Leuconostoc mesenteroides subsp. cremoris is an obligate heterolactic fermentative lactic acid bacterium that is mostly used in industrial dairy fermentations. The phosphoketolase pathway (PKP) is a unique feature of the obligate heterolactic fermentation, which leads to the production of lactate, ethanol, and/or acetate, and the final product profile of PKP highly depends on the energetics and redox state of the organism. Another characteristic of the L. mesenteroides subsp. cremoris is the production of aroma compounds in dairy fermentation, such as in cheese production, through the utilization of citrate. Considering its importance in dairy fermentation, a detailed metabolic characterization of the organism is necessary for its more efficient use in the industry. To this aim, a genome-scale metabolic model of dairy-origin L. mesenteroides subsp. cremoris ATCC 19254 (iLM.c559) was reconstructed to explain the energetics and redox state mechanisms of the organism in full detail. The model includes 559 genes governing 1088 reactions between 1129 metabolites, and the reactions cover citrate utilization and citraterelated flavor metabolism. The model was validated by simulating co-metabolism of glucose and citrate and comparing the in silico results to our experimental results. Model simulations further showed that, in co-metabolism of citrate and glucose, no flavor compounds were produced when citrate could stimulate the formation of biomass. Significant amounts of flavor metabolites (e.g., diacetyl and acetoin) were only produced when citrate could not enhance growth, which suggests that flavor formation only occurs under carbon and ATP excess. The effects of aerobic conditions and different carbon sources on product profiles and growth were also investigated using the reconstructed model. The analyses provided further insights for the growth stimulation and flavor formation mechanisms of the organism.
Considering the need for systematic studies on levan based hydrogels to widen their use in drug delivery systems and biomedical applications, this study is mainly focused on the synthesis and comprehensive characterization as well as drug release properties of hydrogels based on Halomonas levan (HL) and its chemical derivatives. For this, hydrolyzed and phosphonated HL derivatives were chemically synthesized and then cross-linked with 1,4-Butanediol diglycidyl ether (BDDE) and the obtained hydrogels were characterized in terms of their swelling, adhesivity, and rheological properties. Both native and phosphonated HL hydrogels retained their rigid gel like structure with increasing shear stress levels and tack test analysis showed superior adhesive properties of the phosphonated HL hydrogels. Moreover, hydrogels were loaded with resveratrol and entrapment and release studies as well as cell culture studies with human keratinocytes were performed. Biocompatible and adhesive features of the hydrogels confirmed their suitability for tissue engineering and drug delivery applications.
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