High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration

Abstract Xylanase is a widely-used additive in baking industry for enhancing dough and bread quality. Several xylanases used in baking industry were expressed in different systems, but their expression in antibiotic free vector system is highly essential and safe. In the present study, an alternativ...

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Autores principales:	Cheng Fang, Qinhong Wang, Jonathan Nimal Selvaraj, Yuling Zhou, Lixin Ma, Guimin Zhang, Yanhe Ma
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8b62d55a369e4150a69b5b55f1b3e856
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spelling	oai:doaj.org-article:8b62d55a369e4150a69b5b55f1b3e8562021-12-02T11:40:13ZHigh copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration10.1038/s41598-017-08647-x2045-2322https://doaj.org/article/8b62d55a369e4150a69b5b55f1b3e8562017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08647-xhttps://doaj.org/toc/2045-2322Abstract Xylanase is a widely-used additive in baking industry for enhancing dough and bread quality. Several xylanases used in baking industry were expressed in different systems, but their expression in antibiotic free vector system is highly essential and safe. In the present study, an alternative rDNA-mediated technology was developed to increase the copy number of target gene by integrating it into Saccharomyces cerevisiae genome. A xylanase-encoding gene xynHB from Bacillus sp. was cloned into pHBM367H and integrated into S. cerevisiae genome through rDNA-mediated recombination. Exogenous XynHB expressed by recombinant S. cerevisiae strain A13 exhibited higher degradation activity towards xylan than other transformants. The real-time PCR analysis on A13 genome revealed the presence of 13.64 copies of xynHB gene. Though no antibiotics have been used, the genetic stability and the xylanase activity of xynHB remained stable up to 1,011 generations of cultivation. S. cerevisiae strain A13 expressing xylanase reduced the required kneading time and increased the height and diameter of the dough size, which would be safe and effective in baking industry as no antibiotics-resistance risk. The new effective rDNA-mediated technology without using antibiotics here provides a way to clone other food related industrial enzymes for applications.Cheng FangQinhong WangJonathan Nimal SelvarajYuling ZhouLixin MaGuimin ZhangYanhe MaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Cheng Fang Qinhong Wang Jonathan Nimal Selvaraj Yuling Zhou Lixin Ma Guimin Zhang Yanhe Ma High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration
description	Abstract Xylanase is a widely-used additive in baking industry for enhancing dough and bread quality. Several xylanases used in baking industry were expressed in different systems, but their expression in antibiotic free vector system is highly essential and safe. In the present study, an alternative rDNA-mediated technology was developed to increase the copy number of target gene by integrating it into Saccharomyces cerevisiae genome. A xylanase-encoding gene xynHB from Bacillus sp. was cloned into pHBM367H and integrated into S. cerevisiae genome through rDNA-mediated recombination. Exogenous XynHB expressed by recombinant S. cerevisiae strain A13 exhibited higher degradation activity towards xylan than other transformants. The real-time PCR analysis on A13 genome revealed the presence of 13.64 copies of xynHB gene. Though no antibiotics have been used, the genetic stability and the xylanase activity of xynHB remained stable up to 1,011 generations of cultivation. S. cerevisiae strain A13 expressing xylanase reduced the required kneading time and increased the height and diameter of the dough size, which would be safe and effective in baking industry as no antibiotics-resistance risk. The new effective rDNA-mediated technology without using antibiotics here provides a way to clone other food related industrial enzymes for applications.
format	article
author	Cheng Fang Qinhong Wang Jonathan Nimal Selvaraj Yuling Zhou Lixin Ma Guimin Zhang Yanhe Ma
author_facet	Cheng Fang Qinhong Wang Jonathan Nimal Selvaraj Yuling Zhou Lixin Ma Guimin Zhang Yanhe Ma
author_sort	Cheng Fang
title	High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration
title_short	High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration
title_full	High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration
title_fullStr	High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration
title_full_unstemmed	High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration
title_sort	high copy and stable expression of the xylanase xynhb in saccharomyces cerevisiae by rdna-mediated integration
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/8b62d55a369e4150a69b5b55f1b3e856
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High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration

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