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人源发酵乳杆菌SD11产生的细菌素Fermencin SD11的纯化、特性及最佳条件

Purification, Characterization, and Optimum Conditions of Fermencin SD11, a Bacteriocin Produced by Human Orally Lactobacillus fermentum SD11.

作者信息

Wannun Phirawat, Piwat Supatcharin, Teanpaisan Rawee

机构信息

Common Oral Diseases and Epidemiology Research Center and the Department of Stomatology, Faculty of Dentistry, Prince of Songkla University, Hat-Yai, 90112, Thailand.

Common Oral Diseases and Epidemiology Research Center and the Department of Preventive Dentistry, Faculty of Dentistry, Prince of Songkla University, Hat-Yai, 90112, Thailand.

出版信息

Appl Biochem Biotechnol. 2016 Jun;179(4):572-82. doi: 10.1007/s12010-016-2014-y. Epub 2016 Feb 18.

Abstract

Fermencin SD11, a bacteriocin produced by human orally Lactobacillus fermentum SD11, was purified, characterized, and optimized in conditions for bacterial growth and bacteriocin production. Fermencin SD11 was purified using three steps of ammonium sulfate precipitation, gel filtration chromatography, and reverse-phase high-performance liquid chromatography. The molecular weight was found to be 33,000 Da using SDS-PAGE and confirmed as 33,593.4 Da by liquid chromatography-mass spectrometry. Fermencin SD11 exhibited activity against a wide range of oral pathogens including cariogenic and periodontogenic pathogens and Candida. The active activity was stable between 60 - 80 °C in a pH range of 3.0 to 7.0. It was sensitive to proteolytic enzymes (proteinase K and trypsin), but it was not affected by α-amylase, catalase, lysozyme, and saliva. The optimum conditions for growth and bacteriocin production of L. fermentum SD11 were cultured at acidic with pH of 5.0-6.0 at 37 or 40 °C under aerobic or anaerobic conditions for 12 h. It is promising that L. fermentum SD11 and its bacteriocin may be an alternative approach for promoting oral health or prevention of oral diseases, e.g., dental caries and periodontitis, which would require further clinical trials.

摘要

Fermencin SD11是由人源口服发酵乳杆菌SD11产生的一种细菌素,对其进行了纯化、表征,并对细菌生长和细菌素产生的条件进行了优化。Fermencin SD11通过硫酸铵沉淀、凝胶过滤色谱和反相高效液相色谱三步进行纯化。使用SDS-PAGE测得其分子量为33,000 Da,液相色谱-质谱法确认为33,593.4 Da。Fermencin SD11对多种口腔病原体具有活性,包括致龋和致牙周病的病原体以及念珠菌。其活性在60 - 80°C、pH值为3.0至7.0的范围内稳定。它对蛋白水解酶(蛋白酶K和胰蛋白酶)敏感,但不受α-淀粉酶、过氧化氢酶、溶菌酶和唾液的影响。发酵乳杆菌SD11生长和产生细菌素的最佳条件是在37或40°C、pH值为5.0 - 6.0的酸性条件下,需氧或厌氧培养12小时。发酵乳杆菌SD11及其细菌素有望成为促进口腔健康或预防口腔疾病(如龋齿和牙周炎)的替代方法,这需要进一步的临床试验。

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