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Characterization of the first bacterial and thermostable GDP-mannose 3,5-epimerase

Ophelia Gevaert (UGent) , Stevie Van Overtveldt (UGent) , Koen Beerens (UGent) and Tom Desmet (UGent)
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Abstract
GDP-mannose 3,5-epimerase (GM35E) catalyzes the conversion of GDP-mannose towards GDP-L-galactose and GDP-L-gulose. Although this reaction represents one of the few enzymatic routes towards the production of L-sugars and derivatives, it has not yet been exploited for that purpose. One of the reasons is that so far only GM35Es from plants have been characterized, yielding biocatalysts that are relatively unstable and difficult to express heterologously. Through the mining of sequence databases, we succeeded in identifying a promising bacterial homologue. The gene from the thermophilic organism Methylacidiphilum fumariolicum was codon optimized for expression in Escherichia coli, resulting in the production of 40 mg/L of recombinant protein. The enzyme was found to act as a self-sufficient GM35E, performing three chemical reactions in the same active site. Furthermore, the biocatalyst was highly stable at temperatures up to 55 degrees C, making it well suited for the synthesis of new carbohydrate products with application in the pharma industry.
Keywords
Methylacidiphilum fumariolicum, biocatalysis, GDP-mannose 3, 5-epimerase, L-sugars, recombinant expression, L-GALACTOSE, CARBOHYDRATE EPIMERASES, DISACCHARIDE MOIETY, ESCHERICHIA-COLI, L-GULOSE, BIOSYNTHESIS, ENZYME, STABILITY, MECHANISM, LIPOPOLYSACCHARIDE

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MLA
Gevaert, Ophelia, et al. “Characterization of the First Bacterial and Thermostable GDP-Mannose 3,5-Epimerase.” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 20, no. 14, 2019.
APA
Gevaert, O., Van Overtveldt, S., Beerens, K., & Desmet, T. (2019). Characterization of the first bacterial and thermostable GDP-mannose 3,5-epimerase. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 20(14).
Chicago author-date
Gevaert, Ophelia, Stevie Van Overtveldt, Koen Beerens, and Tom Desmet. 2019. “Characterization of the First Bacterial and Thermostable GDP-Mannose 3,5-Epimerase.” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 20 (14).
Chicago author-date (all authors)
Gevaert, Ophelia, Stevie Van Overtveldt, Koen Beerens, and Tom Desmet. 2019. “Characterization of the First Bacterial and Thermostable GDP-Mannose 3,5-Epimerase.” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 20 (14).
Vancouver
1.
Gevaert O, Van Overtveldt S, Beerens K, Desmet T. Characterization of the first bacterial and thermostable GDP-mannose 3,5-epimerase. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES. 2019;20(14).
IEEE
[1]
O. Gevaert, S. Van Overtveldt, K. Beerens, and T. Desmet, “Characterization of the first bacterial and thermostable GDP-mannose 3,5-epimerase,” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 20, no. 14, 2019.
@article{8633983,
  abstract     = {GDP-mannose 3,5-epimerase (GM35E) catalyzes the conversion of GDP-mannose towards GDP-L-galactose and GDP-L-gulose. Although this reaction represents one of the few enzymatic routes towards the production of L-sugars and derivatives, it has not yet been exploited for that purpose. One of the reasons is that so far only GM35Es from plants have been characterized, yielding biocatalysts that are relatively unstable and difficult to express heterologously. Through the mining of sequence databases, we succeeded in identifying a promising bacterial homologue. The gene from the thermophilic organism Methylacidiphilum fumariolicum was codon optimized for expression in Escherichia coli, resulting in the production of 40 mg/L of recombinant protein. The enzyme was found to act as a self-sufficient GM35E, performing three chemical reactions in the same active site. Furthermore, the biocatalyst was highly stable at temperatures up to 55 degrees C, making it well suited for the synthesis of new carbohydrate products with application in the pharma industry.},
  articleno    = {3530},
  author       = {Gevaert, Ophelia and Van Overtveldt, Stevie and Beerens, Koen and Desmet, Tom},
  issn         = {1661-6596},
  journal      = {INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES},
  keywords     = {Methylacidiphilum fumariolicum,biocatalysis,GDP-mannose 3,5-epimerase,L-sugars,recombinant expression,L-GALACTOSE,CARBOHYDRATE EPIMERASES,DISACCHARIDE MOIETY,ESCHERICHIA-COLI,L-GULOSE,BIOSYNTHESIS,ENZYME,STABILITY,MECHANISM,LIPOPOLYSACCHARIDE},
  language     = {eng},
  number       = {14},
  pages        = {13},
  title        = {Characterization of the first bacterial and thermostable GDP-mannose 3,5-epimerase},
  url          = {http://dx.doi.org/10.3390/ijms20143530},
  volume       = {20},
  year         = {2019},
}

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