PCR-DGGE分析木瓜酵素自然发酵过程中微生物的多样性

Polymerase Chain Reaction-denaturing Gradient Gel Electrophoresis Analysis of Microbial Diversity of Papaya Enzyme Preparation During Natural Fermentation

为探究木瓜酵素自然发酵过程中的微生物多样性及变化规律,采用聚合酶链式反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)技术分析木瓜酵素自然发酵过程中细菌和酵母的多样性及变化规律。结果表明,木瓜酵素自然发酵过程中主要细菌有植物乳杆菌(Lactobacillus plantarum)、假肠膜明串珠菌(Leuconostoc pseudomesenteroides)、类肠膜魏斯氏菌(Weissella paramesenteroides)及不可培养的丙酸菌(Uncultured Propionibacterium sp.),其中植物乳杆菌为主要优势细菌;主要酵母有:酿酒酵母(Saccharomyces cerevisiae)、假丝酵母(Candida xestobii、Candida intermedia)、毕赤酵母(Pichia guilliermondii、Komagataella phaffii、Pichia punctispora、Pichia galeiformis)、棒孢酵母(Clavispora sp.)及Cyberlindnera fabianii,其中酿酒酵母和毕赤酵母为主要优势酵母;Lac plantarum、Pic galeiformis分别与Uncultured Propionibacterium sp.、Cyb fabianii之间的亲缘性较高,与其他菌之间的亲缘性较小。木瓜酵素自然发酵过程中菌群交替生长,有一定的亲缘性,菌落结构变化较小,分布较为均匀,这为进一步的研究提供理论基础。

In order to investigate the microbial diversity in a papaya enzyme preparation during the natural fermentation process, polymerase chain reaction-denaturing gradient gel electrophoresis（PCR-DGGE） was used to analyze the diversity and the pattern of changes of bacteria and yeast during natural fermentation. The results indicated that the main bacterial species were Lactobacillus plantarum, Leuconostoc pseudomesenteroides, Weissella paramesenteroides, and uncultured Propionibacterium sp.; the dominant bacterium was Lactobacillus plantarum. The main yeast species were Saccharomyces cerevisiae, Candida xestobii, Candida intermedia, Pichia guilliermondii, Komagataella phaffii, Pichia punctispora, Pichia galeiformis, Clavispora sp., and Cyberlindnera fabianii. The dominant yeast species were Saccharomyces cerevisiae and Pichia guilliermondii. Moreover, Lactobacillus plantarum and Pichia galeiformis shared a high affinity with uncultured Propionibacterium sp. and Cyberlindnera fabianii, respectively, and a low affinity with other microorganisms. During natural fermentation, the microflora populations exhibited alternating growth patterns and showed a certain affinity; the colony structure changed slightly with uniform distribution.