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Kim, Taesung
Microfluidics & Nanomechatronics Lab (μFNM)
Research Interests
  • Micro-/Nanofluidics

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High-Throughput Screening of Acyl-CoA Thioesterase I Mutants Using a Fluid Array Platform

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Title
High-Throughput Screening of Acyl-CoA Thioesterase I Mutants Using a Fluid Array Platform
Author
Lim, Ji WonShin, Kwang SooRyu, Young ShinLee, YongjooLee, Sung KukKim, Taesung
Issue Date
2019-12
Publisher
AMER CHEMICAL SOC
Citation
ACS OMEGA, v.4, no.26, pp.21848 - 21854
Abstract
Screening target microorganisms from a mutated recombinant library plays a crucial role in advancing synthetic biology and metabolic engineering. However, conventional screening tools have several limitations regarding throughput, cost, and labor. Here, we used the fluid array platform to conduct high-throughput screening (HTS) that identified Escherichia coli ‘TesA thioesterase mutants producing elevated yields of free fatty acids (FFAs) from a large (106) mutant library. A growth-based screening method using a TetA-RFP fusion sensing mechanism and a reporter-based screening method using high-level FFA producing mutants were employed to identify these mutants via HTS. The platform was able to cover >95% of the mutation library, and it screened target cells from many arrays of the fluid array platform so that a post-analysis could be conducted by gas chromatography. The ‘TesA mutation of each isolated mutant showing improved FFA production in E. coli was characterized, and its enhanced FFA production capability was confirmed.
URI
https://scholarworks.unist.ac.kr/handle/201301/30760
URL
https://pubs.acs.org/doi/10.1021/acsomega.9b02826
DOI
10.1021/acsomega.9b02826
ISSN
2470-1343
Appears in Collections:
MNE_Journal Papers
ECHE_Journal Papers
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