A Convenient Fluorescence-Based Assay for the Detection of Sucrose Transport and the Introduction of a Sucrose Transporter from Potato into <i>Clostridium</i> Strains
Zhikai Zhang,
Lihua Lin,
Hongchi Tang,
Shaowei Zeng,
Yuan Guo,
Yutuo Wei,
Ribo Huang,
Hao Pang,
Liqin Du
Affiliations
Zhikai Zhang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi Research Center for Microbial and Enzymatic Technology, College of Life Science and Technology, Guangxi University, Daxue Road No. 100, Nanning 530005, China
Lihua Lin
National Engineering Research Center for Non-Food Biorefinery, State Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Bio-refinery, Guangxi Academy of Sciences, Daling Road No. 98, Nanning 530007, China
Hongchi Tang
National Engineering Research Center for Non-Food Biorefinery, State Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Bio-refinery, Guangxi Academy of Sciences, Daling Road No. 98, Nanning 530007, China
Shaowei Zeng
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi Research Center for Microbial and Enzymatic Technology, College of Life Science and Technology, Guangxi University, Daxue Road No. 100, Nanning 530005, China
Yuan Guo
National Engineering Research Center for Non-Food Biorefinery, State Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Bio-refinery, Guangxi Academy of Sciences, Daling Road No. 98, Nanning 530007, China
Yutuo Wei
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi Research Center for Microbial and Enzymatic Technology, College of Life Science and Technology, Guangxi University, Daxue Road No. 100, Nanning 530005, China
Ribo Huang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi Research Center for Microbial and Enzymatic Technology, College of Life Science and Technology, Guangxi University, Daxue Road No. 100, Nanning 530005, China
Hao Pang
National Engineering Research Center for Non-Food Biorefinery, State Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Bio-refinery, Guangxi Academy of Sciences, Daling Road No. 98, Nanning 530007, China
Liqin Du
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi Research Center for Microbial and Enzymatic Technology, College of Life Science and Technology, Guangxi University, Daxue Road No. 100, Nanning 530005, China
A convenient and effective sucrose transport assay for Clostridium strains is needed. Traditional methods, such as 14C-sucrose isotope labelling, use radioactive materials and are not convenient for many laboratories. Here, a sucrose transporter from potato was introduced into Clostridium, and a fluorescence assay based on esculin was used for the analysis of sucrose transport in Clostridium strains. This showed that the heterologously expressed potato sucrose transporter is functional in Clostridium. Recombinant engineering of high-level sucrose transport would aid sucrose fermentation in Clostridium strains. The assay described herein provides an important technological platform for studying sucrose transporter function following heterologous expression in Clostridium.
