Expression, purification and use of the soluble domain of Lactobacillus paracasei β-fructosidase to optimise production of bioethanol from grass fructans

C. M. Martel, A. G. S. Warrilow, Colin John Jackson, J. G. L. Mullins, R. C. Togawa, J. E. Parker, Stephen Michael Morris, Iain Simon Donnison, Diane Elizabeth Kelly, Steven Lewis Kelly

Research output: Contribution to journalArticlepeer-review

24 Citations (SciVal)

Abstract

Microbial inulinases find application in food, pharmaceutical and biofuel industries. Here, a novel Lactobacillus paracasei β-fructosidase was overexpressed as truncated cytosolic protein (tfosEp) in Escherichia coli. Purified tfosEp was thermostable (10–50 °C) with a pH optimum of 5; it showed highest affinity for bacterial levan (β[2-6] linked fructose) followed by nystose, chicory inulin, 1-kestose (β[2-1] linkages) and sucrose (Km values of 0.5, 15, 15.6, 49 and 398 mM, respectively). Hydrolysis of polyfructose moieties in agriculturally-sourced grass juice (GJ) with tfosEp resulted in the release of >13 mg/ml more bioavailable fructose than was measured in untreated GJ. Bioethanol yields from fermentation experiments with Brewer’s yeast and GJ + tfosEp were >25% higher than those achieved using untreated GJ feedstock (36.5[±4.3] and 28.2[±2.7] mg ethanol/ml, respectively). This constitutes the first specific study of the potential to ferment ethanol from grass juice and the utility of a novel core domain of β-fructosidase from L. paracasei.
Original languageEnglish
Pages (from-to)4395-4402
Number of pages8
JournalBioresource Technology
Volume101
Issue number12
Early online date12 Feb 2010
DOIs
Publication statusPublished - Jun 2010

Keywords

  • Bioethanol
  • Fructan
  • Fructanase
  • Lactobacillus paracasei
  • Perennial ryegrass

Fingerprint

Dive into the research topics of 'Expression, purification and use of the soluble domain of Lactobacillus paracasei β-fructosidase to optimise production of bioethanol from grass fructans'. Together they form a unique fingerprint.

Cite this