Analysis of L-glutamic acid fermentation by using a dynamic metabolic simulation model of Escherichia coli.
BACKGROUND
Understanding the process of amino acid fermentation as a comprehensive system is a challenging task. Previously, we developed a literature-based dynamic simulation model, which included transcriptional regulation, transcription, translation, and enzymatic reactions related to glycolysis, the pentose phosphate pathway, the tricarboxylic acid (TCA) cycle, and the anaplerotic pathway of Escherichia coli. During simulation, cell growth was defined such as to reproduce the experimental cell growth profile of fed-batch cultivation in jar fermenters. However, to confirm the biological appropriateness of our model, sensitivity analysis and experimental validation were required.


RESULTS
We constructed an L-glutamic acid fermentation simulation model by removing sucAB, a gene encoding α-ketoglutarate dehydrogenase. We then performed systematic sensitivity analysis for L-glutamic acid production; the results of this process corresponded with previous experimental data regarding L-glutamic acid fermentation. Furthermore, it allowed us to predicted the possibility that accumulation of 3-phosphoglycerate in the cell would regulate the carbon flux into the TCA cycle and lead to an increase in the yield of L-glutamic acid via fermentation. We validated this hypothesis through a fermentation experiment involving a model L-glutamic acid-production strain, E. coli MG1655 ΔsucA in which the phosphoglycerate kinase gene had been amplified to cause accumulation of 3-phosphoglycerate. The observed increase in L-glutamic acid production verified the biologically meaningful predictive power of our dynamic metabolic simulation model.


CONCLUSIONS
In this study, dynamic simulation using a literature-based model was shown to be useful for elucidating the precise mechanisms involved in fermentation processes inside the cell. Further exhaustive sensitivity analysis will facilitate identification of novel factors involved in the metabolic regulation of amino acid fermentation.
Institute for Innovation, Ajinomoto Co, Inc,, Suzuki-cho 1-1, Kawasaki-ku, Kawasaki City, Kanagawa 210-8681, Japan. yousuke_nishio@ajinomoto.com.
Yousuke
Y
Nishio
Soichi
S
Ogishima
Masao
M
Ichikawa
Yohei
Y
Yamada
Yoshihiro
Y
Usuda
Tadashi
T
Masuda
Hiroshi
H
Tanaka
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D005784:Gene Amplification; D005798:Genes, Bacterial / Q000235:genetics; D018698:Glutamic Acid / Q000096:biosynthesis*; D008954:Models, Biological; D015203:Reproducibility of Results
D018698:Glutamic Acid