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Characterization of the enzymatic polymerization of 2,6-linked fructan by leaf extracts from timothy grass (Phleum pratense) by ANDREW J. CAIRNS; ROBERT NASH; MARIA-ANGELA MACHADO DE CARVALHO; IAN M. SIMS is a Agricultural and Biological Sciences article available to read on EtoBox.

What is Characterization of the enzymatic polymerization of 2,6-linked fructan by leaf extracts from timothy grass (Phleum pratense) about?

A fructan polymerase activity was partially purified and concentrated by sequential acid and salt precipitation from extracts of excised, illuminated leaves of timothy grass (__Phleum pratense__). The polymerase catalysed the __de novo__ synthesis of oligo‐ and polyfructan from sucrose as sole substrate at near‐physiological rates (0.5–0.9 mg g^−1^ fresh mass h^−1^; 0.9–1.5 nkat g^−1^). Rates of __in vitro__ polymerisation were high, at up to 4.1 mg cm^−3^ h^−1^ (7.1 nkat cm^−3^) of total products of degree of polymerization greater than 2 (DP>2). The trisaccharides 1‐kestose and 6‐kestose together with oligosaccharides of up to DP__c.__ 10 were synthesized in under 2 h at 30°C. In longer incubations, ethanol‐precipitable polymers of DP=__c.__ 10–35 (1.6–5.7 kDa) were detected by anion‐exchange chromatography and pulsed amperometry. When this polymeric product was used as a primer and re‐incubated with fresh enzyme and sucrose, abundant polymers of up to DP=50 (8.1 kDa) were formed. The structure of the polymeric enzyme product was compared with native fructan from timothy leaves and with standard inulin, using glycosyl‐linkage analysis followed by identification of partially methy

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Author

ANDREW J. CAIRNS; ROBERT NASH; MARIA-ANGELA MACHADO DE CARVALHO; IAN M. SIMS

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Cambridge University Press (CUP); Wiley (ISSN 0028-646X)

Published

1999

Language

EN

Field

Agricultural and Biological Sciences (Life Sciences)