Evaluation of oil palm front hydrolysate as a novel substrate for 2,3-butanediol production using a novel isolate Enterobacter cloacae SG1

The present work deals the production of 2,3-butanediol, an industrially important chemical, through biological route using a novel bacterial isolate. Batch fermentation trials for the production of 2,3-butanediol were carried out using the isolated strain Enterobacter cloacae SG-1. The study resulted 14.67 g/l of 2,3-butanediol with 48.9% yield using glucose as the carbon source. In order to replace the expensive glucose in the production media, non-detoxified oil palm frond hydrolysate was used as the carbon source and it resulted 2,3-butanediol yield of 7.67 g/l. Process parameters like pH, temperature and initial sugar concentration were optimized. The ability of strain E. cloacae SG-1 for utilization various pentoses and hexoses were evaluated and found that the strain can utilize both arabinose and glucose with a comparable 2,3-butanediol yield.     

Fuel ethanol production from steam-pretreated corn stover using SSF at higher dry matter content

Replacing fossil fuels by bio-fuels has many advantages, such as the reduction of CO₂-emission to the atmosphere, the possibility for non-oil-producing countries to be self-sufficient in fuel, and increased local job opportunities. Bio-ethanol is such a promising renewable fuel. However, today it is produced from sugar or starch—raw materials that are relatively expensive. To lower the production cost of bio-ethanol the cost of the raw material must be reduced and the production process made more efficient. The production of bio-ethanol from corn stover using simultaneous saccharification and fermentation (SSF) at high dry matter content addresses both issues. Corn stover is an agricultural by-product and thus has a low economic value. SSF at high dry matter content results in a high ethanol concentration in the fermented slurry, thereby decreasing the energy demand in the subsequent distillation step.In this study, SSF was performed on steam-pretreated corn stover at 5, 7.5 and 10% water-insoluble solids (WIS) with 2 g/L hexose-fermenting Saccharomyces cerevisiae (ordinary compressed baker's yeast). SSF at 10% WIS resulted in an ethanol yield of 74% based on the glucose content in the raw material and an ethanol concentration of 25 g/L. Neither higher yeast concentration (5 g/L) nor yeast cultivated on the liquid after the pretreatment resulted, under these conditions, in a higher overall ethanol yield.     

Feasibility of biohydrogen production from Gelidium amansii

The feasibility of hydrogen production from red algae was investigated. Galactose, the main sugar monomer of red algae, was readily converted to hydrogen by dark fermentation. The maximum hydrogen production rate and yield of galactose were 2.46 L H₂/g VSS/d and 2.03 mol H₂/mol galactose_added, respectively, which were higher than those for glucose (0.914 L H₂/g VSS/d and 1.48 mol H₂/mol galactose_added). The distribution of soluble byproducts showed that H₂ production was the main pathway of galactose uptake. 5-HMF, the main byproduct of acid hydrolysis of red algae causes noncompetitive inhibition of H₂ fermentation. 1.37 g/L of 5-HMF decreased hydrogen production rate by 50% compared to the control. When red algae was hydrolyzed at 150 °C for 15 min and detoxified by activated carbon, 53.5 mL of H₂ was produced from 1 g of dry algae with a hydrogen production rate of 0.518 L H₂/g VSS/d. Red algae, cultivable on vast tracts of sea by sunlight without any nitrogen-based fertilizer, could be a suitable substrate for biohydrogen production.     

Fermentative hydrogen production by a new alkaliphilic Clostridium sp. (strain PROH2) isolated from a shallow submarine hydrothermal chimney in Prony Bay,New Caledonia

The hydrogen-producing strain PROH2 pertaining to the genus Clostridium was successfully isolated from a shallow submarine hydrothermal chimney (Prony Bay, New Caledonia) driven by serpentinization processes. Cell biomass and hydrogen production performances during fermentation by strain PROH2 were studied in a series of batch experiments under various conditions of pH, temperature, NaCl and glucose concentrations. The highest hydrogen yield, 2.71 mol H₂/mol glucose, was observed at initial pH 9.5, 37 °C, and glucose concentration 2 g/L, and was comparable to that reported for neutrophilic clostridial species. Hydrogen production by strain PROH2 reached the maximum production rate (0.55 mM-H₂/h) at the late exponential phase. Yeast extract was required for growth of strain PROH2 and improved significantly its hydrogen production performances. The isolate could utilize various energy sources including cellobiose, galactose, glucose, maltose, sucrose and trehalose to produce hydrogen. The pattern of end-products of metabolism was also affected by the type of energy sources and culture conditions used. These results indicate that Clostridium sp. strain PROH2 is a good candidate for producing hydrogen under alkaline and mesothermic conditions.     

Cellulase production using biomass feed stock and its application in lignocellulose saccharification for bio-ethanol production

A major constraint in the enzymatic saccharification of biomass for ethanol production is the cost of cellulase enzymes. Production cost of cellulases may be brought down by multifaceted approaches which include the use of cheap lignocellulosic substrates for fermentation production of the enzyme, and the use of cost efficient fermentation strategies like solid state fermentation (SSF). In the present study, cellulolytic enzymes for biomass hydrolysis were produced using solid state fermentation on wheat bran as substrate. Crude cellulase and a relatively glucose tolerant BGL were produced using fungi Trichoderma reesei RUT C30 and Aspergillus niger MTCC 7956, respectively. Saccharification of three different feed stock, i.e. sugar cane bagasse, rice straw and water hyacinth biomass was studied using the enzymes. Saccharification was performed with 50 FPU of cellulase and 10 U of β-glucosidase per gram of pretreated biomass. Highest yield of reducing sugars (26.3 g/L) was obtained from rice straw followed by sugar cane bagasse (17.79 g/L). The enzymatic hydrolysate of rice straw was used as substrate for ethanol production by Saccharomyces cerevisiae. The yield of ethanol was 0.093 g per gram of pretreated rice straw.     

Pretreatment and hydrolysis of cellulosic agricultural wastes with a cellulase-producing Streptomyces for bioethanol production

Production of reducing sugar by hydrolysis of corncob material with Streptomyces sp. cellulase and ethanol fermentation of cellulosic hydrolysate was investigated. Cultures of Streptomyces sp. T3-1 improved reducing sugar yields with the production of CMCase, Avicelase and ??-glucosidase activity of 3.8, 3.9 and 3.8 IU/ml, respectively. CMCase, Avicelase, and ??-glucosidase produced by the Streptomyces sp. T3-1 favored the conversion of cellulose to glucose. It was recognized that the synergistic interaction of endoglucanase, exoglucanase and ??-glucosidase resulted in efficient hydrolysis of cellulosic substrate. After 5 d of incubation, the overall reducing sugar yield reached 53.1 g/100 g dried substrate. Further fermentation of cellulosic hydrolysate containing 40.5 g/l glucose was performed using Saccharomyces cerevisiae BCRC 21812, 14.6 g/l biomass and 24.6 g/l ethanol was obtained within 3 d. The results have significant implications and future applications regarding to production of fuel ethanol from agricultural cellulosic waste.     

Bioethanol production from mahula (Madhuca latifolia L.) flowers by solid-state fermentation

There is a growing interest worldwide to find out new and cheap carbohydrate sources for production of bioethanol. In this context, the production of ethanol from mahula (Madhuca latifolia L.) flowers by Saccharomyces cerevisiae in solid-state fermentation was investigated. The moisture level of 70%, pH of 6.0 and temperature of 30 °C were found optimum for maximum ethanol concentration (225.0 ± 4.0 g/kg flower) obtained from mahula flowers after 72 h of fermentation. Concomitant with highest ethanol concentration, the maximum ethanol productivity (3.13 g/kg flower/h), yeast biomass (18.5 × 10⁸ CFU/g flower), the ethanol yield (58.44 g/100 g sugar consumed) and the fermentation efficiency (77.1%) were also obtained at these parametric levels.     

Biological hydrogen production by extremely thermophilic novel bacterium Thermoanaerobacter mathranii A3N isolated from oil producing well

Hydrogen producing novel bacterial strain was isolated from formation water from oil producing well. It was identified as Thermoanaerobacter mathranii A3N by 16S rRNA gene sequencing. Hydrogen production by novel strain was pH and substrate dependent and favored pH 8.0 for starch, pH 7.5 for xylose and sucrose, pH 8.0–9.0 for glucose fermentation at 70 °C. The highest H₂ yield was 2.64 ± 0.40 mol H₂ mol glucose at 10 g/L, 5.36 ± 0.41 mol H₂ mol – sucrose at 10 g/L, 17.91 ± 0.16 mmol H₂ g – starch at 5 g/L and 2.09 ± 0.21 mol H₂ mol xylose at 5 g/L. The maximum specific hydrogen production rates 6.29 (starch), 9.34 (sucrose), 5.76 (xylose) and 4.89 (glucose) mmol/g cell/h. Acetate-type fermentation pathway (approximately 97%) was found to be dominant in strain A3N, whereas butyrate formation was found in sucrose and xylose fermentation. Lactate production increased with high xylose concentrations above 10 g/L.     

Optimization of bioethanol production from glycerol by Escherichia coli SS1

Bioethanol is a promising biofuel and has a lot of great prospective and could become an alternative to fossil fuels. Ethanol fermentation using glycerol as carbon source was carried out by local isolate, ethanologenic bacterium Escherichia coli SS1 in a close system. Factors affecting bioethanol production from pure glycerol were optimized via response surface methodology (RSM) with central composite design (CCD). Four significant variables were found to influence bioethanol yield; initial pH of fermentation medium, substrate concentration, salt content and organic nitrogen concentration with statistically significant effect (p ≤ 0.05) on bioethanol production. The significant factor was then analyzed using central composite design (CCD). The optimum conditions for bioethanol production were substrate concentration at 34.5 g/L, pH 7.61, and organic nitrogen concentration at 6.42 g/L in which giving ethanol yield approximately 1.00 mol/mol. In addition, batch ethanol fermentation in a 2 L bioreactor was performed at the glycerol concentration of 20 g/L, 35 g/L and 45 g/L, respectively. The ethanol yields obtained from all tested glycerol concentrations were approaching theoretical yield when the batch fermentation was performed at optimized conditions.     

Bioproduction of hydrogen by simultaneous saccharification and fermentation of cassava starch with 2-deoxyglucose-resistant mutant strains of Clostridium tyrobutyricum

Laboratory mutagenesis of microorganisms offers the possibility of relating acquired mutations to improve the quality of microbial cultures. In the present study, a mutant strain, Clostridium tyrobutyricum ATCC 25755 DG-8, with significantly elevated α-amylase activity as well as resistant to the non-metabolizable and toxic glucose analog 2-deoxyglucose (2-DG) was obtained by implanting the low-energy nitrogen ion beam. DG-8 was further developed to produce hydrogen by simultaneous saccharification and fermentation (SSF) directly form cassava starch in batch fermentation mode, which to our knowledge is at the first attempt in genus Clostridium. Our results demonstrated that the increased activity of α-amylase would be attributed to the hydrogen over-producing. Higher hydrogen yield (3.2 mol/mol glucose) was achieved with the volumetric productivity of 0.41 L/h/L when the initial total sugar concentration of cassava starch rise up to 100 g/L. The present work will help to decrease the cost of hydrogen fermentation process and stimulate its industrial application in the near future.     

Ethanol production by repeated batch and continuous fermentations of blackstrap molasses using immobilized yeast cells on thin-shell silk cocoons

A thin-shell silk cocoon (TSC), a residual from the silk industry, is used as a support material for the immobilization of Saccharomyces cerevisiae M30 in ethanol fermentation because of its properties such as high mechanical strength, light weight, biocompatibility and high surface area. In batch fermentation with blackstrap molasses as the main fermentation substrate, an optimal ethanol concentration of 98.6 g/L was obtained using a TSC-immobilized cell system at an initial reducing sugar concentration of 240 g/L. The ethanol concentration produced by the immobilized cells was 11.5% higher than that produced by the free cells. Ethanol production in five-cycle repeated batch fermentation demonstrated the enhanced stability of the immobilized yeast cells. Under continuous fermentation in a packed-bed reactor, a maximum ethanol productivity of 19.0 g/(L h) with an ethanol concentration of 52.8 g/L was observed at a 0.36 h⁻¹ dilution rate.     

Ethanol fermentation of sugar beet thick juice diluted with crude cheese whey by the flex yeast Kluyveromyces marxianus KD-15

Kluyveromyces marxianus KD-15, called flex yeast, is a strain that is insensitive to catabolite repression and has the capacity to produce ethanol efficiently from a mixture of beet molasses and whey powder. When a fermentation test was conducted in 50 mL of a medium containing 200 mg mL^?1 of sugar as sugar beet thick juice diluted with an arbitrary amount of crude whey, strain KD-15 produced over 99 mg mL^?1 ethanol in all the media tested, and ethanol formation decreased in proportion to the volume of whey by K. marxianus NBRC 1963, the parental strain of KD-15, and Saccharomyces cerevisiae NBRC 0224, the reference strain for conventional ethanol production. Fermentation of thick juice diluted with whey alone by strain KD-15 at 30 °C initially proceeded slower than that at 33 °C–37 °C but finally bore the highest level of ethanol. The maximum ethanol concentration obtained in 1.5 L of a medium using a 2-L fermentor was elevated by aeration of 15–50 mL min^?1and reduced by that in excess of 100 mL min^?1. Under optimized conditions in 72 h, strain KD-15 converted all of the sugars derived from thick juice and whey to ethanol at 102 mg mL^?1, corresponding to 92.9% of the theoretical yield.     

Isolation and characterization of a new hydrogen-producing strain Bacillus sp. FS2011

A new fermentative hydrogen-producing strain FS2011 was isolated from an effluent of bio-hydrogen production reactor, and identified as Bacillus amyloliquefaciens on the basis of 16S rDNA gene sequence. The strain could utilize various carbon and nitrogen sources to produce hydrogen in a broad range of initial pH (5.29–7.38). Phosphate buffer concentration and fermentation temperature significantly affected hydrogen production and cell growth. The maximum hydrogen yield of 2.26 mol/mol was observed at glucose concentration of 10 g/l, beef extract concentration of 2 g/l, initial pH 6.98, phosphate buffer of 20 mmol/l, and 35 °C, indicating FS2011 was a high-efficiency hydrogen-producing bacterium.     

The utilization of acid-tolerant bacteria on ethanol production from kitchen garbage

In order to achieve ethanol production from kitchen garbage under non-sterilized fermentation, the acid-tolerant Zymomonas mobilis named GZNS1 was selected and applied in the fermentation system. Ethanol production from kitchen garbage under non-sterilized fermentation with GZNS1 was proved to be feasible. The utilization of control strain and acid-tolerant strain under different conditions demonstrated that the sequence of ethanol yield was followed: sterilized garbage with control strain inoculated under pH of 6 (52 g/L) ≈ sterilized garbage with GZNS1 inoculated under pH of 4 (48 g/L) > non-sterilized garbage with GZNS1 inoculated under pH of 4 (46 g/L). Furthermore, the distillery waste during fermentation was adopted to recycle fermentation and acquired 50 g/L ethanol, higher than those adjusted with tap water. The utilization of acid-tolerant bacteria combing with the utilization of distillery waste associated with the process can increase ethanol production, save energy and reduce the cost of ethanol production.     

Hydrogen fermentation of different galactose–glucose compositions during various hydraulic retention times (HRTs)

This study investigated the effects of sugar composition and hydraulic retention time (HRT) on continuous hydrogen fermentation. Continuously-stirred tank reactors (CSTRs) were inoculated with heat-treated digester sludge and fed with 15 g/L of glucose, galactose and galactose: glucose mixture (8:2 ratio-simulating the hydrolysate composition of macroalgae) at HRTs of 6–24 h. Peak hydrogen production rate (HPR) and hydrogen yield (HY) of 4.49 L/L/d and 1.62 mol/mol glucose_added were attained while using glucose as feedstock at HRTs of 6 and 18 h, respectively. Meanwhile, galactose provided a peak HPR and HY of 2.35 L/L/d and 1.00 mol/mol galactose_added during the HRTs of 12 and 24 h, respectively. In case of mixed sugars (galactose 8: glucose 2) the production performances were inferior to the sole sugar conditions due to the low substrate utilization of less than 65%, which showed a maximal HPR and HY of 2.75 L/L/d and 0.48 mol/mol carbohydrate_added at the HRTs of 6 and 18 h, respectively.     

Evaluating biohydrogen production by Clostridium hydrogenum sp. nov. strain CUEA01 isolated from mangrove sediments in Thailand

The hydrogen (H₂) fermentative Clostridium hydrogenum sp. nov. strain CUEA01 was isolated from a mangrove sediment in Thailand. Genome sequencing and analysis revealed a genome size of 5,501,482 bp that encoded for 3,292 predicted protein coding genes with annotated functional assignments and many genes associated with carbon utilization and H₂ evolution. The H₂ production performance was evaluated in batch fermentation, and revealed that this strain can grow and produce H₂ at a broad range of temperatures (15–40 °C), pH (4–10), and initial glucose concentrations (5–60 g/L). The maximum H₂ yield (3.11 mol_H2/mol_glucose) was obtained at 37 °C, pH 8, and an initial glucose concentration of 10 g/L. Furthermore, this strain could utilize various carbon sources, including xylose, xylan, starch, mannose, glycerol, and avicel cellulose, amongst others. Additionally, CUEA01 was compatible with agro-industrial wastes and could achieve a maximum CHP of 4639 mL/L and 4024 mL/L from sugarcane molasses and cassava pulp, respectively. This demonstrates that CUEA01 has a potential for H₂ fermentation from complex organic wastes since it can secrete enzyme cocktails that consolidate the fermentation process.     

Biohydrogen production from sugarcane bagasse hydrolysate by elephant dung: Effects of initial pH and substrate concentration

Pre-heated elephant dung was used as inoculum to produce hydrogen from sugarcane bagasse (SCB) hydrolysate. SCB was hydrolyzed by H₂SO₄ or NaOH at various concentrations (0.25-5% volume) and reaction time of 60 min at 121 °C, 1.5 kg/cm² in the autoclave. The optimal condition for the pretreatment was obtained when SCB was hydrolyzed by H₂SO₄ at 1% volume which yielded 11.28 g/L of total sugar (1.46 g glucose/L; 9.10 g xylose/L; 0.72 g arabinose/L). The maximum hydrogen yield of 0.84 mol H₂/mol total sugar and the hydrogen production rate of 109.55 mL H₂/L day were obtained at the initial pH 6.5 and initial total sugar concentration 10 g/L. Hydrogen-producing bacterium (Clostridium pasteurianum) and non hydrogen-producing bacterium (Flavobacterium sp.) were dominating species in the elephant dung and in hydrogen fermentation broth. Sporolactobacillus sp. was found to be responsible for a low hydrogen yield obtained.     

Optimization of fermentative hydrogen production by Enterococcus faecium INET2 using response surface methodology

A newly isolated strain Enterococcus faecium INET2 was used as inoculum for biohydrogen production through dark fermentation. The individual and interactive effect of initial pH, operation temperature, glucose concentration and inoculation amount on the accumulation of hydrogen during fermentation was examined by a Box–Behnken Design (BBD), and hydrogen production process was analyzed at the optimal condition. A significant interactive effect between glucose concentration and pH was observed, the optimal condition was initial pH 7.1, operation temperature 34.8 °C, glucose concentration 11.3 g/L and inoculation amount 10.4%. Hydrogen yield, maximum hydrogen production rate and hydrogen production potential were determined to be 1.29 mol H₂/mol glucose, 86.7 L H₂/L/h and 1.35 L H₂/L. Metabolites analysis showed that E. faecium INET2 followed the pyruvate: formate lyase (Pfl) pathway in first 16 h, followed by the acetate-type fermentation and then shifted to butyrate-type fermentation. Maximum hydrogen production rate was accompanied with a quick formation of acetic acid.     

Biohydrogen production by Ethanoligenens harbinense B49: Nutrient optimization

A new hydrogen-producing bacterial strain Ethanoligenens harbinense B49 was examined for its capability of H₂ production with glucose as sole carbon source. The H₂ production was significantly affected by the concentration of the yeast powder and phosphate in the synthetic medium. The optimized concentration of yeast powder was 0.3–0.5 g/L and the maximum hydrogen yield was obtained at the concentration of phosphate about 100–150 mmol/L. The dynamics of hydrogen production showed that rapid evolution of hydrogen appeared to start after the middle-phase of exponential growth (about 8 h). The maximum H₂ yield and specific hydrogen production rate were estimated to be 2.26 mol H₂/mol glucose and 27.74 mmol H₂/g cell, respectively, when 10 g/L of glucose was present in the medium. The possible pathway of hydrogen production by Ethanoligenens sp. B49 during glucose fermentation was oxidative decarboxylation of pyruvate and the NADH pathway.     

The alginate fermentation strain Pantoea sp. F16-PCAi-T3P21 and ethanol production

Bio-ethanol production from algae is a promising way to help solve the energy problem. Alginate is a major component of algae, but it cannot be utilized by existing ethanol fermentation microorganisms. In order to improve the utilization rate of brown algae, high alginate fermentation strains should be obtained. In this research, strains for algae fermentation were got from several experiments. The ethanol yield of strain A was the highest, which was 0.095 g/g (ethanol to alginate). The identification of strain A was carried out and it was 99% identical to Pantoea sp. F16-PCAi-T3P21. Fermentation experiments with different substrates were carried out, such as laminaran, mannitol, L. japonica and acid hydrolysate of L. japonica, and the ethanol yield rate of L. japonica acid hydrolysate was the highest, which reached 0.17 g/g ethanol to L. japonica. It showed that strain A can converse alginate to ethanol in a relatively high yield rate, and might be a promising strain with L. japonica as the substrate, we believe more research should be carried out on this strain.