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Metabolic engineering of Agrobacterium sp. for UDP-galactose regeneration and oligosaccharide synthesis

Paper ID Volume ID Publish Year Pages File Format Full-Text
31981 44864 2006 9 PDF Available
Title
Metabolic engineering of Agrobacterium sp. for UDP-galactose regeneration and oligosaccharide synthesis
Abstract

Curdlan-producing Agrobacterium sp. is unique in possessing a highly efficient UDP-glucose regeneration system. A broad-host-range expression strategy was successfully developed to exploit the unique metabolic capability for UDP-galactose regeneration during oligosaccharide synthesis. The engineered Agrobacterium cells functioned as a UDP-galactose regeneration system, allowing galactose-containing disaccharides to be synthesized from glucose or other simple sugars. Unexpectedly, a lag period of 24 h preceded the active synthesis, which could be eliminated with rifampicin. An intracellular nucleotide profiling revealed that the UMP level was elevated by 3.8 fold in the presence of rifampicin, suggesting that rifampicin simulated a nitrogen-limitation condition that triggered the metabolic change. Product selectivity was improved nearly 40-fold by using high acceptor concentration and restricting glucose supply. N-acetyllactosamine concentration near 20 mM (7.5 g/l) was obtained, demonstrating the effectiveness of the engineered strain in UDP-galactose regeneration. This organism could be engineered to regenerate other UDP-sugar nucleotides using the same strategy as illustrated here.

Keywords
Agrobacterium sp.; Curdlan; Sugar nucleotide; Oligosaccharide synthesis; Cofactor regeneration; Whole-cell biocatalysis; Metabolic engineering
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Metabolic engineering of Agrobacterium sp. for UDP-galactose regeneration and oligosaccharide synthesis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 8, Issue 5, September 2006, Pages 465–473
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
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Any Questions? feel free to contact us