Intraoral administration of probiotics and postbiotics: An overview of microorganisms and formulation strategies

Abstract The last decade provided significant advances in the understanding of microbiota and its role in human health. Probiotics are live microorganisms with proven benefits for the host and were mostly studied in the context of gut health, but they can also confer significant benefits for oral health, mainly in the treatment of gingivitis. Postbiotics are cell-free extracts and metabolites of microorganisms which can provide additional preventive and therapeutic value for human health. This opens opportunities for new preventive or therapeutic formulations for oral administration. The microorganisms that colonize the oral cavity, their role in oral health and disease, as well as the probiotics and postbiotics which could have beneficial effects in this complex environment were discussed. The aim of this study was to review, analyse and discuss novel probiotic and postbiotic formulations intended for oral administration that could be of great preventive and therapeutic importance. A special attention has been put on the formulation of the pharmaceutical dosage forms that are expected to provide new benefits for the patients and technological advantages relevant for industry. An adequate dosage form could significantly enhance the efficiency of these products.

Blocking the 2,3-butanediol synthesis pathway of Klebsiella pneumoniae resulted in L-valine production.

Klebsiella pneumoniae is a 2,3-butanediol producing bacterium. Nevertheless, a design and construction of L-valine production strain was studied in this paper. The first step of 2,3-butanediol synthesis and branched-chain amino acid synthesis pathways share the same step of α-acetolactate synthesis from pyruvate. However, the two pathways are existing in parallel and do not interfere with each other in the wild-type strain. A knockout of budA blocked the 2,3-butanediol synthesis pathway and resulted in the L-valine production. The budA coded an α-acetolactate decarboxylase and catalyzed the acetoin formation from α-acetolactate. Furthermore, blocking the lactic acid synthesis by knocking out of ldhA, which is encoding a lactate dehydrogenase, improved the L-valine synthesis. 2-Ketoisovalerate is the precursor of L-valine, it is also an intermediate of the isobutanol synthesis pathway, while indole-3-pyruvate decarboxylase (ipdC) is responsible for isobutyraldehyde formation from 2-ketoisovalerate. Production of L-valine has been improved by knocking out of ipdC. On the other side, the ilvE, encoding a transaminase B, reversibly transfers one amino group from glutamate to α-ketoisovalerate. Overexpression of ilvE exhibited a distinct improvement of L-valine production. The brnQ encodes a branched-chain amino acid transporter, and L-valine production was further improved by disrupting brnQ. It is also revealed that weak acidic and aerobic conditions favor L-valine production. Based on these findings, L-valine production by metabolically engineered K. pneumonia was examined. In fed-batch fermentation, 22.4 g/L of L-valine was produced by the engineered K. pneumoniae ΔbudA-ΔldhA-ΔipdC-ΔbrnQ-ilvE after 55 h of cultivation, with a substrate conversion ratio of 0.27 mol/mol glucose.

Synergistic Effect of Enzyme Hydrolysis and Microwave Reactor Pretreatment as an Efficient Procedure for Gluten Content Reduction.

In this study, we assessed the effects of microwave irradiation of wheat gluten proteins as a pretreatment performed in a microwave reactor that could accurately control process parameters as a function of power and temperature, as well as comparing it with conventional heat treatment. The aim was to identify suitable combinations of partial enzymatic hydrolysis and microwave pretreatment parameters to produce gluten hydrolysates with reduced allergenicity and conserved techno-functional features for food application. FTIR analysis, and total and reactive SH group contents confirmed that the microwave-controlled heating can significantly change the secondary structure and conformation of gluten protein. The microwave treatment had the largest effect at 200 W and 100 °C, at which the content of gluten has been reduced by about 2.5-fold. The microwave pretreatment also accelerated the enzymatic hydrolysis of gluten, changing the kinetic profile. The apparent hydrolysis rate constants (k2) were 1.00, 3.68, 3.48, 4.64 and 4.17 min-1 for untreated gluten, and those pretreated with microwave power of 200, 400, 600 and 800 W, respectively. Compared to the heat treatment, it appeared that microwave specific non-thermal effects had a significant influence on the gluten structure and allergenicity and, in combination with the enzymatic hydrolysis, ultimately yielded protein hydrolysates with enhanced antioxidant and functional properties.

Ethylene glycol and glycolic acid production by wild-type Escherichia coli.

Ethylene glycol and glycolic acid are bulk chemicals with a broad range of applications. The ethylene glycol and glycolic acid biosynthesis pathways have been produced by microorganisms and used as a biological route for their production. Unlike the methods that use xylose or glucose as carbon sources, xylonic acid was used as a carbon source to produce ethylene glycol and glycolic acid in this study. Amounts of 4.2 g/L of ethylene glycol and 0.7 g/L of glycolic acid were produced by a wild-type Escherichia coli W3110 within 10 H of cultivation with a substrate conversion ratio of 0.5 mol/mol. Furthermore, E. coli strains that produce solely ethylene glycol or glycolic acid were constructed. 10.3 g/L of glycolic acid was produced by E. coli ΔyqhD+aldA, and the achieved conversion ratio was 0.56 mol/mol. Similarly, the E. coli ΔaldA+yqhD produced 8.0 g/L of ethylene glycol with a conversion ratio of 0.71 mol/mol. Ethylene glycol and glycolic acid production by E. coli on xylonic acid as a carbon source provides new information on the biosynthesis pathway of these products and opens a novel way of biomass utilization.

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.

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.

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.

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.

Lignin-Degrading Abilities of Novel Autochthonous FungalIsolates Trametes hirsuta F13 and Stereum gausapatum F28.

The aim of this research is to isolate and identify fungi with high lignin-degrading abilities that are autochthonous to southern Serbian region. Two novel fungal isolates identified as Trametes hirsuta F13 and Stereum gausapatum F28 were selected to assess their ligninolytic enzyme activities and the efficiency of lignin removal from beech wood sawdust. Obtained results show that both isolates are good sources of industrially valuable enzymes with a potential for application in various biotechnological and industrial processes. Both isolates showed laccase, manganese-dependent peroxidase, and versatile peroxidase activities, while only S. gausapatum F28 had lignin peroxidase activity. This is the first record of the ability of S. gausapatum species to produce lignin peroxidase. T. hirsuta F13 showed higher laccase activity than S. gausapatum F28, while S. gausapatum F28 had higher manganese peroxidase activity. Also, T. hirsuta F13 exhibited much higher laccase activity under submerged cultivation conditions than solid-state cultivation conditions, which is rare for fungi. This is important for industrial processes since the submerged fermentation is a dominant technique in industry. The test of the efficiency of lignin removal showed that both isolates are efficient lignin decomposers. After five weeks of incubation on beech wood sawdust, the total lignin losses were 33.84% with T. hirsuta F13 and 28.8% with S. gausapatum F28.

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.

NIR photo-driven upconversion in NaYF4:Yb,Er/PLGA particles for in vitro bioimaging of cancer cells.

Lanthanide-doped fluoride up-converting nanoparticles (UCNPs) represent the new class of imaging contrast agents which hold great potential for overcoming existing problems associated with traditionally used dyes, proteins and quantum dots. In this study, a new kind of hybrid NaYF4:Yb,Er/PLGA nanoparticles for efficient biolabeling were prepared through one-pot solvothermal synthesis route. Morphological and structural characteristics of the as-designed particles were obtained using X-ray powder diffraction (XRPD), scanning and transmission electron microscopy (SEM/TEM), energy dispersive spectroscopy (EDS), Fourier transform infrared (FTIR) and photoluminescence (PL) spectroscopy, while their cytotoxicity as well as up-conversion (UC) labeling capability were tested in vitro toward human gingival cells (HGC) and oral squamous cell carcinoma (OSCC). The results revealed coexistence of the cubic (Fm-3m) and hexagonal (P63/m) phase in spherical and irregularly shaped nanoparticles, respectively. PLGA [Poly(lactic-co-glycolic acid)] ligands attached at the surface of UCNPs particles provide their enhanced cellular uptake and enable high-quality cells imaging through a near-infrared (NIR) laser scanning microscopy (λex = 980 nm). Moreover, the fact that NaYF4:Yb,Er/PLGA UCNPs show low cytotoxicity against HGC over the whole concentration range (10-50 µg/mL) while a dose dependent viability of OSCC is obtained indicates that these might be a promising candidates for targeted cancer cell therapy.

One-step synthesis of amino-functionalized up-converting NaYF4:Yb,Er nanoparticles for in vitro cell imaging.

The emerging up-conversion nanoparticles (UCNPs) offer a wide range of biotechnology applications, from biomarkers and deep tissue imaging, to single molecule tracking and drug delivery. Their successful conjugation to biocompatible agents is crucial for specific molecules recognition and usually requires multiple steps which may lead to low reproducibility. Here, we report a simple and rapid one-step procedure for in situ synthesis of biocompatible amino-functionalized NaYF4:Yb,Er UCNPs that could be used for NIR-driven fluorescence cell labeling. X-ray diffraction showed that UCNPs synthesized through chitosan-assisted solvothermal processing are monophasic and crystallize in a cubic α phase. Scanning and transmission electron microscopy revealed that the obtained crystals are spherical in shape with a mean diameter of 120 nm. Photoluminescence spectra indicated weaker green (2H11/2, 4S3/2 → 4I15/2) and stronger red emission (4F9/2 → 4I15/2), as a result of enhanced non-radiative 4I11/2 → 4I13/2 Er3+ relaxation. The presence of chitosan groups at the surface of UCNPs was confirmed by Fourier transform infrared spectroscopy, thermogravimetry and X-ray photoelectron spectroscopy. This provides their enhanced internalization in cells, at low concentration of 10 µg ml-1, without suppression of cell viability after 24 h of exposure. Furthermore, upon 980 nm laser irradiation, the amino-functionalized NaYF4:Yb,Er UCNPs were successfully used in vitro for labeling of two human cell types, normal gingival and oral squamous cell carcinoma.

Production of bioethanol from pre-treated cotton fabrics and waste cotton materials.

This study highlights the potential of cotton fabric as a promising feedstock for the production of bioethanol as renewable biofuel. The effect of corona pre-treatment of non-mercerized and mercerized cotton fabrics on glucose and ethanol yield is discussed. Fermentation kinetics for ethanol production and the basic process parameters were assessed and compared. Corona pre-treatment of cotton fabrics led to an increase in the glucose yield (compared to control sample) during enzymatic hydrolysis, and consequently the ethanol yield during fermentation by yeast Saccharomyces cerevisiae var. ellipsoideus. The system with mercerized cotton fabric was found to be superior obtaining an ethanol productivity of 0.900g/Lh and ethanol yield of 0.94g/g (based on glucose) after 6h of fermentation time. The similar results were obtained during processing of waste cotton materials performed under the same process conditions. The obtained results showed that cotton fabric could become an alternative feedstock for the bioethanol production. For potential industrial implementation the waste mercerized cotton scraps would be the materials of choice.

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.

Two-stage fermentation for 2-Ketogluconic acid production by Klebsiella pneumoniae.

2-Ketogluconic acid production by Klebsiella pneumoniae is a pH-dependent process, strictly proceeding under acidic conditions. Unfortunately, cell growth is inhibited by acidic conditions, resulting in low productivity of 2-ketogluconic acid. To overcome this deficiency, a two-stage fermentation strategy was exploited in the current study. During the first stage, the culture was maintained at neutral pH, favoring cell growth. During the second stage, the culture pH was switched to acidic conditions favoring 2-ketogluconic acid accumulation. Culture parameters, including switching time, dissolved oxygen levels, pH, and temperature were optimized for the fed-batch fermentation. Characteristics of glucose dehydrogenase and gluconate dehydrogenase were revealed in vitro, and the optimal pHs of the two enzymes coincided with the optimum culture pH. Under optimum conditions, a total of 186 g/l 2- ketogluconic acid was produced at 26 h, and the conversion ratio was 0.98 mol/mol. This fermentation strategy has successfully overcome the mismatch between optimum parameters required for cell growth and 2-ketogluconic acid accumulation, and this result has the highest productivity and conversion ratio of 2-ketogluconic and produced by microorganism.

Suitability of some selected maize hybrids from Serbia for the production of bioethanol and dried distillers' grains with solubles.

BACKGROUND: Bioethanol is mostly produced from starchy parts of the corn grain kernel leaving significant amounts of valuable by-products such as dried distillers' grains with solubles (DDGS) which can be used as a substitute for traditional feedstuff. The suitability of six maize hybrids from Serbia was investigated for bioethanol and DDGS production. The correlation between physical and chemical characteristics of the grain, bioethanol yield and quality of the corresponding DDGS was assessed. RESULTS: All hybrids had very different chemical composition and physical characteristics which could allow various applications. The highest bioethanol yield (94.5% of theoretical) and volumetric productivity (2.01 g l(-1) h(-1)) were obtained with hybrid ZP 434 and the lowest with ZP 611k. Regarding chemical composition, all DDGS samples manifested good properties as feed components. Their protein content was higher compared to the kernel. In addition, the samples showed high digestibility and high mineral content, especially of calcium and phosphorus. CONCLUSION: A hybrid ZP 434 was selected as the most promising bioethanol producer. This property is attributed to the highest level of soft endosperm which is more susceptible to starch-hydrolysing enzymes. A high yield potential per hectare makes it the best candidate for commercial bioethanol production.

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.