Improvement of L(+)‐lactic acid production from cassava wastewater by Lactobacillus rhamnosus B 103

BACKGROUND: L (+)‐Lactic acid is used in the pharmaceutical, textile and food industries as well as in the synthesis of biodegradable plastics. The aim of this study was to investigate the effects of different medium components added in cassava wastewater for the production of L (+)‐lactic acid by Lactobacillus rhamnosus B 103. RESULTS: The use of cassava wastewater (50 g L^?1 of reducing sugar) with Tween 80 and corn steep liquor, at concentrations (v/v) of 1.27 mL L^?1 and 65.4 mL L^?1 respectively led to a lactic acid concentration of 41.65 g L^?1 after 48 h of fermentation. The maximum lactic acid concentration produced in the reactor after 36 h of fermentation was 39.00 g L^?1 using the same medium, but the pH was controlled by addition of 10 mol L^?1 NaOH. CONCLUSION: The use of cassava wastewater for cultivation of L. rhamnosus is feasible, with a considerable production of lactic acid. Furthermore, it is an innovative proposal, as no references were found in the scientific literature on the use of this substrate for lactic acid production. Copyright © 2010 Society of Chemical Industry     

Brewers' spent grain and thin stillage as raw materials in l‐(+)‐lactic acid fermentation

Brewer's spent grain (BSG) hydrolysates were used for l ‐(+)‐lactic acid (LA) fermentation by Lactobacillus rhamnosus ATCC 7469. In this study the effect of the addition of various amounts of thin stillage (TS) in BSG hydrolysate on LA fermentation parameters were evaluated. TS addition significantly increased utilization of glucose by up to 43.0%. In batch fermentation the highest LA concentration and volumetric productivity of 31.0 g/L, and 0.93 g/L/h, respectively, were obtained with the addition of 50% TS. L. rhamnosus cell viability also increased with the addition of 50% TS (by 2.4%). TS addition significantly increased free amino nitrogen concentration (by up to 209%) which is important for bacterial growth. A strong positive correlation between free amino nitrogen and LA concentration was noted. Compared with the results obtained in the batch fermentation (50% TS), significantly higher LA concentration, yield and volumetric productivity (54.8, 1.9 and 4.0%, respectively) were achieved in fed‐batch fermentation with glucose and TS addition. The results suggest that the combination of the by‐products of brewing and bioethanol industries could be suitable for LA production. 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     

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     

The effect of inoculation and additives on D(−)‐ and L(+)‐lactic acid production and fermentation quality of guineagrass (Panicum maximum Jacq) silage

Two experiments were carried out to study the influence of storage time, glucose and urea additions ( Experiment 1 ) and lactic acid bacteria inoculation with and without glucose addition ( Experiment 2 ) on the production of lactate isomers and fermentation quality of guineagrass (Panicum maximum Jacq) silage. All silages in both experiments were well preserved, as indicated by lower pH and little or no butyric acid. In Experiment 1 , addition of glucose or urea did not significantly affect the pH of silages (P > 0.05). Urea addition tended to reduce acetic acid content and greatly increased NH₃ − N content. L (+)‐Lactic acid was produced predominantly in the first 3 days of ensiling, but D (−)‐lactic acid increased gradually until 1 month after ensiling. Thereafter all silages became stable. In Experiment 2 , inoculation of Lactobacillus casei or L rhamnosus with or without glucose reduced D (−)‐lactic acid and increased L (+)‐lactic acid of silages. The proportions of L (+)‐lactic acid in these silages were higher than 80% of total lactic acid. L plantarum alone or in combination with glucose promoted D (−)‐lactic acid production and decreased the proportion of L (+)‐lactic acid. Glucose addition alone tended to reduce the proportion of L (+)‐lactic acid in both experiments. © 2000 Society of Chemical Industry     

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).     

Efficient production of L‐lactic acid by Crabtree‐negative yeast Candida boidinii

Industrial production of L ‐lactic acid, which in polymerized form as poly‐lactic acid is widely used as a biodegradable plastic, has been attracting world‐wide attention. By genetic engineering we constructed a strain of the Crabtree‐negative yeast Candida boidinii that efficiently produced a large amount of L ‐lactic acid. The alcohol fermentation pathway of C. boidinii was altered by disruption of the PDC1 gene encoding pyruvate decarboxylase, resulting in an ethanol production that was reduced to 17% of the wild‐type strain. The alcohol fermentation pathway of the PDC1 deletion strain was then successfully utilized for the synthesis of L ‐lactic acid by placing the bovine L ‐lactate dehydrogenase‐encoding gene under the control of the PDC1 promoter by targeted integration. Optimizing the conditions for batch culture in a 5 l jar‐fermenter resulted in an L ‐lactic acid production reaching 85.9 g/l within 48 h. This productivity (1.79 g/l/h) is the highest thus far reported for L ‐lactic acid‐producing yeasts. DDBJ/EMBL/GenBank nucleotide database with Accession Nos. AB440630 and AB440631. Copyright © 2009 John Wiley & Sons, Ltd.     

Optimisation of bambara groundnut water extract and skim milk composition as cryoprotectant for increasing cell viability of Lactobacillus spp. using response surface methodology

Four legume water extracts, that is bambara groundnut, soya bean, red kidney bean and black bean as well as skim milk, were examined for their effectiveness in protecting Lactobacillus rhamnosus GG and Lactobacillus fermentum SK5 during the freeze‐drying and storage. Bambara groundnut water extract (BGWE) showed promising cryoprotective activity that was comparable to skim milk. BGWE and skim milk at 2–10% w/v and 5–20% w/v individually produced survival rates for both strains of 87–88%. To further optimise the synergistic cryoprotective medium, response surface methodology was employed. The optimal combination was 4.93% w/v BGWE and 11.68% w/v skim milk for L. rhamnosus GG and 5.17% w/v BGWE and 11.36% w/v skim milk for L. fermentum SK5 with survival rates of 95.17% and 94.36%, respectively. The storage life of freeze‐dried cells of both probiotics at 4 °C and 30 °C for 6 months was markedly improved when they were produced with these optimal combinations.     

Modeling and Optimization of Lactic Acid Production using Cashew Apple Juice as Substrate

The production of lactic acid by Lactobacillus casei B-442 was studied and modeled. Sugar feedstock was provided using cashew apple juice, an alternative glucose and fructose feedstock that proved to yield high concentrations of lactic acid. The fermentations were carried out in a 1-L fermenter under constant agitation (150 rpm) and controlled pH (6.5). Lactic acid production was evaluated through a dynamic study, varying the initial concentration of sugar in the range of 20 to 60 g/L. Biomass, reducing sugars, and lactic acid concentration were measured throughout the experiments. The highest production of lactic acid (59.3 g/L) was obtained operating the fermentation with 60 g/L of reducing sugar from the cashew apple juice. A rigorous kinetic model was developed for batch fermentation of cashew apple juice for lactic acid production by L. casei B-442. The growth of biomass and lactic acid production were affected by substrate limitation, substrate inhibition and lactic acid inhibition. The model assumed growth- and non-growth-associated lactic acid production and a term for microorganism death was also included in the model. Parameters of the kinetic model were determined based on experimental data by using the least mean squares method and Levenberg–Marquardt algorithm. The model validation was performed and the model was statistically able to fit the profiles for growth of biomass, sugar consumption and lactic acid production. The optimization of the process, using the model, was carried out and the optimum operating conditions aiming highest productivity, lowest production cost and highest gross profit are presented.     

Impact of buffering capacity on the acidification of wort by brewing‐relevant lactic acid bacteria

Acidified wort produced biologically using lactic acid bacteria (LAB) has application during sour beer production and in breweries adhering to the German purity law (Reinheitsgebot ). LAB cultures, however, suffer from end product inhibition and low pH, leading to inefficient lactic acid (LA) yields. Three brewing‐relevant LAB (Pediococcus acidilactici AB39, Lactobacillus amylovorus FST2.11 and Lactobacillus plantarum FST1.7) were examined during batch fermentation of wort possessing increasing buffering capacities (BC). Bacterial growth was progressively impaired when exposed to higher LA concentrations, ceasing in the pH range of 2.9–3.4. The proteolytic rest (50°C) during mashing was found to be a major factor improving the BC of wort. Both a longer mashing profile and the addition of an external protease increased the BC (1.21 and 1.24, respectively) compared with a control wort (1.18), and a positive, linear correlation (R ² = 0.957) between free amino nitrogen and BC was established. Higher levels of BC led to significant greater LA concentration (up to +24%) after 48 h of fermentation, reaching a maximal value of 11.3 g/L. Even higher LA (maximum 12.8 g/L) could be obtained when external buffers were added to wort, while depletion of micronutrient(s) (monosaccharides, amino acids and/or other unidentified compounds) was suggested as the cause of LAB growth cessation. Overall, a significant improvement in LA production during batch fermentation of wort is possible when BC is improved through mashing and/or inclusion of additives (protease and/or external buffers), with further potential for optimization when strain‐dependent nutritional requirements, e.g. sugar and amino acids, are considered. Copyright © 2017 The Institute of Brewing & Distilling     

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     

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 CaCO₃ was evaluated. Maximal lactic acid yield of 73.4%, as well as high biomass production (3 × 10⁸ CFU ml⁻¹) were achieved under selected conditions (41 °C, 5% (v/v) of inoculum, 1% (w/v) of CaCO₃, 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.     

The involvement of ATPase activity in the acid tolerance of Lactobacillus rhamnosus strain GG

This study examined the survival and acid stress response of nine lactic acid bacteria (LAB) including Lactobacillus rhamnosus strain GG in commercially available yoghurt products (low pH environment) during the long‐term storage. Under the storage conditions investigated (52 weeks period at 5 °C), there was a loss of the viability of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium sp. within 35 weeks. However, L. rhamnosus strain GG, one of starter strains in a commercially available yoghurt product, exhibited excellent survival throughout the whole storage period. Our results indicate that the viability of L. rhamnosus strain GG was increased by modulating the stress‐related factors as well as the activity of ATPase with exposure to the low pH conditions.     

Characterization of bacteriocin-like inhibitory substances (BLIS) from lactic acid bacteria isolated from traditional Azerbaijani cheeses

The lactic acid bacteria (LAB) of southern Caucasus region present a special interest due to the diversity of lactic flora used for fermentations by various local populations during thousands of years. Four LAB strains, not identified previously, isolated from Motal and Brunza typical Azerbaijani cheeses were subjected to phenotypic identification and three of them could be identified as Lactobacillus paracasei subsp. paracasei and one as Lactobacillus rhamnosus. Test strains such as Lactobacillus bulgaricus 340 and Saccharomyces cerevisiae were inhibited by the four isolated strains. Antibacterial activity against Escherichia coli HB 101 was detected in Lactobacillus paracasei BN ATS 8w and L. rhamnosus FAZ 16m. L. paracasei BN ATS 5w and 8w, and L. rhamnosus FAZ 16m showed inhibitory effect on Staphylococcus aureus Cip 9973. The inhibition of Candida pseudotropicalis was detected only when using L. paracasei species BN ATS 5w and 7w. Culture of Listeria innocua was insensitive to the antimicrobial substances of all the studied strains. Complete inactivation or significant reduction in antibacterial activity was observed after treatment of cell-free supernatants with pronase E and proteinase K, but not with trypsin (except for L. rhamnosus FAZ 16m), indicating the protein nature of the active agents. Amylase treatment totally inactivated the substances of L. paracasei, what implies the importance of glycosylation for the activity. The activities of all the bacteriocin-like substances from studied LABs were stable over a wide pH range from 3 to 11.     

Production of Citric Acid from Brewery Wastes by Surface Fermentation Using Aspergillus niger

The possible use of spent grain liquor and lager tank sediment as fermentation media for the production of citric acid by various Aspergillus niger strains has been investigated. A strain of A. niger ATCC 9142, on spent grain liquor, gave a citric acid concentration of 19 g/L, representing a yield of 78.5% (w/w) based on total reducing sugars consumed. On lager tank sediment 11.5 g/L citric acid were produced. The yield of citric acid, based on the total reducing sugars consumed, was 57.5% (w/w).     

Viability of the Lactobacillus rhamnosus HN001 Probiotic Strain in Swiss‐ and Dutch‐Type Cheese and Cheese‐Like Products

The objective of this study was to determine the viability of the probiotic Lactobacillus rhamnosus HN001 in Swiss‐type and Dutch‐type cheese and cheese‐like products (milk fat is substituted by stearin fraction of palm fat) during manufacture, ripening, and storage. The use of the probiotic L. rhamnosus HN001 in Dutch‐type cheese and cheese‐like products significantly (P = 0.1) changed their chemical composition (protein and fat content) and an insignificant increase (approximately 1.6% in cheese‐like products and approximately 0.3% in cheese) in yield. L. rhamnosus HN001 did not affect the rate of changes in the pH of ripened cheese and cheese‐like products. A minor increase in probiotic counts was observed in initial stages of production and were partially removed with whey. Ripened cheese and cheese‐like products were characterized by high survival rates of probiotic bacteria which exceeded 8 log CFU/g after ripening. An insignificant reduction in the number of viable probiotic cells was noted during storage of Swiss‐type and Dutch‐type cheese, whereas a significant increase in probiotic cell counts was observed in cheese‐like products during storage.     

Study on the Influence of Tea Extract on Probiotics in Skim Milk: From Probiotics Propagation to Metabolite

In this study, the influence of tea extract (TE) on the growth of probiotics in skim milk was examined. Lactobacillus plantarum ST‐III, Bifidobacterium bifidum Bb02, Lactobacillus acidophilus NCFM, and Lactobacillus rhamnosus GG were used in this study. The introduction of TE in milk significantly stimulated the propagation and acidification of L. rhamnosus GG and L. acidophilus NCFM. The antioxidant capacities and the total free amino acid contents of all fermented milk products were enhanced by the addition of TE; however, there were different antioxidant properties and free amino acid contents of fermented milk samples fermented by different bacteria. With a 9% (w/w) level, the fermentation with L. rhamnosus GG and L. acidophilus NCFM showed larger numbers of viable cells and faster acidifying rates, as well as excellent antioxidant capacity and abundant free amino acids.  The stimulative effects of TE on probiotics can be considered for industrial purposes and has practical implications for commercial applications.     

Cashew Apple Juice as Substrate for Lactic Acid Production

The aim of the present work was to investigate the use of cashew apple juice as a low cost substrate for Lactobacillus casei B-442 cultivation and lactic acid production. Ammonium sulfate was employed as the only exogenous nitrogen source. The effect of cashew apple juice reducing sugars and ammonium sulfate concentration and the fermentation pH and temperature on biomass formation, lactic acid production, and productivity were evaluated. The highest productivity (2.36 g/L.h) was obtained applying 50 g/L of reducing sugar from the cashew apple juice supplemented with 6 g/L of ammonium sulfate. The process yield was about 95% when fermentation was carried out at 37 °C with pH controlled at pH 6.5 using NaOH (120 g/L).     

The influence of calcium-carbonate and yeast extract addition on lactic acid fermentation of brewer's spent grain hydrolysate

Brewer's spent grain (BSG) is the major by-product of the brewing industry, representing around 85% of the total by-products generated. In this study BSG hydrolysate was produced using optimal conditions. Hydrolysates were used for lactic acid (LA) fermentation by Lactobacillus fermentum (PL-1) and Lactobacillus rhamnosus (ATCC 7469). The aim of this study was to evaluate possibilities of the BSG hydrolysate utilization as a substrate for LA fermentation. The effect of calcium-carbonate (2%) and yeast extract (0.5 to 5%) addition in hydrolysate on LA fermentation were investigated. The LA production by L. fermentum and L. rhamnosus in BSG hydrolysate was influenced by calcium-carbonate and yeast extract supplementation. L. fermentum produced a racemic mixture of L-(+)- and D-(−)-LA while L. rhamnosus produced mostly L-(+)-LA (95–98%) in all fermentations. Calcium-carbonate addition increased total LA yield by 13% in L. fermentum fermentations and by 17% in L. rhamnosus fermentations. Yeast extract addition increased total LA yield by 4–26% in L. fermentum fermentations and by 6–8% in L. rhamnosus fermentations.     

Alpha‐Cyclodextrin Production using Cyclodextrin Glycosyltransferase from Klebsiella pneumoniae AS‐22

Alpha‐cyclodextrin (α‐CD) production, using cyclodextrin glycosyltransferase (CGTase) from Klebsiella pneumoniae AS‐22, was investigated in the presence of various complexing agents using raw wheat starch and dextrin as substrates. The addition of many alcohols resulted in increased conversion of both raw wheat starch and dextrin to α‐CD. With 125 g/L raw wheat starch, 42.5% (w/w) conversion to CDs was obtained in the presence of 2% (v/v) butan‐1‐ol. The ratio of α:β‐CD formed was 97:3, with negligible formation of γ‐CD and malto‐oligosaccharides. The production of α‐CD was optimized using two‐level factorial designs in three variables: dextrin, hexan‐1‐ol and enzyme concentrations. Under optimum conditions, 12.1% (w/w) conversion of 500 g/L dextrin to total‐CDs was achieved. The ratio of α:β:γ‐CD formed was 91:3:6, with negligible production of malto‐oligosaccharides. This CGTase was strongly inhibited by α:‐CD; 50% inhibition of reaction rate was observed at an initial concentration of 4 g/L α:‐CD. The effectiveness of an ultrafiltration membrane bioreactor for continuous removal of product was also tested.