Xylitol production from DEO hydrolysate of corn stover by Pichia stipitis YS-30

Corn stover that had been treated with vapor-phase diethyl oxalate released a mixture of mono- and oligosaccharides consisting mainly of xylose and glucose. Following overliming and neutralization, a d-xylulokinase mutant of Pichia stipitis, FPL-YS30 (xyl3-a dagger 1), converted the stover hydrolysate into xylitol. This research examined the effects of phosphoric or gluconic acids used for neutralization and urea or ammonium sulfate used as nitrogen sources. Phosphoric acid improved color and removal of phenolic compounds. d-Gluconic acid enhanced cell growth. Ammonium sulfate increased cell yield and maximum specific cell growth rate independently of the acid used for neutralization. The highest xylitol yield (0.61 g(xylitol)/g(xylose)) and volumetric productivity (0.18 g(xylitol)/g(xylose) l) were obtained in hydrolysate neutralized with phosphoric acid. However, when urea was the nitrogen source the cell yield was less than half of that obtained with ammonium sulfate.

Fermentation of cellulosic hydrolysates obtained by enzymatic saccharification of sugarcane bagasse pretreated by hydrothermal processing

This work aims to evaluate the fermentability of cellulosic hydrolysates obtained by enzymatic saccharification of sugarcane bagasse pretreated by hydrothermal processing using Candida guilliermondii FTI 20037 yeast. The inoculum was obtained from yeast culture in a medium containing glucose as a carbon source supplemented with rice bran extract, CaCl(2)center dot 2H(2)O and (NH(4))(2)SO(4) in 50 mL Erlenmeyer flasks, containing 20 mL of medium, initial 5.5 pH under agitation of an orbital shaker (200 rpm) at 30A degrees C for 24 h. The cellulosic hydrolysates, prior to being used as a fermentation medium, were autoclaved for 15 min at 0.5 atm and supplemented with the same nutrients employed for the inoculum, except the glucose, using the same conditions for the inoculum, but with a period of 48 h. Preliminary results showed the highest consumption of glucose (97%) for all the hydrolysates, at 28 h of fermentation. The highest concentration of ethanol (20.5 g/L) was found in the procedure of sugarcane bagasse pretreated by hydrothermal processing (195A degrees C/10 min in 20 L reactor) and delignificated with NaOH 1.0% (w/v), 100A degrees C, 1 h in 500 mL stainless steel ampoules immersed in an oil bath.

Characterization of commercial cellulases and their use in the saccharification of a sugarcane bagasse sample pretreated with dilute sulfuric acid

This study aimed to correlate the efficiency of enzymatic hydrolysis of the cellulose contained in a sugarcane bagasse sample pretreated with dilute H(2)SO(4) with the levels of independent variables such as initial content of solids and loadings of enzymes and surfactant (Tween 20), for two cellulolytic commercial preparations. The preparations, designated cellulase I and cellulase II, were characterized regarding the activities of total cellulases, endoglucanase, cellobiohydrolase, cellobiase, beta-glucosidase, xylanase, and phenoloxidases (laccase, manganese and lignin peroxidases), as well as protein contents. Both extracts showed complete cellulolytic complexes and considerable activities of xylanases, without activities of phenoloxidases. For the enzymatic hydrolyses, two 2(3) central composite full factorial designs were employed to evaluate the effects caused by the initial content of solids (1.19-4.81%, w/w) and loadings of enzymes (1.9-38.1 FPU/g bagasse) and Tween 20 (0.0-0.1 g/g bagasse) on the cellulose digestibility. Within 24 h of enzymatic hydrolysis, all three independent variables influenced the conversion of cellulose by cellulase I. Using cellulase II, only enzyme and surfactant loadings showed significant effects on cellulose conversion. An additional experiment demonstrated the possibility of increasing the initial content of solids to values much higher than 4.81% (w/w) without compromising the efficiency of cellulose conversion, consequently improving the glucose concentration in the hydrolysate.

A study on the pretreatment of a sugarcane bagasse sample with dilute sulfuric acid

Experiments based on a 2(3) central composite full factorial design were carried out in 200-ml stainless-steel containers to study the pretreatment, with dilute sulfuric acid, of a sugarcane bagasse sample obtained from a local sugar-alcohol mill. The independent variables selected for study were temperature, varied from 112.5A degrees C to 157.5A degrees C, residence time, varied from 5.0 to 35.0 min, and sulfuric acid concentration, varied from 0.0% to 3.0% (w/v). Bagasse loading of 15% (w/w) was used in all experiments. Statistical analysis of the experimental results showed that all three independent variables significantly influenced the response variables, namely the bagasse solubilization, efficiency of xylose recovery in the hemicellulosic hydrolysate, efficiency of cellulose enzymatic saccharification, and percentages of cellulose, hemicellulose, and lignin in the pretreated solids. Temperature was the factor that influenced the response variables the most, followed by acid concentration and residence time, in that order. Although harsher pretreatment conditions promoted almost complete removal of the hemicellulosic fraction, the amount of xylose recovered in the hemicellulosic hydrolysate did not exceed 61.8% of the maximum theoretical value. Cellulose enzymatic saccharification was favored by more efficient removal of hemicellulose during the pretreatment. However, detoxification of the hemicellulosic hydrolysate was necessary for better bioconversion of the sugars to ethanol.

ETHANOL PRODUCTION FROM XYLOSE BY Pichia stipitis NRRL Y-7124 IN A STIRRED TANK BIOREACTOR

The ethanol production by Pichia stipitis was evaluated in a stirred tank bioreactor using semi-defined medium containing xylose (90.0 g/l) as the main carbon source. Experimental assays were performed according to a 2(2) full factorial design to evaluate the influence of aeration (0.25 to 0.75 vvm) and agitation (150 to 250 rpm) conditions on ethanol production. In the studied range of values, the agitation increase and aeration decrease favored ethanol production, which was maximum (26.7 g/l) using 250 rpm and 0.25 vvm, conditions that gave a volumetric oxygen transfer coefficient (k(L)a value) of 4.9 h(-1). Under these conditions, the ethanol yield factor, ethanol productivity, and the process efficiency were 0.32 g/g, 0.32 g/l.h, and 63%, respectively. These results are promising and contribute to the development of a suitable process for ethanol production from xylose by Pichia stipitis.

Production of fibrolytic enzymes by Aspergillus japonicus C03 using agro-industrial residues with potential application as additives in animal feed

Solid-state fermentation obtained from different and low-cost carbon sources was evaluated to endocellulases and endoxylanases production by Aspergillus japonicus C03. Regarding the enzymatic production the highest levels were observed at 30 degrees C, using soy bran added to crushed corncob or wheat bran added to sugarcane bagasse, humidified with salt solutions, and incubated for 3 days (xylanase) or 6 days (cellulase) with 70% relative humidity. Peptone improved the xylanase and cellulase activities in 12 and 29%, respectively. The optimum temperature corresponded to 60 degrees C and 50-55 degrees C for xylanase and cellulase, respectively, both having 4.0 as optimum pH. Xylanase was fully stable up to 40 degrees C, which is close to the rumen temperature. The enzymes were stable in pH 4.0-7.0. Cu(++) and Mn(++) increased xylanase and cellulase activities by 10 and 64%, respectively. A. japonicus C03 xylanase was greatly stable in goat rumen fluid for 4 h during in vivo and in vitro experiments.

Enzymatic Hydrolysis of Chemithermomechanically Pretreated Sugarcane Bagasse and Samples with Reduced Initial Lignin Content

Chemithermomechanical (CTM) processing was used to pretreat sugarcane bagasse with the aim of increasing cell wall accessibility to hydrolytic enzymes. Yields of the pretreated samples were in the range of 75-94%. Disk refining and alkaline-CTM and alkaline/sulfite-CTM pretreatments yielded pretreated materials with 21.7, 17.8, and 15.3% of lignin, respectively. Hemicellulose content was also decreased to some extent. Fibers of the pretreated materials presented some external fibrillation, fiber curling, increased swelling, and high water retention capacity. Cellulose conversion of the alkaline-CTM- and alkaline/sulfite-CTM-pretreated samples reached 50 and 85%, respectively, after 96 h of enzymatic hydrolysis. Two samples with low initial lignin content were also evaluated after the mildest alkaline-CTM pretreatment. One sample was a partially delignified mill-processed bagasse. The other was a sugarcane hybrid selected in a breeding program. Samples with lower initial lignin content were hydrolyzed considerably faster in the first 24 h of enzymatic digestion. For example, enzymatic hydrolysis of the sample with the lowest initial lignin content (14.2%) reached 64% cellulose conversion after only 24 h of hydrolysis when compared with the 30% observed for the mill-processed bagasse containing an initial lignin content of 24.4%. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 27: 395-401, 2011

Nutritional characteristics and in vitro digestibility of silages from different corn cultivars harvested at two cutting heights

The objective of this study was to evaluate the nutritional traits and in vitro digestibility of silages from different corn cultivars harvested at two cutting heights. It was evaluated 11 cultivars (Dina 766, Dina 657, Dina 1000, P 3021, P 3041, C 805, C 333, AG 5011, FO 01, Dina co 9621 and BR 205) harvest 5 cm above ground (low) and 5 cm below the intersection of the first ear (high). It was used a random block design (three blocks), arranged in a 11 × 2 factorial scheme. Silages from plants harvested at high cutting height presented average content of dry matter significantly superior to silages from plants harvested at low height. Cultivars FO 01, AG 5011, Dina co 9621 and Dina 766 presented greater content of crude protein than cultivars C 805, P 3041 and P 3021, which presented the lowest contents of this nutrient. The raise in the cut height increased in vitro dry matter true digestibility coefficients and in vitro dry matter digestibility of silage evaluated. The increase in cut height improved nutritive value of silages by decreasing concentrations of fibrous fractions and increasing in vitro dry matter digestibility.

Evaluation of corn cultivars harvested at two cutting heights for ensilage

The objective of this study was to evaluate the agronomic characteristics, bromatological-chemical composition and digestibility of 11 corn cultivars (Zea mays) harvested at two cutting heights. Cultivars D 766, D 657, D 1000, P 3021, P 3041, C 805, C 333, AG 5011, FO 01, CO 9621 and BR 205 were evaluated when they were harvested 5 cm above ground (low) and 5 cm below the insertion of the first ear (high). The experiment was designed as random blocks, with three replicates, arranged in an 11 x 2 factorial scheme. Cultivars presented similar productions of forage dry matter and grains. Percentages of stalk, leaf, straw, cob and kernel fractions were different among cultivars, as well as dry matter content of the whole plant at harvest. Considering the whole plant, only the contents of gross energy, nitrogen in neutral detergent fiber, and in vitro neutral and acid detergent fiber digestibility did not differ among cultivars. Increase on the cutting height improved forage quality due to the reduction of stalk and leaf fractions and contents of cell wall constituents.

Storage effect on the acceptability of snacks made of pure amaranth and blends of amaranth and corn or chickpea

Snacks made by extrusion cooking of pure amaranth flour or mixtures of 80 per cent amaranth flour and 20 per centcorn grits or chickpea flour were developed to replace the traditional commercial ones with improved nutritional and functional quality. Pure amaranth snacks and the blended ones were flavored with salty and sweet flavors and evaluated for acceptability using a 9-point hedonic scale. The good acceptance observed for either salty or sweet flavored snacks indicated that they have characteristics to compete with similar commercial products. Acceptability of salty snacks increased with storage time at room temperature in BOPP (polypropylene bi-guided) packs whereas slightly decreased for the sweet ones. This type of storage proved to be very efficient for the conservation of the salty product and also suitable for the sweet ones

Effect of fructans-based fat replacer on chemical composition, starch digestibility and sensory acceptability of corn snacks

To develop a convenience healthy food snack the partially hydrogenated vegetable fat, used as the flavour fixative agent, was replaced by a non-fat-flavouring solution enriched with inulin and oligofructose. The effects of this replacement on chemical composition, in vitro rate of starch digestion and sensory acceptability were assessed. The new snack presented low-fat levels (0.1 per cent) and around a sevenfold increase in dietary fibre (15.3 per cent of dietary fibre, being 13.3 per cent of fructans) when compared with the traditional ones. The enrichment with fructans reduced the predicted Glycaemic Index by 25 per cent, thus indicating that this dietary fibre contributes effectively towards delaying the in vitro glycaemic response. Fructans-enriched snack presented overall acceptability score (6.6 ± 1.7) similar to the traditional one, flavoured with fatty fixative agent (7.4 ± 1.4). The healthy low-fat fibre-enriched snack produced presented the high sensory acceptability typical for this food product type

Mycoflora and Co-Occurrence of Fumonisins and Aflatoxins in Freshly Harvested Corn in Different Regions of Brazil

Natural mycoflora and co-occurrence of fumonisins (FB(1), FB(2)) and aflatoxins (AFB(1), AFB(2), AFG(1) and AFG(2)) in freshly harvested corn grain samples from four regions of Brazil were investigated. Fusarium verticillioides was predominant in all samples. Analysis of fumonisins showed that 98% of the samples were contaminated with FB(1) and 74.5% with FB(1) + FB(2), with toxin levels ranging from 0.015 to 9.67 mu g/g for FB(1) and from 0.015 to 3.16 mu g/g for FB(2). Twenty-one (10.5%) samples were contaminated with AFB(1), seven (3.5%) with AFB(2) and only one (0.5%) with AFG(1) and AFG(2). Co-contamination with aflatoxins and fumonisins was observed in 7% of the samples. The highest contamination of fumonisins and aflatoxins was observed in Nova Odessa (SP) and Varzea Grande (MT), respectively. The lowest contamination of these mycotoxins was found in Varzea Grande and Nova Odessa, respectively.

Economic viability of doses and split-applications of nitrogen fertilization in corn crop in a eutrophic Red Latosol

Nitrogen is the nutrient that is most absorbed by the corn crop, with the most complex management, and has the highest share on the cost of corn production. The objective of this work was to evaluate the economic viability of different rates and split-applications of nitrogen fertilization, as such as urea, in the corn crop in a eutrophic Red Latosol (Oxisol). The study was carried out in the Experimental Station of the Regional Pole of the Sao Paulo Northwest Agribusiness Development (APTA), in Votuporanga, State of Sao Paulo, Brazil. The experimental design was randomized complete blocks with nine treatments and four replications, consisting of five N rates: 0, 55, 95, 135 and 175 kg ha(-1), 15 kg ha-l applied in the seeding and the remainder in top dressing: 40 and 80 kg ha(-1) N at forty days after seeding (DAS), or 1/2 + 1/2 at 20 and 40 DAS; 120 kg ha-1 N split in 1/2 + 1/2 or 1/3 + 1/3 + 1/3 at 20, 40 or 60 DAS; 160 kg ha(-1) N split in 1/4 + 3/8 + 3/8 or 114 + 1/4 + 1/4 + 1/4 at 20, 40, 60 and 80 DAS. The application of 135 kg ha-l of N split in three times provided the best benefit/cost ratio. The non-application of N provided the lowest economic return, proving to be unviable.

Xylooligosaccharides Production from Alkali-Pretreated Sugarcane Bagasse Using Xylanases from Thermoascus aurantiacus

Sugarcane bagasse hemicellulose was isolated in a one-step chemical extraction using hydrogen peroxide in alkaline media. The polysaccharide containing 80.9% xylose and small amounts of L-arabinose, 4-O-methyl-D-glucuronic acid and glucose, was hydrolyzed by crude enzymatic extracts from Thermoascus aurantiacus at 50 degrees C. Conditions of enzymatic hydrolysis leading to the best yields of xylose and xylooligosaccharides (DP 2-5) were investigated using substrate concentration in the range 0.5-3.5% (w/v), enzyme load 40-80 U/g of the substrate, and reaction time from 3 to 96 h, applying a 22 factorial design. The maximum conversion to xylooligosaccharides (37.1%) was obtained with 2.6% of substrate and xylanase load of 60 U/g. The predicted maximum yield of xylobiose by a polynomial model was 41.6%. Crude enzymatic extract of T. aurantiacus generate from sugarcane bagasse hemicellulose 39% of xylose, 59% of xylobiose, and 2% of other xylooligosaccharides.

The roles of xylan and lignin in oxalic acid pretreated corncob during separate enzymatic hydrolysis and ethanol fermentation

High yields of hemicellulosic and cellulosic sugars are critical in obtaining economical conversion of agricultural residues to ethanol. To optimize pretreatment conditions, we evaluated oxalic acid loading rates, treatment temperatures and times in a 2(3) full factorial design. Response-surface analysis revealed an optimal oxalic acid pretreatment condition to release sugar from the cob of Zea mays L ssp. and for Pichia stipitis CBS 6054. To ferment the residual cellulosic sugars to ethanol following enzymatic hydrolysis, highest saccharification and fermentation yields were obtained following pretreatment at 180 degrees C for 50 min with 0.024 g oxalic acid/g substrate. Under these conditions, only 7.5% hemicellulose remained in the pretreated substrate. The rate of cellulose degradation was significantly less than that of hemicellulose and its hydrolysis was not as extensive. Subsequent enzymatic saccharification of the residual cellulose was strongly affected by the pretreatment condition with cellulose hydrolysis ranging between 26.0% and 76.2%. The residual xylan/lignin ratio ranged from 0.31 to 1.85 depending on the pretreatment condition. Fermentable sugar and ethanol were maximal at the lowest ratio of xylan/lignin and at high glucan contents. The model predicts optimal condition of oxalic acid pretreatment at 168 degrees C, 74 min and 0.027 g/g of oxalic acid. From these findings, we surmised that low residual xylan was critical in obtaining maximal glucose yields from saccharification. Published by Elsevier Ltd.