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Improvement of n-butanol tolerance in Escherichia coli by membrane-targeted tilapia metallothionein

By Wei-Chih Chin, Kuo-Hsing Lin, Jui-Jen Chang and Chieh-Chen Huang

Abstract

Background: Though n-butanol has been proposed as a potential transportation biofuel, its toxicity oftencauses oxidative stress in the host microorganism and is considered one of the bottlenecks preventing itsefficient mass production.Results: To relieve the oxidative stress in the host cell, metallothioneins (MTs), which are known as scavengersfor reactive oxygen species (ROS), were engineered in E. coli hosts for both cytosolic and outer-membrane-targeted (osmoregulatory membrane protein OmpC fused) expression. Metallothioneins from human (HMT),mouse (MMT), and tilapia fish (TMT) were tested. The host strain expressing membrane-targeted TMT showed thegreatest ability to reduce oxidative stresses induced by n-butanol, ethanol, furfural, hydroxymethylfurfural, andnickel. The same strain also allowed for an increased growth rate of recombinant E. coli under n-butanol stress.Further experiments indicated that the TMT-fused OmpC protein could not only function in ROS scavenging butalso regulate either glycine betaine (GB) or glucose uptake via osmosis, and the dual functional fusion proteincould contribute in an enhancement of the host microorganism’s growth rate.Conclusions: The abilities of scavenging intracellular or extracellular ROS by these engineering E. coli wereexamined, and TMT show the best ability among three MTs. Additionally, the membrane-targeted fusion protein,OmpC-TMT, improved host tolerance up to 1.5% n-butanol above that of TMT which is only 1%. These resultspresented indicate potential novel approaches for engineering stress tolerant microorganism strains

Topics: Metallothionein, n-butanol, OmpC, Tilapia, E. coli, Oxidative stress
Year: 2014
DOI identifier: 10.1186/1754-6834-6-130
OAI identifier: oai:ir.lib.nchu.edu.tw:11455/85105

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