Purification and characterization of a novel β-carotene-9',10'-oxygenase from Saccharomyces cerevisiae ULI3

Biotechnol Lett. 2015 Oct;37(10):1993-8. doi: 10.1007/s10529-015-1872-7. Epub 2015 May 31.

Abstract

Objectives: A novel β-carotene-9,10'-oxygenase (ScBCO2) has been characterized from Saccharomyces cerevisiae ULI3 to convert β-carotene to β-apo-10'-carotenal, which is a precursor of the plant hormone strigolactone.

Results: The ScBCO2 enzyme was purified to homogeneity by ammonium sulfate precipitation, Q sepharose and Superdex-200 chromatography. The molecular mass of the enzyme was ~50 kDa by SDS-PAGE. The purified ScBCO2 enzyme displayed optimal activity at 45 °C and pH 8. Tween 20 (1%, w/v), Trition X-100 (1%, w/v), Mg(2+) (5 mM), Zn(2+) (5 mM), Cu(2+) (5 mM), Ca(2+) (5 mM) or DTT (5 mM) increased in the activity by 3, 7, 14, 17, 23, 26 and 27%, respectively. ScBCO2 only exhibited cleavage activity towards carotenoid substrates containing two β-ionone rings and its catalytic efficiency (kcat/Km) followed the order β-carotene > α-carotene > lutein.

Conclusion: ScBCO2 could be used as a potential candidate for the enzymatic biotransformation of β-carotene to β-apo-10'-carotenal in biotechnological applications.

Keywords: Saccharomyces cerevisiae; Strigolactone; β-Apo-10′-carotenal; β-Carotene; β-Carotene-9,10′-oxygenase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carotenoids / metabolism*
  • Chemical Precipitation
  • Chromatography, Liquid
  • Dioxygenases / chemistry
  • Dioxygenases / isolation & purification*
  • Dioxygenases / metabolism*
  • Enzyme Activators / analysis
  • Humans
  • Hydrogen-Ion Concentration
  • Infant, Newborn
  • Molecular Weight
  • Oxidation-Reduction
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / isolation & purification*
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Substrate Specificity
  • Temperature
  • beta Carotene / metabolism*

Substances

  • Enzyme Activators
  • Saccharomyces cerevisiae Proteins
  • beta-apo-10'-carotenal
  • beta Carotene
  • Carotenoids
  • BCO2 protein, S cerevisiae
  • Dioxygenases