Expression of Phosphofructokinase Is Not Sufficient to Enable Embden-Meyerhof-Parnas Glycolysis in Zymomonas mobilis ZM4

Felczak, Magdalena M.; Jacobson, Tyler B.; Ong, Wai Kit; Amador-Noguez, Daniel; TerAvest, Michaela A.

doi:10.3389/fmicb.2019.02270

Expression of Phosphofructokinase Is Not Sufficient to Enable Embden-Meyerhof-Parnas Glycolysis in Zymomonas mobilis ZM4

Journal Article · Fri Sep 27 00:00:00 EDT 2019 · Frontiers in Microbiology

DOI:https://doi.org/10.3389/fmicb.2019.02270· OSTI ID:1579344

Felczak, Magdalena M. ^[1]; Jacobson, Tyler B. ^[2]; Ong, Wai Kit ^[2]; Amador-Noguez, Daniel ^[2]; TerAvest, Michaela A. ^[3]

Michigan State Univ., East Lansing, MI (United States); Michigan State University Department of Biochemistry and Molecular Biology
Univ. of Wisconsin, Madison, WI (United States)
Michigan State Univ., East Lansing, MI (United States)

Zymomonas mobilis is a bacterium that produces ethanol from glucose at up to 97% of theoretical efficiency on a carbon basis. One factor contributing to the high efficiency of ethanol production is that Z. mobilis has a low biomass yield. The low biomass yield may be caused partly by the low ATP yield of the Entner-Doudoroff (ED) glycolytic pathway used by Z. mobilis, which produces only one ATP per glucose consumed. To test the hypothesis that ATP yield limits biomass yield in Z. mobilis, we attempted to introduce the Embden-Meyerhof-Parnas (EMP) glycolytic pathway (with double the ATP yield) by expressing phosphofructokinase (Pfk I) from Escherichia coli. Expression of Pfk I caused growth inhibition and resulted in accumulation of mutations in the pfkA gene. Co-expression of additional EMP enzymes, fructose bisphosphate aldolase (Fba) and triose phosphate isomerase (Tpi), with Pfk I did not enable EMP flux, and resulted in production of glycerol as a side product. Further analysis indicated that heterologous reactions may have operated in the reverse direction because of native metabolite concentrations. This study reveals how the metabolomic context of a chassis organism influences the range of pathways that can be added by heterologous expression.

View Accepted Manuscript (DOE)

Research Organization:: Great Lakes Bioenergy Research Center (GLBRC), Madison, WI (United States); USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: SC0018409; FC02-07ER64494; AC02-05CH11231

OSTI ID:: 1579344

Journal Information:: Frontiers in Microbiology, Journal Name: Frontiers in Microbiology Vol. 10; ISSN 1664-302X

Publisher:: Frontiers Research FoundationCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Embden-Meyerhof-Parnas pathway
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Zymomonas mobilis
glycolysis
metabolic engineering

Expression of Phosphofructokinase Is Not Sufficient to Enable Embden-Meyerhof-Parnas Glycolysis in Zymomonas mobilis ZM4

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References (23)

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