Sago starch saccharification and simultaneous ethanol fermentation by amyloglucosidase and Zymomonas mobilis

Simultaneous saccharification and ethanol fermentation (SSF) of sago starch was studied using amyloglucosidase (AMG) and Zymomonas mobilis. The optimal concentration of AMG and operating temperature for the SSF process were found to be 0.5% (v/w) and 35°C, respectively. Under these conditions with 150 g dm^?3 sago starch as a substrate, the final ethanol concentration obtained was 69.2 g dm^?3 and ethanol yield, Y_P/S, 0.50 g g^?1 (97% of theoretical yield). Sago starch in the concentration range of 100–200 g dm^?3 was efficiently converted into ethanol. When compared to a two-step process involving separate saccharification and fermentation stages, the SSF reduced the total process time by half.     

Mathematical model for growth process of a recombinant yeast having saccharification and fermentation activities

A mathematical model for direct alcohol fermentation from starch was proposed using an amylase‐producing recombinant yeast, Saccharomyces cerevisiae SR93. This model consisted of the reaction rate equations for glucoamylase synthesis in the recombinant yeast, starch degradation by a glucoamylase, cell growth, production of glucose, and production of ethanol. The rate of glucoamylase synthesis was expressed on the basis of the diauxic growth model that represents catabolite repression and enzyme induction. The rate of starch degradation was expressed on the basis of the enzymatic hydrolysis model representing the change of structure resulting from starch degradation. The calculated values were in satisfactory agreement with the experimental data in a batch culture of direct alcohol fermentation from starch using S cerevisiae SR93. Furthermore, the calculated values obtained by changing only one parameter concerning the synthesis rate of glucoamylase were in satisfactory agreement with the experimental data using another recombinant yeast, S cerevisiae SR96. Copyright © 2003 Society of Chemical Industry     

Fermentative production of L(+)‐lactic acid from starch hydrolyzate and corn steep liquor as inexpensive nutrients by batch culture of Enterococcus faecalis RKY1

BACKGROUND: Attempts were made to determine the lactic acid production efficiency of novel isolate, Enterococcus faecalis RKY1 using four different starches (corn, tapioca, potato, and wheat starch) with different concentrations (50, 75, 100, and 125 g L^?1) and corn steep liquor as an inexpensive nitrogen source. RESULTS: The yield of lactic acid from each starch was higher than 95% based on initial starch concentrations. High lactic acid concentration (129.9 g L^?1) and yield (1.04 g‐lactic acid g^?1‐starch) were achieved faster (84 h) from 125 g L^?1 of corn starch. Among the starches used, tapioca starch fermentation usually completed in a shorter incubation period. The final dry cell weight was highest (7.0 g L^?1) for the medium containing 75 g L^?1 of corn starch, which resulted in maximum volumetric productivity of lactic acid (3.6 g L^?1 h^?1). The addition of 30 g L^?1 corn steep liquor supplemented with a minimal amount of yeast extract supported both cell growth and lactic acid fermentation. CONCLUSION: Enterococcus faecalis RKY1 was found to be capable of growing well on inexpensive nutrients and producing maximum lactic acid from starches and corn steep liquor as lower‐cost raw materials than conventionally‐used refined sugars such as glucose, and yeast extract as an organic nitrogen source in laboratory‐scale studies. These fermentation characteristics are prerequisites for the industrial scale production of lactic acid. Copyright © 2008 Society of Chemical Industry     

Continuous ethanol fermentation using immobilized yeast in a fluidized bed reactor

Continuous ethanol fermentation of glucose using fluidized bed technology was studied. Saccharomyces cerevisiae were immobilized and retained on porous microcarriers. Over two-thirds of the total reactor yeast cell mass was immobilized. Ethanol productivity was examined as dilution rate was varied, keeping all other experimental parameters constant. Ethanol yield remained high at an average of 0.36 g ethanol g^?1 glucose (71% of theoretical yield) as the dilution rate was increased stepwise from 0.04 h^?1 to 0.14 h^?1. At a dilution rate of 0.15 h^?1, the ethanol yield steeply declined to 0.22 g ethanol g^?1 glucose (44% of theoretical yield). The low maximum percentage of theoretical yield is primarily due to an extended mean cell residence time, and possibly due to the inhibitory effect of a high dissolved carbon dioxide concentration, enhanced by the probable intermittent levels of low pH in the reactor. Constant ethanol production was possible at a high glucose loading rate of 840 g dm^?3 day^?1 (attained at a dilution rate of 0.14 h^?1). Although the highest average ethanol concentration (97.14 g dm^?3) occurred at the initial dilution rate of 0.04 h^?1, the peak average ethanol production rate (2.87 g (g yeast)^?1 day^?1) was reached at a greater dilution rate of 0.11 ^h-1. Thus, the optimal dilution rate was determined to be between 0.11 h^?1 and 0.14 h^?1. Ethanol inhibition on yeast cells was absent in the reactor at average bulk-liquid ethanol concentrations as high as 97.14 g dm^?3. In addition, zero-order kinetics on ethanol production and glucose utilization was evident.     

Fate of free amino nitrogen during liquefaction and yeast fermentation of maize and sorghums differing in endosperm texture

Biofuels demands have motivated the increasing research in production technology as well as effective raw material utilization. Fusel alcohol production had been related with the free amino nitrogen (FAN) content produced during hydrolyses stages and also as a stress response of Saccharomyces cerevisiae. This paper analyses the effect of grain type and endosperm texture on FAN production during starch liquefaction and yeast fermentation of maize, white and red sorghums bioconverted into ethanol. The white sorghum had a harder endosperm texture and upon milling produced a coarser granulation compared to the red counterpart. Therefore, the red sorghum produced higher amounts of FAN during liquefaction and the kinetics of these nitrogenous compounds were metabolized faster during the first hours of yeast fermentation. The white and red sorghums yielded 13.6% and 2.7% lower ethanol compared to the maize counterpart. The white sorghum yielded the lowest amount of ethanol likely to its relatively harder endosperm texture that was less susceptible to biocatalysis and yeast fermentation.     

Fermentation of glucose and starch particles using an inexpensive medium

BACKGROUND: In North America, compared with sugar, starch is a more economical raw material to produce ethanol. Recently low temperature processing of starch granules directly to ethanol has been commercialized. With a view to minimizing process costs while maintaining a satisfactory ethanol yield, an inexpensive medium that does not require the addition of commercial α‐amylase has been developed for low temperature, direct fermentation of starch particles. RESULTS: Compared with an expensive medium that contained both yeast extract and α‐amylase, the inexpensive medium resulted in identical and 10% higher ethanol yields using glucose and starch granules as the raw substrates, respectively, but required longer processing times. Based on the different ingredient prices for expensive and inexpensive media, the operating cost to produce 10 million litres per year of ethanol utilizing simultaneous raw starch hydrolysis and fermentation is reduced over $ 2 million (Cdn) per year when using inexpensive medium. CONCLUSION: The new inexpensive medium and simultaneous raw starch hydrolysis and fermentation is considered the best strategy to produce ethanol directly from starch particles in North America. Copyright © 2009 Society of Chemical Industry     

Direct ethanol production from dextran industrial waste water by Zymomonas mobilis

The direct production of ethanol from dextran industrial waste water was investigated by using Zymomonas mobilis via batch and semi-continuous fermentation mode. In batch fermentation, pretreated waste water (unsterilized and sterilized), pH value (3.8 and 6.0), and Mg²⁺ (with and without) was compared with OD₆₀₀, sugar and ethanol concentration. After 24 h fermentation, sugar in the dextran waste water was almost exhausted, and the amount of ethanol accumulated reached 24.33–29.92 g/l, which is nearly 99% of the theoretical yield of ethanol. Kinetic parameters of Z. mobilis in batch fermentation were also investigated. The raw dextran waste water was also used in semi-continuous fermentation. After 48 h fermentation, the production of ethanol was 28.65 g/l. These results indicated that dextran waste water may be used as a candidate substrate and Z. mobilis could convert the raw material into ethanol directly.     

The Effect of Ozonation on the Size Fractions of Manganese

Manganese is a commonly found substance in groundwater in Finland. As a powerful oxidant, ozone can be used for the oxidizing of manganese even without raising the pH. The SFS (Finnish Standards Association) standard has set the accepted limit for soluble manganese to 0.45 μm. However, some research papers have used the limit of 30 kD (kilodalton) for soluble manganese. This research concentrates on the size fractions of manganese in four samples of untreated groundwater and in four samples of ozonized groundwater when treated with 0.45 μm, 0.20 μm, 100 kD, 30 kD, and 10 kD filters. In all tests, nearly all manganese contained in raw water penetrated all filters. There were slight variations in the flocculation of manganese in ozonized groundwater; nevertheless, hardly any reduction in manganese levels took place beyond 100 kD. After ozonation, there were two water samples which surpassed the manganese limit of 50 μgl^?1 set for domestic water when the filtration was 0.20 μm and another two samples when the filtration was 100 kD.     

Ozone treatment of water-soluble polymers. III. Ozone degradation of polyacrylamide in water

Polyacrylamide (MW 350,000) was ozonized at pH 2 and pH 10 buffer solution. There was little ozonization at pH 2, but at pH 10 the COD and the viscosity of the solution decreased upon ozonization, and a linear relationship existed between the ozone consumed and the number of breaks calculated from the viscosity. This relationship apparently indicated that 45 molecules ozone were consumed for one cleavage of the polymer chain. Such random cleavage was confirmed also by the observation of molecular weight distribution by means of gel filtration chromatography. The amide group decreased scarcely by the ozonization, while a small amount of aldehyde was observed in the ozonized solution. No remarkable change was observed in the IR spectra of the ozonized sample, except for a weak absorption band at 1725 cm^?1 which arose from the carbonyl of aldehyde or ketone. Although no variation was observed in the ¹³C NMR spectra either, a strong absorption peak at 266 nm appeared in the UV spectra of the ozonized solution and increased with the ozonization time. This phenomenon was presumed to be due to the formation of a certain ring structure between the amide group and the small amount of ketone produced in the main chain. However, the details of the ozonization mechanism could not be ascertained.     

Effect of simultaneous saccharification and fermentation conditions of native triticale starch on the dynamics and efficiency of process and composition of the distillates obtained

BACKGROUND: Among ethanol production technologies, attention should be focused towards simultaneous saccharification and fermentation of native starch with enzymes capable of its degradation without prior gelatinization. Selection of process conditions makes it possible to achieve high efficiency of the process and to reduce the costs of the production of ethanol. RESULTS: This study determined the effect of hydrolysis conditions of native triticale starch based on the results of their fermentation and concentration of by‐products in the distillates obtained. The pre‐activation of starch with acid α‐amylase is not necessary for the proper conduct and efficiency of saccharification and fermentation of native triticale starch. Beneficial impact on the synthesis of ethanol was afforded by the treatment of mashes with protease preparation (EC 3.4.21.62) (Bacillus licheniformis). Raising the pH of mashes from 3.6 to 4.8 did not improve the ethanol yield. Process conditions had a diversified impact on concentration of by‐products in the raw spirits obtained. CONCLUSION: Under the optimal conditions found in these experiments 63.01 ± 0.33–63.56 ± 0.33 L of absolute ethanol was obtained from 100 kg of starch. Simultaneous saccharification and fermentation of the native starch raw materials, especially without pre‐activation of starch, can simplify the process and improve the economic index in the alcohol‐distilling industry. © 2012 Society of Chemical Industry     

Ozone Processing of Peach Juice: Impact on Physicochemical Parameters,Color, and Viscosity

The impact of gaseous ozone on pH, °Brix, titratable acidity, and optical and rheological properties of fresh squeezed peach juice was investigated. Peach juice was exposed to ozone (0.06–2.48 g. L^?1) in a bubble column at 20 ± 1 °C. Nonsignificant or slight changes in pH, °Brix, and titratable acidity were found during ozonation. Lightness (L*) slightly decreased in the first minute of O₃ exposure and then remained practically constant, while a* parameter slightly increased in all treated samples. L* and a* parameters as well as Browning Index values reflected a slight increase in browning in ozonized juices. All juices, treated and untreated, exhibited non-Newtonian flow characteristics with pseudoplastic behavior. Significant reductions in apparent viscosity and a trend toward Newtonian flow as O₃ treatment time increased were observed for ozonized juices. The Power Law model was suitable to fit rheological data.     

Continuous alcohol production from whey permeate using immobilised cell reactor systems

The present paper reports the performance of a bioreactor packed with alginate-entrapped Kluyveromyces marxianus NCYC179 for continuous fermentation of whey permeate to ethanol. A maximum ethanol productivity as 28.21 gl^?1 h^?1 was attained at D=0.42h^?1 and 75% lactose consumption (substrate feed rate in the inflowing medium was 200 g lactose I^?1). However, the higher dilution rates (0.6-1.Oh^?1) resulted in poor productivities and higher substrate washout in the effluent samples. The maximum specific ethanol production (qpi) and maximum specific lactose uptake (qsi) of the immobilised Kluyveromyces marxianus NCYC179 was found to be 3.88g ethanol/g immobilised cell/hx10^?2 and 8.75g lactose consumed/g immobilised cell/hx10^?2 respectively. A bead size of 2.5 mm in diameter and activation period of 24h of alginate beads in lactose solution (10%) prior to their packing in column reactor were found to support the efficient working of the bioreactor. The immobilised cell bioreactor system was operated continuously at a constant dilution rate of 0.15h^?1 and 10% lactose for 562 h without any significant change in the efficiency (varied from 84 to 88% of theoretical) and viability of the entrapped yeast cells (dropped from 84 to 81%).     

DEGRADATION RATE OF WATER-SOLUBLE POLYMERS BY OZONE

Water-soluble polymers were degraded by ozone, and the degradation rate defined by the change of weight average polymerization degree per unit time was obtained. The degradation rate of poly(oxyethylene) (PEG) was proportional to the 1.5 to 2.0 power of the polymerization degree, and was 20 to 60 times that of poly(acrylamide) (PAAm). The experimental results of PEG and PAAm were correlated as a function of polymerization degree, temperature, dissolved ozone concentration and OH^? ion concentration. The degradation rate of poly(sodium acrylate)(PANa) was proportional to the 2.0 power of the polymerization degree when no other electrolyte was added. However, the degradation rate of PANa decreased with the increase in NaCl concentration, and approached that of PAAm in the range of high NaCl concentrations.     

Ethanol production by solid state fermentation of sweet sorghum using thermotolerant yeast strain

Solid state fermentation of chopped sweet sorghum particles to produce ethanol was studied statically using thermotolerant yeast. The influence of various process parameters, such as yeast cell concentration, particle size and moisture content, on the ethanol yield was investigated. Optimal values of these parameters were 4 × 10⁶ cells/g raw sorghum, Dp = 1.5 mm and 75%, respectively. Addition of reducing agent H₂SO₃ into the fermentation medium provided anaerobic condition, and obtained the maximum ethanol yield of 7.9 g ethanol per 100 g fresh stalks or 0.46 g ethanol/g total sugar, which was 91% of the theoretic yield.     

Production of L‐lactic acid with very high gravity distillery wastewater from ethanol fermentation by a newly isolated Enterococcus hawaiiensis

BACKGROUND: A great amount of wastewater with high contents of chemical oxygen demand (COD) are produced by ethanol production. It would be useful to utilize distillery wastewater to produce L‐lactic acid, which could be a high additional value byproduct of ethanol production. The fermentation process of L‐lactic acid production by a newly isolated Enterococcus hawaiiensis CICIM‐CU B0114 is reported for the first time. RESULTS: The strain produced 56 g L^?1 of L‐lactic acid after cultivation for 48 h in optimized medium consisting of (g L^?1) 80 glucose, 10 peptone, 10 yeast extract, 1.5 Na₂HPO₄ and 0.2 MgSO₄. E. hawaiiensis CICIM‐CU B0114 was isolated and purified by subculture for growing and producing L‐lactic acid in distillery wastewater of very high gravity (VHG) from ethanol fermentation. L‐lactic acid fermentation was further studied with distillery wastewater substrate in 7 L and 15 L fermentors. The results showed that L‐lactic acid concentrations of 52 g L^?1 and 68 g L^?1 was achieved in 7 L and 15 L fermentors with the initial sugar concentrations of 67 g L^?1 and 87 g L^?1, respectively. CONCLUSION: The production of L‐lactic acid by the newly isolated E. hawaiiensis CICIM‐CU B0114 was carried out and the fermentation medium was optimized by orthogonal experimental design. This new strain holds the promise of L‐lactic acid production utilizing distillery wastewater from VHG ethanol fermentation. Copyright © 2010 Society of Chemical Industry     

Effect of Saccharomyces cerevisiae fermentation conditions on expanded bed adsorption of heterologous cutinase

The effect of culture media composition, and fermentation conditions and strategy on the growth and cutinase production of recombinant Saccharomyces cerevisiae and subsequent cutinase purification by expanded bed adsorption (EBA) was studied. The reduction in the amount of yeast extract used as nitrogen source from 20 g dm^?3 to 10 g dm^?3 in batch cultures led to a 29% decrease in the heterologous cutinase production, while the 5% cutinase dynamic adsorption capacity (q5%) on the cation Streamline SP XL was increased 6.7‐fold. By dilution of the whole fermentation broth, performed with the lowest yeast extract content, which reduces conductivity, the q5% was additionally increased by 1.9‐fold. After implementation of a fed‐batch strategy the cutinase concentration, cutinase yield on carbon source, cutinase yield on nitrogen source and productivity were increased by 10.8‐, 2.9‐, 5.3‐ and 6.4‐fold, respectively, in relation to the previously‐mentioned batch fermentation. However, the increased cutinase production was compromised by heterologous protein loss during the EBA recovery operation and the cutinase dynamic adsorption capacity and purification productivity decreased by 90% and 75%, respectively. Thus, target protein production by S cerevisiae fermentation and a downstream process with EBA cannot be considered as separate entities, where the understanding of the factors that affect the interactions among them are crucial towards optimization of the overall production process of heterologous proteins. © 2002 Society of Chemical Industry     

Study on the substrate specificity of α-amylases that contribute to soil removal in detergents

The actions of α-amylases in the degradation of amylose and amylopectin were investigated, and the contribution of the enzymatic hydrolysis of high-viscosity gelatinized starch to the removal of food stains that contain starch was characterized. α-Amylases from Bacillus amyloliquefaciens, B. licheniformis, Aspergillus oryzae, and porcine pancreas were used to test the removal of curry stains, which are hard to remove under low-temperature washing conditions and include large amounts of starch. In a detergent, the enzyme from B. amyloliquefaciens was the most effective in removing stains and hydrolyzed amylopectin more efficiently than amylose as a result of the more efficient formation of an enzyme-substrate complex in the former case. It is suggested that the cleaning power of α-amylases is due to their hydrolytic action, with endo-degradation of amylopectin, which results in an efficient reduction in the degree of polymerization. The liquefaction of amylopectin then accelerates the removal of food stains.     

Synthesis and characterization of ozonized polyethylene

Polyolefins (LDPE and HDPE) can be ozonized with ozone/air gas flow to create peroxide and hydroperoxide groups on polymers. These reactive groups can be used to initiate radical polymerization of vinylic monomers and produce graft copolymers. In this study, the optimization of the experimental conditions and the explanation of the phenomena occurring during the ozonization of polyolefins in a fluidized bed have been studied. The most reproducible methods for placing peroxide and hydroperoxide groups onto polyolefins are the iodometric method and indirect titration by thiol. This work shows that it is necessary to control the temperature of the fluidized bed to avoid the acceleration of the reaction and the degradation of the polymers. Then, we studied the effect of different parameters such as the crystallinity and the granulometry of polymers, the time of ozonization and the effect of the load of the reactor on the quality of ozonization. Infrared spectroscopy and steric exclusion chromatography were used to characterize the ozonized polymers. We show that double bonds of PE, particularly vinyl groups, are very reactive to ozone and that ozonized HDPE leads essentially to the formation of hydroperoxide groups, whereas ozonized LDPE leads to the formation of peroxide groups.     

Continuous simultaneous saccharification and fermentation of raw starch in a membrane reactor

A kinetic model for simultaneous saccharification and fermentation (SSF) of raw starch is proposed. The model includes the effect of ethanol on an active site for saccharification and the decay of a raw starch affinity site. The kinetic parameters were determined by using the experimental results of a batch saccharification and a long-term repeated-batch SSF of raw sweet potato starch. From analysis of the experimental results it is concluded that two subsites took part in ethanol inhibition, and that the inactivation of the raw starch affinity site was induced by adsorption of glucoamylase onto raw starch. The proposed kinetic model successfully predicted the progress of continuous SSF in a membrane reactor.     

生料淀粉制乙醇研究及产业化进展

燃料乙醇作为一种高效的液体燃料已在全世界范围推广使用。目前,广泛使用的干法制乙醇工艺已十分成熟。然而,生产过程更为简化、条件更为温和、可实现更高水平浓醪发酵的生料淀粉制乙醇工艺已成为燃料乙醇工业进一步迈向绿色减排、高效节能的重要发展方向。生料淀粉工艺经历了几十年实验室研究,并且也已在产业化方面有巨大发展。尤其以美国POET公司为代表,其BPX生淀粉水解专利技术已在24套燃料乙醇工业装置上得到应用。综述了生料淀粉工艺研究进展,并对已经产业化的案例作了介绍。开发具有自主知识产权的生料淀粉制乙醇工艺、关键技术及装备也是我国未来燃料乙醇产业发展的一个重要方向。