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

被引:17
|
作者
Mao, Shuhong [1 ]
Liu, Yanna [2 ]
Yang, Juanjuan [1 ]
Ma, Xiaoyu [2 ]
Zeng, Fang [2 ]
Zhang, Zhaohui [2 ]
Wang, Shan [3 ]
Han, Haichao [1 ]
Qin, Hui-Min [1 ,3 ]
Lu, Fuping [1 ,2 ]
机构
[1] Tianjin Univ Sci & Technol, Minist Educ, Key Lab Ind Fermentat Microbiol, Tianjin 300457, Peoples R China
[2] Tianjin Univ Sci & Technol, State Key Lab Food Nutr & Safety, Tianjin 300457, Peoples R China
[3] Tianjin Univ Sci & Technol, Coll Biotechnol, Tianjin Key Lab Ind Microbiol, Tianjin Econ & Technol Dev Zone, 29,13 Main St, Tianjin 300457, Peoples R China
基金
国家重点研发计划; 中国国家自然科学基金;
关键词
BETA-FRUCTOFURANOSIDASE; INVERTASE GENE; PURIFICATION; ENZYMES; CELL; INSIGHTS;
D O I
10.1039/c9ra02520k
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
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 degrees C and pH 6.0, and it was stable below 50 degrees 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.
引用
收藏
页码:23856 / 23863
页数:8
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