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Employing bacterial microcompartment technology to engineer a shell-free enzyme-aggregate for enhanced 1,2-propanediol production in Escherichia coli

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논문

Employing bacterial microcompartment technology to engineer a shell-free enzyme-aggregate for enhanced 1,2-propanediol production in Escherichia coli

학술지

Metabolic engineering

저자명

Lee, Matthew J.; Brown, Ian R.; Juodeikis, Rokas; Frank, Stefanie; Warren, Martin J.

초록

<▼1><P>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.</P></▼1><▼2><P><B>Highlights</B></P><P>&#x2022;<P>Fusion of BMC targeting peptides to enzymes has a variable effect on activity.</P>&#x2022;<P>Tagged enzymes for 1,2-propanediol synthesis are localised to a BMC.</P>&#x2022;<P>BMC-targeted proteins localised within the BMC are protected from proteases.</P>&#x2022;<P>TEM reveals tagged enzymes form large intracellular protein aggregates.</P>&#x2022;<P>Strains with enzyme aggregates are shown to have enhanced 1,2-propanediol production.</P></P></▼2>

발행연도

2016

발행기관

Academic Press

라이선스

cc-by

ISSN

1096-7176

ISSN

1096-7184

36

페이지

pp.48-56

주제어

BMC, bacterial microcompartment; Pdu, 1,2-propanediol utilisation; 1,2-PD, 1,2-propanediol; D18, First 18 amino acids of PduD; P18, First 18 amino acids of PduP; Synthetic biology; Metabolic engineering; Compartmentalisation; Protein aggregation; Biotechnology

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논문; 2016-07-01

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