Bioprocessing analysis of Pyrococcus furiosus strains engineered for CO₂-based 3-hydroxypropionate production

Biotechnology and Bioengineering
Aaron S HawkinsR M Kelly

Abstract

Metabolically engineered strains of the hyperthermophile Pyrococcus furiosus (T(opt) 95-100°C), designed to produce 3-hydroxypropionate (3HP) from maltose and CO2 using enzymes from the Metallosphaera sedula (T(opt) 73°C) carbon fixation cycle, were examined with respect to the impact of heterologous gene expression on metabolic activity, fitness at optimal and sub-optimal temperatures, gas-liquid mass transfer in gas-intensive bioreactors, and potential bottlenecks arising from product formation. Transcriptomic comparisons of wild-type P. furiosus, a genetically-tractable, naturally-competent mutant (COM1), and COM1-based strains engineered for 3HP production revealed numerous differences after being shifted from 95°C to 72°C, where product formation catalyzed by the heterologously-produced M. sedula enzymes occurred. At 72°C, significantly higher levels of metabolic activity and a stress response were evident in 3HP-forming strains compared to the non-producing parent strain (COM1). Gas-liquid mass transfer limitations were apparent, given that 3HP titers and volumetric productivity in stirred bioreactors could be increased over 10-fold by increased agitation and higher CO2 sparging rates, from 18 mg/L to 276 mg/L and from 0....Continue Reading

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Citations

Dec 9, 2016·Microbial Biotechnology·Michael W W Adams, Robert M Kelly
Jan 13, 2017·Critical Reviews in Biotechnology·Changshui LiuGuang Zhao
Feb 22, 2017·Microbial Biotechnology·Servé W M Kengen
Feb 17, 2017·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·James A CountsRobert M Kelly
Aug 11, 2020·Environmental Microbiology·James A CountsRobert M Kelly
Mar 20, 2019·Applied Microbiology and Biotechnology·Gyu Bi LeeJung-Hyun Lee
Jun 27, 2018·FEMS Microbiology Reviews·Christopher T StraubRobert M Kelly
Dec 4, 2020·Biotechnology Advances·Kevin PfeiferSimon K-M R Rittmann
Mar 21, 2017·Current Opinion in Biotechnology·Christopher T StraubRobert M Kelly

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