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
相关论文
共 50 条
  • [41] Synthesis of Fructooligosaccharides from Aspergillus niger Commercial Inulinase Immobilized in Montmorillonite Pretreated in Pressurized Propane and LPG
    Kuhn, Graciele de Oliveira
    Dalla Rosa, Clarissa
    Silva, Marceli Fernandes
    Treichel, Helen
    de Oliveira, Debora
    Vladimir Oliveira, J.
    APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 2013, 169 (03) : 750 - 760
  • [42] Purification, Characterization and Application of Polygalacturonase from Aspergillus niger CSTRF
    Juwon, Arotupin Daniel
    Akinyosoye, F. A.
    Kayode, Onifade Anthony
    MALAYSIAN JOURNAL OF MICROBIOLOGY, 2012, 8 (03) : 175 - 183
  • [43] CLONING, CHARACTERIZATION, AND EXPRESSION OF 2 ALPHA-AMYLASE GENES FROM ASPERGILLUS-NIGER VAR AWAMORI
    KORMAN, DR
    BAYLISS, FT
    BARNETT, CC
    CARMONA, CL
    KODAMA, KH
    ROYER, TJ
    THOMPSON, SA
    WARD, M
    WILSON, LJ
    BERKA, RM
    CURRENT GENETICS, 1990, 17 (03) : 203 - 212
  • [44] Screening of microbes for novel acidic cutinases and cloning and expression of an acidic cutinase from Aspergillus niger CBS 513.88
    Nyyssola, Antti
    Pihlajaniemi, Ville
    Jarvinen, Riikka
    Mikander, Saara
    Kontkanen, Hanna
    Kruus, Kristiina
    Kallio, Heikki
    Buchert, Johanna
    ENZYME AND MICROBIAL TECHNOLOGY, 2013, 52 (4-5) : 272 - 278
  • [45] Purification and characterization of a novel prolyl endopeptidase from Aspergillus niger
    Kubota, K
    Tanokura, M
    Takahashi, K
    PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES, 2005, 81 (10): : 447 - 453
  • [46] Isolation and characterization of a novel antifungal peptide from Aspergillus niger
    Lee, DG
    Shin, SY
    Maeng, CY
    Jin, ZZ
    Kim, KL
    Hahm, KS
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 263 (03) : 646 - 651
  • [47] Purification and kinetic characterization of a fructosyltransferase from Aspergillus aculeatus
    Ghazi, Iraj
    Fernandez-Arrojo, Lucia
    Garcia-Arellano, Humberto
    Ferrer, Manuel
    Ballesteros, Antonio
    Plou, Francisco J.
    JOURNAL OF BIOTECHNOLOGY, 2007, 128 (01) : 204 - 211
  • [48] Expression and Activity Analysis of Fructosyltransferase from Aspergillus oryzae
    Guan, Lihong
    Chen, Liping
    Chen, Yongsen
    Zhang, Nu
    Han, Yawei
    PROTEIN JOURNAL, 2017, 36 (04): : 352 - 360
  • [49] Expression and Activity Analysis of Fructosyltransferase from Aspergillus oryzae
    Lihong Guan
    Liping Chen
    Yongsen Chen
    Nu Zhang
    Yawei Han
    The Protein Journal, 2017, 36 : 352 - 360
  • [50] Molecular cloning and heterologous expression of the isopullulanase gene from Aspergillus niger ATCC 9642
    Aoki, H
    Yopi
    Sakano, Y
    BIOCHEMICAL JOURNAL, 1997, 323 : 757 - 764