The biosynthesis of erythroascorbate in saccharomyces cerevisiae and its role as an antioxidant

C.M. Spickett, N. Smirnoff, A.R. Pitt

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

This study investigated the ability of the yeast Saccharomyces cerevisiae to synthesize ascorbate and its 5-carbon analogue erythroascorbate from a variety of precursors, and their importance as antioxidants in this organism. Studies of ascorbate and analogues in micro-organisms have been reported previously, but their function as antioxidants have been largely ignored. Ascorbate and erythroascorbate concentrations in yeast extracts were measured spectrophotometrically, and their levels and identity were checked using liquid chromatography-electrospray mass spectrometry. The yeast was readily able to synthesize ascorbate from galactono-1,4-lactone or erythroascorbate from -arabinose and -arabino-1,4-lactone, whereas -gulono-1,4-lactone was a much poorer substrate for ascorbate biosynthesis. In untreated cells, the concentration of ascorbate-like compounds was below the level of detection of the methods of analysis used in this study (approximately 0.1 mM). Intracellular ascorbate and erythroascorbate were oxidized at high concentrations of tert-butylhydroperoxide, but not hydrogen peroxide. Their synthesis was not increased in response to low levels of stress, however, and preloading with erythroascorbate did not protect glutathione levels during oxidative stress. This study provides new information on the metabolism of ascorbate and erythroascorbate in S. cerevisiae, and suggests that erythroascorbate is of limited importance as an antioxidant in S. cerevisiae.
Original languageEnglish
Pages (from-to)183-192
Number of pages9
JournalFree Radical Biology and Medicine
Volume28
Issue number2
DOIs
Publication statusPublished - 15 Jan 2000

Fingerprint

Biosynthesis
Yeast
Saccharomyces cerevisiae
Antioxidants
Yeasts
Lactones
tert-Butylhydroperoxide
Arabinose
Liquid Chromatography
Hydrogen Peroxide
Glutathione
Mass Spectrometry
Oxidative Stress
Carbon
Oxidative stress
Liquid chromatography
Metabolism
Mass spectrometry
Substrates

Keywords

  • antioxidant
  • ascorbate
  • electrospray mass spectrometry
  • erythroascorbate
  • glutathione
  • oxidative stress
  • saccharomyces cerevisiae
  • yeast
  • free radicals

Cite this

Spickett, C.M. ; Smirnoff, N. ; Pitt, A.R. / The biosynthesis of erythroascorbate in saccharomyces cerevisiae and its role as an antioxidant. In: Free Radical Biology and Medicine. 2000 ; Vol. 28, No. 2. pp. 183-192.
@article{df8fc93a921b4edcb723f5381253a5bd,
title = "The biosynthesis of erythroascorbate in saccharomyces cerevisiae and its role as an antioxidant",
abstract = "This study investigated the ability of the yeast Saccharomyces cerevisiae to synthesize ascorbate and its 5-carbon analogue erythroascorbate from a variety of precursors, and their importance as antioxidants in this organism. Studies of ascorbate and analogues in micro-organisms have been reported previously, but their function as antioxidants have been largely ignored. Ascorbate and erythroascorbate concentrations in yeast extracts were measured spectrophotometrically, and their levels and identity were checked using liquid chromatography-electrospray mass spectrometry. The yeast was readily able to synthesize ascorbate from galactono-1,4-lactone or erythroascorbate from -arabinose and -arabino-1,4-lactone, whereas -gulono-1,4-lactone was a much poorer substrate for ascorbate biosynthesis. In untreated cells, the concentration of ascorbate-like compounds was below the level of detection of the methods of analysis used in this study (approximately 0.1 mM). Intracellular ascorbate and erythroascorbate were oxidized at high concentrations of tert-butylhydroperoxide, but not hydrogen peroxide. Their synthesis was not increased in response to low levels of stress, however, and preloading with erythroascorbate did not protect glutathione levels during oxidative stress. This study provides new information on the metabolism of ascorbate and erythroascorbate in S. cerevisiae, and suggests that erythroascorbate is of limited importance as an antioxidant in S. cerevisiae.",
keywords = "antioxidant, ascorbate, electrospray mass spectrometry, erythroascorbate, glutathione, oxidative stress, saccharomyces cerevisiae, yeast, free radicals",
author = "C.M. Spickett and N. Smirnoff and A.R. Pitt",
year = "2000",
month = "1",
day = "15",
doi = "10.1016/S0891-5849(99)00214-2",
language = "English",
volume = "28",
pages = "183--192",
journal = "Free Radical Biology and Medicine",
issn = "0891-5849",
number = "2",

}

The biosynthesis of erythroascorbate in saccharomyces cerevisiae and its role as an antioxidant. / Spickett, C.M.; Smirnoff, N.; Pitt, A.R.

In: Free Radical Biology and Medicine, Vol. 28, No. 2, 15.01.2000, p. 183-192.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The biosynthesis of erythroascorbate in saccharomyces cerevisiae and its role as an antioxidant

AU - Spickett, C.M.

AU - Smirnoff, N.

AU - Pitt, A.R.

PY - 2000/1/15

Y1 - 2000/1/15

N2 - This study investigated the ability of the yeast Saccharomyces cerevisiae to synthesize ascorbate and its 5-carbon analogue erythroascorbate from a variety of precursors, and their importance as antioxidants in this organism. Studies of ascorbate and analogues in micro-organisms have been reported previously, but their function as antioxidants have been largely ignored. Ascorbate and erythroascorbate concentrations in yeast extracts were measured spectrophotometrically, and their levels and identity were checked using liquid chromatography-electrospray mass spectrometry. The yeast was readily able to synthesize ascorbate from galactono-1,4-lactone or erythroascorbate from -arabinose and -arabino-1,4-lactone, whereas -gulono-1,4-lactone was a much poorer substrate for ascorbate biosynthesis. In untreated cells, the concentration of ascorbate-like compounds was below the level of detection of the methods of analysis used in this study (approximately 0.1 mM). Intracellular ascorbate and erythroascorbate were oxidized at high concentrations of tert-butylhydroperoxide, but not hydrogen peroxide. Their synthesis was not increased in response to low levels of stress, however, and preloading with erythroascorbate did not protect glutathione levels during oxidative stress. This study provides new information on the metabolism of ascorbate and erythroascorbate in S. cerevisiae, and suggests that erythroascorbate is of limited importance as an antioxidant in S. cerevisiae.

AB - This study investigated the ability of the yeast Saccharomyces cerevisiae to synthesize ascorbate and its 5-carbon analogue erythroascorbate from a variety of precursors, and their importance as antioxidants in this organism. Studies of ascorbate and analogues in micro-organisms have been reported previously, but their function as antioxidants have been largely ignored. Ascorbate and erythroascorbate concentrations in yeast extracts were measured spectrophotometrically, and their levels and identity were checked using liquid chromatography-electrospray mass spectrometry. The yeast was readily able to synthesize ascorbate from galactono-1,4-lactone or erythroascorbate from -arabinose and -arabino-1,4-lactone, whereas -gulono-1,4-lactone was a much poorer substrate for ascorbate biosynthesis. In untreated cells, the concentration of ascorbate-like compounds was below the level of detection of the methods of analysis used in this study (approximately 0.1 mM). Intracellular ascorbate and erythroascorbate were oxidized at high concentrations of tert-butylhydroperoxide, but not hydrogen peroxide. Their synthesis was not increased in response to low levels of stress, however, and preloading with erythroascorbate did not protect glutathione levels during oxidative stress. This study provides new information on the metabolism of ascorbate and erythroascorbate in S. cerevisiae, and suggests that erythroascorbate is of limited importance as an antioxidant in S. cerevisiae.

KW - antioxidant

KW - ascorbate

KW - electrospray mass spectrometry

KW - erythroascorbate

KW - glutathione

KW - oxidative stress

KW - saccharomyces cerevisiae

KW - yeast

KW - free radicals

UR - http://dx.doi.org/10.1016/S0891-5849(99)00214-2

U2 - 10.1016/S0891-5849(99)00214-2

DO - 10.1016/S0891-5849(99)00214-2

M3 - Article

VL - 28

SP - 183

EP - 192

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

SN - 0891-5849

IS - 2

ER -