Engineering redox-balanced ethanol production in the cellulolytic and extremely thermophilic bacterium, Caldicellulosiruptor bescii

Metabolic Engineering Communications
Amanda M Williams-RhaesaMichael W W Adams

Abstract

Caldicellulosiruptor bescii is an extremely thermophilic cellulolytic bacterium with great potential for consolidated bioprocessing of renewable plant biomass. Since it does not natively produce ethanol, metabolic engineering is required to create strains with this capability. Previous efforts involved the heterologous expression of the gene encoding a bifunctional alcohol dehydrogenase, AdhE, which uses NADH as the electron donor to reduce acetyl-CoA to ethanol. Acetyl-CoA produced from sugar oxidation also generates reduced ferredoxin but there is no known pathway for the transfer of electrons from reduced ferredoxin to NAD in C. bescii. Herein, we engineered a strain of C. bescii using a more stable genetic background than previously reported and heterologously-expressed adhE from Clostridium thermocellum (which grows optimally (Topt) at 60 °C) with and without co-expression of the membrane-bound Rnf complex from Thermoanaerobacter sp. X514 (Topt 60 °C). Rnf is an energy-conserving, reduced ferredoxin NAD oxidoreductase encoded by six genes (rnfCDGEAB). It was produced in a catalytically active form in C. bescii that utilized the largest DNA construct to be expressed in this organism. The new genetic lineage containing AdhE ...Continue Reading

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Citations

Aug 10, 2020·Biotechnology and Bioengineering·Christopher T StraubRobert M Kelly
Nov 30, 2019·Frontiers in Bioengineering and Biotechnology·Uldis KalnenieksReinis Rutkis
Jul 31, 2019·Extremophiles : Life Under Extreme Conditions·Laura L LeeRobert M Kelly
Mar 19, 2019·Annual Review of Chemical and Biomolecular Engineering·Igor A PodolskyMichelle A O'Malley
Jul 1, 2020·Frontiers in Bioengineering and Biotechnology·Daochen ZhuJianzhong Sun
Aug 9, 2019·Nature Communications·Christopher T StraubRobert M Kelly
Mar 16, 2019·Current Opinion in Biotechnology·James R CrosbyRobert M Kelly
Mar 9, 2021·Environmental Microbiology Reports·Ryan G BingRobert M Kelly
Apr 30, 2021·Biotechnology and Bioengineering·Sajjad Ghaffarinasab, Ehsan Motamedian
Aug 10, 2021·International Journal of Biological Macromolecules·Anica DadwalTulasi Satyanarayana

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Methods Mentioned

BETA
PCR

Software Mentioned

MxPro
GeneWiz

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