Ivna Vrana Špoljarić

Mathematical modeling of poly((R)-3-hydroxyalkanoate) synthesis by Cupriavidus necator DSM 545 on substrates stemming from biodiesel production

Two low structured mathematical models for fed-batch production of polyhydroxybutyrate and poly[hydroxybutyrate-co-hydroxyvalerate] by Cupriavidus necator DSM 545 on renewable substrates (glycerol and fatty acid methyl esters-FAME) combined with glucose and valeric acid, were established. The models were used for development/optimization of feeding strategies of carbon and nitrogen sources concerning PHA content and polymer/copolymer composition. Glycerol/glucose fermentation featured a max. specific growth rate of 0.171 h(-1), a max. specific production rate of 0.038 h(-1) and a PHB content of 64.5%, whereas the FAME/valeric acid fermentation resulted in a max. specific growth rate of 0.046 h(-1), a max. specific production rate of 0.07 h(-1) and 63.6% PHBV content with 4.3% of 3-hydroxyvalerate (3HV) in PHBV. A strong inhibition of glycerol consumption by glucose was confirmed (inhibition constant ki,G=4.28×10(-4) g L(-1)). Applied concentration of FAME (10-12 g L(-1)) positively influenced on PHBV synthesis. HV/PHBV ratio depends on applied VA concentration.

In silico optimization and low structured kinetic model of poly[(R)-3-hydroxybutyrate] synthesis by Cupriavidus necator DSM 545 by fed-batch cultivation on glycerol.

Glycerol was utilized by Cupriavidus necator DSM 545 for production of poly-3-hydroxybutyrate (PHB) in fed-batch fermentation. Maximal specific growth rates (0.12 and 0.3h(-1)) and maximal specific non-growth PHB production rate (0.16 g g(-1)h(-1)) were determined from two experiments (inocula from exponential and stationary phase). Saturation constants for nitrogen (0.107 and 0.016 g L(-1)), glycerol (0.05 g L(-1)), non-growth related PHB synthesis (0.011 g L(-1)) and nitrogen/PHB related inhibition constant (0.405 g L(-1)), were estimated. Five relations for specific growth rate were tested using mathematical models. In silico performed optimization procedures (varied glycerol/nitrogen ratio and feeding) has resulted in a PHB content of 70.9%, shorter cultivation time (23 h) and better PHB yield (0.347 g g(-1)). Initial concentration of biomass 16.8 g L(-1) and glycerol concentration in broth between 3 and 5 g L(-1) were decisive factors for increasing of productivity.

Study of metabolic network of Cupriavidus necator DSM 545 growing on glycerol by applying elementary flux modes and yield space analysis

A metabolic network consisting of 48 reactions was established to describe intracellular processes during growth and poly-3-hydroxybutyrate (PHB) production for Cupriavidus necator DSM 545. Glycerol acted as the sole carbon source during exponential, steady-state cultivation conditions. Elementary flux modes were obtained by the program Metatool and analyzed by using yield space analysis. Four sets of elementary modes were obtained, depending on whether the pair NAD/NADH or FAD/FADH2 contributes to the reaction of glycerol-3-phosphate dehydrogenase (GLY-3-P DH), and whether 6-phosphogluconate dehydrogenase (6-PG DH) is present or not. Established metabolic network and the related system of equations provide multiple solutions for the simultaneous synthesis of PHB and biomass; this number of solutions can be further increased if NAD/NADH or FAD/FADH2 were assumed to contribute in the reaction of GLY-3-P DH. As a major outcome, it was demonstrated that experimentally determined yields for biomass and PHB with respect to glycerol fit well to the values obtained in silico when the Entner–Doudoroff pathway (ED) dominates over the glycolytic pathway; this is also the case if the Embden–Meyerhof–Parnas pathway dominates over the ED.

Influence of Barley Sourdough and Vacuum Cooling on Shelf-Life Quality of Partially Baked Bread

Driven by the bakery industry urge to satisfy consumer demand for fresh, diverse and high quality bakery products, we investigated the influence of barley sourdough and vacuum cooling on shelf life quality of partially baked bread stored in modified atmosphere packaging at ambient conditions. Barley sourdough was fermented with Lactobacillus reuteri (DSM 20016, F275). Partially baked bread with sourdough was microbiologically acceptable during 30 days of storage, while bread without sourdough had detectable mould on the 30th day. Stored bread samples were rebaked after 1, 8, 15, 22 and 30 days to determine moisture content, physical and sensorial properties. Moisture loss (5%) was detected on the 15th day, after which it remained stable until the end of investigated storage period. Nevertheless, textural quality of stored bread continuously declined due to crumb firming. Bread flavour did not change during mould-free storage time. The principal component analysis identified major differences in the flavour of sour and control bread, also in crumb firmness and moisture content of samples. This study indicates the positive role of barley sourdough fermented with L. reuteri in improving crumb texture for at least 15 days, and ensuring mould- and bacteria-free partially baked bread for 30 days. Vacuum cooling combined with sourdough improved bread shape, porosity, and reduced sour taste, crust colouring and crumbliness. Hence, it can successfully extend shelf life quality of partially baked bread in modified atmosphere packaging.

Frozen stored barley sourdough: stability and application to wheat bread

The primary aim of the industrial use of sourdough is to improve bread quality and to replace additives. This study investigates the applicability of frozen stored barley sourdough in wheat bread. Sourdough was fermented with Weissella cibaria in the presence of sucrose and xylanase, and stored frozen for 4, 8, or 12 weeks. During storage, the number of viable cells in the sourdough declined. Softer dough and crumb were obtained after the sourdough was stored for a longer period (4 weeks), while bread specific volume was unaffected by the sourdough storage time. The highest bread specific volume, shape, crumb softness, and sensory liking were obtained with 20% of the sourdough. This study demonstrates the possibility for the industrial application of frozen barley sourdough, since it can improve the quality of bread compared to the native barley flour.