Identification of Potential Citrate Metabolism Pathways in Carnobacterium maltaromaticum

In the present study, we describe the identification of potential citrate metabolism pathways for the lactic acid bacterium (LAB) Carnobacterium maltaromaticum. A phenotypic assay indicated that four of six C. maltaromaticum strains showed weak (Cm 6-1 and ATCC 35586) or even delayed (Cm 3-1 and Cm 5-1) citrate utilization activity. The remaining two strains, Cm 4-1 and Cm 1-2 gave negative results. Additional analysis showed no or very limited utilization of citrate in media containing 1% glucose and 22 or 30 mM citrate and inoculated with Cm 6-1 or ATCC 35586. Two potential pathways of citrate metabolism were identified by bioinformatics analyses in C. maltaromaticum including either oxaloacetate (pathway 1) or tricarboxylic compounds such as isocitrate and α-ketoglutarate (pathway 2) as intermediates. Genes encoding pathway 1 were present in two out of six strains while pathway 2 included genes present in all six strains. The two potential citrate metabolism pathways in C. maltaromaticum may potentially affect the sensory profiles of milk and soft cheeses subjected to growth with this species.

KRAS, A Prime Mediator in Pancreatic Lipid Synthesis through Extra Mitochondrial Glutamine and Citrate Metabolism

Kirsten rat sarcoma viral oncogene homolog (KRAS)-driven pancreatic cancer is very lethal, with a five-year survival rate of <9%, irrespective of therapeutic advances. Different treatment modalities including chemotherapy, radiotherapy, and immunotherapy demonstrated only marginal efficacies because of pancreatic tumor specificities. Surgery at the early stage of the disease remains the only curative option, although only in 20% of patients with early stage disease. Clinical trials targeting the main oncogenic driver, KRAS, have largely been unsuccessful. Recently, global metabolic reprogramming has been identified in patients with pancreatic cancer and oncogenic KRAS mouse models. The newly reprogrammed metabolic pathways and oncometabolites affect the tumorigenic environment. The development of methods modulating metabolic reprogramming in pancreatic cancer cells might constitute a new approach to its therapy. In this review, we describe the major metabolic pathways providing acetyl-CoA and NADPH essential to sustain lipid synthesis and cell proliferation in pancreatic cancer cells.

Regional Citrate Anticoagulation Protocol For Patients With Presumed Absent Citrate Metabolism

Background: Regional citrate anticoagulation (RCA) is not recommended in patients with shock or severe liver failure. We designed a protocol with personalized pre-calculated flow settings for patients with absent citrate metabolism that abrogates risk of citrate toxicity, maintains neutral CKRT circuit calcium mass balance and normal systemic ionized calcium levels. Methods: Single center prospective cohort study of patients in five Adult Intensive Care Units triaged to the CVVHDF-RCA "Shock" protocol. Results: Of 31 patients included in the study, 30 (97%) had acute kidney injury, 16 (52%) had acute liver failure, and 5 (16%) had cirrhosis at the start of CKRT. The median (interquartile range (IQR)) lactate was 5 (3.2 to 10.7), AST 822 (122 to 2950), ALT 352 (41 to 2238), total bilirubin 2.7 (1.0 to 5.1), INR 2 (1.5 to 2.6). The median first hemofilter life censored for causes other than clotting exceeded 70 hours. The cumulative incidence of hypernatremia (Na >148 mM), metabolic alkalosis (HCO3- >30 mM) and hypophosphatemia (P< 2 mg/dL) were 1/26 (4%), 0/30 (0%), 1/30 (3%) respectively and were not clinically significant. Mild hypocalcemia occurred in the first 4 hours in 2/31 patients and corrected by hour 6 with no additional Ca-supplementation beyond the per-protocol administered Ca-infusion. The maximum systemic total Ca (tCa; mM)/ionized Ca (iCa; mM) ratio never exceeded 2.5. Conclusions: The "Shock" protocol can be used without contra-indications and is effective in maintaining circuit patency with a high, fixed ACDA infusion rate to blood flow ratio. Keeping single-pass citrate extraction on the dialyzer >0.75 minimizes the risk of citrate toxicity even in patients with absent citrate metabolism. Pre-calculated, personalized dosing of the initial Ca-infusion rate from a table based on the patient's albumin level and the filter effluent flow rate maintains neutral CKRT circuit calcium mass balance and a normal systemic iCa level.

Selection of Lactic Acid Bacteria Isolated from Fresh Fruits and Vegetables Based on Their Antimicrobial and Enzymatic Activities

Lactic acid bacteria (LAB) are an important source of bioactive metabolites and enzymes. LAB isolates from fresh vegetable sources were evaluated to determine their antimicrobial, enzymatic, and adhesion activities. A saline solution from the rinse of each sample was inoculated in De Man, Rogosa and Sharpe Agar (MRS Agar) for isolates recovery. Antimicrobial activity of cell-free supernatants from presumptive LAB isolates was evaluated by microtitration against Gram-positive, Gram-negative, LAB, mold, and yeast strains. Protease, lipase, amylase, citrate metabolism and adhesion activities were also evaluated. Data were grouped using cluster analysis, with 85% of similarity. A total of 76 LAB isolates were recovered, and 13 clusters were formed based on growth inhibition of the tested microorganisms. One cluster had antimicrobial activity against Gram-positive bacteria, molds and yeasts. Several LAB strains, PIM4, ELO8, PIM5 and CAL14 strongly inhibited the growth of L. monocytogenes and JAV15 and TOV9 strongly inhibited the growth of F. oxysporum. Based on enzymatic activities, 5 clusters were formed. Seven isolates hydrolyzed starch, 46 proteins, 14 lipids, and 36 metabolized citrate. LAB isolates with the best activities were molecularly identified as Leuconostoc mesenteroides, Enterococcus mundtii and Enterococcus faecium. Overall, LAB isolated from vegetables showed potential technological applications and should be further evaluated.

Complete Genome Sequence of Lactococcus lactis subsp. lactis bv. diacetylactis SD96

The genome of Lactococcus lactis subsp. lactis bv. diacetylactis SD96, a strain used for cheese production, is presented. SD96 is refractory to phage attack, which is a desired property for starter bacteria. Its 10 plasmids provide industrially important traits, such as lactose and citrate metabolism, proteolytic activity, and phage resistance.

Nutritional requirements for Lactobacillus vini growth in sugarcane derivative substrate of ethanol fermentation

ABSTRACT Lactobacillus vini is a bacterial contaminant found in industrial environments of winemaking and fuel-ethanol fermentation. However, there has been no standard analysis of its physiology that can pinpoint its adaptive traits to these kinds of environments. In view of this lack of information, the aim of this study is to determine the nutritional factors that lead to the growth of L. vini in the industrial plants of fuel-ethanol. First of all, the limited growth of this bacterium was studied in the industrial substrate, which was improved by nutritional supplementation with amino acids, and its homofermentative status was confirmed. Metabolite analysis showed that citrate is a growth factor of paramount importance for this bacterium in industrial processes through pyruvate metabolization, and increases ATP production and biomass formation. Furthermore,e acetate uptake, either from the medium or generated from citrate metabolism, was assimilated for biomass production. Hence, a metabolic model was designed to describe the role of citrate and acetate in the growth of L. vini that could be tested on other lactobacilli.