Enhancement of malate production through engineering of the periplasmic rTCA pathway in Escherichia coli

Biotechnology and Bioengineering
Liang GuoLiming Liu

Abstract

The compartmentalization of enzymes into organelles is a promising strategy for limiting metabolic crosstalk and improving pathway efficiency; however, prokaryotes are unicellular organisms that lack membrane-bound organelles. To mimic this natural compartmentalization, we present here the targeting of the reductive tricarboxylic acid (rTCA) pathway to the periplasm to enhance the production of malate. A multigene combination knockout strategy was used to construct a phosphoenolpyruvate (PEP) pool. Then, the genes encoding phosphoenolpyruvate carboxykinase and malate dehydrogenase were combinatorially overexpressed to construct a cytoplasmic rTCA pathway for malate biosynthesis; however, the efficiency of malate production was low. To further enhance malate production, the rTCA pathway was targeted to the periplasm, which led to a 100% increase in malate production to 18.8 mM. Next, dual metabolic engineering regulation was adopted to balance the cytoplasmic and periplasmic pathways, leading to an increase in malate production to 58.8 mM. The final engineered strain, GL2306, produced 193 mM malate with a yield of 0.53 mol/mol in 5 L of pH-stat fed-batch culture. The strategy described here paves the way for the development of m...Continue Reading

References

Aug 15, 1995·Proceedings of the National Academy of Sciences of the United States of America·B Kenny, B B Finlay
Jun 1, 1994·Molecular Microbiology·C Wülfing, A Plückthun
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Feb 13, 2001·Biochemical and Biophysical Research Communications·M A KaderbhaiS L Kelly
Apr 25, 2001·Applied and Environmental Microbiology·M A KaderbhaiD C Lamb
Jun 28, 2005·Molecular Microbiology·Jesper E Mogensen, Daniel E Otzen
May 18, 2007·Annual Review of Microbiology·Martine P BosJan Tommassen
Oct 24, 2007·Methods in Molecular Biology·Filipe J Mergulhão, Gabriel A Monteiro
Jun 17, 2008·Molecular Microbiology·Julia HullmannVolkmar Braun
Apr 22, 2009·Journal of the American Chemical Society·Travis S BayerChristopher A Voigt
Nov 18, 2009·Proceedings of the National Academy of Sciences of the United States of America·Xueli ZhangLonnie O Ingram
Jan 1, 2009·Advances in Protein Chemistry and Structural Biology·William J AllenGabriel Waksman
Nov 26, 2010·Applied and Environmental Microbiology·X ZhangL O Ingram
Sep 29, 2011·Metabolic Engineering·Hanson LeeJohn E Dueber
Jun 19, 2012·Bioresource Technology·Hyun-Dong ShinRachel R Chen
Jul 24, 2012·Journal of Pediatric Nursing·Gaye Iannelli-Madigan
May 28, 2013·Metabolic Engineering·Xiulai ChenJian Chen
Nov 18, 2015·Scientific Reports·Suguru MurakamiKouichi Funato
Mar 22, 2016·Trends in Biotechnology·Gang WuMattheos A G Koffas
Mar 31, 2016·Nature Communications·William C DeLoacheJohn E Dueber
Aug 30, 2016·Nature·Markus JeschekThomas R Ward
Sep 27, 2016·Biotechnology and Bioengineering·Xiaoxiang DongLiming Liu
Nov 26, 2016·Science·Justin R MeyerRichard E Lenski
Jan 28, 2017·Nature Communications·Christopher P LongMaciek R Antoniewicz

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Citations

Jul 11, 2020·Biotechnology and Bioengineering·Chao YeXiulai Chen
Aug 23, 2019·Nature Communications·Cong GaoLiming Liu
May 10, 2020·Nature Communications·Qiang DingXiulai Chen
Feb 15, 2021·Biotechnology Advances·Wenna LiQipeng Yuan
Apr 6, 2021·Applied Microbiology and Biotechnology·Lihao JiLiming Liu
Jul 28, 2021·Nature Chemical Biology·Wei JiangRodrigo Ledesma-Amaro
Dec 23, 2021·Biotechnology and Bioengineering·Liang GuoLiming Liu

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