Sugarcane Bagasse Hemicellulosic Hydrolysate Research Articles

Enhanced Xylitol Production by Precultivation of Candida guilliermondii Cells in Sugarcane Bagasse Hemicellulosic Hydrolysate

The present work evaluated the key enzymes involved in xylitol production (xylose reductase [XR] and xylitol dehydrogenase [XDH]) and their correlation with xylose, arabinose, and acetic acid assimilation during cultivation of Candida guilliermondii FTI 20037 cells in sugarcane bagasse hemicellulosic hydrolysate. For this purpose, inocula previously grown either in sugarcane bagasse hemicellulosic hydrolysate (SBHH) or in semidefined medium (xylose as a substrate) were used. The highest xylose/acetic acid consumption ratio (1.78) and the lowest arabinose consumption (13%) were attained in the fermentation using inoculum previously grown in semidefined medium (without acetic acid and arabinose). In this case, the highest values of XR (1.37 U mg prot(-1)) and XDH (0.91 U mg prot(-1)) activities were observed. The highest xylitol yield (approximately 0.55 g g(-1)) and byproducts (ethanol and glycerol) formation were not influenced by inoculum procedure. However, the cell previously grown in the hydrolysate was effective in enhancing xylitol production by keeping the XR enzyme activity at high levels (around 0.99 U.mg(prot) (-1)), reducing the XDH activity (34.0%) and increasing xylitol volumetric productivity (26.5%) with respect to the inoculum cultivated in semidefined medium. Therefore, inoculum adaptation to SBHH was shown to be an important strategy to improve xylitol productivity.

Effect of glucose:xylose ratio on xylose reductase and xylitol dehydrogenase activities from Candida guilliermondii in sugarcane bagasse hydrolysate

AbstractThe influence of glucose on xylose reductase (XR) and xylitol dehydrogenase (XDH) enzyme activity was evaluated from sugarcane bagasse hydrolysate fermentations with different glucose:xylose ratios (1:25, 1:12, 1:5 and 1:2.5) by employing an inoculum of Candida guilliermondii grown in media containing glucose, a mixture of glucose and xylose, or only xylose as carbon sources. According to the results, the glucose:xylose ratio affected positively this bioconversion and a correlation was not observed between the favourable conditions for xylitol production and the XR and XDH activities. Also, the results were influenced not only by the glucose:xylose ratio in the fermentation medium, but also by the carbon source employed in the growth medium of the inoculum. The optimum condition for xylitol production by C. guilliermondii in sugarcane bagasse hemicellulosic hydrolysate should use hydrolysate with a 1:5 glucose:xylose ratio and inoculum grown in medium containing xylose as the only carbon source. Copyright © 2006 Society of Chemical Industry

Yeast Immobilization in LentiKats®: A New Strategy for Xylitol Bioproduction from Sugarcane Bagasse

A new PVA-hydrogel matrix for yeast cell immobilization for xylitol bioproduction from sugarcane bagasse was studied. Five repeated-batch fermentation runs were carried out in medium based on sugarcane bagasse hemicellulosic hydrolysate with reuse of the entrapped biocatalyst. The system performance as well as the metabolic behaviour of cells entrapped into the matrix were evaluated. The biocatalyst remained stable and exhibited a similar fermentative profile in all the successive batches, demonstrating the viability of the system. At the end of the run, an average xylitol production was observed of 35.1 g l−1 and average xylitol yield and productivity of 0.58 g g−1 and 0.49 g l−1 h−1, respectively.

Purification of Xylitol from Fermented Hemicellulosic Hydrolyzate Using Liquid–Liquid Extraction and Precipitation Techniques

Xylitol was produced by Candida guilliermondii by fermentation of sugarcane bagasse hemicellulosic hydrolysate. Undesirable impurities were extracted from the broth using either ethyl acetate, chloroform or dichloromethane. The best results on clarification of the broth without xylitol loss were obtained with ethyl acetate. When ethanol, acetone or tetrahydrofuran were used for precipitation of impurities, only tetrahydrofuran clarified the fermented broth, but a high xylitol loss (approximately 30%) was observed.

Simultaneous Saccharification and Co‐Fermentation of Crystalline Cellulose and Sugar Cane Bagasse Hemicellulose Hydrolysate to Lactate by a Thermotolerant Acidophilic Bacillus sp.

Polylactides produced from renewable feedstocks, such as corn starch, are being developed as alternatives to plastics derived from petroleum. In addition to corn, other less expensive biomass resources can be readily converted to component sugars (glucose, xylose, etc.) by enzyme and/or chemical treatment for fermentation to optically pure lactic acid to reduce the cost of lactic acid. Lactic acid bacteria used by the industry lack the ability to ferment pentoses (hemicellulose-derived xylose and arabinose), and their growth and fermentation optima also differ from the optimal conditions for the activity of fungal cellulases required for depolymerization of cellulose. To reduce the overall cost of simultaneous saccharification and fermentation (SSF) of cellulose, we have isolated bacterial biocatalysts that can grow and ferment all sugars in the biomass at conditions that are also optimal for fungal cellulases. SSF of Solka Floc cellulose by one such isolate, Bacillus sp. strain 36D1, yielded l(+)-lactic acid at an optical purity higher than 95% with cellulase (Spezyme CE; Genencor International) added at about 10 FPU/g cellulose, with a product yield of about 90% of the expected maximum. Volumetric productivity of SSF to lactic acid was optimal between culture pH values of 4.5 and 5.5 at 50 degrees C. At a constant pH of 5.0, volumetric productivity of lactic acid was maximal at 55 degrees C. Strain 36D1 also co-fermented cellulose-derived glucose and sugar cane bagasse hemicellulose-derived xylose simultaneously (SSCF). In a batch SSCF of 40% acid-treated hemicellulose hydrolysate (over-limed) and 20 g/L Solka Floc cellulose, strain 36D1 produced about 35 g/L lactic acid in about 144 h with 15 FPU of Spezyme CE/g cellulose. The maximum volumetric productivity of lactic acid in this SSCF was 6.7 mmol/L (h). Cellulose-derived lactic acid contributed to about 30% of this total lactic acid. These results show that Bacillus sp. strain 36D1 is well-suited for simultaneous saccharification and co-fermentation of all of the biomass-derived sugars to lactic acid.

Evaluation of porous glass and zeolite as cells carriers for xylitol production from sugarcane bagasse hydrolysate

Adsorbing carriers for immobilisation of Candida guilliermondii cells to use for xylitol production from sugarcane bagasse hemicellulose hydrolysate were tested. Biomass was immobilised in situ by natural adsorption, i.e. through direct contact between cells for the inoculum and carrier particles at the beginning of fermentations. The carriers employed were: Syrane porous glass beads with 2.53 mm diameter in average and pore diameter in the range 60–300 μm, supplied by Bioengineering (Wald, Switzerland), and NaX zeolite UOP WE 894 purchased from Plury Química S.A. (Diadema, SP, Brazil). At the end of the run with free cells taken as a reference test, xylitol concentration ( P f) achieved 35.5 g/l, corresponding to a xylose-to-xylitol yield factor ( Y P/S) of 0.72 g/g and a volumetric productivity ( Q P) of 0.49 g/l h, while final cell concentration ( X f) and productivity ( Q x) were only 5.32 g/l and 0.048 g/l h, respectively. Both systems with immobilised cells exhibited lower xylitol productions ( P f = 28.8–29.5 g/l, Y P/S = 0.52–0.53 g/g, Q P = 0.32–0.33 g/l h) and higher cell growth, with particular concern to porous glass ( X f = 10.5 g/l, Q x = 0.10 g/l h). Electronic microscopy observations demonstrated that the excellent performance of porous glass as cell support was due to the development of a thick extracellular matrix either within the large pores or on the surface of this material. As a consequence, almost 50% of the cells resulted to be adsorbed to the carrier at the end of the run. This growth was also responsible for a decrease in the fraction of xylose available for xylitol production. Employing zeolite, a material with pore size smaller than cell size, immobilised cells represented only 30% of the final population and immobilisation was just observed on the carrier surface. The low cell attachment on this material can be explained by the stress exerted on the outer immobilised cells by the friction among beads.

Fermentation of sugar cane bagasse hemicellulose hydrolysate to L(+)-lactic acid by a thermotolerant acidophilic Bacillus sp.

Sugar cane bagasse hemicellulose, hydrolyzed by dilute H2SO4, supplemented with mineral salts and 0.5% corn steep liquor, was fermented to L(+)-lactic acid using a newly isolated strain of Bacillus sp. In batch fermentations at 50 degrees C and pH 5, over 5.5% (w/v) L(+)-lactic acid was produced (89% theoretical yield; 0.9 g lactate per g sugar) with an optical purity of 99.5%.

Detoxification of sugarcane bagasse hemicellulosic hydrolysate with ion‐exchange resins for xylitol production by calcium alginate‐entrapped cells

AbstractThis study describes the performance of four different resins, in sequence, to detoxify sugarcane bagasse hemicellulosic hydrolysate and to improve xylitol production by calcium alginate‐entrapped Candida guilliermondii FTI20037 cells under conditions of low oxygen concentration. The treatment resulted in a removal of 82.1% furfural, 66.5% hydroxymethylfurfural, 61.9% phenolic compounds derived from lignin degradation, 100% chromium, 46.1% zinc, 28.5% iron, 14.7% sodium and 3.5% nickel. On the other hand, the removal of acetic acid was not significant. A xylitol yield factor (YP/S) of 0.62 g g−1 and a volumetric productivity (Qp) of 0.24 g dm−3 h−1 were attained in the fermentation process for xylitol production from detoxified hydrolysate. Copyright © 2004 Society of Chemical Industry

Optimation xilitol production with variation of sugar cane bagasse hemicellulose hydrolysate concentration by Candida tropicalis

The aims of this research were to study the growth of C. tropicalis, the optimation of xilitol production and the efficiency of xilitol production by varying the concentration of sugar cane bagasse hemicellulose hydrolysate. The frame work of this research was bioconversion xilosa into xilitol by C. tropicalis influenced substrate concentration in production medium. By using different sugar cane bagasse hemicellulose hydrolysate concentration could be known the optimum sugar cane bagasse hemicellulose hydrolysate concentration on xilitol production by introducing C. tropicalis. The methods used in this experiment were as follow: sugar cane bagasse was hydrolyzed by H2SO4 0.035 M in autoclave at 121ï‚°C, 2 atm for 20 minutes. The liquid fraction was concentrated at 50ï‚°C using rotary evaporator. The hydrolysate was neutralized with Ca(OH)2 to a pH of 10 and then removed by centrifugation at 2000 rpm for 20 minutes. The hydrolysate was added with H2SO4 to a pH 6.5 and then removed by centrifugation at 2000 rpm for 20 minutes and sterilized by autoclaving at 121ï‚°C for 15 minutes. Sugar cane bagasse hemicellulose hydrolysate concentration was variated from 10%, 20%, and 30% and used as substrate in production medium. Bioconversion process by C. tropicalis lasted for 4 days. Parameters used in this experiment were biomass of C. tropicalis measured by hemacytometer, xilosa and xilitol concentration in the sugar cane bagasse hemicellulose hydrolysate and production medium was analyzed by High Performance Liquid Chromatographic (HPLC). The results showed that: the growth of C. tropicalis lasted for 4 days of cultivation increased in every variation of sugar cane hemicellulose hydrolysate concentration. The optimum xilitol production founded in production medium contained 20% sugar cane bagasse hemicellulose hydrolysate in the third days cultivation with xilitol production 10.258 g/l, yield 0.22 g/g and biomass of C. tropicalis 2.9 x 108 cell/ml; The optimum efficiency of xilitol production from sugar cane bagasse hemicellulose hydrolysate by C. tropicalis was 24.21% in 20% sugar cane bagasse hemicellulose hydrolysate in the third days cultivation.

A study on xylitol production from sugarcane bagasse hemicellulosic hydrolysate by Ca-alginate entrapped cells in a stirred tank reactor

Candida guilliermondii FTI 20037 cells were entrapped in Ca-alginate beads and used for xylitol production from sugarcane bagasse hemicellulosic hydrolysate in a stirred tank reactor (STR). Screening design and response surface methodologies were used to determine adequate cultivation conditions for this fermentation system. Quadratic models were fitted to the experimental data by regression analysis, considering the yield ( Y P/S) and the productivity ( Q P) of the xylose-to-xylitol bioconversion as dependent variables. Using a five-fold concentrated hydrolysate, air flowrate of 1.30 l/min, agitation speed of 300 rpm, initial cell concentration of 1.4 g/l and value 6.0 for the initial pH of the fermentation medium resulted in a xylitol production of 47.5 g/l after 120 h of fermentation, corresponding to a Y P/S of 0.81 g/g and to a Q P of 0.40 g/l h.

Xylitol production by Ca-alginate entrapped cells: comparison of different fermentation systems

Candida guilliermondii cells were entrapped in Ca-alginate beads and used for xylitol production from concentrated sugarcane bagasse hemicellulosic hydrolysate in three different fermentation systems, namely 125-ml Erlenmeyer flasks (EF), 2.4-l stirred tank reactor (STR) and 2.4-l basket-type stirred tank reactor (BSTR). The EF system provided a xylitol production of 21.0 g P l −1, a product yield based on xylose consumption of 0.54 g P g S −1, and an overall production rate of 0.44 g P l −1 h −1 after 48 h of fermentation. By the STR system, 23.5 g P l −1 was produced after 60 h of fermentation, corresponding to a yield of 0.58 g P g S −1 and a production rate of 0.39 g P l −1 h −1. As the average volume of the beads decreased by 10.7% ( p<0.05) during this fermentation, a basket was fitted into the reactor vessel to prevent the beads abrasion and thus maintain their integrity. However, the xylitol yield (0.46 g P g S −1), production rate (0.21 g P l −1 h −1) and concentration (15.0 g P l −1) attained by this system were affected by mass-transfer limitations.

The influence of pH and dilution rate on continuous production of xylitol from sugarcane bagasse hemicellulosic hydrolysate by C. guilliermondii

Continuous fermentation of sugarcane bagasse hemicellulosic hydrolysate by the yeast Candida guilliermondii FTI 20037 was used for xylitol production from xylose. Experiments were carried out in a reactor with 1.25 l of treated hydrolysate, at 30 °C and 300 rpm. A 2 2 full-factorial central composite design was employed for experimental study and analysis of the results. A statistical analysis of the results showed that the effects of the pH and dilution rate ( D ), the interactions between these variables and the second-order effect of D on the xylitol volumetric productivity ( Q p ) were significant at a 95% confidence level. The second-order effect of pH was also significant at a 90% confidence level. The k L a effect on the Q p was not significant. A volumetric productivity of 0.68 g/l h, representing 95.8% of the predicted value (0.72 g/l h), was obtained.

Xylose reductase and xylitol dehydrogenase activities of Candida guilliermondii as a function of different treatments of sugarcane bagasse hemicellulosic hydrolysate employing experimental design.

The sugarcane bagasse hydrolysate, which is rich in xylose, can be used as culture medium for Candida guilliermondii in xylitol production. However, the hydrolysate obtained from bagasse by acid hydrolysis at 120 degrees C for 20 min has by-products (acetic acid and furfural, among others), which are toxic to the yeast over certain concentrations. So, the hydrolysate must be pretreated before using in fermentation. The pretreatment variables considered were: adsorption time (15,37.5, and 60 min), type of acid used (H2So4 and H3Po4), hydrolysate concentration (original, twofold, and fourfold concentrated), and active charcoal (0.5, 1.75 and 3.0%). The suitability of the pretreatment was followed by measuring the xylose reductase (XR) and xylitol dehydrogenase (XD) activity of yeast grown in each treated hydrolysate. The response surface methodology (2(4) full factorial design with a centered face) indicated that the hydrolysate might be concentrated fourfold and the pH adjusted to 7.0 with CaO, followed by reduction to 5.5 with H3PO4. After that it was treated with active charcoal (3.0%) by 60 min. This pretreated hydrolysate attained the high XR/XD ratio of 4.5.

Use of immobilized Candida cells on xylitol production from sugarcane bagasse.

In this study we used the yeast Candida guilliermondii FTI 20037 immobilized by entrapment in Ca-alginate beads (2.5-3 mm diameter) for xylitol production from concentrated sugarcane bagasse hemicellulosic hydrolysate in a repeated batch system. The fermentation runs were carried out in 125- and 250-ml Erlenmeyer flasks placed in an orbital shaker at 30 degrees C and 200 rpm during 72 h, keeping constant the proportion between work volume and flask total volume. According to the results, cell viability was substantially high (98%) in all fermentative cycles. The values of parameters xylitol yield and volumetric productivity increased significantly with the reutilization of the immobilized biocatalysts. The highest values of xylitol final concentration (11.05 g/l), yield factor (0.47 g/g) and volumetric productivity (0.22 g/lh) were obtained in 250-ml Erlenmeyer flasks containing 80 ml of medium plus 20 ml of immobilized biocatalysts. The support used in this study (Ca-alginate) presented stability in the experimental conditions used. The results show that the use of immobilized cells is a promising approach for increasing the xylitol production rates.

Fermentation of sugar cane bagasse hemicellulosic hydrolysate and sugar mixtures to ethanol by recombinant Escherichia coli KO11

Escherichia coli KO11, carrying the ethanol pathway genes pdc (pyruvate decarboxylase) and adh (alcohol dehydrogenase) from Zymomonas mobilis integrated into its chromosome, has the ability to metabolize pentoses and hexoses to ethanol, both in synthetic medium and in hemicellulosic hydrolysates. In the fermentation of sugar mixtures simulating hemicellulose hydrolysate sugar composition (10.0 g of glucose/l and 40.0 g of xylose/l) and supplemented with tryptone and yeast extract, recombinant bacteria produced 24.58 g of ethanol/l, equivalent to 96.4% of the maximum theoretical yield. Corn steep powder (CSP), a byproduct of the corn starch-processing industry, was used to replace tryptone and yeast extract. At a concentration of 12.5 g/l, it was able to support the fermentation of glucose (80.0 g/l) to ethanol, with both ethanol yield and volumetric productivity comparable to those obtained with fermentation media containing tryptone and yeast extract. Hemicellulose hydrolysate of sugar cane bagasse supplemented with tryptone and yeast extract was also readily fermented to ethanol within 48 h, and ethanol yield achieved 91.5% of the theoretical maximum conversion efficiency. However, fermentation of bagasse hydrolysate supplemented with 12.5 g of CSP/l took twice as long to complete.

Effect of the oxygen transfer coefficient on xylitol production from sugarcane bagasse hydrolysate by continuous stirred-tank reactor fermentation.

The effect of the oxygen transfer coefficient on the production of xylitol by bioconversion of xylose present in sugarcane bagasse hemicellulosic hydrolysate using the yeast Candida guilliermondii was investigated. Continuous cultivation was carried out in a 1.25-L fermentor at 30 degrees C, pH 5.5, 300 rpm, and a dilution rate of 0.03/h, using oxygen transfer coefficients of 10, 20, and 30/h. The results showed that the microbial xylitol production (11 g/L) increased by 108% with the decrease in the oxygen volumetric transfer coefficient from 30 to 20/h. The maximum values of xylitol productivity (0.7 g/[L.h]) and yield (0.58 g/g) were obtained akLa 20/h.

Inhibition of microbial xylitol production by acetic acid and its relation with fermentative parameters.

Precipitated sugarcane bagasse hemicellulosic hydrolysate containing acetic acid was fermented by Candida guilliermondii FTI20037 under different operational conditions (pH 4.0 and 7.0, three aeration rates). At pH 7.0 and kLa of 10 (0.75 vvm) and 22.5/h (3.0 vvm) the acetic acid had not been consumed until the end of the fermentations, whereas at the same pH and kLa of 35/h (4.5 vvm) the acid was rapidly consumed and acetic acid inhibition was not important. On the other hand, fermentations at an initial pH of 4.0 and kLa of 22.5 and 35/h required less time for the acid uptake than fermentations at kLa of 10/h. The acetic acid assimilation by the yeast indicates the ability of this strain to ferment in partially detoxified medium, making possible the utilization of the sugarcane bagasse hydrolysate in this bio-process. The effects on xylitol yield and production are reported.

Adaptation and reutilization of Candida guilliermondii cells for xylitol production in bagasse hydrolysate.

The xylitol productivity increased by about 15% with the use of cells of Candida guilliermondii FTI 20037 previously recycled through four consecutive batch cultures and adapted to the sugar cane bagasse hemicellulosic hydrolysate. Furthermore, the more concentrated the hydrolysate, the more necessary was the adaptation of the cells, owing to the presence of toxic substances at high concentration which inhibited the xylose-xylitol conversion by the yeast.

Xylanase recovery by ethanol and Na2SO4 precipitation

Xylans are the major components of the hemicellulosic fraction of lignocellulosic biomass and their hydrolysis can be obtained using xylanases from Penicillium janthinellum. In this work, sugarcane bagasse hemicellulosic hydrolysate was used as the substrate for producing xylanase. The precipitation of these enzymes was studied using ethanol and Na2SO4 as precipitating agents. Ethanol precipitation experiments were performed batchwise in concentrations ranging from 10 to 80%, pH 4.0 to 7.0, at 4 degrees C. The concentrations used in the precipitations with Na2SO4 were from 5 to 60% at pH 5.5 and 25 degrees C. Solubility curves as a function of xylanase activity and total protein for both precipitating agents were made. According to the results, Na2SO4 is not appropriate for precipitating xylanases in this medium since at salt concentrations higher than 25%, the enzyme was denaturated and at this concentration less than 80% of the enzyme and total protein were precipitated. Because of differences in xylanase and total protein solubility, a fractionated precipitation using ethanol can be performed, since with 40% ethanol, 49% of the total protein was precipitated and more than 95% of the enzyme was kept in solution. On the other hand approx 100% of the xylanases were recovered by precipitation after adding 80% ethanol.

Maximizing the xylitol production from sugar cane bagasse hydrolysate by controlling the aeration rate

Batch fermentations of sugar cane bagasse hemicellulosic hydrolysate treated for removing the inhibitors of the fermentation were performed by Candida guilliermondii FTI20037 for xylitol production. The fermentative parameters agitation and aeration rate were studied aiming the maximization of xylitol production from this agroindustrial residue. The maximal xylitol volumetric productivity (0.87 g/L h) and yield (0.67 g/g) were attained at 400/min and 0.45 v.v.m. (K(L)a 27/h). According to the results, a suitable control of the oxygen input permitting the xylitol formation from sugar cane bagasse hydrolysate is required for the development of an efficient fermentation process for large-scale applications.