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Cloning, expression and characterization of a novel fructosyltransferase from Aspergillus niger and its application in the synthesis of fructooligosaccharides

Fructosyltransferases have been used in the industrial production of fructooligosaccharides (FOS). However, it is still not possible to explain the difference in FOS production based on the variations observed in the FOS synthesizing enzymes of A. niger. In the present study, a novel fructosyltransf...

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Autores principales: Mao, Shuhong, Liu, Yanna, Yang, Juanjuan, Ma, Xiaoyu, Zeng, Fang, Zhang, Zhaohui, Wang, Shan, Han, Haichao, Qin, Hui-Min, Lu, Fuping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9069702/
https://www.ncbi.nlm.nih.gov/pubmed/35530578
http://dx.doi.org/10.1039/c9ra02520k
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author Mao, Shuhong
Liu, Yanna
Yang, Juanjuan
Ma, Xiaoyu
Zeng, Fang
Zhang, Zhaohui
Wang, Shan
Han, Haichao
Qin, Hui-Min
Lu, Fuping
author_facet Mao, Shuhong
Liu, Yanna
Yang, Juanjuan
Ma, Xiaoyu
Zeng, Fang
Zhang, Zhaohui
Wang, Shan
Han, Haichao
Qin, Hui-Min
Lu, Fuping
author_sort Mao, Shuhong
collection PubMed
description Fructosyltransferases have been used in the industrial production of fructooligosaccharides (FOS). However, it is still not possible to explain the difference in FOS production based on the variations observed in the FOS synthesizing enzymes of A. niger. In the present study, a novel fructosyltransferase (FT-A) from A. niger TCCC41686 with high FOS synthesis ability was characterized. The FT-A gene was obtained and expressed in Pichia pastoris. A homology model of FT-A showed that the changes of residues identified outside the conserved domains were found to have an effect on its characteristics and its FOS-synthesis capacity. The optimal activity of the recombinant FT-A was observed at 50 °C and pH 6.0, and it was stable below 50 °C and over the range of pH 3.0–11.0. The K(m) and V(max) values of FT-A were 151.13 g L(−1) and 6.55 g L(−1) min(−1), respectively. The production of FOS using the recombinant FT-A remained above 60% during 50–80 min of synthesis based on sucrose as substrate. The novel fructosyltransferase (FT-A) investigated in this study can potentially be applied for the efficient industrial production of FOS. The results also provide more valuable information for explaining the relationship between the structure and function of the FT-A.
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spelling pubmed-90697022022-05-05 Cloning, expression and characterization of a novel fructosyltransferase from Aspergillus niger and its application in the synthesis of fructooligosaccharides Mao, Shuhong Liu, Yanna Yang, Juanjuan Ma, Xiaoyu Zeng, Fang Zhang, Zhaohui Wang, Shan Han, Haichao Qin, Hui-Min Lu, Fuping RSC Adv Chemistry Fructosyltransferases have been used in the industrial production of fructooligosaccharides (FOS). However, it is still not possible to explain the difference in FOS production based on the variations observed in the FOS synthesizing enzymes of A. niger. In the present study, a novel fructosyltransferase (FT-A) from A. niger TCCC41686 with high FOS synthesis ability was characterized. The FT-A gene was obtained and expressed in Pichia pastoris. A homology model of FT-A showed that the changes of residues identified outside the conserved domains were found to have an effect on its characteristics and its FOS-synthesis capacity. The optimal activity of the recombinant FT-A was observed at 50 °C and pH 6.0, and it was stable below 50 °C and over the range of pH 3.0–11.0. The K(m) and V(max) values of FT-A were 151.13 g L(−1) and 6.55 g L(−1) min(−1), respectively. The production of FOS using the recombinant FT-A remained above 60% during 50–80 min of synthesis based on sucrose as substrate. The novel fructosyltransferase (FT-A) investigated in this study can potentially be applied for the efficient industrial production of FOS. The results also provide more valuable information for explaining the relationship between the structure and function of the FT-A. The Royal Society of Chemistry 2019-07-31 /pmc/articles/PMC9069702/ /pubmed/35530578 http://dx.doi.org/10.1039/c9ra02520k Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/
spellingShingle Chemistry
Mao, Shuhong
Liu, Yanna
Yang, Juanjuan
Ma, Xiaoyu
Zeng, Fang
Zhang, Zhaohui
Wang, Shan
Han, Haichao
Qin, Hui-Min
Lu, Fuping
Cloning, expression and characterization of a novel fructosyltransferase from Aspergillus niger and its application in the synthesis of fructooligosaccharides
title Cloning, expression and characterization of a novel fructosyltransferase from Aspergillus niger and its application in the synthesis of fructooligosaccharides
title_full Cloning, expression and characterization of a novel fructosyltransferase from Aspergillus niger and its application in the synthesis of fructooligosaccharides
title_fullStr Cloning, expression and characterization of a novel fructosyltransferase from Aspergillus niger and its application in the synthesis of fructooligosaccharides
title_full_unstemmed Cloning, expression and characterization of a novel fructosyltransferase from Aspergillus niger and its application in the synthesis of fructooligosaccharides
title_short Cloning, expression and characterization of a novel fructosyltransferase from Aspergillus niger and its application in the synthesis of fructooligosaccharides
title_sort cloning, expression and characterization of a novel fructosyltransferase from aspergillus niger and its application in the synthesis of fructooligosaccharides
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9069702/
https://www.ncbi.nlm.nih.gov/pubmed/35530578
http://dx.doi.org/10.1039/c9ra02520k
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