in <i>Pichia pastoris</i>
title_full_unstemmed	Improved Production of Recombinant Myrosinase in <i>Pichia pastoris</i>
title_sort	improved production of recombinant myrosinase in <i>pichia pastoris</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f9a3e6dad4c4487384b229e48f5920b5
work_keys_str_mv	AT zuzanarosenbergova improvedproductionofrecombinantmyrosinaseinipichiapastorisi AT zuzanahegyi improvedproductionofrecombinantmyrosinaseinipichiapastorisi AT miroslavferko improvedproductionofrecombinantmyrosinaseinipichiapastorisi AT nataliaandelova improvedproductionofrecombinantmyrosinaseinipichiapastorisi AT martinrebros improvedproductionofrecombinantmyrosinaseinipichiapastorisi
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Improved Production of Recombinant Myrosinase in <i>Pichia pastoris</i>

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