Enzymatic synthesis of oligosaccharides, alkyl and terpenyl glucosides, by recombinant Escherichia coli-expressed Pichia etchellsii β-glucosidase II

  • Preeti Bachhawat
  • Saroj Mishra
  • Yukti Bhatia
  • V. S. Bisaria


The biosynthetic activity of yeast Pichia etchellsii β-glucosidase II (BglII) expressed in recombinant Escherichia coli was utilized for synthesis of cellooligosaccharides, alkyl and terpene glucosides. Cellooligosaccharides with a degree of polymerization of 3 and greater were resolved by thin-layer chromatography (TLC) using an ethyl acetate: 1-propanol:2-propanol:water (8:5:1:1) solvent system followed by visualization with 0.2% naphthoresorcinol reagent. Using 2M cellobiose and 15 IU of partially purified BglII, 57 mmol/L of oligosaccharides (comprising mostly cellotriose and cellopentaose) was synthesized in 16 h. Similarly, alkyl glucosides with chain lengths from 6 to 10 carbons were synthesized and products extracted to near purity by ethylacetate extraction. The same extraction method was employed to separate, to near purity, various monoterpenyl (nerol, geraniol, citronellol) glucosides. A reliable and simple method for separation of cellooligosaccharides using a combination of Bio-Gel P-2 gel filtration and charcoal celite adsorption chromatography was developed. The cellooligosaccharides were separated to purity as confirmed by TLC. The enzyme was among the very few that could synthesize a wide variety of glycoconjugates.

Index Entries

β-Glucosidase Pichia etchellsii cellooligosaccharides alkyl glucosides monoterpenyl glucosides 


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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Preeti Bachhawat
    • 1
  • Saroj Mishra
    • 1
  • Yukti Bhatia
    • 1
  • V. S. Bisaria
    • 1
  1. 1.Department of Biochemical Engineering and BiotechnologyIndian Institute of Technology DelhiHauz-KhasIndia

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