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Enzymatic production of β-D-glucose-1-phosphate from trehalose

Jef Van der Borght (UGent) , Tom Desmet (UGent) and Wim Soetaert (UGent)
(2010) BIOTECHNOLOGY JOURNAL. 5(9). p.986-993
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Abstract
beta-D-Glucose-1-phosphate (beta Glc1P) is an efficient glucosyl donor for both enzymatic and chemical glycosylation reactions but is currently very costly and not available in large amounts. This article provides an efficient production method of beta Glc1P from trehalose and phosphate using the thermostable trehalose phosphorylase from Thermoanaerobacter brockii. At the process temperature of 60 C, Escherichia coli expression host cells are lysed and cell treatment prior to the reaction is, therefore, not required. In this way, the theoretical maximum yield of 26% could be easily achieved. Two different purification strategies have been compared, anion exchange chromatography or carbohydrate removal by treatment with trehalase and yeast, followed by chemical phosphate precipitation. In a next step, beta Glc1P was precipitated with ethanol but this did not induce crystallization, in contrast to what ist observed with other glycosylphosphates. After conversion of the product to its cyclohexylammonium salt, however, crystals could be readily obtained. Although both purification methods were quantitative (>99% recovery), a large amount of product (50%) was lost during crystallization. Nevertheless, a production process for crystalline beta Glc1P is now available from the cheap substrates trehalose and inorganic phosphate.
Keywords
Downstream processing, Phosphorolysis, Beta-D-glucose-1-phosphate, Bioconversion, Trehalose phosphorylase, ESCHERICHIA-COLI, PHOSPHORYLASE, CELLS, GLUCOSE-1-PHOSPHATE, 1-PHOSPHATE, PHOSPHATASE, RELEASE, ACID

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Citation

Please use this url to cite or link to this publication:

Chicago
Van der Borght, Jef, Tom Desmet, and Wim Soetaert. 2010. “Enzymatic Production of β-D-glucose-1-phosphate from Trehalose.” Biotechnology Journal 5 (9): 986–993.
APA
Van der Borght, J., Desmet, T., & Soetaert, W. (2010). Enzymatic production of β-D-glucose-1-phosphate from trehalose. BIOTECHNOLOGY JOURNAL, 5(9), 986–993.
Vancouver
1.
Van der Borght J, Desmet T, Soetaert W. Enzymatic production of β-D-glucose-1-phosphate from trehalose. BIOTECHNOLOGY JOURNAL. 2010;5(9):986–93.
MLA
Van der Borght, Jef, Tom Desmet, and Wim Soetaert. “Enzymatic Production of β-D-glucose-1-phosphate from Trehalose.” BIOTECHNOLOGY JOURNAL 5.9 (2010): 986–993. Print.
@article{1089221,
  abstract     = {beta-D-Glucose-1-phosphate (beta Glc1P) is an efficient glucosyl donor for both enzymatic and chemical glycosylation reactions but is currently very costly and not available in large amounts. This article provides an efficient production method of beta Glc1P from trehalose and phosphate using the thermostable trehalose phosphorylase from Thermoanaerobacter brockii. At the process temperature of 60 C, Escherichia coli expression host cells are lysed and cell treatment prior to the reaction is, therefore, not required. In this way, the theoretical maximum yield of 26\% could be easily achieved. Two different purification strategies have been compared, anion exchange chromatography or carbohydrate removal by treatment with trehalase and yeast, followed by chemical phosphate precipitation. In a next step, beta Glc1P was precipitated with ethanol but this did not induce crystallization, in contrast to what ist observed with other glycosylphosphates. After conversion of the product to its cyclohexylammonium salt, however, crystals could be readily obtained. Although both purification methods were quantitative ({\textrangle}99\% recovery), a large amount of product (50\%) was lost during crystallization. Nevertheless, a production process for crystalline beta Glc1P is now available from the cheap substrates trehalose and inorganic phosphate.},
  author       = {Van der Borght, Jef and Desmet, Tom and Soetaert, Wim},
  issn         = {1860-6768},
  journal      = {BIOTECHNOLOGY JOURNAL},
  keyword      = {Downstream processing,Phosphorolysis,Beta-D-glucose-1-phosphate,Bioconversion,Trehalose phosphorylase,ESCHERICHIA-COLI,PHOSPHORYLASE,CELLS,GLUCOSE-1-PHOSPHATE,1-PHOSPHATE,PHOSPHATASE,RELEASE,ACID},
  language     = {eng},
  number       = {9},
  pages        = {986--993},
  title        = {Enzymatic production of \ensuremath{\beta}-D-glucose-1-phosphate from trehalose},
  url          = {http://dx.doi.org/10.1002/biot.201000203},
  volume       = {5},
  year         = {2010},
}

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