Filannino Pasquale, Gobbetti Marco, De Angelis Maria, Di Cagno Raffaella
Department of Soil, Plant and Food Science, University of Bari Aldo Moro, Bari, Italy.
Department of Soil, Plant and Food Science, University of Bari Aldo Moro, Bari, Italy
Appl Environ Microbiol. 2014 Dec;80(24):7574-82. doi: 10.1128/AEM.02413-14. Epub 2014 Sep 26.
The metabolism of hydroxycinnamic acids by strictly heterofermentative lactic acid bacteria (19 strains) was investigated as a potential alternative energy route. Lactobacillus curvatus PE5 was the most tolerant to hydroxycinnamic acids, followed by strains of Weissella spp., Lactobacillus brevis, Lactobacillus fermentum, and Leuconostoc mesenteroides, for which the MIC values were the same. The highest sensitivity was found for Lactobacillus rossiae strains. During growth in MRS broth, lactic acid bacteria reduced caffeic, p-coumaric, and ferulic acids into dihydrocaffeic, phloretic, and dihydroferulic acids, respectively, or decarboxylated hydroxycinnamic acids into the corresponding vinyl derivatives and then reduced the latter compounds to ethyl compounds. Reductase activities mainly emerged, and the activities of selected strains were further investigated in chemically defined basal medium (CDM) under anaerobic conditions. The end products of carbon metabolism were quantified, as were the levels of intracellular ATP and the NAD(+)/NADH ratio. Electron and carbon balances and theoretical ATP/glucose yields were also estimated. When CDM was supplemented with hydroxycinnamic acids, the synthesis of ethanol decreased and the concentration of acetic acid increased. The levels of these metabolites reflected on the alcohol dehydrogenase and acetate kinase activities. Overall, some biochemical traits distinguished the common metabolism of strictly heterofermentative strains: main reductase activity toward hydroxycinnamic acids, a shift from alcohol dehydrogenase to acetate kinase activities, an increase in the NAD(+)/NADH ratio, and the accumulation of supplementary intracellular ATP. Taken together, the above-described metabolic responses suggest that strictly heterofermentative lactic acid bacteria mainly use hydroxycinnamic acids as external acceptors of electrons.
研究了19株严格异型发酵乳酸菌对羟基肉桂酸的代谢情况,将其作为一种潜在的替代能量途径。弯曲乳杆菌PE5对羟基肉桂酸的耐受性最强,其次是魏斯氏菌属菌株、短乳杆菌、发酵乳杆菌和肠系膜明串珠菌,它们的最低抑菌浓度(MIC)值相同。罗氏乳杆菌菌株的敏感性最高。在MRS肉汤中生长期间,乳酸菌分别将咖啡酸、对香豆酸和阿魏酸还原为二氢咖啡酸、根皮酸和二氢阿魏酸,或将羟基肉桂酸脱羧为相应的乙烯基衍生物,然后将后者化合物还原为乙基化合物。主要出现还原酶活性,并在化学限定的基础培养基(CDM)中在厌氧条件下进一步研究了选定菌株的活性。对碳代谢的终产物、细胞内ATP水平和NAD(+)/NADH比率进行了定量。还估计了电子和碳平衡以及理论ATP/葡萄糖产量。当CDM中添加羟基肉桂酸时,乙醇的合成减少,乙酸浓度增加。这些代谢物的水平反映在乙醇脱氢酶和乙酸激酶活性上。总体而言,一些生化特征区分了严格异型发酵菌株的共同代谢:对羟基肉桂酸的主要还原酶活性、从乙醇脱氢酶活性向乙酸激酶活性的转变、NAD(+)/NADH比率的增加以及细胞内补充ATP的积累。综上所述,上述代谢反应表明,严格异型发酵乳酸菌主要将羟基肉桂酸用作外部电子受体。