Kinetics and isotope patterns of ethanol and acetaldehyde emissions from yeast fermentations of glucose and glucose-6,6-d2 using selected ion flow tube mass spectrometry: a case study.
As a prelude to investigations of the emission of metabolites from human cell lines in vitro, we have conducted a study using selected ion flow tube mass spectrometry (SIFT-MS) of the acetaldehyde and ethanol that appear in the headspace above a fermenting yeast/glucose/water mixture in sealed glass bottles at a temperature of 30 degrees C. A fixed quantity of yeast (10 mg) and varying amounts (2, 4, 8 and 16 mg) of both non-deuterated glucose and glucose-6,6-d2 in 5 mL of water were used and the emission of the acetaldehyde and the ethanol were observed as a function of time. The ethanol and acetaldehyde concentrations in the headspace were obtained from the magnitudes of their characteristic ions on the accumulated SIFT mass spectra and, when the deuterated glucose was used, characteristic singly and doubly deuterated ions were obvious. This study indicates, as expected, that ethanol is the major species generated and that acetaldehyde is a relatively minor component of the headspace and a very minor component of the liquid phase. We estimate that about 10(8) ethanol molecules are produced per minute per cell in this yeast fermentation process. The distribution of the non-deuterated and partially deuterated ethanol under these fermentation conditions is observed to be C2H5OH (66 +/- 4)%, C2H4DOH(6 +/- 1)%, C2H3D2OH(28 +/- 4)%, and the analogous distribution for the acetaldehyde is the same, within error. These results indicate that the D atoms in the glucose-6,6-d2 are mostly retained by the 6-C atom, but the appearance of the singly deuterated ethanol and acetaldehyde indicates that some D/H mixing must be occurring in the enzymatic reactions. The results of this study illustrate the potential and power of on-line SIFT-MS analysis in this area of research.
Smith
David
D
Centre for Science and Technology in Medicine, School of Postgraduate Medicine, Keele University, Thornburrow Drive, Hartshill, Stoke-on-Trent ST4 7QB, UK. d.smith@bemp.keele.ac.uk
Wang
Tianshu
T
Spanel
Patrik
P
D000079:Acetaldehyde / Q000032:analysis* / Q000378:metabolism; D003903:Deuterium; D000431:Ethanol / Q000032:analysis* / Q000378:metabolism; D005285:Fermentation; D005947:Glucose / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D013058:Mass Spectrometry; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism*; D014835:Volatilization
D000431:Ethanol; D003903:Deuterium; D000079:Acetaldehyde; D005947:Glucose