Molecular species of phosphatidylethanolamine from continuous cultures of Saccharomyces pastorianus syn. carlsbergensis strains.
Saccharomyces pastorianus syn. carlsbergensis strain 34/70 is well known to be the most used strain for lager beer production. The difference between this strain and very closely related strain 34/78 is the latter's greater flocculating character. This single physiological trait can cause technical difficulties in beer production. The aim of this study was to determine whether lipid analysis by a combination of thin layer chromatography (TLC) with electrospray ionization mass spectrometry (ESI-MS) could be used as a strain-typing technique in order to distinguish S. pastorianus syn. carlsbergensis strain 34/70 from strain 34/78. Both strains (34/70 and 34/78) were harvested after continuous culture under standard conditions. Polar lipids were then extracted from lyophilized cultures and analysed by TLC in order to separate phospholipid families. Phosphatidylethanolamine (PE) was extracted and investigated using ESI-MS, to gain further information on individual molecular species. Using TLC analysis, lipids were separated corresponding to standards for PE, phosphatidylcholine (PC), phosphatidylglycerol (PG), cardiolipin (CL), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidic acid (PA) and sphingomyelin (SM). ESI-MS of the PE band, separated by TLC, showed that electrospray mass spectra were highly reproducible for repeat cultures. Novel findings were that both brewing strains displayed major phospholipid peaks with m/z 714, PE (34 : 2) m/z 742, PE (36 : 2) and m/z 758, PE (37 : 1). However, strain 34/78 had additional peaks of m/z 700, PE (33 : 2) and m/z 728, PE (35 : 2). Strain 34/70 had an extra peak with m/z 686 PE (32 : 2). We conclude that combined TLC/ESI-MS can distinguish between S. pastorianus syn. carlsbergensis 34/70 and 34/78 and may be a useful typing technique for differentiation of closely related yeast strains. This novel approach may aid quality assurance and could be suitable for yeast collections and larger industrial companies.
Tosch
W
W
Faculty of Life Sciences, University of Manchester, Manchester, UK.
Lanthaler
K
K
Boote
V
V
Stretz
D
D
Robson
G D
GD
Geiger
E
E
Drucker
D B
DB
D002308:Cardiolipins / Q000737:chemistry / Q000302:isolation & purification; D002855:Chromatography, Thin Layer; D010712:Phosphatidic Acids / Q000737:chemistry / Q000302:isolation & purification; D010713:Phosphatidylcholines / Q000737:chemistry / Q000302:isolation & purification; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000737:chemistry / Q000302:isolation & purification; D010716:Phosphatidylinositols / Q000737:chemistry / Q000302:isolation & purification; D010718:Phosphatidylserines / Q000737:chemistry / Q000302:isolation & purification; D012440:Saccharomyces / Q000145:classification* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D013109:Sphingomyelins / Q000737:chemistry / Q000302:isolation & purification
D002308:Cardiolipins; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D010716:Phosphatidylinositols; D010718:Phosphatidylserines; D013109:Sphingomyelins; C483858:phosphatidylethanolamine