Bae S, Fleet G H, Heard G M
Food Science and Technology, School of Chemical Engineering and Industrial Chemistry, The University of New South Wales, Sydney, New South Wales, Australia.
J Appl Microbiol. 2006 Apr;100(4):712-27. doi: 10.1111/j.1365-2672.2006.02890.x.
The detection and isolation of lactic acid bacteria by enrichment methods from wine grapes cultivated in vineyards located in New South Wales, Australia.
Enrichment cultures in de Man, Rogosa and Sharpe (MRS) broth, MRS + ethanol (5%), MRS broth supplemented with 15% (v/v) tomato juice (MRST), pH 5.5 and 3.5 and autoenrichment in grape juice homogenate were used to detect lactic acid bacteria on wine grapes. Bacteria were isolated from enrichment cultures by plating onto MRS and MRST agar and identified by 16S rDNA sequence analysis and phenotypical methods. A molecular method, PCR-denaturing gradient gel electrophoresis (DGGE) was also used to examine the bacteria that developed in enrichment cultures. Species of Lactobacillus, Enterococcus, Lactococcus and Weissella were detected in enrichments by plating and PCR-DGGE. Other bacteria (Sporolactobacillus, Asaia, Bacillus ssp.) were also found in some enrichment cultures. The principal malolactic bacterium, Oenococcus oeni, was not isolated.
The incidence and populations of lactic acid bacteria on wine grapes were very low. Damaged grape berries showed a greater presence of these bacteria than undamaged berries. The diversity of bacterial species isolated from the grapes was greater than those previously reported and represented both lactic acid bacteria and nonlactic acid bacteria. Some of these bacteria (i.e. Lactobacillus lindneri, Lactobacillus kunkeei) could be detrimental to wine production. Oenococcus oeni was not found on grapes, but its recovery could be obscured by overgrowth from other species.
Lactic acid bacteria are significant in wine production because they conduct the malolactic fermentation and cause stuck or sluggish alcoholic fermentation and wine spoilage. This study investigates wine grapes as a potential source of these bacteria.
采用富集培养法从澳大利亚新南威尔士州葡萄园种植的酿酒葡萄中检测和分离乳酸菌。
使用德氏、罗格斯和夏普(MRS)肉汤、MRS + 乙醇(5%)、添加15%(v/v)番茄汁的MRS肉汤(MRST)、pH 5.5和3.5的培养基以及葡萄汁匀浆自动富集培养法来检测酿酒葡萄上的乳酸菌。通过将富集培养物接种到MRS和MRST琼脂平板上进行细菌分离,并采用16S rDNA序列分析和表型方法进行鉴定。还使用了一种分子方法,即聚合酶链反应 - 变性梯度凝胶电泳(PCR - DGGE)来检测富集培养物中生长的细菌。通过平板培养和PCR - DGGE在富集培养物中检测到了乳酸杆菌属、肠球菌属、乳球菌属和魏斯氏菌属的菌种。在一些富集培养物中还发现了其他细菌(芽孢乳酸杆菌属(Sporolactobacillus)、亚细亚菌属(Asaia)、芽孢杆菌属(Bacillus ssp.))。未分离到主要的苹果酸 - 乳酸细菌——酒酒球菌(Oenococcus oeni)。
酿酒葡萄上乳酸菌的发生率和数量非常低。受损的葡萄浆果中这些细菌的存在量比未受损的浆果更多。从葡萄中分离出的细菌种类比之前报道的更多,包括乳酸菌和非乳酸菌。其中一些细菌(如林氏乳酸杆菌(Lactobacillus lindneri)、库氏乳酸杆菌(Lactobacillus kunkeei))可能对葡萄酒生产有害。在葡萄上未发现酒酒球菌,但其他菌种的过度生长可能会掩盖其检出。
乳酸菌在葡萄酒生产中很重要,因为它们进行苹果酸 - 乳酸发酵,并导致酒精发酵停滞或迟缓以及葡萄酒变质。本研究调查了酿酒葡萄作为这些细菌潜在来源的情况。