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A Natural Sterilant (Reuterin) Fermented from Glycerol Using Lactobacillus Reuteri: Fermentation Conditions

以Lactobacillus reuteri乳酸菌發酵Glycerol生成天然滅菌劑Reuterin及其發酵最適化條件的探討

摘要


Reuterin是一種新發現的滅菌物質,對於gram-positive菌、gram-negative菌、酵母菌、黴菌及原蟲等皆有抑制作用。本研究分別探討了reuterin的生成及分離純化和reuterin生成的最適化發酵條件,包括了:發酵使用的菌量、發酵時間、發酵溫度和甘油起始濃度等。另外,我們也測試了reuterin對臨床上常感染的致病菌的最低抑菌濃度和最低滅菌濃度。實驗的對照組為臨床上生物組織材料最常使用的滅菌劑glutaraldehyde。實驗結果顯示,由高效能液相層析儀的分離純化可以得到高純度的reuterin,而reuterin生成的最適化發酵條件為配製含有Lactobacillus reutei 100 mg菌重/ml的300 mM甘油溶液,在37℃下反應1小時。另外,滅菌的結果顯示,reuterin所需的最低滅菌濃度要比glutaraldehyde低約2〜3倍。

並列摘要


Cultures of Lactobacillus reuteri have been shown to accumulate a large quantity of reuterin during anaerobic growth in the presence of glycerol. It is known that reuterin has broad-spectrum antibacterial, antimycotic, and antiprotozoal activities. Therefore, it may be feasible using reuterin as a sterilant for the sterilization of biological tissues. The objective of the study was to investigate the optimal conditions (cell mass, fermentation time, fermentation temperature, and initial glycerol concentration) to produce reuterin fermented form glycerol using Lactobacillus reuteri. Additionally, the antimicrobial activity of reuterin was examined. Glutaraldehyde, the most commonly used sterilant for the sterilization of biological materials, was employed as a control. In the study, reuterin was successfully isolated from the fermented mixtures by a high-performance liquid chromatographer. The results revealed that the optimal fermentation condition was: 100 mg Lactobacillus reuteri cells/ml suspended in 300 mM glycerol in distilled water incubated under anaerobic condition at 37ºC for 1 h. Additionally, as per the minimal inhibitory and bactericidal concentrations, the antimicrobial activity of reuterin is significantly superior to glutaraldehyde.

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