Linked Life Data

20600382

Source:http://linkedlifedata.com/resource/pubmed/id/20600382

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-2
pubmed:dateCreated
2010-8-11
pubmed:abstractText
Thymidine is a commercially important precursor in the production of antiviral drugs, including azidothymidine for the treatment of AIDS. In order to produce thymidine in a large scale, we previously developed a thymidine-overproducing Escherichia coli strain BLdtug24 by engineering pathways. To further enhance thymidine yield, we increased the availability of a cofactor (NADPH) in thymidine biosynthesis by disrupting phosphoglucose isomerase in BLdtug24 to construct BLdtugp24. Additionally, NAD(+) kinase or soluble transhydrogenase was overexpressed in BLdtugp24, which can reroute glucose metabolic flow from the EMP pathway to the PP pathway, to construct BLdtugp34N or BLdtugp34U, respectively. In chemostat cultures, BLdtugp24 had an increased NADPH availability and a 4-fold enhancement in thymidine yield for glucose compared with BLdtug24. BLdtugp34N and BLdtugp34U had increased thymidine yields for glucose by 1.2- and 2-fold compared with BLdtugp24, respectively. The NADPH/NADP(+) ratios at steady-state had overall positive correlations with thymidine yields in these strains. Real-time RT-PCR analysis revealed that the transcriptional regulations of NAD(P)H-related enzymes and transhydrogenases affected the redox balance and shifted reaction equilibrium toward increasing NADPH. In fed-batch fermentation, BLdtugp34U with the highest NADPH/NADP(+) ratio in chemostat experiment produced 1.9gl(-1) of thymidine with 29.7mgl(-1)h(-1) of thymidine productivity.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1873-4863
pubmed:author
pubmed:copyrightInfo
Copyright 2010 Elsevier B.V. All rights reserved.
pubmed:issnType
Electronic
pubmed:day
20
pubmed:volume
149
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
24-32
pubmed:meshHeading
pubmed:year
2010
pubmed:articleTitle
High NADPH/NADP+ ratio improves thymidine production by a metabolically engineered Escherichia coli strain.
pubmed:affiliation
BioNgene Co., Ltd. 10-1, 1Ka, Myungryun-Dong, Jongro-Ku, Seoul 110-521, Republic of Korea.
pubmed:publicationType
Journal Article