Qi Zhengliang, Yang Hailin, Xia Xiaole, Wang Wu, Leng Yunwei, Yu Xiaobin, Quan Wu
Wei Sheng Wu Xue Bao. 2014 Mar 4;54(3):299-308.
The aim of the study is to propose a dynamic acetic acid resistance mechanism through analysis on response of cellular morphology, physiology and metabolism of A. pasteurianus CICIM B7003 during vinegar fermentation.
Vinegar fermentation was carried out in a Frings 9 L acetator by strain B7003 and cultures were sampled at different cellular growth phases. Simultaneously, percentage of capsular polysaccharide versus dry cells weight, ratio of unsaturated fatty acids to saturated fatty acids, transcription of acetic acid resistance genes, activity of alcohol respiratory chain enzymes and ATPase were detected for these samples to assay the responses of bacterial morphology, physiology and metabolism.
When acetic acid was existed, no obvious capsular polysaccharide was secreted by cells. As vinegar fermentation proceeding, percentage of capsular polysaccharide versus dry cells weight was reduced from 2.5% to 0.89%. Ratio of unsaturated fatty acids to saturated fatty acids was increased obviously which can improve membrane fluidity. Also transcription level of acetic acid resistance genes was promoted. Interestingly, activity of alcohol respiratory chain and ATPase was not inhibited but promoted obviously with acetic acid accumulation which could provide enough energy for acetic acid resistance mechanism.
On the basis of the results obtained from the experiment, A. pasteurianus CICIM B7003 relies mainly on the cooperation of changes of extracellular capsular polysaccharide and membrane fatty acids, activation of acid resistance genes transcription, enhancement of activity of alcohol respiratory chain and rapid energy production to tolerate acidic environment.
本研究旨在通过分析巴氏醋酸杆菌CICIM B7003在醋酸发酵过程中细胞形态、生理和代谢的响应,提出一种动态的耐醋酸机制。
利用菌株B7003在Frings 9 L醋酸发酵罐中进行醋酸发酵,并在不同细胞生长阶段对培养物进行取样。同时,检测这些样品中荚膜多糖占干细胞重量的百分比、不饱和脂肪酸与饱和脂肪酸的比例、耐醋酸基因的转录、酒精呼吸链酶和ATP酶的活性,以分析细菌形态、生理和代谢的响应。
当存在醋酸时,细胞未分泌明显的荚膜多糖。随着醋酸发酵的进行,荚膜多糖占干细胞重量的百分比从2.5%降至0.89%。不饱和脂肪酸与饱和脂肪酸的比例明显增加,这可以提高膜的流动性。此外,耐醋酸基因的转录水平也得到促进。有趣的是,随着醋酸积累,酒精呼吸链和ATP酶的活性未受到抑制,反而明显增强,这可为耐醋酸机制提供足够的能量。
基于实验结果,巴氏醋酸杆菌CICIM B7003主要依靠细胞外荚膜多糖和膜脂肪酸变化的协同作用、耐酸基因转录的激活、酒精呼吸链活性的增强以及快速的能量产生来耐受酸性环境。