Coenzyme A-transferase-independent butyrate re-assimilation in Clostridium acetobutylicum - evidence from a mathematical model

The hetero-dimeric CoA-transferase CtfA/B is believed to be crucial for the metabolic transition from acidogenesis to solventogenesis in Clostridium acetobutylicum as part of the industrial-relevant acetone-butanol-ethanol (ABE) fermentation. Here, the enzyme is assumed to mediate re-assimilation of acetate and butyrate during a pH-induced metabolic shift and to faciliate the first step of acetone formation from acetoacetyl-CoA. However, recent investigations using phosphate-limited continuous cultures have questioned this common dogma. To address the emerging experimental discrepancies, we investigated the mutant strain Cac-ctfA398s::CT using chemostat cultures. As a consequence of this mutation, the cells are unable to express functional ctfA and are thus lacking CoA-transferase activity. A mathematical model of the pH-induced metabolic shift, which was recently developed for the wild type, is used to analyse the observed behaviour of the mutant strain with a focus on re-assimilation activities for the two produced acids. Our theoretical analysis reveals that the ctfA mutant still re-assimilates butyrate, but not acetate. Based upon this finding, we conclude that C. acetobutylicum possesses a CoA-tranferase-independent butyrate uptake mechanism that is activated by decreasing pH levels. Furthermore, we observe that butanol formation is not inhibited under our experimental conditions, as suggested by previous batch culture experiments. In concordance with recent batch experiments, acetone formation is abolished in chemostat cultures using the ctfa mutant

Mathematical modelling of clostridial acetone-butanol-ethanol fermentation

Clostridial acetone-butanol-ethanol (ABE) fermentation features a remarkable shift in the cellular metabolic activity from acid formation, acidogenesis, to the production of industrial-relevant solvents, solventogensis. In recent decades, mathematical models have been employed to elucidate the complex interlinked regulation and conditions that determine these two distinct metabolic states and govern the transition between them. In this review, we discuss these models with a focus on the mechanisms controlling intra- and extracellular changes between acidogenesis and solventogenesis. In particular, we critically evaluate underlying model assumptions and predictions in the light of current experimental knowledge. Towards this end, we briefly introduce key ideas and assumptions applied in the discussed modelling approaches, but waive a comprehensive mathematical presentation. We distinguish between structural and dynamical models, which will be discussed in their chronological order to illustrate how new biological information facilitates the ‘evolution’ of mathematical models. Mathematical models and their analysis have significantly contributed to our knowledge of ABE fermentation and the underlying regulatory network which spans all levels of biological organization. However, the ties between the different levels of cellular regulation are not well understood. Furthermore, contradictory experimental and theoretical results challenge our current notion of ABE metabolic network structure. Thus, clostridial ABE fermentation still poses theoretical as well as experimental challenges which are best approached in close collaboration between modellers and experimentalists

Food Matrix Design for Effective Lactic Acid Bacteria Delivery

International audienceThe range of foods featuring lactic acid bacteria (LAB) with potential associated health benefits has expanded over the years from traditional dairy products to meat, cereals, vegetables and fruits, chocolate, etc. All these new carriers need to be compared for their efficacy to protect, carry, and deliver LAB, but because of their profusion and the diversity of methods this remains difficult. This review points out the advantages and disadvantages of the main food matrix types, and an additional distinction between dairy and nondairy foods is made. The food matrix impact on LAB viability during food manufacturing, storage, and digestion is also discussed. The authors propose an ideal hypothetical food matrix that includes structural and physicochemical characteristics such as pH, water activity, and buffering capacities, all of which need to be taken into account when performing LAB food matrix design. Guidelines are finally provided to optimize food matrix design in terms of effective LAB delivery

Fluorescent labeling of nisin Z and assessment of anti-listerial action

Biomolecule labeling by fluorescent markers has emerged as an innovative methodology for bio-analytical purposes in foodmicrobiology, medicine and pharmaceutics due to the great advantages of thismethod such as precision, wide detection limits, and in vivo recognition. Fluorescent nisin Z was synthesized by linking the carboxyl group and amino group of nisin Z and 5-aminoacetamido fluorescein (AAA-flu). This new structure was fully characterized by mass spectrometry with a molecular weight of 3717.3 Da. Intracellular K+ leakage and transmembrane electrical potential (Δψ)were used to evaluate the antibacterial action of the labeledmolecule against three listerial strains and demonstrated that nisin Z endured the labeling processwithout any activity loss. In vivo activity of labeled nisin was observed by confocal laser microscope which revealed its localization at the septum of listerial cell division sitewhere themembrane-bound cellwall precursor lipid II ismaximal. Fluorescent nisin Z showed its great potential as a tool to study antibacterial mechanism of action of nisin in biological system

Microbiome engineering for biopreservation of food

International audienc

Surface properties of bacteria sensitive and resistant to the class IIa carnobacteriocin Cbn BM1

International audienc

Fluorescent labeling of nisin Z and assessment of anti-listerial action

Biomolecule labeling by fluorescent markers has emerged as an innovative methodology for bio-analytical purposes in foodmicrobiology, medicine and pharmaceutics due to the great advantages of thismethod such as precision, wide detection limits, and in vivo recognition. Fluorescent nisin Z was synthesized by linking the carboxyl group and amino group of nisin Z and 5-aminoacetamido fluorescein (AAA-flu). This new structure was fully characterized by mass spectrometry with a molecular weight of 3717.3 Da. Intracellular K+ leakage and transmembrane electrical potential (Δψ)were used to evaluate the antibacterial action of the labeledmolecule against three listerial strains and demonstrated that nisin Z endured the labeling processwithout any activity loss. In vivo activity of labeled nisin was observed by confocal laser microscope which revealed its localization at the septum of listerial cell division sitewhere themembrane-bound cellwall precursor lipid II ismaximal. Fluorescent nisin Z showed its great potential as a tool to study antibacterial mechanism of action of nisin in biological system

Comparative genomic analysis reveals ecological differentiation in the genus carnobacterium

Lactic acid bacteria (LAB) differ in their ability to colonize food and animal-associated habitats: while some species are specialized and colonize a limited number of habitats, other are generalist and are able to colonize multiple animal-linked habitats. In the current study, Carnobacterium was used as a model genus to elucidate the genetic basis of these colonization differences. Analyses of 16S rRNA gene meta-barcoding data showed that C. maltaromaticum followed by C. divergens are the most prevalent species in foods derived from animals (meat, fish, dairy products), and in the gut. According to phylogenetic analyses, these two animal-adapted species belong to one of two deeply branched lineages. The second lineage contains species isolated from habitats where contact with animal is rare. Genome analyses revealed that members of the animal-adapted lineage harbor a larger secretome than members of the other lineage. The predicted cell-surface proteome is highly diversified in C. maltaromaticum and C. divergens with genes involved in adaptation to the animal milieu such as those encoding biopolymer hydrolytic enzymes, a heme uptake system, and biopolymer-binding adhesins. These species also exhibit genes for gut adaptation and respiration. In contrast, Carnobacterium species belonging to the second lineage encode a poorly diversified cell-surface proteome, lack genes for gut adaptation and are unable to respire. These results shed light on the important genomics traits required for adaptation to animal-linked habitats in generalist Carnobacterium