A new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity

Abstract A new serine carboxypeptidase gene, capA, was identified in Aspergillus niger CBS 513.88 by reading genomic information and performing sequence alignment, and the gene was cloned and expressed in Pichia pastoris GS115. In a shake flask, the enzyme activity of the recombinant strain GS115 (p...

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Autores principales:	Peng Song, Wei Xu, Yang Zhang, Fei Wang, Xiuling Zhou, Haiying Shi, Wei Feng
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ee35c0874d634abfad8c52b63da73129
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spelling	oai:doaj.org-article:ee35c0874d634abfad8c52b63da731292021-12-02T18:48:23ZA new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity10.1038/s41598-021-98003-x2045-2322https://doaj.org/article/ee35c0874d634abfad8c52b63da731292021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-98003-xhttps://doaj.org/toc/2045-2322Abstract A new serine carboxypeptidase gene, capA, was identified in Aspergillus niger CBS 513.88 by reading genomic information and performing sequence alignment, and the gene was cloned and expressed in Pichia pastoris GS115. In a shake flask, the enzyme activity of the recombinant strain GS115 (pPIC9K-capA) reached 209.3 U mg−1. The optimal temperature and pH for enzyme activity were determined to be 45 °C and 6.0, respectively. After incubation at 40–50 °C or at pH 4.0–8.0 for 1 h, the enzyme retained more than 80% or 60% of its initial activity. The presence of 1–10 mmol L−1 Mg2+ enhanced the activity of CapA, whereas 1–10 mmol L−1 Cu2+, Fe2+, or Co2+, 10 mmol L−1 Mn2+, or 1–10 mmol L−1 phenylmethylsulfonyl fluoride (PMSF) significantly inhibited its activity. CapA had a broad substrate specificity and preferred the hydrophobic amino acids Leu and Lys at the C terminus of proteins, and N-benzyloxycarbonyl-l-phenylalanyl-l-leucine (Cbz-Phe-Leu) was the optimal substrate, for which CapA exhibited K m 0.063 mmol L−1 and k cat /K m 186.35 mmol L−1 s−1. The good thermostability, pH stability and hydrolysis characteristics of CapA provide a solid foundation for application in the food and biotechnology fields.Peng SongWei XuYang ZhangFei WangXiuling ZhouHaiying ShiWei FengNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Peng Song Wei Xu Yang Zhang Fei Wang Xiuling Zhou Haiying Shi Wei Feng A new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity
description	Abstract A new serine carboxypeptidase gene, capA, was identified in Aspergillus niger CBS 513.88 by reading genomic information and performing sequence alignment, and the gene was cloned and expressed in Pichia pastoris GS115. In a shake flask, the enzyme activity of the recombinant strain GS115 (pPIC9K-capA) reached 209.3 U mg−1. The optimal temperature and pH for enzyme activity were determined to be 45 °C and 6.0, respectively. After incubation at 40–50 °C or at pH 4.0–8.0 for 1 h, the enzyme retained more than 80% or 60% of its initial activity. The presence of 1–10 mmol L−1 Mg2+ enhanced the activity of CapA, whereas 1–10 mmol L−1 Cu2+, Fe2+, or Co2+, 10 mmol L−1 Mn2+, or 1–10 mmol L−1 phenylmethylsulfonyl fluoride (PMSF) significantly inhibited its activity. CapA had a broad substrate specificity and preferred the hydrophobic amino acids Leu and Lys at the C terminus of proteins, and N-benzyloxycarbonyl-l-phenylalanyl-l-leucine (Cbz-Phe-Leu) was the optimal substrate, for which CapA exhibited K m 0.063 mmol L−1 and k cat /K m 186.35 mmol L−1 s−1. The good thermostability, pH stability and hydrolysis characteristics of CapA provide a solid foundation for application in the food and biotechnology fields.
format	article
author	Peng Song Wei Xu Yang Zhang Fei Wang Xiuling Zhou Haiying Shi Wei Feng
author_facet	Peng Song Wei Xu Yang Zhang Fei Wang Xiuling Zhou Haiying Shi Wei Feng
author_sort	Peng Song
title	A new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity
title_short	A new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity
title_full	A new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity
title_fullStr	A new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity
title_full_unstemmed	A new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity
title_sort	new carboxypeptidase from aspergillus niger with good thermostability, ph stability and broad substrate specificity
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/ee35c0874d634abfad8c52b63da73129
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A new carboxypeptidase from Aspergillus niger with good thermostability, pH stability and broad substrate specificity

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