Modeling and Optimization of Triticale Wort Production Using an Artificial Neural Network and a Genetic Algorithm.

Triticale grain, a wheat-rye hybrid, has been reported to comply very well with the requirements for modern brewing adjuncts. In this study, two triticale varieties, in both unmalted and malted forms, were investigated at various ratios in the grist, applying different mashing regimes and concentrations of the commercial enzyme Shearzyme® 500 L with the aim of evaluating their impact on wort production. In order to capture the complex relationships between the input (triticale ratio, enzyme ratio, mashing regime, and triticale variety) and output variables (wort extract content, wort viscosity, and free amino nitrogen (FAN) content in wort), the study aimed to implement the use of artificial neural networks (ANNs) to model the mashing process. Also, a genetic algorithm (GA) was integrated to minimize a specified multi-objective function, optimizing the mashing process represented by the ANN model. Among the solutions on the Pareto front, one notable set of solutions was found with objective function values of 0.0949, 0.0131, and 1.6812 for the three conflicting objectives, respectively. These values represent a trade-off that optimally balances the different aspects of the optimization problem. The optimized input variables had values of 23%, 9%, 1, and 3 for the respective input variables of triticale ratio, enzyme ratio, mashing regime, and triticale variety. The results derived from the ANN model, applying the GA-optimized input values, were 8.65% w/w for wort extract content, 1.52 mPa·s for wort viscosity, and 148.32 mg/L for FAN content in wort. Comparatively, the results conducted from the real laboratory mashing were 8.63% w/w for wort extract content, 1.51 mPa·s for wort viscosity, and 148.88 mg/L for FAN content in wort applying same input values. The presented data from the optimization process using the GA and the subsequent experimental verification on the real mashing process have demonstrated the practical applicability of the proposed approach which confirms the potential to enhance the quality and efficiency of triticale wort production.

Reduction of sterigmatocystin biosynthesis and growth of food-borne fungi by lactic acid.

Food contamination by fungi and mycotoxins presents a problem for food safety even today. Since lactic acid (LA) has Generally Recognized As Safe (GRAS) status, the aim of this research was to determine its potential in protection of food against mycological and mycotoxicological contamination. In this study, LA showed an inhibitory effect on the growth of food-borne fungi (Penicillium aurantiogriseum K51, Aspergillus parasiticus KB31, Aspergillus versicolor S72, and Aspergillus niger K95) and on biosynthesis of sterigmatocystin (STE). For the antifungal effect of LA on the growth of food-borne fungi, the disc diffusion and microdilution methods were performed. The effect of LA on the STE biosynthesis by A. versicolor was determined using an LC-MS/MS technique. The largest inhibition zone was observed for A. versicolor (inhibition zone of 24 ± 0.35 mm), while there were no inhibition zones for A. niger and A. parasiticus at all tested LA concentrations. The minimal inhibitory concentration (MIC) of LA on fungi ranged from 25.0 mg/mL to 50.0 mg/mL, while the minimum fungicidal concentrations (MFCs) ranged from 50.0 mg/mL to 100.0 mg/mL. Complete inhibition of STE biosynthesis by A. versicolor was observed at an LA concentration of 50.0 mg/mL. The obtained results showed that LA could be efficient for protection of food against mycological and STE contamination.

Immobilization of Lactobacillus rhamnosus in polyvinyl alcohol/calcium alginate matrix for production of lactic acid.

Immobilization of Lactobacillus rhamnosus ATCC7469 in poly(vinyl alcohol)/calcium alginate (PVA/Ca-alginate) matrix using "freezing-thawing" technique for application in lactic acid (LA) fermentation was studied in this paper. PVA/Ca-alginate beads were made from sterile and non-sterile PVA and sodium alginate solutions. According to mechanical properties, the PVA/Ca-alginate beads expressed a strong elastic character. Obtained PVA/Ca-alginate beads were further applied in batch and repeated batch LA fermentations. Regarding cell viability, L. rhamnosus cells survived well rather sharp immobilization procedure and significant cell proliferation was observed in further fermentation studies achieving high cell viability (up to 10.7 log CFU g-1) in sterile beads. In batch LA fermentation, the immobilized biocatalyst was superior to free cell fermentation system (by 37.1%), while the highest LA yield and volumetric productivity of 97.6% and 0.8 g L-1 h-1, respectively, were attained in repeated batch fermentation. During seven consecutive batch fermentations, the biocatalyst showed high mechanical and operational stability reaching an overall productivity of 0.78 g L-1 h-1. This study suggested that the "freezing-thawing" technique can be successfully used for immobilization of L. rhamnosus in PVA/Ca-alginate matrix without loss of either viability or LA fermentation capability.

Non-thermal plasma and ultrasound-assisted open lactic acid fermentation of distillery stillage.

Stillage is the main by-product of bioethanol production and the cost of its treatment significantly affects the economy of bioethanol production. A process of thermal sterilization before lactic acid fermentation (LAF) is energy demanding and is causing deterioration of valuable compounds in stillage. In this study, ultrasound (UT) and plasma (PT) treatments were used for microbial inactivation, and a significant reduction in the number of viable microorganisms in the stillage after PT and UT was observed. After application of treatment, LAF by Lactobacillus rhamnosus ATCC 7469 was initiated. The concentration of LA is used to quantify the efficiency of the stillage revalorization. The highest LA productivity of 1.21 g/Lh and yield of 0.82 g/g were obtained after PT, while UT of 10 min provided productivity of 1.02 g/Lh and LA yield of 0.69 g/g. The results were benchmarked against closed LAF. Around 20% better revalorization of stillage by PT was achieved when compared with conventional sterilization. In addition, an excellent L (+) LA stereoselectivity of 95.5% was attained after PT. From the aspect of energy efficiency, that of PT was three times lower than UT and almost ten times lower than thermal sterilization, but it is the most expensive due to the high consumption of gas which could reduce application of closed Ar atmosphere on larger scales. This way, a simpler and energy efficient process for LA production on stillage was accomplished by "open" fermentation.

Bioprocessing of agro-industrial residues into lactic acid and probiotic enriched livestock feed.

BACKGROUND: Growing challenges of resource depletion, food security and environmental protection are putting stress on the development of biorefinery processes for bioprocessing of residues from food and agro-industry into value-added products. In this study, the simultaneous production of lactic acid (LA) and livestock feed on a combined substrate based on molasses and potato stillage by Lactobacillus paracasei NRRL B-4564 immobilized onto sunflower seed hull (SSH), brewer's spent grain (BSG) and sugar beet pulp (SBP) was studied. RESULTS: The highest total LA concentration of 399 g L-1 with overall productivity of 1.27 g L-1  h-1 was achieved in repeated batch fermentation by SBP-immobilized biocatalyst, followed by BSG- and SSH-immobilized cells. Fermentation improved the content of proteins and ash, and decreased the content of fibers in all three support materials. In addition, the fermentation had favorable effect on in vitro dry matter digestibility and energy values of SSH and BSG. According to assessment of probiotic potential, L. paracasei demonstrated a favorable probiotic profile, exhibiting high resistance to simulated ruminant digestive tract and significant antioxidant and antimicrobial activity. CONCLUSIONS: The proposed strategy enables valorization of agro-industrial residues as value-added ruminant feed and simultaneous LA production. Following principles of circular economy, the developed process combines different raw materials and integrates them into a biorefinery process, improving the overall profitability and productivity. © 2019 Society of Chemical Industry.

Utilization of brewing and malting by-products as carrier and raw materials in l-(+)-lactic acid production and feed application.

Application of agro-industrial by-products for the production of lactic acid was studied in this paper. Brewer's spent grain (BSG), malt rootlets (MR), brewer's yeast (BY), and soy lecithin (SL) were used as raw materials in L-(+)-LA fermentation by free and immobilized Lactobacillus rhamnosus ATCC 7469. The BSG, solid remains after BSG and MR hydrolysis (BSGMRSR), and MR were evaluated as carriers for batch and repeated batch fermentations with immobilized cells. During batch fermentations with immobilized cells, high cell viability (10 to 11 log CFU/g) was achieved on all carriers. In batch fermentation with BSG as a carrier, the highest LA yield of 93.79% and volumetric productivity of 1.15 g/L/h were obtained. Furthermore, very high LA yield (95.46%), volumetric productivity (1.98 g/L/h) and L. rhamnosus viability (11.5 log CFU/g) were achieved in repeated batch fermentations with the cells immobilized on this carrier. The immobilized cells showed high survival rate (94-95%) during exposure to simulated gut condition. Based on the analysis of BSGMRSR, and BY solid remains, and on in vitro evaluation of the probiotic characteristics of immobilized cells, it was observed that they could satisfy the recommendations for high-quality feed preparation.

Enhanced Lactic Acid Production by Adaptive Evolution of Lactobacillus paracasei on Agro-industrial Substrate.

The aim of this study was to perform the adaptation of Lactobacillus paracasei NRRL B-4564 to substrate through adaptive evolution in order to ensure intensive substrate utilization and enhanced L (+)-lactic acid (LA) production on molasses-enriched potato stillage. To evaluate the strain response to environmental conditions exposed during the adaptation process and to select the best adapted cells, the antioxidant activity and LA-producing capability were assessed in batch fermentation. The most promising adapted strain was further used in a pulsed fed-batch mode. Among three selected adapted strains, L. paracasei A-22 showed considerably improved antioxidant capacity, demonstrating more than onefold higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging rates compared to parent strain. This strain also exhibited superior LA production in batch fermentation and reached 89.4 g L-1 of LA, with a yield of 0.89 g g-1, a productivity of 1.49 g L-1 h-1, and an optical purity greater than 99%. Furthermore, in fed-batch mode L. paracasei A-22 resulted in 59% higher LA concentration (169.9 g L-1) compared to parent strain (107.1 g L-1). The strain adaptation to molasses environment, performed in this study, is a rather simple and promising method for enhancement of LA production on the complex agro-industrial substrate.

Possibility of L-(+)-lactic acid fermentation using malting, brewing, and oil production by-products.

Industrial by-products such as brewer's spent grain (BSG) hydrolysate, malt rootlets extract (MRE) and soybean meal extract (SME) were used for L-(+) lactic acid (LA) production by a pure L. rhamnosus ATCC 7469 strain. The effect of the addition of MRE (10-50%) or SME (10-50%) in BSG hydrolysate on batch and fed-batch LA fermentation was evaluated. The addition of MRE and SME increased the concentration of free amino nitrogen (FAN) and essential minerals (Fe, Mg, Mn, and Zn), which had a positive effect on the fermentation. Also, the MRE addition significantly lowered C/N ration to a more favorable level for the efficient LA fermentation. In batch fermentation, the highest LA concentration (25.73 g/L), yield (86.31%), and volumetric productivity (0.95 g/L h-1), were obtained with the addition of 50% MRE. Further increase in LA concentration to 58.01 g/L, yield to 88.54%, and volumetric productivity to 1.19 g/L h-1 was achieved in fed-batch fermentation with addition of 50% MRE. A high optical purity of LA with 99.7% of L-(+)-isomer was obtained on the substrate based on industrial by-products. In addition, solid remains after BSG hydrolysis and MRE and SME preparation, together with the biomass of L. rhamnosus separated after the fermentation could be a good base for feed preparation.

Wastes from bioethanol and beer productions as substrates for l(+) lactic acid production - A comparative study.

Waste substrates from bioethanol and beer productions are cheap, abundant and renewable substrates for biorefinery production of lactic acid (LA) and variability in their chemical composition presents a challenge in their valorisation. Three types of waste substrates, wasted bread and wasted potato stillage from bioethanol production and brewers' spent grain hydrolysate from beer production were studied as substrates for the production of l(+) LA and probiotic biomass by Lactobacillus rhamnosus ATCC 7469. The correlation of the content of free alpha amino nitrogen and the production of LA was determined as a critical characteristic of the waste media for efficient LA production by L. rhamnosus on the substrates which contained equal amount of fermentable sugars. A maximal LA productivity of 1.54gL(-1)h(-1) was obtained on wasted bread stillage media, whilst maximal productivities achieved on the potato stillage and brewers' spent grain hydrolysate media were 1.28gL(-1)h(-1)and 0.48gL(-1)h(-1), respectively. A highest LA yield of 0.91gg(-1) was achieved on wasted bread stillage media, followed by the yield of 0.81gg(-1) on wasted potato stillage and 0.34gg(-1) on brewers' spent grain hydrolysate media. The kinetics of sugar consumption in the two stillage substrates were similar while the sugar conversion in brewers' spent grain hydrolysate was slower and less efficient due to significantly lower content of free alpha amino nitrogen. The lignocellulosic hydrolysate from beer production required additional supplementation with nitrogen.

Integrated production of lactic acid and biomass on distillery stillage.

The possibilities of parallel lactic acid and biomass production in batch and fed-batch fermentation on distillery stillage from bioethanol production were studied. The highest lactic acid yield and productivity of 92.3 % and 1.49 g L(-1) h(-1) were achieved in batch fermentation with initial sugar concentration of 55 g L(-1). A significant improvement of the process was achieved in fed-batch fermentation where the concentration of lactic acid was increased to 47.6 % and volumetric productivity for 21 % over the batch process. A high number of Lactobacillus rhamnosus ATCC 7469 viable cells of 10(9) CFU ml(-1) was attained at the end of fed-batch fermentation. The survival of 92.9 % of L. rhamnosus cells after 3 h of incubation at pH 2.5 validated that the fermentation media remained after lactic acid removal could be used as a biomass-enriched animal feed thus making an additional value to the process.

Lactic acid production on liquid distillery stillage by Lactobacillus rhamnosus immobilized onto zeolite.

In this study, lactic acid and biomass production on liquid distillery stillage from bioethanol production with Lactobacillus rhamnosus ATCC 7469 was studied. The cells were immobilized onto zeolite, a microporous aluminosilicate mineral and the lactic acid production with free and immobilized cells was compared. The immobilization allowed simple cell separation from the fermentation media and their reuse in repeated batch cycles. A number of viable cells of over 10(10) CFU g(-1) of zeolite was achieved at the end of fourth fermentation cycle. A maximal process productivity of 1.69 g L(-1), maximal lactic acid concentration of 42.19 g L(-1) and average yield coefficient of 0.96 g g(-1) were achieved in repeated batch fermentation on the liquid stillage without mineral or nitrogen supplementation.

Effect of different fermentation parameters on L-lactic acid production from liquid distillery stillage.

Expansion of lactic acid applications, predominantly for the preparation of biodegradable polymers increased the research interest for new, economically favourable production processes. Liquid stillage from bioethanol production can be an inexpensive, valuable source of nutrients for growth of lactic acid bacteria. Utilisation of residual biomass with spent fermentation media as a functional animal feed can greatly influence the process value and its ecological aspect. In this paper, the kinetics of lactic acid and biomass production on liquid stillage by Lactobacillus rhamnosus ATCC 7469 was studied. In addition, the impact of temperature, inoculum concentration, shaking and pH control by addition of CaCO(3) was evaluated. Maximal lactic acid yield of 73.4%, as well as high biomass production (3×10(8) CFU ml(-1)) were achieved under selected conditions (41°C, 5% (v/v) of inoculum, 1% (w/v) of CaCO(3), initial pH of 6.5 and shaking rate of 90 rpm). These results were achieved without supplementation of the stillage with nitrogen or mineral sources.