Optimization of the fermentation conditions and partial characterization for acido-thermophilic <Emphasis Type="Italic">α</Emphasis>-amylase from <Emphasis Type="Italic">Aspergillus niger</Emphasis> NCIM 548

A high performance thermostable α-amylase at low pH values has been synthesized. Sugarcane bagasse was hydrolyzed in a dilute acid solution and utilized as carbon source for the growth of Aspergillus niger strain NCIM 548. Glucose, xylose and arabinose with the ratio of 1.0: 0.9: 0.3 (w/w/w) were detected in the hydrolyzate by HPLC analysis. Optimization of the fermentation conditions for α-amylase production was performed by varying four influential parameters such as Sugarcane bagasse hydrolyzate (SBH), NH₄Cl, pH and incubation time using a central composite design (CCD) under response surface methodology (RSM). The optimum values of SBH, NH₄Cl, pH and incubation time were 20.49, 2.34 g/l, 5.65 and 76.67 h, respectively. The acido-thermophilic enzyme showed maximum stability at 70°C and pH value of 4. The rate constant, K _m and maximum reaction rate, V _max were 18.79 g/l and 15.85 g/l·min, respectively.     

Sequential hydrolysis of hemicellulose and lignin in lignocellulosic biomass by two-stage percolation process using dilute sulfuric acid and ammonium hydroxide

To obtain the total fractionation and pretreatment of the corn stover, two-stage percolation process was investigated. This process consists of two steps: use of 0.07 wt% sulfuric acid for hemicellulose recovery in first stage and ARP (ammonia recycled percolation) in the following stage for lignin recovery. Among tested conditions, the best conditions of two-stage process were as follows: 1^st stage; 170 °C, 2.5 ml/min, 30 minutes using 0.07 wt% sulfuric acid and 2^nd stage; 170 °C, 5.0 ml/min, 60minutes using 15 wt% ammonium hydroxide. At above two-stage treatment conditions, the hemicellulose in corn stover was easily hydrolyzed (95%) and recovered with high yields (86%) and the extent of the lignin removal was 81%. After two-stage process, the treated biomass contained nearly pure glucan (85%). Two-stage treatment brought about enzymatic digestibility of 90% and 89% with 60 and 15 FPU/g glucan cellulase enzyme loadings, respectively.     

Production of fermentable sugars from corn fiber using soaking in aqueous ammonia (SAA) pretreatment and fermentation to succinic acid using <Emphasis Type="Italic">Escherichia coli</Emphasis> AFP184

Conversion of corn fiber (CF), a by-product from the corn-to-ethanol conversion process, into fermentable sugar and succinic acid was investigated using soaking in aqueous ammonia (SAA) pretreatment followed by biological conversions, including enzymatic hydrolysis and fermentation using genetically engineered E. coli (AFP184). The SAA pretreatment (using a 15% w/w NH₄OH solution at a solid-to-liquid ratio of 1: 10 at 60 °C for 24 h) removed 20-38% of lignin and significantly improved the digestibility of the treated solid (85-99% of glucan digestibility). Following the enzymatic hydrolysis, the sugar-rich hydrolysate was subjected to dilute sulfuric acid treatment (1 wt% sulfuric acid and 120 °C for 1 h), which hydrolyzed the oligosaccharides in the hydrolysate into fermentable monomeric sugars. The mixed sugar hydrolysates containing hexose and pentose obtained from the two-step hydrolysis and SAA pretreatment were fermented to succinic acid using a genetically engineered microorganism, Escherichia coli AFP184, for evaluating the fermentability. Engineered E. coli AFP184 effectively converted soluble sugars in the hydrolysate to succinic acid (20.7 g/L), and the production rate and yield were further enhanced with additional nutrients; the highest concentration of succinic acid was 26.3 g/L for 48 h of fermentation.     

Mild autohydrolysis: an environmentally friendly technology for xylooligosaccharide production from wood

Eucalyptus globulus wood samples were subjected to hydrothermal treatments under mild operational conditions (145–190 ° C, liquor to solid ratio 6–10 g g⁻¹, reaction times up to 7.5 h). Residual xylan, xylooligosaccharides, other sugars, furfural, glucan and lignin contents were determined. Negligible effects were caused by hydrothermal treatments on both cellulose and lignin. Kinetic models were developed which describe the hydrolysis of hemicelluloses. Xylan degradation, xylooligosaccharide and xylose generation, and xylose dehydration to furfural were accurately described by models based on pseudohomogeneous, first‐order kinetics with Arrhenius‐type temperature dependence. These models are useful for a technical evaluation of this environmentally friendly technology. © 1999 Society of Chemical Industry     

Optimization the process variables for the fractionation of Saccharina japonica to enhance glucan content

Saccharina japonica was fractionated by dilute sulfuric acid to increase the glucan content. The optimal fractionation conditions were determined as follows: reaction temperature 141.14 °C, reaction time 27.85 min and catalyst concentration 0.30%. The CCD model predicted 32.83% glucan content under these conditions. Experiments confirmed the maximum glucan content of 32.67% under the optimal reaction conditions, which was 4.7-fold higher than that of the raw S. japonica (6.95%). With the residual solid, an enzymatic digestibility of 89.38% was obtained using 15 FPU/g-glucan of cellulase enzyme loading, which was 2.6-fold higher than that of the raw S. japonica (34.85%).     

Chemicals Generated During Oxygen-Organosolv Pulping of Wood

In this paper, we have characterized the black liquors from oxygen-organosolv pulping of Eucalyptus globulus wood in ethanol/water solutions (50:50, V/V) and acetic acid/water solutions (80:20, V/V).

After pulp separation, the organic solvent has been distilled and recycled; the precipitated lignin has been recovered by filtration. The degradation products in solution have been easily separated by simple extraction techniques. Five main groups of compounds have been isolated: solid lignin (11–12% o.d.w.), sugars (10–17% o.d.w.), aliphatic acids (12,5%), aromatic acids (2% o.d.w.) and low molecular weight phenols (1% o.d.w.). Most of these compounds were identified by HPLC and GC. Solid lignins have been analyzed by FTIR and ¹³C NMR spectroscopy.

It has been çoncluded that oxygen-organosolv pulping process can be a new and interesting way for pulp preparation with little pollution and possibility of total biomass recovery; alternatively, it can be used for the fractionation and valorization of the components of lignocellulosics, including agriculture residues.     

Enhanced ethanol production from enzymatically treated steam‐exploded rice straw using extractive fermentation

Alcohol fermentation of an enzymatic hydrolyzate of exploded rice straw was studied experimentally. Rice straw was treated under variable conditions, such as steam pressure and steaming time. The exploded rice straw was separated into water‐soluble material, methanol‐soluble lignin, Klason lignin, and a mixture of cellulose and a low molecular weight substance. The effects of steam explosion on the characteristics of the exploded rice straw were clarified from the point of view of the amounts of extractive components. Steam explosion was found to be effective for the delignification of rice straw and for increasing its susceptibility to enzyme hydrolysis and alcohol fermentation. The polysaccharides (cellulose and hemicellulose) in the rice straw treated at a steam pressure of 3.5 MPa with a steaming time of 2 min were hydrolyzed almost completely into monosaccharides, (ie glucose and xylose) by a mixture of Trichoderma viride cellulase (Meicelase) and Aspergillus aculeatus cellulase (Acucelase). The enzymatic hydrolyzate of exploded rice straw was converted into ethanol efficiently by Pichia stipitis and the ethanol yield from sugar was about 86%(w/w) of the theoretical value. The ethanol concentration in a membrane bioreactor coupled with a pervaporation system reached 50 gdm^?3 and was about five times higher than that in the culture broth. The energy efficiency (ratio of combustion energy of ethanol produced to energy for steam explosion) reached a maximum value at a pressure of 3.5 MPa for 2 min. © 2001 Society of Chemical Industry     

Separation and Determination of Functional Complex Lipids from Chicken Skin

Complex lipids including sphingolipid and plasmalogens were expected to be used as functional supplement, although their physiological activities have not been fully demonstrated. Although these complex lipids exist voluminously in brain and nervous tissues, hardly any animal resources of these lipids have been used since the outbreak of bovine spongiform encephalopathy. Thus, the chemical composition and concentration method of complex lipids from the skin of mature laying hens, a huge amount of which is wasted every year, has been investigated. Total lipid yield (32 g/100 g) prepared from chicken skin contained 2% complex lipids. Total lipids predominantly consisted of triacylglycerol (TAG), with phosphatidylcholine, sphingomyelin (SM) and phosphatidylethanolamine (PE) generally predominant as complex lipids. PE was primarily plasmalogens (62 mol%), of which arachidonic acid (47.6 mol%) and docosahexaenoic acid (11.2 mol%) were the predominant fatty acids. The component sphingoid base of sphingomyelin was almost totally 4-trans sphingenine (sphingosine). The complex lipids were able to be separated from an ethanol extract of minced skin in good yield by solvent fractionation with a hexane/ethanol system. Moreover, highly purified SM (>95 wt%) was prepared by a combination of solvent fractionation and alkaline/acidic hydrolysis from the ethanol extract. Thus, it was shown that culled chicken skin could be a potential resource of the antioxidant phospholipid plasmalogens and human-type sphingolipid.     

Synthesis and characterization of polyethoxylate surfactants derived from phenolic lipids

Polyethoxylates from cardanol and from cardol, the main component phenolic lipids in technical cashew nutshell liquid derived from the replenishable source Anacardium occidentale, were obtained by reaction with ethylene oxide under base-catalyzed conditions. Oligomeric mixtures resulted. In the cardanol series, the first six members of the mixture were independently synthesized for characterization purposes by a variety of reactions. The members in the oligomeric mixture were separated by HPLC, and the composition was determined by ¹H NMR spectroscopy and by chromatography. These studies were preparatory to surfactant and biodegradation studies involving comparison with commercially available polyethoxylates derived from petrochemical nonylphenol.     

Enzymatic fractionation and enrichment of n−9 PUFA

A single-cell oil from a Mortierella alpina mutant (TGM17 oil) contains n−9 PUFA: 14.3 wt% 6,9-octadecadienoic acid (18∶2n−9; n−9 LnA) and 17.1 wt% Mead acid (20∶3n−9; MA). Lipase screening indicated that Pseudomonas aeruginosa lipase acted strongly on n−9 LnA and weakly on MA, and Candida rugosa lipase acted weakly on the two PUFA. Hence, fractionation and enrichment of the two FA were conducted with the lipases. The first step was selective hydrolysis of IGM17 oil with P. aeruginosa lipase. The hydrolysis fractionated the oil into FFA containing 20.4 wt% n−9 LnA and 6.3 wt% MA, and acylglycerols containing 10.7 wt% n−9 LnA and 23.7 wt% MA. The FFA fraction was used for preparation of n−9 LnA-rich FFA. After removal of saturated FA, the FFA were esterified with lauryl alcohol (LauOH) using C. rugosa lipase. Two selective esterifications increased the n−9 LnA content to 54.0 wt% with 38.2% recovery of the initial content of TGM17 oil. The acylglycerol fraction obtained in the hydrolysis with P. aeruginosa lipase was used for preparation of MA-rich FFA. The acylglycerol fraction was hydrolyzed under alkaline conditions, and saturated FA were eliminated by urea adduct fractionation. Two selective esterifications of the FFA with LauOH increased the MA content to 60.2 wt% with 53.5% recovery. Thus, the two-step enzymatic process was effective for fractionation and enrichment of n−9 LnA and MA.     

Fractionation of barley straw with dilute sulfuric acid for improving hemicellulose recovery

Dilute acid fractionation of barley straw improves dissolving hemicellulose fraction of the straw, while leaving the cellulose more reactive and accessible to enzyme as a strategy of pretreatment. To characterize the fractionation process, the effects of the acid concentration, temperature and reaction time on the hemicellulose removal as well as on the formation of by-products (furfural, 5-hydroxymethylfurfural and acetic acid) were investigated. The optimum fractionation conditions of barley straw were 1% (w/v) concentration of sulfuric acid, 158 °C of reaction temperature and 15 min of reaction time. Under the optimum conditions, 87% of xylan was hydrolyzed and recovered in liquid hydrolyzate, which was 7% higher than that of the predicted yield. The hydrolyzate contained glucose 2.44 g/L, arabinose 1.70 g/L, xylose 13.41 g/L, acetic acid 1.55 g/L, levulinic acid 0.03 g/L, 5-HMF 0.03 g/L and furfural 0.75 g/L.     

Comparison of alkali and steam (acid) pretreatments of lignocellulosic materials to increase enzymic susceptibility: Evaluation under optimised pretreatment conditions

The alkali pretreatment of sugar cane bagasse was optimised and compared with an optimised steam (acid) pretreatment. The optimised alkali pretreatment gave an overall sugar yield of 37.4 g sugars per 100 g bagasse accounting for 43.8% of the initial xylan and 64.5% of the glucan. Potential effluent problems were identified with the alkali treatment and several strategies were suggested to minimise them, with liquor recycling showing some promise. The optimised steam (acid) pretreatment gave an overall sugar yield of 55.7 g sugars per 100 g bagasse accounting for 76.5% of the xylan and 89.5% of the glucan. The superiority of the steam (acid) pretreatment over the alkali pretreatment depends on the ability to utilise the xylose produced.     

Conversions of olive mill wastewater‐based media by Saccharomyces cerevisiae through sterile and non‐sterile bioprocesses

BACKGROUND: Olive mill wastewaters (OMWs) are an important residue and several methods have been proposed for their treatment. RESULTS: Remarkable decolorization (~63%) and phenol removal (~34% w/w) from OMW was achieved. In glucose‐based flask sterile cultures, enrichment with OMWs increased ethanol and biomass production compared with cultures without OMWs added. Flask sterile and un‐sterilized cultures demonstrated similar kinetic results. Batch‐bioreactor trials performed showed higher ethanol and lower biomass quantities compared with the respective shake‐flask experiments, while cultures used under un‐sterilized conditions revealed equivalent results to the sterile ones. In non‐sterile bioreactor cultures, OMWs addition enhanced biomass production in comparison with culture with no OMWs added, whereas ethanol biosynthesis was not affected. The maximum ethanol quantity achieved was 52 g L^?1 (conversion yield per sugar consumed of 0.46 g g^?1) in a batch bioreactor non‐sterilized trial with OMW–glucose enriched medium used as substrate, that presented initial reducing sugars concentration at ~115 g L^?1. Fatty acid analysis of cellular lipids demonstrated that in OMW‐based media, cellular lipids containing increased concentrations of oleic and linoleic acid were produced in comparison with cultures with no OMWs added. CONCLUSIONS: S. cerevisiae simultaneously produced bio‐ethanol and biomass and detoxified OMWs, under non‐sterile conditions. © 2012 Society of Chemical Industry     

Comparison of bioethanol production of simultaneous saccharification & fermentation and separation hydrolysis & fermentation from cellulose-rich barley straw

Cellulose rich barley straw, which has a glucan content of 62.5%, followed by dilute acid pretreatment, was converted to bioethanol by simultaneous saccharification and fermentation (SSF). The optimum fractionation conditions for barley straw were an acid concentration of 1% (w/v), a reaction temperature of 158 °C and a reaction time of 15 min. The maximum saccharification of glucan in the fractionated barley straw was 70.8% in 72 h at 60 FPU/gglucan, while the maximum digestibility of the untreated straw was only 18.9%. With 6% content WIS (water insoluble solid) for the fractionated barley straw, 70.5 and 83.2% of the saccharification yield were in SHF and SSF (representing with glucose equivalent), respectively, and a final ethanol concentration of 18.46 g/L was obtained under the optimized SSF conditions: 34 °C with 15 FPU/g-glucan enzyme loading and 1 g dry yeast cells/L. The results demonstrate that the SSF process is more effective than SHF for bioethanol production by around 18%.     

Cationic graft copolymerization of tetrahydrofuran onto bromodeoxycellulose

Cationic graft copolymerization of tetrahydrofuran onto halodeoxycelluloses, which were prepared under homogeneous conditions, was studied with silver tetrafluoroborate as an initiator. 6-Bromo-6-deoxycelluloses gave graft copolymers, insoluble but swollen in tetrahydrofuran, whereas chlorodeoxycelluloses gave no graft copolymer. The graft copolymers were hydrolyzed in 2N HCl to give hydrolyzate solutions and insoluble solids. The hydrolyzate was found to be composed of 6-O-(4-hydroxybutyl) glucose and 6-O-(9-hydroxy-5-oxanonyl) glucose as well as oligomers of tetrahydrofuran by gas chromatography and gas chromatography–mass spectrometry after trifluoroacetylation. The insoluble solid was cross-linked polyoxytetramethylene, which could be hydrolyzed only very slowly in 2N HCl to yield oligomers of tetrahydrofuran. Thermal behavior of the graft copolymer was compared with that of cellulose and of homopolymer of tetrahydrofuran. © 1994 John Wiley & Sons, Inc.     

Exploring long-range connectivity of the hard segment phase in model tri-segment oligomeric polyurethanes via lithium chloride

The influence of the extent of hydrogen bonding in mediating the long-range connectivity and percolation of the hard segment phase in model tri-segment oligomeric polyurethanes (PU) was explored by using LiCl as a molecular probe. A 22 wt% hard segment containing model PU plaque based on a mono-functional oligomeric polyether, 80:20 2,4:2,6 isomeric mixture of toluene diisocyanate, and water as a chain extender was employed. Samples cast from 20 wt% solutions in dimethyl acetamide were utilized. The tapping-mode atomic force microscopy (AFM) phase image of the solution cast film sample (soft segment T_g −63 °C) without LiCl exhibited the presence of long interconnected ribbon-like hard domains. The long-range connectivity and percolation of the hard phase that arose during plaque formation gave rise to a brittle rigid solid. A systematic break-up of the hard domains was also observed by AFM when the concentration of LiCl was increased from 0.1 to 1.5 wt%. DSC analysis indicated that the samples were able, however, to maintain a microphase separated morphology even at the highest LiCl concentration utilized in the study. FT-IR data confirmed that LiCl interacts with the hard domains of the model PU samples by disrupting the hydrogen bonding capability of the urea hard segments. A systematic softening of the samples was observed with increasing LiCl content as confirmed by thermomechanical analysis. Thus, this study indicates that hydrogen bonding plays an important role in assisting the hard segments in PU to develop long-range connectivity and percolation of this phase through the soft matrix.     

Influence of Octasilane POSS on Non-Isothermal Crystallization,Thermal Stability and Conductivity of Poly(vinyl alcohol) Nanocomposite

Nonisothermal crystallization kinetics and properties of poly(vinyl alcohol)/octasilane polyhedral oligomeric silsesquioxanes nanocomposite were investigated by differential scanning calorimetry and compared with pure poly(vinyl alcohol). The effects of octasilane polyhedral oligomeric silsesquioxanes content and cooling rate on the degree of crystallinity (X_c), crystallization half time (t_1/2), Ozawa exponent (m), and Mo kinetics parameters were studied. The role of thermal treatment on the morphology and structure of the samples was studied by X-ray diffraction and scanning electron microscopy analyses. UV–visible analysis was used to show the polyhedral oligomeric silsesquioxanes effect on the nanocomposite structure. Electrochemical impedance spectroscopy analysis demonstrated that the best conductivity was for the nanocomposite with 1 wt% of octasilane polyhedral oligomeric silsesquioxanes.     

Enzymatic hydrolysis and fermentation of lime pretreated wheat straw to ethanol

BACKGROUND: The objective of this work is to develop an efficient pretreatment method that can help enzymes break down the complex carbohydrates present in wheat straw to sugars, and to then ferment of all these sugars to ethanol. RESULTS: The yield of sugars from wheat straw (8.6%, w/v) by lime pretreatment (100 mg g^?1 straw, 121 °C, 1 h) and enzymatic hydrolysis (45 °C, pH 5.0, 120 h) using a cocktail of three commercial enzyme preparations (cellulase, β‐glucosidase, and xylanase) at the dose level of 0.15 mL of each enzyme preparation g^?1 straw was 568 ± 13 mg g^?1 (82% yield). The concentration of ethanol from lime pretreated enzyme saccharified wheat straw (78 g) hydrolyzate by recombinant Escherichia coli strain FBR5 at pH 6.5 and 35 °C in 24 h was 22.5 ± 0.6 g L^?1 with a yield of 0.50 g g^?1 available sugars (0.29 g g^?1 straw). The ethanol concentration was 20.6 ± 0.4 g L^?1 with a yield of 0.26 g g^?1 straw in the case of simultaneous saccharification and fermentation by the E. coli strain at pH 6.0 and 35 °C in 72 h. CONCLUSION: The results are important in choosing a suitable pretreatment option for developing bioprocess technologies for conversion of wheat straw to fuel ethanol. Copyright © 2007 Society of Chemical Industry     

Oil‐Fat Mixtures with Low Solid Fat Concentration: Influence of Fat Concentration and Cooling Conditions

Uniform suspension of particulates (salt or spices) in oil‐based marinades requires a gel behavior of the matrix. This can be achieved by adding a solid fat to the liquid oil. Besides rheology, appearance and thermal stability are important for the utilization as marinades. The influence of solid fat concentration (c_fat = 2.5–5.5 wt%) and average cooling speed (1.4, 2.6, and 4.7 °C/min) on the functional properties of oil‐fat gels from palm fat and canola oil was investigated. Oil‐fat mixtures showed complex physiochemical behavior depending on the solid fat concentration and cooling rate. All samples had a shear‐thinning behavior. Yield stresses and apparent viscosities increased at a constant cooling rate with increasing solid fat concentration. Frequency dependence of storage and loss modulus showed a transition from a viscous solution to a weak gel at c_fat > 3.5 wt%. Samples at increasing cooling rates transitioned to weak gels at lower fat concentration (2.5 wt%). Mixtures became turbid and increasingly whiter as both solid fat concentration and cooling rates increased, which was explained by increased light‐scattering by fat crystal aggregates. Results show the critical importance of proper formulation and preparation conditions on the functionality of oil‐based marinades.     

Blends of epoxy resins and polyphenylene oxide as processing aids and toughening agents 2: Curing kinetics,rheology, structure and properties

The curing behaviour, chemorheology, morphology and dynamic mechanical properties of epoxy ? polyphenylene oxide (PPO) blends were investigated over a wide range of compositions. Two bisphenol A based di‐epoxides ? pure and oligomeric DGEBA ? were used and their cure with primary, tertiary and quaternary amines was studied. 4,4′‐methylenebis(3‐chloro‐2,6‐diethylaniline) (MCDEA) showed high levels of cure and gave the highest exotherm peak temperature, and so was chosen for blending studies. Similarly pure DGEBA was selected for blending due to its slower reaction rate because of the absence of accelerating hydroxyl groups. For the PPO:DGEBA340/MCDEA system, the reaction rate was reduced with increasing PPO content due to a dilution effect but the heat of reaction were not significantly affected. The rheological behaviour during cure indicated that phase separation occurred prior to gelation, followed by vitrification. The times for phase separation, gelation and vitrification increased with higher PPO levels due to a reduction in the rate of polymerization. Dynamic mechanical thermal analysis of PPO:DGEBA340/MCDEA clearly showed two glass transitions due to the presence of phase separated regions where the lower T_g corresponded to an epoxy‐rich phase and the higher T_g represented the PPO‐rich phase. SEM observations of the cured PPO:DGEBA340/MCDEA blends revealed PPO particles in an epoxy matrix for blends with 10 wt% PPO, co‐continuous morphology for the blend with 30 wt% PPO and epoxy‐rich particles dispersed in a PPO‐rich matrix for 40wt% and more PPO. © 2014 Society of Chemical Industry