Employing bacterial microcompartment technology to engineer a shell-free enzyme-aggregate for enhanced 1,2-propanediol production in Escherichia coli.
Employing bacterial microcompartment technology to engineer a shell-free enzyme-aggregate for enhanced 1,2-propanediol production in Escherichia coli.
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DOI：
10.1016/j.ymben.2016.02.007
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发表时间：
2016-07
影响因子：
8.4
通讯作者：
Warren MJ
中科院分区：
文献类型：
--
作者：
Lee MJ;Brown IR;Juodeikis R;Frank S;Warren MJ
Bacterial microcompartments (BMCs) enhance the breakdown of metabolites such as 1,2-propanediol (1,2-PD) to propionic acid. The encapsulation of proteins within the BMC is mediated by the presence of targeting sequences. In an attempt to redesign the Pdu BMC into a 1,2-PD synthesising factory using glycerol as the starting material we added N-terminal targeting peptides to glycerol dehydrogenase, dihydroxyacetone kinase, methylglyoxal synthase and 1,2-propanediol oxidoreductase to allow their inclusion into an empty BMC. 1,2-PD producing strains containing the fused enzymes exhibit a 245% increase in product formation in comparison to un-tagged enzymes, irrespective of the presence of BMCs. Tagging of enzymes with targeting peptides results in the formation of dense protein aggregates within the cell that are shown by immuno-labelling to contain the vast majority of tagged proteins. It can therefore be concluded that these protein inclusions are metabolically active and facilitate the significant increase in product formation. Fusion of BMC targeting peptides to enzymes has a variable effect on activity. Tagged enzymes for 1,2-propanediol synthesis are localised to a BMC. BMC-targeted proteins localised within the BMC are protected from proteases. TEM reveals tagged enzymes form large intracellular protein aggregates. Strains with enzyme aggregates are shown to have enhanced 1,2-propanediol production.
