The effect of NADP-dependent malic enzyme expression and anaerobic C4 metabolism in Escherichia coli compared with other anaplerotic enzymes.
OBJECTIVE
To understand the modification of C4-metabolism under anaerobic glycolysis condition by overexpressing anaplerotic enzymes, which mediating carboxylation of C3 into C4 metabolites, in Escherichia coli.


RESULTS
Anaplerotic NADP-dependent malic enzyme (MaeB), as well as the other anaplerotic enzymes, including phosphoenolpyruvate carboxylase (Ppc), phosphoenolpyruvate carboxykinase (Pck) and NAD-dependent malic enzyme (MaeA), were artificially expressed and their C4 metabolism was compared in E. coli. Increasing MaeB expression enhanced the production of C4 metabolites by 2.4 times compared to the wild-type strain in anaerobic glucose medium with bicarbonate supplementation. In MaeB expression, C4 metabolism by supplementing 10 g l(-1) of NaHCO(3) was three times than that by no supplementation, which showed the greatest response to increased CO(2) availability among the tested anaplerotic enzyme expressions.


CONCLUSIONS
The higher C4 metabolism was achieved in E. coli expressing increased levels of the NADPH-dependent MaeB. The greatest increase in the C4 metabolite ratio compared to the other tested enzymes were also found in E. coli with enhanced MaeB expression as CO(2) availability increased.


CONCLUSIONS
The higher C4 metabolites and related biomolecule productions can be accomplished by MaeB overexpression in metabolically engineered E. coli.
Department of Biotechnology, The Catholic University of Korea, Bucheon, Gyunggi, Korea.
Y-D
YD
Kwon
O-H
OH
Kwon
H-S
HS
Lee
P
P
Kim
D000693:Anaerobiosis; D002244:Carbon / Q000378:metabolism*; D002245:Carbon Dioxide / Q000378:metabolism; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006019:Glycolysis; D007218:Industrial Microbiology; D008291:Malate Dehydrogenase / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D050538:Malate Dehydrogenase (NADP+) / Q000235:genetics / Q000378:metabolism*; D019764:Phosphoenolpyruvate Carboxykinase (ATP) / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D010730:Phosphoenolpyruvate Carboxylase / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000008:administration & dosage; D016133:Polymerase Chain Reaction / Q000379:methods; D017693:Sodium Bicarbonate / Q000378:metabolism / Q000494:pharmacology
D002245:Carbon Dioxide; D002244:Carbon; D017693:Sodium Bicarbonate; D008291:Malate Dehydrogenase; C046446:malate dehydrogenase-(oxaloacetate-decarboxylating) (NAD+); D050538:Malate Dehydrogenase (NADP+); D010730:Phosphoenolpyruvate Carboxylase; D019764:Phosphoenolpyruvate Carboxykinase (ATP)