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KMID : 0545120150250071093
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Journal of Microbiology and Biotechnology
2015 Volume.25 No. 7 p.1093 ~ p.1100
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Reduction of Acetate and Lactate Contributed to Enhancement of a Recombinant Protein Production in E. coli BL21
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Kim Tae-Su
Jung Hyung-Moo
Kim Sang-Yong
Liaoyuan Zhang
Jinglin Li
Sujan Sigdel
Park Ji-Hyun
Haw Jung-Rim
Lee Jung-Kul
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Abstract
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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.
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KEYWORD
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large-scale bioreactor, maltogenic amylase, metabolically engineered, production, recombinant protein
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