de las Rivas Blanca, Rodríguez Héctor, Curiel José Antonio, Landete José María, Muñoz Rosario
Departamento de Microbiología, Instituto de Fermentaciones Industriales, Madrid, Spain.
J Agric Food Chem. 2009 Jan 28;57(2):490-4. doi: 10.1021/jf803016p.
The potential to produce volatile phenols from hydroxycinnamic acids was investigated for lactic acid bacteria (LAB) isolated from Spanish grape must and wine. A PCR assay was developed for the detection of LAB that potentially produce volatile phenols. Synthetic degenerate oligonucleotides for the specific detection of the pdc gene encoding a phenolic acid decarboxylase were designed. The pdc PCR assay amplifies a 321 bp DNA fragment from phenolic acid decarboxylase. The pdc PCR method was applied to 85 strains belonging to the 6 main wine LAB species. Lactobacillus plantarum, Lactobacillus brevis, and Pediococcus pentosaceus strains produce a positive response in the pdc PCR assay, whereas Oenococcus oeni, Lactobacillus hilgardii, and Leuconostoc mesenteroides strains did not produce the expected PCR product. The production of vinyl and ethyl derivatives from hydroxycinnamic acids in culture media was determined by high-performance liquid chromatography. A relationship was found between pdc PCR amplification and volatile phenol production, so that the LAB strains that gave a positive pdc PCR response produce volatile phenols, whereas strains that did not produce a PCR amplicon did not produce volatile phenols. The proposed method could be useful for a preliminary identification of LAB strains able to produce volatile phenols in wine.
对从西班牙葡萄汁和葡萄酒中分离出的乳酸菌（LAB）产生挥发性酚类物质的潜力进行了研究。开发了一种PCR检测方法，用于检测可能产生挥发性酚类物质的乳酸菌。设计了用于特异性检测编码酚酸脱羧酶的pdc基因的合成简并寡核苷酸。pdc PCR检测可从酚酸脱羧酶扩增出一个321 bp的DNA片段。将pdc PCR方法应用于属于6种主要葡萄酒乳酸菌的85个菌株。植物乳杆菌、短乳杆菌和戊糖片球菌菌株在pdc PCR检测中呈阳性反应，而酒类酒球菌、希氏乳杆菌和肠系膜明串珠菌菌株未产生预期的PCR产物。通过高效液相色谱法测定了培养基中羟基肉桂酸的乙烯基和乙基衍生物的产生情况。发现pdc PCR扩增与挥发性酚类物质的产生之间存在关联，因此，pdc PCR反应呈阳性的乳酸菌菌株会产生挥发性酚类物质，而未产生PCR扩增产物的菌株则不会产生挥发性酚类物质。所提出的方法可用于初步鉴定葡萄酒中能够产生挥发性酚类物质的乳酸菌菌株。
