| Improving protein secretion of a transglutaminase-secreting Corynebacterium glutamicum recombinant strain on the basis of (13)C metabolic flux analysis. | |
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MedLine Citation:
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PMID: 21903468 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Corynebacterium glutamicum is known as a host species for amino acid production. This microorganism was recently noticed as a host that produces secreted proteins. In this study, we performed (13)C metabolic flux analysis ((13)C-MFA) on a recombinant C. glutamicum strain that secretes a heterologous transglutaminase (TGase) to improve TGase secretion. For the (13)C-MFA of a TGase-secreting C. glutamicum strain in batch cultivation, a (13)C-labeling experiment and measurement of mass isotopomer distributions of proteinogenic amino acids were performed, and metabolic fluxes were determined considering the changes in fractional (13)C-labeling of proteinogenic amino acids with respect to culture time. The TGase yield increased at the stationary phase but decreased toward its end. The results of (13)C-MFA revealed that the flux from glycolysis to the TCA cycle gradually increased during TGase secretion. We speculate that the NADH/NAD(+) ratio in the cells increases and that as a result, the specific glucose uptake rate decreases in the stationary phase because of the increased flux of the TCA cycle. Since it is expected that a decrease in the NADH/NAD(+) ratio would improve the TGase yield, we tried to enhance lactate production in a TGase-secreting C. glutamicum strain to decrease cellular NADH levels by increasing the pH level in the culture. The TGase yield increased in 1.4-fold by increasing the pH from 6.7 to 7.2, indicating that the TGase yield was successfully improved on the basis of the (13)C-MFA. |
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Authors:
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Motoki Umakoshi; Takashi Hirasawa; Chikara Furusawa; Yasuhiro Takenaka; Yoshimi Kikuchi; Hiroshi Shimizu |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-9-6 |
Journal Detail:
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Title: Journal of bioscience and bioengineering Volume: - ISSN: 1347-4421 ISO Abbreviation: - Publication Date: 2011 Sep |
Date Detail:
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Created Date: 2011-9-9 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 100888800 Medline TA: J Biosci Bioeng Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved. |
Affiliation:
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Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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