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Engineering Bacteria to Catabolize the Carbonaceous Component of Sarin: Teaching E. coli to Eat Isopropanol.

Engineering Bacteria to Catabolize the Carbonaceous Component of Sarin: Teaching E. coli to Eat Isopropanol.

Authors :
Brown, Margaret E
Brown, Margaret E
Mukhopadhyay, Aindrila
Keasling, Jay D
Brown, Margaret E
Brown, Margaret E
Mukhopadhyay, Aindrila
Keasling, Jay D
Source :
ACS synthetic biology; vol 5, iss 12, 1485-1496; 2161-5063
Publication Year :
2016

Abstract

We report an engineered strain of Escherichia coli that catabolizes the carbonaceous component of the extremely toxic chemical warfare agent sarin. Enzymatic decomposition of sarin generates isopropanol waste that, with this engineered strain, is then transformed into acetyl-CoA by enzymatic conversion with a key reaction performed by the acetone carboxylase complex (ACX). We engineered the heterologous expression of the ACX complex from Xanthobacter autotrophicus PY2 to match the naturally occurring subunit stoichiometry and purified the recombinant complex from E. coli for biochemical analysis. Incorporating this ACX complex and enzymes from diverse organisms, we introduced an isopropanol degradation pathway in E. coli, optimized induction conditions, and decoupled enzyme expression to probe pathway bottlenecks. Our engineered E. coli consumed 65% of isopropanol compared to no-cell controls and was able to grow on isopropanol as a sole carbon source. In the process, reconstitution of this large ACX complex (370 kDa) in a system naìˆve to its structural and mechanistic requirements allowed us to study this otherwise cryptic enzyme in more detail than would have been possible in the less genetically tractable native Xanthobacter system.

Details

Database :
OAIster
Journal :
ACS synthetic biology; vol 5, iss 12, 1485-1496; 2161-5063
Notes :
ACS synthetic biology vol 5, iss 12, 1485-1496 2161-5063
Publication Type :
Electronic Resource
Accession number :
edsoai.on1287414745
Document Type :
Electronic Resource