Lucas Patrick M, Claisse Olivier, Lonvaud-Funel Aline
UMR 1219, INRA-Université Victor Segalen Bordeaux 2, Talence, France.
Appl Environ Microbiol. 2008 Feb;74(3):811-7. doi: 10.1128/AEM.01496-07. Epub 2007 Dec 7.
Lactic acid bacteria contribute to wine transformation during malolactic fermentation. They generally improve the sensorial properties of wine, but some strains produce histamine, a toxic substance that causes health issues. Histamine-producing strains belong to species of the genera Oenococcus, Lactobacillus, and Pediococcus. All carry an hdcA gene coding for a histidine decarboxylase that converts histidine into histamine. For this study, a method based on quantitative PCR and targeting hdcA was developed to enumerate these bacteria in wine. This method was efficient for determining populations of 1 to 10(7) CFU per ml. An analysis of 264 samples collected from 116 wineries of the same region during malolactic fermentation revealed that these bacteria were present in almost all wines and at important levels, exceeding 10(3) CFU per ml in 70% of the samples. Histamine occurred at an often important level in wines containing populations of the above-mentioned bacteria. Fifty-four colonies of histamine producers isolated from four wines were characterized at the genetic level. All were strains of Oenococcus oeni that grouped into eight strain types by randomly amplified polymorphic DNA analysis. Some strains were isolated from wines collected in distant wineries. Moreover, hdcA was detected on a large and possibly unstable plasmid in these strains of O. oeni. Taken together, the results suggest that the risk of histamine production exists in almost all wines and is important when the population of histamine-producing bacteria exceeds 10(3) per ml. Strains of O. oeni producing histamine are frequent in wine during malolactic fermentation, but they may lose this capacity during subcultures in the laboratory.
乳酸菌在苹果酸 - 乳酸发酵过程中对葡萄酒的转化有作用。它们通常会改善葡萄酒的感官特性,但有些菌株会产生组胺,一种会引发健康问题的有毒物质。产生组胺的菌株属于酒球菌属、乳杆菌属和片球菌属。所有这些菌株都携带一个编码组氨酸脱羧酶的hdcA基因,该酶可将组氨酸转化为组胺。在本研究中,开发了一种基于定量PCR并靶向hdcA的方法来对葡萄酒中的这些细菌进行计数。该方法对于测定每毫升1至10⁷CFU的菌量很有效。对在同一地区116个酒庄的苹果酸 - 乳酸发酵过程中收集的264个样品进行分析发现,这些细菌几乎存在于所有葡萄酒中,且含量较高,70%的样品中每毫升超过10³CFU。在含有上述细菌群体的葡萄酒中,组胺含量通常较高。从四种葡萄酒中分离出的54个组胺产生菌菌落进行了基因水平的鉴定。所有菌株均为酒酒球菌,通过随机扩增多态性DNA分析可分为八种菌株类型。有些菌株是从相距较远的酒庄收集的葡萄酒中分离出来的。此外,在这些酒酒球菌菌株的一个大的且可能不稳定的质粒上检测到了hdcA。综合来看,结果表明几乎所有葡萄酒都存在产生组胺的风险,当产生组胺的细菌数量超过每毫升10³时风险较大。在苹果酸 - 乳酸发酵过程中,酒酒球菌产生组胺的菌株在葡萄酒中很常见,但它们在实验室传代培养过程中可能会失去这种能力。