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在铜绿假单胞菌 PAO1 的分批培养过程中,N-丁酰高丝氨酸内酯浓度随时间变化的动力学模型。

Kinetic modeling of the time course of N-butyryl-homoserine lactone concentration during batch cultivations of Pseudomonas aeruginosa PAO1.

机构信息

Institute of Process Engineering in Life Sciences, Section II: Technical Biology, Karlsruhe Institute of Technology (KIT), Engler-Bunte-Ring 1, 76131, Karlsruhe, Germany.

出版信息

Appl Microbiol Biotechnol. 2013 Sep;97(17):7607-16. doi: 10.1007/s00253-013-5024-5. Epub 2013 Jun 19.

Abstract

Quorum sensing affects the regulation of more than 300 genes in Pseudomonas aeruginosa, influencing growth, biofilm formation, and the biosynthesis of several products. The quorum sensing regulation mechanisms are mostly described in a qualitative character. Particularly, in this study, the kinetics of N-butyryl-homoserine lactone (C4-HSL) and rhamnolipid formation in P. aeruginosa PAO1 were of interest. In this system, the expression of the rhamnolipid biosynthesis genes rhlAB is directly coupled to the C4-HSL concentration via the rhl system. Batch cultivations in a bioreactor with sunflower oil have been used for these investigations. 3-oxo-dodecanoyl-homoserine lactone (3o-C12-HSL) displayed a lipophilic character and accumulated in the hydrophobic phase. Degradation of C4-HSL has been found to occur in the aqueous supernatant of the culture by yet unknown extracellular mechanisms, and production was found to be proportional to biomass concentration rather than by autoinduction mechanisms. Rhamnolipid production rates, as determined experimentally, were shown to correlate linearly with the concentration of autoinducer C4-HSL. These findings were used to derive a simple model, wherein a putative, extracellular protein with C4-HSL degrading activity was assumed (putative C4-HSL acylase). The model is based on data for catalytic efficiency of HSL-acylases extracted from literature (k cat/K m), experimentally determined basal C4-HSL production rates (q C4 - HSL (basal)), and two fitted parameters which describe the formation of the putative acylase and is therefore comparatively simple.

摘要

群体感应会影响铜绿假单胞菌中超过 300 个基因的调控,影响其生长、生物膜形成和多种产物的生物合成。群体感应的调控机制主要以定性特征来描述。特别是在这项研究中,我们对铜绿假单胞菌 PAO1 中 N-丁酰高丝氨酸内酯 (C4-HSL) 和鼠李糖脂形成的动力学很感兴趣。在这个系统中,鼠李糖脂生物合成基因 rhlAB 的表达通过 rhl 系统与 C4-HSL 浓度直接偶联。我们在带有向日葵油的生物反应器中进行了分批培养,以进行这些研究。3-氧代-十二烷酰高丝氨酸内酯 (3o-C12-HSL) 表现出亲脂性,积累在疏水区。通过未知的细胞外机制,在培养物的水相上清液中发现 C4-HSL 的降解,并且发现产量与生物量浓度成正比,而不是通过自动诱导机制。通过实验确定的鼠李糖脂生产速率与自动诱导物 C4-HSL 的浓度呈线性相关。这些发现被用来推导一个简单的模型,其中假设存在一种具有 C4-HSL 降解活性的假定细胞外蛋白(假定的 C4-HSL 酰基酶)。该模型基于从文献中提取的 HSL 酰基酶的催化效率 (k cat/K m)、实验确定的基础 C4-HSL 产生速率 (q C4-HSL (basal)) 和两个拟合参数,这些参数描述了假定的酰基酶的形成,因此相对简单。

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