2005-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/686004摘要：克弗爾粒由複合乳酸菌及酵母菌叢所組成，為製造克弗爾乳製品之天然菌元，其所包含乳酸菌不僅在克弗爾黏稠質地、酸味與機能特性上扮演重要角色，同時也與克弗爾粒形成與增生有密切關聯，而酵母菌則是製品香氣及風味生成之關鍵微生物。由於不同來源克弗爾粒之菌相組成複雜多變，遂製造出各種地方特色之克弗爾，若能精準掌握克弗爾粒中所有菌種之身份，並純化分離之，將有助於純菌株合成克弗爾粒之研究。本試驗乃針對台灣地區四種克弗爾粒，進行微生物之鑑定，以Harrison’s disc法自各組克弗爾粒中挑出具代表性之菌株，除了以傳統鑑定方式觀察菌株形態、生化特性外，同時利用PCR-DGGE技術及DNA定序等分子生物技術，透過菌種間DNA序列不同，來鑑定菌株身分，結果顯示，四組克弗爾粒中有三組菌相相同，其中之新竹組克弗爾粒鑑定出Lactobacillus kefiranofaciens為優勢菌株 ，其次為Lactobacillus kefiri, Leuconostoc mesenteroides, Lactococcus lactis strain LA35，而酵母菌優勢菌株為乳糖利用型的Kluyveromyces marxianus, 其次為 Saccharomyces sp. KPU，另外不具克弗爾粒形式之台北組，其菌元中包含Lactococcus lactis subsp. cremoris , Leuconostoc mesenteroides subsp. mesenteroides兩株乳酸菌及Kluyveromyces marxianus, Saccharomyces unisporus, 和 Pichia fermentans strain VTT C-04567三株酵母菌。 <br> Abstract: Kefir grains, consisting of a variety of microorganisms, are the natural starter for kefir production. Specific uniqueness of kefir is attributed to the presence of a complex microbial population. Among those microorganisms, both lactic acid bacteria and yeasts play important roles not only in health-promoting and functional properties but also in grain formation and flavor production. The kefir - specific yeasts play a key role in the formation of flavour and aroma. Characterization of microorganisms at the species level will be helpful in the selection of starter cultures to standardize the microorganisms in kefir products. Thus, our objective was to identify the complex microbial compositions present in four different Taiwanese kefir starter. Lactic acid bacteria and yeast in kefir starter were purified by the Harrison disc method and identified by traditional and molecular characteristic. API analysis , alive strains were identified. Molecular characteristic was using the denaturing gradient gel electrophoresis (DGGE). In addition, sequence analysis of PCR products was performed to confirm the DGGE results. Results indicated that Lactobacillus kefiranofaciens, Lactobacillus kefiri, Leuconostoc mesenteroides, Lactococcus lactis strain LA35, Kluyveromyces marxianus, and Saccharomyces sp. KPU were identified from Hsin-Cho grains. Whereas, Lactococcus lactis subsp. cremoris , Kluyveromyces marxianus, Saccharomyces unisporus, and Pichia fermentans strain VTT C-04567 were found in Taipei starter.克弗爾粒變性梯度膠體電泳kefir grainsDGGE