Reduction of Acetate and Lactate Contributed to Enhancement of a Recombinant Protein Production in E. coli BL21

Journal of Microbiology and Biotechnology
Tae-Su KimJung-Kul Lee

Abstract

Acetate and lactate in growth media are detrimental to the production of Thermus maltogenic amylase (ThMA), a heterologous protein, as well as to the growth of recombinant Escherichia coli. Only 50 mM of acetate or 10 mM of lactate reduced 90% of specific ThMA activity. In this study, mutant E. coli strains blocked in the ackA-pta or ackA-pta and ldh pathways were created, characterized, and assessed for their culture performace in 300 L-scale fermentation. The ackApta and ldh double-mutant strain formed significantly less lactate and acetate, and produced a concomitant increase in the excretion of pyruvate (17.8 mM) under anaerobic conditions. The ackA-pta mutant strain accumulated significant acetate but had an approximately 2-fold increase in the formation of lactate. The ackA-pta and ldh double-mutant strain had superior overall performance in large-scale culture under suboptimal conditions, giving 67% higher cell density and 66% higher ThMA activity compared with those of the control strain. The doublemutant strain also achieved a 179% improvement in volumetric ThMA production.

Citations

Sep 21, 2016·Applied Microbiology and Biotechnology·Vicente BernalManuel Cánovas
Aug 2, 2018·Frontiers in Chemistry·Désirée L RöhlenMichael J Schöning
May 13, 2019·Bioprocess and Biosystems Engineering·Karim E JaénAlvaro R Lara
Jul 24, 2020·Journal of Microbiology and Biotechnology·Alvaro R LaraJuan-Carlos Sigala
Oct 30, 2020·Frontiers in Microbiology·Fabrizio CarteniStefano Mazzoleni
Jul 9, 2021·Frontiers in Microbiology·Gema Lozano TerolTeresa de Diego Puente

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