β-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.

Mushroom β-glucans are potent immunological stimulators in medicine, but their productivities are very low. In this study, we successfully improved its production by promoter engineering in Pleurotus ostreatus. The promoter for β-1,3-glucan synthase gene (GLS) was replaced by the promoter of glycera...

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Autores principales:	Ran Chai, Cuiwei Qiu, Dongren Liu, Yuancheng Qi, Yuqian Gao, Jinwen Shen, Liyou Qiu
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2359d390504b4884bf52f5ff35bb0ce1
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spelling	oai:doaj.org-article:2359d390504b4884bf52f5ff35bb0ce12021-11-18T07:48:04Zβ-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.1932-620310.1371/journal.pone.0061693https://doaj.org/article/2359d390504b4884bf52f5ff35bb0ce12013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23637884/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Mushroom β-glucans are potent immunological stimulators in medicine, but their productivities are very low. In this study, we successfully improved its production by promoter engineering in Pleurotus ostreatus. The promoter for β-1,3-glucan synthase gene (GLS) was replaced by the promoter of glyceraldehyde-3-phosphate dehydrogenase gene of Aspergillus nidulans. The homologous recombination fragment for swapping GLS promoter comprised five segments, which were fused by two rounds of combined touchdown PCR and overlap extension PCR (TD-OE PCR), and was introduced into P. ostreatus through PEG/CaCl2-mediated protoplast transformation. The transformants exhibited one to three fold higher transcription of GLS gene and produced 32% to 131% higher yield of β-glucans than the wild type. The polysaccharide yields had a significant positive correlation to the GLS gene expression. The infrared spectra of the polysaccharides all displayed the typical absorption peaks of β-glucans. This is the first report of successful swapping of promoters in filamentous fungi.Ran ChaiCuiwei QiuDongren LiuYuancheng QiYuqian GaoJinwen ShenLiyou QiuPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 4, p e61693 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ran Chai Cuiwei Qiu Dongren Liu Yuancheng Qi Yuqian Gao Jinwen Shen Liyou Qiu β-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.
description	Mushroom β-glucans are potent immunological stimulators in medicine, but their productivities are very low. In this study, we successfully improved its production by promoter engineering in Pleurotus ostreatus. The promoter for β-1,3-glucan synthase gene (GLS) was replaced by the promoter of glyceraldehyde-3-phosphate dehydrogenase gene of Aspergillus nidulans. The homologous recombination fragment for swapping GLS promoter comprised five segments, which were fused by two rounds of combined touchdown PCR and overlap extension PCR (TD-OE PCR), and was introduced into P. ostreatus through PEG/CaCl2-mediated protoplast transformation. The transformants exhibited one to three fold higher transcription of GLS gene and produced 32% to 131% higher yield of β-glucans than the wild type. The polysaccharide yields had a significant positive correlation to the GLS gene expression. The infrared spectra of the polysaccharides all displayed the typical absorption peaks of β-glucans. This is the first report of successful swapping of promoters in filamentous fungi.
format	article
author	Ran Chai Cuiwei Qiu Dongren Liu Yuancheng Qi Yuqian Gao Jinwen Shen Liyou Qiu
author_facet	Ran Chai Cuiwei Qiu Dongren Liu Yuancheng Qi Yuqian Gao Jinwen Shen Liyou Qiu
author_sort	Ran Chai
title	β-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.
title_short	β-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.
title_full	β-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.
title_fullStr	β-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.
title_full_unstemmed	β-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.
title_sort	β-glucan synthase gene overexpression and β-glucans overproduction in pleurotus ostreatus using promoter swapping.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/2359d390504b4884bf52f5ff35bb0ce1
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β-Glucan synthase gene overexpression and β-glucans overproduction in Pleurotus ostreatus using promoter swapping.

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