Fed‐batch l‐(+)‐lactic acid fermentation of brewer's spent grain hydrolysate

Brewer's spent grain (BSG) hydrolysates were used for l ‐(+)‐lactic acid (LA) fermentation by Lactobacillus rhamnosus ATCC 7469. The aim of this study was to evaluate fed‐batch LA fermentation of BSG hydrolysate with the addition of glucose, glucose and yeast extract, and wort during LA fermentation and its effect on fermentation parameters such as LA concentration, its volumetric productivity and yield, and L. rhamnosus cell viability. The highest LA yield, volumetric productivity and concentration of 93.3%, 2.0 g/L/h, and 116.1 g/L, respectively, were achieved with glucose and yeast extract addition during fermentation. In fed‐batch fermentation with glucose and yeast extract addition significantly higher LA concentration, yield and volumetric productivity (by 194.8; 2.2, and 20.7%, respectively) were achieved compared with batch fermentation. The results indicated that fed‐batch fermentation could be used to increase LA fermentation efficiency. Copyright © 2017 The Institute of Brewing & Distilling     

Use of spent brewer's yeast in L‐(+) lactic acid fermentation

The application of by‐products from the brewing industry in lactic acid (LA) production was investigated in order to replace expensive nitrogen sources (such as yeast extract) with cheaper and renewable nitrogenous materials such as brewer's yeast (BY). In this study, brewer's spent grain (BSG) hydrolysate was used for L‐(+)‐LA fermentation by Lactobacillus rhamnosus ATCC 7469. The effect of pH control during the fermentation and the addition of various BY contents (5–50 g/L) in BSG hydrolysate on fermentation parameters was evaluated. BY addition significantly increased free amino nitrogen (FAN) concentration (by 25.2% at 5 g/L to 616% at 50 g/L). A strong positive correlation between FAN concentration in the hydrolysate and concentration of L‐(+)‐LA produced was observed (correlation coefficient of 0.913). A high cell viability of L. rhamnosus ATCC 7469 (1.95–3.32 × 10⁹ CFU/mL at the end of fermentation) was achieved in all fermentations with the addition of brewer's yeast. The addition of BY increased L‐(+)‐lactic acid yield and volumetric productivity up to 8.4% (5 g/L) and 48.3% (50 g/L). The highest L‐(+)‐LA yield (89%) and volumetric productivity (0.89 g/L h^?1) were achieved in fermentation of BSG hydrolysate with 50 g/L of BY. © 2019 The Institute of Brewing & Distilling     

Lactic acid fermentation of brewer's spent grain hydrolysate by Lactobacillus rhamnosus with yeast extract addition and pH control

Lactic acid (LA) is a versatile chemical with a wide range of applications in food, pharmaceutical, cosmetic, textile and polymer industries. Brewer's spent grain (BSG) is the most abundant brewing by‐product. In this study BSG hydrolysates were used for LA fermentation by Lactobacillus rhamnosus ATCC 7469. The aim of this study was to evaluate the effects of pH control during fermentation, reducing sugar content and yeast extract content in BSG hydrolysate on LA fermentation parameters. The pH control greatly increased reducing sugar utilization, l ‐(+)‐LA content, yield and volumetric productivity. The highest l ‐(+)‐LA yield and volumetric productivity were achieved with the reducing sugar content of 54 g/L. Yeast extract addition significantly increased reducing sugar utilization, l ‐(+)‐LA content, L. rhamnosus cell viability, l ‐(+)‐LA yield and volumetric productivity. The highest l ‐(+)‐LA content (39.38 g/L), L. rhamnosus cell viability (9.67 log CFU/mL), l ‐(+)‐LA yield (91.29%) and volumetric productivity (1.69 g/L/h) were achieved with the reducing sugar content of 54 g/L and yeast extract content of 50 g/L. Copyright © 2017 The Institute of Brewing & Distilling     

Bioconversion of agro‐industrial by‐products to lactic acid using Lactobacillus sakei and two Pediococcus spp. strains

The aim of this study was to evaluate the bioconversion efficiency of rich in cellulose agro‐industrial by‐products such as wheat bran (WB), spent distiller's grain with solids (DGS), brewer's spent grain (BSG) and lupin (Lupinus angustifolius L.) wholemeal fraction (LF) to lactic acid (LA) using acid tolerant lactic acid bacteria (LAB) strains Lactobacillus sakei KTU05‐06, Pediococcus acidilactici KTU05‐7 and P. pentosaceus KTU05‐9. Carbohydrase preparation Depol^? 692L was used for the hydrolysis of non‐starch polysaccharides. Analysed raw materials were suitable substrates for LAB propagation and L‐lactic acid production. The lowest pH (3.6) was found in LF medium after 48 h fermentation with P. acidilactici and P. pentosaceus strains. The lowest pH (3.86) was measured in WB fermented with L. sakei, and in DGS and BSG (pH 3.8 and 3.9 respectively) fermented with P. acidilactici. The highest endoxylanase activity was excreted by the P. acidilactici and P. pentosaceus (84 and 69 XU g^?1 respectively), and the highest α‐amylase activity was of L. sakei (255.6 AU g^?1) after 24 h incubation in WB medium. The L‐lactic acid concentration of 86.11 g kg^?1 was reached after the bioconversion of hydrolysed WB in combination with 48 h fermentation by P. pentosaceus KTU05‐9 strain. LA contents between 222 and 282 mg kg^?1 was produced from lupin processing residues via fermentation using P. acidilactici and P. pentosaceus KTU05‐9 strains. The major challenge within the presented study is the viability of tested LAB in cereal waste media and effective LA production at a low pH (3.6–3.8).     

Brewers' spent grain: a review with an emphasis on food and health

Brewers' spent grain (BSG) is the most abundant by‐product generated in the beer‐brewing process. This material consists of the barley grain husks obtained as solid residue after the production of wort. BSG is rich in fibre and protein and, to date, the main use for the elimination of this by‐product has been as an animal feed. However, because of its nutritional content, BSG is of interest for application and fortification of human food products, particularly in view of its low cost and availability in large amounts. In addition, the importance of BSG as an ingredient and potential source of health‐promoting bioactive components is beginning to be recognised. The investigation of alternative uses of BSG is pertinent, not only from the perspective of the brewer who can benefit from valorisation of this by‐product, but also from an environmental perspective as the recycling and re‐use of industrial wastes and by‐products has become increasingly important. This review presents the current knowledge on BSG, covering its production, composition and methods for the release of valuable components, and focuses on the potential health benefits attributed to its constituents and the use of this brewer by‐product in food applications. Copyright © 2016 The Institute of Brewing & Distilling     

Acid hydrolysis and fermentation of brewer's spent grain to produce xylitol

The hemicellulosic fraction of brewer's spent grain (BSG) was hydrolysed with diluted acid under different conditions of liquid/solid ratio (8–12 g g⁻¹), sulfuric acid concentration (100–140 mg g⁻¹ dry matter) and reaction time (17–37 min) in order to produce a liquor with a large amount of xylose and good fermentability to produce xylitol. Results showed that all the evaluated reaction conditions were able to hydrolyse xylan and arabinan with efficiencies higher than 85.8 and 95.7% respectively, and even under the mildest reaction condition a considerable amount (92.7%) of the hemicellulosic fraction could be extracted. The hydrolysates presented different fermentabilities when used as fermentation media for xylitol production by Candida guilliermondii yeast, owing to the differences in their composition. Based on statistical analysis, the best condition for BSG acid hydrolysis was the use of a liquid/solid ratio of 8 g g⁻¹, 100 mg H₂SO₄ g⁻¹ dry matter and a reaction time of 17 min. Under this condition a high extraction efficiency of hemicellulosic sugars (92.7%) and good fermentation results (Y_P/S = 0.70 g g⁻¹ and Q_P = 0.45 g dm⁻³ h⁻¹) were attained. Copyright © 2005 Society of Chemical Industry     

Production of lactic acid from vine‐trimming wastes and viticulture lees using a simultaneous saccharification fermentation method

An effective process for the chemical–biotechnological utilization of trimming wastes of vineshoots, an agricultural waste with little use, is reported. Initial treatment with sulfuric acid (prehydrolysis) allowed the solubilization of hemicelluloses to give xylose and glucose‐containing liquors (suitable to make fermentation media for lactic acid production with Lactobacillus pentosus) and a solid phase containing cellulose and lignin. The solid residues from prehydrolysis were treated with NaOH in order to increase their cellulase digestibility. In the alkaline treatments, the effects of temperature (in the range, 50–130 °C), reaction time (30–120 min) and NaOH concentration (4–12 wt% of solution) on the composition and susceptibility to enzymatic hydrolysis of solid residues were assessed by means of an experimental plan with factorial structure. The lignin content decreased, whereas the susceptibility towards the enzymatic hydrolysis increased with temperature, reaction time and NaOH concentration within the tested range. Using the cellulosic residues achieved under the harsher conditions, favorable fermentation kinetics during simultaneous saccharification and fermentation carried out by L rhamnosus for lactic acid production were observed. The nutrients employed were the complete MRS broth and a cheaper medium developed using viticulture lees coming from the white wine making technology. In all cases the final lactic acid concentration achieved was similar, although the volumetric productivity was lower when using lees due to inhibitory effects over the enzymatic hydrolysis. Copyright © 2004 Society of Chemical Industry     

Feeding barley grain steeped in lactic acid modulates rumen fermentation patterns and increases milk fat content in dairy cows

The objectives of the present in vivo and in situ trials were to evaluate whether feeding barley grain steeped in lactic acid (LA) would affect rumen fermentation patterns, in situ dry matter (DM) degradation kinetics, and milk production and composition in lactating dairy cows. The in vivo trial involved 8 rumen-fistulated Holstein cows fed once daily a total mixed ration containing rolled barley grain (27% in DM) steeped for 48 h in an equal quantity of tap water (CTR) or in 0.5% LA (TRT) in a 2 × 2 crossover design. The in situ trials consisted of incubation of untreated rolled barley grain in cows fed CTR or TRT diets and of incubation of 3 different substrates including CTR or barley grain steeped in 0.5% or 1.0% LA (TRT1 and TRT2, respectively) up to 72 h in the rumen. Results of the in vivo trial indicated that cows fed the TRT diet had greater rumen pH during most intensive fermentation phases at 10 and 12 h post-feeding. The latter effect was associated with a shorter duration in which rumen pH was below 5.8 for cows fed the TRT diet (2.4 h) compared with CTR diet (3.9 h). Furthermore, cows fed the TRT diet had lower concentrations of volatile fatty acids at 2 and 4 h post-feeding. In addition, concentrations of preprandial volatile fatty acids were lower in the rumen fluid of cows fed the TRT diet. Results also showed that molar proportion of acetate was lower, whereas propionate tended to increase by feeding cows the TRT diet. Cows fed the TRT diet demonstrated greater rumen in situ lag time of substrate DM degradation and a tendency to lower the fractional degradation rate. Other in situ results indicated a quadratic effect of LA on the effective rumen degradability of substrates whereby the latter variable was decreased from CTR to TRT1 but increased for TRT2 substrate. Although the diet did not affect actual milk yield, fat-corrected milk, percentages of milk protein, and lactose and concentration of milk urea nitrogen, cows fed the TRT diet increased milk fat content and tended to increase fat:protein ratio in the milk. In conclusion, results demonstrated that treatment of barley grain with LA lowered the risk of subacute rumen acidosis and maintained high milk fat content in late-lactating Holstein cows fed diets based on barley grain.     

Direct starch conversion into L‐(+)‐lactic acid by a novel amylolytic strain of Lactobacillus paracasei B41

A new amylolytic strain of Lactobacillus paracasei able to convert starch directly into L ‐(+)‐lactic acid (LA) was isolated. The identification of the by 16S rDNA sequencing proved that this strain, B41, is the first amylolytic representative of Lactobacillus casei group. The amylase activity assay revealed that L. paracasei B41 produced extracellular amylolytic enzyme, reaching 62 U/mL in the cell‐free supernatants. The optimal conditions for its action were pH 5.0 and temperature 45°C. The gene amy1 (1779 bp) encoding the putative B41 amylopullulanase was cloned, sequenced, and analyzed. The deduced protein contained a leader peptide of 28 amino acids and a mature peptide of 564 amino acids. Compared to the amylases of closely related species, B41 enzyme had several amino acid substitutions. An inducible control at amy1 expression was demonstrated. The starch fermentation abilities of L. paracasei B41 were studied in batch processes performed with and without pH control. The highest amount of LA from starch was obtained during 48 h fermentation from 40 g/L substrate at pH maintained at 5.0–37.3 g/L. In addition, 93.3% starch conversion into LA and the highest reported productivity for 24 h were achieved – 1.30 g/L/h.     

Production of α‐amylase by Aspergillus oryzae As 3951 in solid state fermentation using spent brewing grains as substrate

BACKGROUND: The optimisation of nutrient levels for the production of α‐amylase by Aspergillus oryzae As 3951 in solid state fermentation (SSF) with spent brewing grains (SBG), an inexpensive substrate and solid support, was carried out using response surface methodology (RSM) based on Plackett–Burman design (PBD) and Box–Behnken design (BBD). RESULTS: In the first optimisation step a PBD was used to evaluate the influences of related factors. Corn steep liquor, CaCl₂ and MgSO₄ were found to be the most compatible supplements to the substrate of SBG and influenced α‐amylase activity positively. In the second step the concentrations of these three nutrients were optimised using a BBD. The final concentrations (g/g dry substrate basis) in the medium optimised with RSM were 1.8% corn steep liquor, 0.22% CaCl₂ and 0.2% MgSO₄ · 7H₂O using SBG as the solid substrate. The average α‐amylase activity reached 6186 U g^?1 dry substrate under the optimised conditions at 30 °C after 96 h. Under the optimised conditions of SSF an approximately 17.5% increase in enzyme yield was observed. CONCLUSION: SBG was found to be a good substrate for the production of α‐amylase by A. oryzae As 3951 under SSF. Copyright © 2007 Society of Chemical Industry     

Influence of malting and lactic acid fermentation on functional bioactive components in cereal‐based raw materials: a review paper

Driven by increased health awareness among consumers, the production of foods and beverages enriched with functional bioactive components is gaining more attention. Malting and lactic acid fermentation are biotechnological processes having potential for producing functional foods and beverages. Due to various biochemical and enzymatic induced changes in raw materials, malting of cereal grains and probiotic lactic acid fermentation of plant‐based media increases the nutritional quality of treated raw materials. The improved nutritional quality is attributed to the accumulation of functional bioactive components along with the degradation of anti‐nutritional components. The selection of raw materials and process parameters are important factors to be considered for increasing the functional bioactive components such as dietary fibres, antioxidants and probiotics. This review article reports the current knowledge on the changes of bioactive components during malting and lactic acid fermentation using probiotic bacterial strains. Process parameters which affect the concentration of bioactive components in raw materials will also be described.     

Interaction between lactic acid bacteria and food‐borne pathogens on putrescine production in ornithine‐enriched broth

Interactions of mixed cultures [lactic acid bacteria (LAB) and food‐borne pathogens (FBP)] on putrescine (PUT) as well as other biogenic amines (BAs) production were investigated in ornithine‐enriched broth. Significant differences in BAs production were found among the bacterial strains (P < 0.05). Conversion of ornithine into PUT by Salmonella Paratyphi A and Aeromonas hydrophila as well as Listeria monocytogenes and Staphylococcus aureus was high (>75 mg L^?1), whereas other bacterial strains yielded below 50 mg L^?1 of PUT. LAB strains resulted in significant reduction in PUT by Pseudomonas aeruginosa and Enterobacteriaceae, except for Escherichia coli, which was stimulated more than two‐fold PUT in the presence of Lactococcus lactis subsp. lactis. Lactobacillus plantarum had generally inhibition effect on histamine (HIS) and tyramine production by FBP, whereas Lc. lactic subsp. lactic slightly stimulated HIS by E. coli and A. hydrophila. Streptococcus thermophilus resulted in 1.5‐fold higher HIS formation by bacteria (10 mg L^?1). Consequently, the interaction between LAB and specific FBP might result in significant inhibition of amine accumulation, if the correct LAB strains are used.     

Selection of γ‐aminobutyric acid‐producing lactic acid bacteria and their potential as probiotics for use as starter cultures in Thai fermented sausages (Nham)

To produce a novel product of Thai fermented meats, such as healthy Nham, 602 lactic acid bacteria were isolated from fermented foods and fourteen isolates that produced high amounts of γ‐aminobutyric acid (GABA) were also tested for inhibiting the seven foodborne bacteria. Four selected isolates produced GABA in the range of 7339–9060 mg L^?1 (NH2 < NH102 < NH116 < HN8) and their culture filtrates with a pH of 3.8, except 4.2 for NH2 inhibited all target organisms. Selected strains were investigated for their physiological and functional properties of probiotics in vitro. Strain NH102 survived best in the gastrointestinal tract with only a one log cell decrease over 4 h, whereas strain NH116 was the best cholesterol removal (43%). None showed any haemolysis and all hydrolased bile salts. Strain NH2 was identified as Lactobacillus namurensis and all the others were Pediococcus pentosaceus. Strains HN8 and NH2 were potential starters for fermenting meats.     

Determination of γ‐aminobutyric acid in Chinese rice wines and its evolution during fermentation

γ‐Aminobutyric acid (GABA) is a functional amino acid that is widely present in Chinese rice wine. In this study, high‐performance liquid chromatography coupled with ultraviolet detection (HPLC‐UV) was established for the determination of γ‐aminobutyric acid in 22 Chinese rice wines collected from the Shaoxing region of China. Furthermore, the evolution of GABA was studied in Chinese rice wine during primary and post‐fermentation process. Results showed that the HPLC method was reliable with good linearity, accuracy, precision and stability. Additionally, the GABA content varied significantly in the 22 Chinese rice wines, and the content was much higher in wine samples with long aging periods. Regarding the evolution of GABA in Chinese rice wine during the brewing process, the level slowly increased during primary fermentation. A decrease in GABA was observed in the wine at the early stage of the post‐fermentation process. However, a marked increase on the GABA content occurred in wine at the late stage of post‐fermentation. The findings from this study are that HPLC can be successfully applied to determine GABA in Shaoxing brewed rice wines, and further provide useful information on quality control of such wines. Copyright © 2017 The Institute of Brewing & Distilling     

Different response to acetic acid stress in Saccharomyces cerevisiae wild‐type and l‐ascorbic acid‐producing strains

Biotechnological processes are of increasing significance for industrial production of fine and bulk chemicals, including biofuels. Unfortunately, under operative conditions microorganisms meet multiple stresses, such as non‐optimal pH, temperature, oxygenation and osmotic stress. Moreover, they have to face inhibitory compounds released during the pretreatment of lignocellulosic biomasses, which constitute the preferential substrate for second‐generation processes. Inhibitors include furan derivatives, phenolic compounds and weak organic acids, among which acetic acid is one of the most abundant and detrimental for cells. They impair cellular metabolism and growth, reducing the productivity of the process: therefore, the development of robust cell factories with improved production rates and resistance is of crucial importance. Here we show that a yeast strain engineered to endogenously produce vitamin C exhibits an increased tolerance compared to the parental strain when exposed to acetic acid at moderately toxic concentrations, measured as viability on plates. Starting from this evidence, we investigated more deeply: (a) the nature and levels of reactive oxygen species (ROS); (b) the activation of enzymes that act directly as detoxifiers of reactive oxygen species, such as superoxide dismutase (SOD) and catalase, in parental and engineered strains during acetic acid stress. The data indicate that the engineered strain can better recover from stress by limiting ROS accumulation, independently from SOD activation. The engineered yeast can be proposed as a model for further investigating direct and indirect mechanism(s) by which an antioxidant can rescue cells from organic acid damage; moreover, these studies will possibly provide additional targets for further strain improvements. Copyright © 2013 John Wiley & Sons, Ltd.