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Improved Homologous Expression of the Acidic Lipase from Aspergillus niger

In this study, the acidic lipase from Aspergillus niger (ANL) was homologously expressed in A. niger. The expression of ANL was significantly improved by the expression of the native ANL with the introns, the addition of the Kozak sequence and the optimization of the signal sequences. When the cDNA...

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Autores principales: Zhu, Si-Yuan, Xu, Yan, Yu, Xiao-Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society for Microbiology and Biotechnology 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728306/
https://www.ncbi.nlm.nih.gov/pubmed/31752069
http://dx.doi.org/10.4014/jmb.1906.06028
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author Zhu, Si-Yuan
Xu, Yan
Yu, Xiao-Wei
author_facet Zhu, Si-Yuan
Xu, Yan
Yu, Xiao-Wei
author_sort Zhu, Si-Yuan
collection PubMed
description In this study, the acidic lipase from Aspergillus niger (ANL) was homologously expressed in A. niger. The expression of ANL was significantly improved by the expression of the native ANL with the introns, the addition of the Kozak sequence and the optimization of the signal sequences. When the cDNA sequence of ANL fused with the glaA signal was expressed under the gpdA promoter in A. niger, no lipase activity could be detected. We then tried to improve the expression by using the full-length ANL gene containing three introns, and the lipase activity in the supernatant reached 75.80 U/ml, probably as a result of a more stable mRNA structure. The expression was further improved to 100.60 U/ml by introducing a Kozak sequence around the start codon due to a higher translation efficiency. Finally, the effects of three signal sequences including the cbhI signal, the ANL signal and the glaA signal on the lipase expression were evaluated. The transformant with the cbhI signal showed the highest lipase activity (314.67 U/ml), which was 1.90-fold and 3.13-fold higher than those with the ANL signal and the glaA signal, respectively. The acidic lipase was characterized and its highest activity was detected at pH 3.0 and a temperature of 45ºC. These results provided promising strategies for the production of the acidic lipase from A. niger.
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spelling pubmed-97283062022-12-13 Improved Homologous Expression of the Acidic Lipase from Aspergillus niger Zhu, Si-Yuan Xu, Yan Yu, Xiao-Wei J Microbiol Biotechnol Research article In this study, the acidic lipase from Aspergillus niger (ANL) was homologously expressed in A. niger. The expression of ANL was significantly improved by the expression of the native ANL with the introns, the addition of the Kozak sequence and the optimization of the signal sequences. When the cDNA sequence of ANL fused with the glaA signal was expressed under the gpdA promoter in A. niger, no lipase activity could be detected. We then tried to improve the expression by using the full-length ANL gene containing three introns, and the lipase activity in the supernatant reached 75.80 U/ml, probably as a result of a more stable mRNA structure. The expression was further improved to 100.60 U/ml by introducing a Kozak sequence around the start codon due to a higher translation efficiency. Finally, the effects of three signal sequences including the cbhI signal, the ANL signal and the glaA signal on the lipase expression were evaluated. The transformant with the cbhI signal showed the highest lipase activity (314.67 U/ml), which was 1.90-fold and 3.13-fold higher than those with the ANL signal and the glaA signal, respectively. The acidic lipase was characterized and its highest activity was detected at pH 3.0 and a temperature of 45ºC. These results provided promising strategies for the production of the acidic lipase from A. niger. Korean Society for Microbiology and Biotechnology 2020-02-28 2019-11-22 /pmc/articles/PMC9728306/ /pubmed/31752069 http://dx.doi.org/10.4014/jmb.1906.06028 Text en Copyright©2020 by The Korean Society for Microbiology and Biotechnology https://creativecommons.org/licenses/by/4.0/This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research article
Zhu, Si-Yuan
Xu, Yan
Yu, Xiao-Wei
Improved Homologous Expression of the Acidic Lipase from Aspergillus niger
title Improved Homologous Expression of the Acidic Lipase from Aspergillus niger
title_full Improved Homologous Expression of the Acidic Lipase from Aspergillus niger
title_fullStr Improved Homologous Expression of the Acidic Lipase from Aspergillus niger
title_full_unstemmed Improved Homologous Expression of the Acidic Lipase from Aspergillus niger
title_short Improved Homologous Expression of the Acidic Lipase from Aspergillus niger
title_sort improved homologous expression of the acidic lipase from aspergillus niger
topic Research article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728306/
https://www.ncbi.nlm.nih.gov/pubmed/31752069
http://dx.doi.org/10.4014/jmb.1906.06028
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