Removal of residual methylene chloride from homoharringtonine by pre-treatment with ethanol

In this study, methylene chloride, which is a residual solvent in final purified homoharringtonine, was removed effectively through pre-treatment with ethanol. When the final HPLC-purified sample was concentrated using a rotary evaporator, the residual methanol easily met the ICH-specified value (3000 ppm), but methylene chloride did not meet the ICH-specified value (600 ppm). However, when the sample (methylene chloride: 10,000 ppm, methanol: 500 ppm) was concentrated through pre-treatment with 95% ethanol using a rotary evaporator, the residual methylene chloride easily met the ICH-specified value. Also, the residual ethanol (concentration > 10,000 ppm) was removed effectively below the ICH-specified value (5000 ppm) through microwave-assisted drying (microwave power: 400 W).     

HTST-extrusion cooking in ethanol production from starchy materials

Starch syrup for ethanol fermentation is conventionally produced by acid or enzymatic hydrolysis. Recently, however, promising results have been obtained using HTST-extrusion cooking in starch liquefaction. The starchy material was pregelatinized and preliquefied in a Creusot-Loire BC45 twin-screw HTST-extrusion cooker before simultaneous saccharification by amyloglucosidase and fermentation by Saccharomyces cerevisiae or Zymomonas mobilis. With pretreatment of milled whole grain or starch by HTST-extrusion cooking a significantly shorter fermentation time could be achieved. Maximum ethanol yield was obtained in 45 h using conventional yeast and amyloglucosidase (1,4-α-d-glucan glucohydrolase, EC 3.2.1.3) dosage, even without addition of Termamyl α-amylase (1,4-α-d-glucan glucanohydrolase, EC 3.2.1.1) during thermomechanical liquefaction. Immobilized yeast could also be used to produce ethanol both by a batch or continuous process. In this case, for a continuous process the DE-value of the syrup should be sufficiently high. A model for ethanol production as a function of dry matter, fermentation time, and yeast and Termamyl quantities has been developed.     

Glycerol accumulation while recycling thin stillage in corn fermentations to ethanol

Summary By succesive recycling of the thin stillage in mashing and fermenting fresh corn, the glycerol content in each fermentation increased by about 0.4% and accumulated to a high of 2.1% in the beer of the fifth recycle. Glycerol concentration declined after the fifth recycle. The original fermentation contained 0.8% glycerol.Presented in part at the Society for Industrial Microbiology Annual Meeting, August 7–12, 1988, Chicago, IL.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

光合菌群发酵玉米秸秆水解液产氢

以玉米秸秆水解液作为产氢底物,研究光合菌群产氢性能。考察了硫酸浓度、固液比、水解时间、水解温度等秸秆水解条件对产氢的影响,确定了最佳水解条件,并对不同脱毒方法进行了对比研究。结果表明,最佳的水解条件为硫酸浓度1%,固液比1:12,水解时间0.5 h,水解温度为110°C。采用Ca(OH)2脱毒方法的产氢效果要优于其他2种脱毒方法;NH4+在一定浓度范围内对该光合菌群产氢有促进作用。     

Changes of the surface structure of corn stalk in the cooking process with active oxygen and MgO-based solid alkali as a pretreatment of its biomass conversion

This study presents a novel, efficient and environmentally friendly process for the cooking of corn stalk that uses active oxygen (O₂ and H₂O₂) and a recoverable solid alkali (MgO). The structural changes on the surface of corn stalk before and after cooking were characterized by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) techniques. The results showed that lignin and extractives were effectively removed, especially those on the surface of corn stalk. Additionally, the changes included becoming fibrillar, the exposure of cellulose and hemi-cellulose and the pitting corrosion on the surface, etc. The results also showed that the removal reaction is from outside to inside, but the main reaction is possibly on the surface. Furthermore, the results of active oxygen cooking with a solid alkali are compared with those of alkaline cooking in the paper.     

Evaluation of Saccharomyces cerevisiae Y5 for ethanol production from enzymatic hydrolysate of non-detoxified steam-exploded corn stover

Saccharomyces cerevisiae Y5 was used to produce ethanol from enzymatic hydrolysate of non-detoxified steam-exploded corn stover, with and without a nitrogen source, and decreasing inoculum size. The results indicated that the ethanol concentration of 44.55 g/L, corresponding to 94.5% of the theoretical yield was obtained after 24 h, with an inoculum size of 10% (v/v) and nitrogen source (corn steep liquor, CSL) of 40 mL/L. With the same inoculum size, and without CSL, the ethanol concentration was 43.21 g/L, corresponding to 91.7% of the theoretical value after 60 h. With a decreased inoculum size of 5% (v/v), and without CSL, the ethanol concentration was 40.00 g/L, corresponding to 85.8% of the theoretical value after 72 h. The strain offers the potential to improve the economy of cellulosic ethanol production by simplifying the production process and reducing the costs associated with the process such as water, capital equipment and nutrient supplementation.     

Industrial yeast strain engineered to ferment ethanol from lignocellulosic biomass

In this study an industrial Saccharomyces cerevisiae yeast strain capable of fermenting ethanol from pretreated lignocellulosic material was engineered. Genes encoding cellulases (endoglucanase, exoglucanase and β-glucosidase) were integrated into the chromosomal ribosomal DNA and delta regions of a derivative of the K1-V1116 wine yeast strain. The engineered cellulolytic yeast produces ethanol in one step through simultaneous saccharification and fermentation of pretreated biomass without the addition of exogenously produced enzymes. When ethanol fermentation was performed with 10% dry weight of pretreated corn stover, the recombinant strain fermented 63% of the cellulose in 96 h and the ethanol titer reached 2.6% v/v. These results demonstrate that cellulolytic S. cerevisiae strains can be used as a platform for developing an economical advanced biofuel process.     

Efficient production of ethanol from crude glycerol by a Klebsiella pneumoniae mutant strain

A mutant strain of Klebsiella pneumoniae, termed GEM167, was obtained by γ irradiation, in which glycerol metabolism was dramatically affected on exposure to γ rays. Levels of metabolites of the glycerol reductive pathway, 1,3-propanediol (1,3-PD) and 3-hydroxypropionic acid (3-HP), were decreased in the GEM167 strain compared to a control strain, whereas the levels of metabolites derived from the oxidative pathway, 2,3-butanediol (2,3-BD), ethanol, lactate, and succinate, were increased. Notably, ethanol production from glycerol was greatly enhanced upon fermentation by the mutant strain, to a maximum production level of 21.5 g/l, with a productivity of 0.93 g/l/h. Ethanol production level was further improved to 25.0 g/l upon overexpression of Zymomonas mobilispdc and adhII genes encoding pyruvate decarboxylase (Pdc) and aldehyde dehydrogenase (Adh), respectively in the mutant strain GEM167.     

Onsite bio-detoxification of steam-exploded corn stover for cellulosic ethanol production

In the process of ethanol production from steam-exploded corn stover (SECS), a cellulose-degradation strain of Aspergillus nidulans (FLZ10) was investigated whether it could remove the inhibitors released from steam exploded pretreatment , and thereby be used for biological detoxification on Saccharomycescerevisiae. The results showed that FLZ10 removed 75.2% formic acid, 53.6% acetic acid, and 100% hydroxymethyl furfural (5-HMF) and furfural from the hydrolysate washed from SECS after 72 h cultivation. A cellulase activity of 0.49 IU/ml was simultaneously produced while the biological detoxification occurred. An ethanol yield of 0.45 g/g on glucose was obtained in the hydrolysate biodetoxified by FLZ10. The glucose consumption rate of FLZ10 was much lower than that of S. cerevisiae, thereby it had little competition with S. cerevisiae on glucose consumption. Based on SECS to ethanol mass balance analysis, with the onsite bio-detoxification, fermentation using S. cerevisiae effectively converted monomeric glucose with 94.4% ethanol yield.     

Ethanol production from syngas by Clostridium strain P11 using corn steep liquor as a nutrient replacement to yeast extract

The feasibility of replacing yeast extract (YE) by corn steep liquor (CSL), a low cost nutrient source, for syngas fermentation to produce ethanol using Clostridium strain P11 was investigated. About 32% more ethanol (1.7 g L⁻¹) was produced with 20 g L⁻¹ CSL media in 250-mL bottle fermentations compared to media with 1 g L⁻¹ YE after 360 h. Maximum ethanol concentrations after 360 h of fermentation in a 7.5-L fermentor with 10 and 20 g L⁻¹ CSL media were 8.6 and 9.6 g L⁻¹, respectively, which represent 57% and 60% of the theoretical ethanol yields from CO. Only about 6.1 g L⁻¹ of ethanol was obtained in the medium with 1 g L⁻¹ YE after 360 h, which represents 53% of the theoretical ethanol yield from CO. The use of CSL also enhanced butanol production by sevenfold compared to YE in bottle fermentations. These results demonstrate that CSL can replace YE as the primary medium component and significantly enhance ethanol production by Clostridium strain P11.     

Continuous 2-keto-gluconic acid (2KGA) production from corn starch hydrolysate by Pseudomonas fluorescens AR4

A continuous fermentation process for 2-keto-gluconic acid (2KGA) production from cheap raw material corn starch hydrolysate was developed using the strain Pseudomonas fluorescens AR4. The dilution rate and feeding glucose concentration had a significant effect on the cell concentrations, glucose utilization and 2KGA production performance. The optimal operating factors were obtained as: 0.065 h⁻¹ of dilution rate, 180 g/L of feeding glucose concentration, and 16 h of batch fermentation time as the starting point. Under these conditions, the steady state had the 135.92 g/L of produced 2KGA concentration, 8.83 g/L.h of average volumetric productivity, and 0.9510 g/g of yield. In conclusion, the proposed efficient and stable continuous fermentation process for 2KGA production by the strain P. fluorescens AR4 is potentially competitive for industrial production from corn starch hydrolysate in terms of 2KGA productivity and yield.     

Biodiesel production by two-stage transesterification with ethanol

A two-stage process consisting of two reactions steps with glycerin separation and ethanol/catalyst addition in each of them was optimized for ethyl esters production. The optimal reaction temperature was 55 °C. At an ethanol/oil molar ratio of 4.25:1 (25%v/v alcohol with respect to oil), a 99% conversion value was obtained with low ethanol consumption. In contrast to methoxide catalysts, sodium and potassium hydroxide catalysts severely complicate the purification since no phase separation took place under most conditions. With a total sodium methoxide concentration of 1.06 g catalyst/100 g oil, and adding 50% of the catalyst in each reaction step, biodiesel with a total glycerin content of 0.172% was obtained. The optimal conditions found in this study make it possible to use the same industrial facility to produce either methyl or ethyl esters.     

Chemical characterization and in vitro crude protein degradability of thin stillage derived from barley- and wheat-based ethanol production

A study was conducted to characterize the different carbohydrate and protein fractions of wheat- and barley-based thin stillage samples. In vitro crude protein degradability of wheat- and barley-based thin stillage was estimated relative to soyabean (SBM) and canola (CM) meal using a protease enzyme assay. Results of the carbohydrate analysis showed that wheat thin stillage had similar neutral (NDF, average 328.5 g kg⁻¹) and lower (P < 0.05) acid detergent fibre (ADF) than barley-based thin stillage. Relative to barley-based thin stillage, wheat thin stillage had higher (P < 0.05) crude protein (CP) and soluble CP content. However, the amount of CP associated with NDF and ADF was higher (P < 0.05) in barley-based thin stillage than in wheat thin stillage. Fractionation of true protein showed that most of the CP (average 707 g kg⁻¹ of CP) was present in the slowly degradable true protein fraction and was similar in both byproducts. Glutamic acid was the main amino acid in thin stillage and was higher (P < 0.05) in wheat than in barley-based thin stillage. However, barley-based thin stillage had higher (P < 0.05) levels of lysine, methionine, arginine, threonine, leucine and isoleucine than wheat thin stillage. Results of the in vitro trial indicated that effective degradability of CP (g kg⁻¹ of CP) followed the order (P < 0.05): SBM (665.0) > wheat thin stillage (614.0) > CM (531.0) > barley-based thin stillage (493.0). It was concluded that barley-based thin stillage had different chemical characteristics than wheat thin stillage. The reduced CP degradability of barley-based thin stillage relative to wheat thin stillage was attributed to a lower CP and a higher acid detergent in soluble CP level.     

Optimization of fermentation parameters for production of ethanol from kinnow waste and banana peels by simultaneous saccharification and fermentation

A study was taken up to evaluate the role of some fermentation parameters like inoculum concentration, temperature, incubation period and agitation time on ethanol production from kinnow waste and banana peels by simultaneous saccharification and fermentation using cellulase and co-culture of Saccharomyces cerevisiae G and Pachysolen tannophilus MTCC 1077. Steam pretreated kinnow waste and banana peels were used as substrate for ethanol production in the ratio 4:6 (kinnow waste: banana peels). Temperature of 30°C, inoculum size of S. cerevisiae G 6% and (v/v) Pachysolen tannophilus MTCC 1077 4% (v/v), incubation period of 48 h and agitation for the first 24 h were found to be best for ethanol production using the combination of two wastes. The pretreated steam exploded biomass after enzymatic saccharification containing 63 gL⁻¹ reducing sugars was fermented with both hexose and pentose fermenting yeast strains under optimized conditions resulting in ethanol production, yield and fermentation efficiency of 26.84 gL⁻¹, 0.426 gg ⁻¹ and 83.52 % respectively. This study could establish the effective utilization of kinnow waste and banana peels for bioethanol production using optimized fermentation parameters.     

Effect of corn preparation methods on dry-grind ethanol production by granular starch hydrolysis and partitioning of spent beer solids

Two corn preparation methods, rollermill flaking and hammermill grinding, were compared for efficient processing of corn into ethanol by granular starch hydrolysis and simultaneous fermentation by yeast Saccharomyces cerevisiae. Corn was either ground in a hammermill with different size screens or crushed in a smooth-surfaced rollermill at different roller gap settings. The partitioning of beer solids and size distribution of solids in the thin stillage were compared. The mean particle diameter d₅₀ for preparations varied with set-ups and ranged between 210 and 340 μm for ground corn, and 1180-1267 μm for flaked corn. The ethanol concentrations in beer were similar (18-19% v/v) for ground and flaked preparations, however, ethanol productivity increased with reduced particle size. Roller versus hammermilling of corn reduced solids in thin stillage by 28%, and doubled the volume percent of fines (d₅₀ ∼ 7 μm)in thin stillage and decreased coarse (d₅₀ ∼ 122 μm) by half compared to hammermilling.     

A consolidated bio-processing of ethanol from cassava pulp accompanied by hydrogen production

A biphasic fermentation approach was undertaken for the production of ethanol and hydrogen from cassava pulp. The glucose generated by co-culture of Clostridium thermocellum and Thermoanaerobacterium aotearoense was 13.65±0.45 g L(-1), which was 1.75 and 1.17-fold greater than that produced by mono-cultures of C. thermocellum and T. aotearoense, respectively. The accumulated glucose could be utilised rapidly by subsequently inoculated Saccharomyces cerevisiae. An inoculum ratio of 1:1, a thermophilic fermentation of 84 h, and a pulp concentration of 4% proved optimal for ethanol production, fermentation efficiency, and productivity. With these conditions, the ethanol level reached 8.83±0.31 g L(-1) with a fermentation efficiency of 64.95±2.71%. Hydrogen production of 4.06 mmol by the co-culture system was 1.54 and 2.09-fold greater than that produced by mono-cultures of C. thermocellum and T. aotearoense, respectively. This sequential co-culture approach provided a consolidated bio-processing means to produce ethanol and hydrogen from cassava pulp.     

Single temperature liquefaction process at different operating pHs to improve ethanol production from Indian rice and corn feedstock

Conventional grain ethanol manufacturing is a high-temperature energy-intensive process comprising of multiple-unit operations when combined with lower ethanol recovery results in higher production cost. In liquefaction, jet cooking accounts for significant energy cost, while strong acid or base used for pH adjustment presents a safety hazard. A need is felt for sustainable ethanol manufacturing process that is less hazardous, consumes lower energy, and operates in a low pH range of 4.50–5.50. A single temperature liquefaction (STL) process that could efficiently operate at lower liquefaction temperature over a pH range of 4.50–5.50 was developed using rice and corn feedstock. Ethanol recovery witnessed at pH 4.5, 5.0, and 5.5 are 481.2?±?1.5, 492.4?±?1.5, and 493.6?±?1.5?L?MT^?1 rice, respectively. Similarly, ethanol recovery witnessed at pH 4.5, 5.0, and 5.5 are 404.6?±?1.3, 413.9?±?0.8, and 412.4?±?1.8?L?MT^?1 corn, respectively. The improvement in ethanol recovery is attributed to higher starch conversion by alpha-amylase even at pH as low as 4.50. Thus, the STL process operated at pH lower than 5.20 is poised to enhance sustainability by offering dual advantage of energy as well as chemical saving.     

A rapid screening method to detect ethanol production by microorganisms

A method for detecting ethanol production by microorganisms on an agar plate is described. The assay is based on the enzymatic determination of ethanol. Yellow zones surround ethanol-producing colonies on a blue background and their diameter is an indication of the amount of ethanol produced.     

Importance of stability study of continuous systems for ethanol production

Fuel ethanol industry presents different problems during bioreactors operation. One of them is the unexpected variation in the output ethanol concentration from the bioreactor or a drastic fall in the productivity. In this paper, a compilation of concepts and relevant results of several experimental and theoretical studies about dynamic behavior of fermentation systems for bioethanol production with Saccharomyces cerevisiae and Zymomonas mobilis is done with the purpose of understanding the stability phenomena that could affect the productivity of industries producing fuel ethanol. It is shown that the design of high scale biochemical processes for fuel ethanol production must be done based on stability studies.     

Factors in acid treated bagasse inhibiting ethanol production from d-xylose by Pachysolen tannophilus

The fermentation of d-xylose, the major sugar-cane bagasse hemicellulose component, to ethanol by Pachysolen tannophilus is inhibited by various factors produced or released during the acid hydrolysis of the bagasse or during the fermentation process. These include ethanol, iron, chromium, copper, nickel, acetic acid and furfural. Ethanol production by P. tannophilus is inhibited by ethanol fconcentrations >24 g l^?1. Furfural and acetic acid concentrations as low as 0.3 and 7 g l^?1, respectively, and iron, chromium, nickel and copper at concentrations of 0.07, 0.01, 0.01 and 0.004 g l^?1, respectively. Similar concentrations may be found in acid-hydrolysed bagasse. The removal of these factors by treatment with ion-exchange resin resulted in the fermentation of the sugars to ethanol. The d-glucose was used rapidly and completely whereas d-xylose utilization was slow and incomplete. An ethanol concentration of 4.1 g l^?1 was produced and an ethanol yield of 0.32 was obtained. Xylitol in significant amounts was produced.