Deep eutectic solvent pretreatment of cork dust - Effects on biomass composition, phenolic extraction and anaerobic degradability.

In this study, phenolic compounds using deep eutectic solvents (DES) were extracted from cork dust, and the biogas production potential of DES-treated cork dust samples was determined. The DES treatment was carried out using choline chloride and formic acid (1:2 M ratio) at various temperatures (90, 110 and 130 °C) and treatment times (20, 40 and 60 min) at a solid-to-solvent ratio of 1:10 g mL-1. The highest total phenolic content (137 mg gallic acid equivalent (GAE) g-1 dry cork dust) was achieved at 110 °C/20 min. The extracts exhibited an antioxidant capacity of up to 56.3 ± 3.1 % 1,1-diphenyl-2-picrylhydazyl (DPPH) inhibition at a dilution rate of 100. DES treatment resulted in minimal sugar solubilization at low temperatures, while approximately 42 % of the xylan fraction in the biomass degraded under severe conditions (e.g., 130 °C/60 min). Catechin, 4-hydroxybenzoic acid and gallic acid were the major phenolics in DES extracts. The biogas yield of DES-treated cork dust increased with treatment severity. The highest biogas yield (115.1mLN gVS-1) was observed at 130 °C/60 min, representing an increase of 125 % compared to the untreated sample. SEM images revealed that the surface structure of the samples became smoother after mild pretreatment and rougher after harsh pretreatment. Compositional and FTIR analyses indicated that a higher biogas formation potential was associated with increased cellulose content in the substrate, which could be attributed to hemicellulose solubilization in the hydrolysate. Overall, DES pretreatment effectively enhanced phenol extraction and anaerobic degradability.

Influence of different culture conditions on exopolysaccharide production by indigenous lactic acid bacteria isolated from pickles.

The objective of this study was to assess the effects of some culture conditions [temperature (20, 30, 37 °C), incubation time (48, 72, 120 h), pH (5.0, 6.0, 7.0), NaCl concentration (0, 3, 6%), carbon (glucose, fructose, lactose), nitrogen (sodium nitrate, ammonium sulfate, bacto-peptone), and mineral sources (calcium carbonate, ferric chloride)] on the exopolysaccharide (EPS) production by lactic acid bacteria (LAB) strains (belonging to Lactobacillus (L.) plantarum, L. namurensis, and Pediococcus (P.) ethanolidurans species) isolated from naturally fermented pickles. The maximum EPS production was determined at 30 °C and pH 6.0. The highest amount of EPS was obtained after 120 h of incubation, with glucose as carbon source, bacto-peptone as nitrogen source and calcium carbonate as mineral source for most of the tested strains. The EPS formation was not stimulated by NaCl, indicating that EPS formation of the tested strains was not a stress response. L. plantarum MF460 produced the highest amount of EPS at 30 °C after 48 h of incubation, which was 515.48 mg/L. One of the most pronounced results of this study was that the EPS production of L. plantarum MF556 strain was increased up to 512.81 mg/L with the addition of calcium carbonate to MRS medium. The effect of different culture conditions, particularly of incubation time, carbon, nitrogen, and mineral sources, on the EPS production often vary depending on the strain. Therefore, these apparent strain specific results demonstrated that the optimum culture conditions for the enhanced EPS production should be specifically determined for each LAB strain.

Ionic liquid pretreatment of yellow pine followed by enzymatic hydrolysis and fermentation.

In this study, it was aimed to enhance the enzymatic hydrolysis of recalcitrant yellow pine wood and subsequently to produce ethanol efficiently through the application of 1-ethyl-3-methylimidazolium acetate (EMIMAc) ionic liquid. The effect of biomass particle size (<2.5 mm, 500-850 µm, 363-500 µm, and <363 µm) prior to pretreatment and pretreatment time (15, 30, and 45 min) on the structural properties and enzymatic hydrolysis yield of EMIMAc pretreated pine wood were also investigated. In general, EMIMAc functioned more efficiently at larger biomass particle sizes compared to smaller ones. Dissolution of solids by pretreatment, amount of lignin extracted from biomass as well as biomass to glucose yield was increased with increasing pretreatment time. The conversion of biomass to glucose was 56% under the selected optimum conditions (500-850 µm particle size; 5% biomass concentration and 140°C-45 min EMIMAc pretreatment). The biomass samples pretreated at the optimal conditions were subjected to simultaneous saccharification and fermentation (SSF) and separate hydrolysis and fermentation (SHF) which resulted in ethanol yields of 86 and 96%, respectively. The results presented in this study give a general framework for reducing the recalcitrance of pine to enzymatic saccharification and microbial fermentation by EMIMAc pretreatment without need for long pretreatment times and size reduction. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018.

Optimization of organic acid pretreatment of wheat straw.

The objective of this study was to determine the effectiveness of different organic acids (maleic, succinic, and oxalic acid) on enzymatic hydrolysis and fermentation yields of wheat straw. It was also aimed to optimize the process conditions (temperature, acid concentration, and pretreatment time) by using response surface methodology (RSM). In line with this objective, the wheat straw samples were pretreated at three different temperatures (170, 190, and 210°C), acid concentrations (1%, 3%, and 5%) and pretreatment time (10, 20, and 30 min). The findings show that at extreme pretreatment conditions, xylose was solubilized in liquid phase, causing an increase in cellulose and lignin content of biomass. Enzymatic hydrolysis experiments revealed that maleic and oxalic acids were quite effective at achieving high sugar yields (>90%) from wheat straw. In contrast, the highest sugar yields were 50-60%, when the samples were pretreated with succinic acid, indicating that succinic acid was not as effective. The optimum process conditions for maleic acid were, 210°C, 1.08% acid concentration, and 19.8 min; for succinic acid 210°C, 5% acid concentration, and 30 min; for oxalic acid 210°C, 3.6% acid concentration, and 16.3 min. The ethanol yields obtained at optimum conditions were 80, 79, and 59% for maleic, oxalic and succinic acid, respectively. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1487-1493, 2016.

In Vitro Properties of Potential Probiotic Indigenous Lactic Acid Bacteria Originating from Traditional Pickles.

The suitable properties of potential probiotic lactic acid bacteria (LAB) strains (preselected among 153 strains on the basis of their potential technological properties) isolated from traditional Çubuk pickles were examined in vitro. For this purpose, these strains (21 Lactobacillus plantarum, 11 Pediococcus ethanolidurans, and 7 Lactobacillus brevis) were tested for the ability to survive at pH 2.5, resistance to bile salts, viability in the presence of pepsin-pancreatin, ability to deconjugate bile salts, cholesterol assimilation, and surface hydrophobicity properties. Most of the properties tested could be assumed to be strain-dependent. However, L. plantarum and L. brevis species were found to possess desirable probiotic properties to a greater extent compared to P. ethanolidurans. In contrast to P. ethanolidurans strains, the tested L. plantarum and L. brevis strains exhibited bile salt tolerance, albeit to different extent. All tested strains showed less resistance to intestinal conditions than gastric juice environment. Based on the survival under gastrointestinal conditions, 22 of the 39 strains were selected for further characterization. The eight strains having the highest cholesterol assimilation and surface hydrophobicity ratios could be taken as promising probiotic candidates for further in vivo studies, because of the strongest variations found among the tested strains with regard to these properties.

Phenotypic and genotypic identification of lactic acid bacteria isolated from traditional pickles of the Çubuk region in Turkey.

A total of 152 lactic acid bacteria (LAB) were isolated from pickles produced in the Ankara-Çubuk region. These isolates were clustered into eight groups on the basis of their phenotypic characteristics including cell morphology, CO2 production from glucose, growth at 10 and 45 °C, growth in 6.5 % NaCl, and growth at pH 9.6. API 50 CH carbohydrate fermentation test, 16S ribosomal RNA (rRNA) sequence analysis, and sodium dodecyl sulfate-acrylamide gel electrophoresis (SDS-PAGE) whole-cell protein profile analysis were also performed for precise identification of the isolates at the species level. Molecular identification revealed that the most prevalent LAB species involved in pickle fermentation were Pediococcus ethanolidurans (46 isolates, 30.3 %), Lactobacillus brevis (37 isolates, 24.3 %), Lactobacillus plantarum (37 isolates, 24.3 %), and Lactobacillus buchneri (15 isolates, 9.9 %). Other LAB were found in minor frequencies such as Pediococcus parvulus (8 isolates, 5.3 %), Lactobacillus namurensis (6 isolates, 3.9 %), Lactobacillus diolivorans (1 isolate, 0.7 %), Lactobacillus parabrevis (1 isolate, 0.7 %), and Enterococcus casseliflavus (1 isolate, 0.7 %). When results of phenotypic and genotypic identification methods were compared, differences in the species distribution of LAB associated with pickles were defined between the API and the 16S rRNA sequencing. The API 50 CHL test coincided with the 16S rRNA results in 71 out of the 152 tested isolates, indicating that API gave unreliable identification results. A clear correlation could not be found between the results of whole-cell SDS profiles and 16S rRNA sequencing. Therefore, molecular characterization by 16S rRNA sequencing was considered to be the most reliable method for identifying isolates. The results presented in this work provide insight in to the LAB population associated with traditional Çubuk pickles and constitute a LAB strain resource for further studies involving the development of starter cultures.

Effectiveness of chitosan against wine-related microorganisms.

The antimicrobial action of chitosan against wine related microorganisms, including Lactobacillus plantarum, Saccharomyces cerevisiae, Oeonococcus oeni, Lactobacillus hilgardii, Brettanomyces bruxellensis, Hanseniaspora uvarum and Zygosaccharomyces bailii was examined in laboratory media. In order to assess the potential applicability of chitosan as a microbial control agent for wine, the effect of chitosan, applied individually and/or in combination with sulphur dioxide (SO2), on the growth of microorganisms involved in various stages of winemaking and on the fermentative performance of S. cerevisiae was investigated. Of the seven wine-related microorganisms studied, S. cerevisiae exhibited the strongest resistance to antimicrobial action of chitosan in laboratory media with a minimum inhibitory concentration (MIC) greater than 2 g/L. L. hilgardii, O. oeni and B. bruxellensis were the most susceptible to chitosan since they were completely inactivated by chitosan at 0.2 g/L. The MIC of chitosan for L. plantarum, H. uvarum and Z. bailii was 2, 0.4 and 0.4 g/L, respectively. In wine experiments, it was found that chitosan had a retarding effect on alcoholic fermentation without significantly altering the viability and the fermentative performance of S. cerevisiae. With regard to non-Saccharomyces yeasts (H. uvarum and Z. bailii) involved in winemaking, the early deaths of these yeasts in mixed cultures with S. cerevisiae were not probably due to the antimicrobial action of chitosan but rather due to ethanol produced by the yeasts. The complex interactions between chitosan and wine ingredients as well as microbial interactions during wine fermentation considerably affect the efficacy of chitosan. It was concluded that chitosan was worthy of further investigation as an alternative or complementary preservative to SO2 in wine industry.

Technological properties of indigenous wine yeast strains isolated from wine production regions of Turkey.

The purpose of this study was to evaluate the important technological and fermentative properties of wine yeast strains previously isolated from different wine producing regions of Turkey. The determination of the following important properties was made: growth at high temperatures; fermentative capability in the presence of high sugar concentration; fermentation rate; hydrogen sulfide production; killer activity; resistance to high ethanol and sulfur dioxide; foam production; and enzymatic profiles. Ten local wine yeast strains belonging to Saccharomyces, and one commercial active dry yeast as a reference strain were evaluated. Fermentation characteristics were evaluated in terms of kinetic parameters, including ethanol yield (YP/S), biomass yield (YX/S), theoretical ethanol yield (%), specific ethanol production rate (qp; g/gh), specific glucose uptake rate (qs; g/gh), and the substrate conversion (%). All tested strains were able to grow at 37 °C and to start fermentation at 30° Brix, and were resistant to high concentrations of sulfur dioxide. 60 % of the strains were weak H2S producers, while the others produced high levels. Foam production was high, and no strains had killer activity. Six of the tested strains had the ability to grow and ferment at concentrations of 14 % ethanol. Except for one strain, all fermented most of the media sugars at a high rate, producing 11.0-12.4 % (v/v) ethanol. Although all but one strain had suitable characteristics for wine production, they possessed poor activities of glycosidase, esterase and proteinase enzymes of oenological interest. Nine of the ten local yeast strains were selected for their good oenological properties and their suitability as a wine starter culture.