Characterization of a highly hop-resistant Lactobacillus brevis strain lacking hop transport.
Resistance to hops is a prerequisite for lactic acid bacteria to spoil beer. In this study we analyzed mechanisms of hop resistance of Lactobacillus brevis at the metabolism, membrane physiology, and cell wall composition levels. The beer-spoiling organism L. brevis TMW 1.465 was adapted to high concentrations of hop compounds and compared to a nonadapted strain. Upon adaptation to hops the metabolism changed to minimize ethanol stress. Fructose was used predominantly as a carbon source by the nonadapted strain but served as an electron acceptor upon adaptation to hops, with concomitant formation of acetate instead of ethanol. Furthermore, hop adaptation resulted in higher levels of lipoteichoic acids (LTA) incorporated into the cell wall and altered composition and fluidity of the cytoplasmic membrane. The putative transport protein HitA and enzymes of the arginine deiminase pathway were overexpressed upon hop adaptation. HorA was not expressed, and the transport of hop compounds from the membrane to the extracellular space did not account for increased resistance to hops upon adaptation. Accordingly, hop resistance is a multifactorial dynamic property, which can develop during adaptation. During hop adaptation, arginine catabolism contributes to energy and generation of the proton motive force until a small fraction of the population has established structural improvements. This acquired hop resistance is energy independent and involves an altered cell wall composition. LTA shields the organism from accompanying stresses and provides a reservoir of divalent cations, which are otherwise scarce as a result of their complexation by hop acids. Some of the mechanisms involved in hop resistance overlap with mechanisms of pH resistance and ethanol tolerance and as a result enable beer spoilage by L. brevis.
Behr
Jürgen
J
Technische Mikrobiologie, Technische Universität München, Weihenstephaner Steig 16, 85350 Freising, Germany.
Gänzle
Michael G
MG
Vogel
Rudi F
RF
D000222:Adaptation, Physiological; D001515:Beer; D001692:Biological Transport; D024881:Drug Resistance, Bacterial; D005516:Food Microbiology; D027582:Humulus / Q000737:chemistry*; D052196:Levilactobacillus brevis / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology; D008070:Lipopolysaccharides / Q000378:metabolism; D008560:Membrane Fluidity / Q000187:drug effects; D010936:Plant Extracts / Q000494:pharmacology*; D011312:Pressure; D012333:RNA, Messenger / Q000378:metabolism; D013682:Teichoic Acids / Q000378:metabolism
D008070:Lipopolysaccharides; D010936:Plant Extracts; D012333:RNA, Messenger; D013682:Teichoic Acids; C009900:lipoteichoic acid