Rice Straw Hemicellulose Hydrolysate Research Articles

Evaluation of Fermentative Xylitol Production Potential of Adapted Strains of Meyerozyma caribbica and Candida tropicalis from Rice Straw Hemicellulosic Hydrolysate

Dilute acid hydrolysis of lignocellulosic biomass generates inhibitors in the hydrolysate which hamper yeast metabolism and the fermentation process. Therefore, understanding the effect of these compounds on the performance of microorganisms becomes essential to achieve improved product yields. In this study, the effect of acetic acid, furfural, and hydroxymethylfurfural was evaluated on yeast growth and fermentation efficiency. Various parameters for the pretreatment of rice straw, such as an acid catalyst, and its concentration and residence time, were optimized for the maximum liberation of sugars in the hydrolysate. Further, the yeast strains Candida tropicalis and Meyerozyma caribbica were adapted for the tolerance of inhibitors at higher concentrations. A comparative analysis was carried out using un-adapted and adapted strains of Candida tropicalis and Meyerozyma caribbica for xylitol production. The findings of this study revealed that sulfuric acid (1.25% v/v) at 121 ரC for 30 min can efficiently convert rice straw xylan to xylose, with the release of 16.07 g/L xylose in the hydrolysate. Further, the adaptation results showed an increase of 76.42% and 69.33% in xylose assimilation by C. tropicalis and M. caribbica, respectively. The xylitol production with the adapted C. tropicalis was increased by 7.54% to 28.03 g/L xylitol. However, the xylitol production with the adapted M. caribbica was increased by 8.33%, yielding 26.02 g/L xylitol in the non-detoxified hydrolysate when compared to the un-adapted strains. Repeated batch fermentation was carried out for seven batches, and xylitol was found to be efficiently produced by the yeasts during five successive batches without any significant loss in the xylitol yield. Moreover, the results suggest that M. caribbica is a promising microorganism for the transformation of rice straw-derived xylose to xylitol.

Xylitol production from rice straw hemicellulosic hydrolysate by Candida tropicalis GS18 immobilized on bacterial cellulose-sodium alginate matrix

Xylitol production from rice straw hemicellulosic hydrolysate by Candida tropicalis GS18 immobilized on bacterial cellulose-sodium alginate matrix

Assessment of advanced oxidative processes based on heterogeneous catalysis as a detoxification method of rice straw hemicellulose hydrolysate and their effect on ethanol production by Pichia stipitis

In the present work, detoxification treatments based on advanced oxidative processes (AOPs) aiming to improve the ethanol production by Pichia stipitis were evaluated. The experimental design was carried out according to a Taguchi L8 orthogonal array in order to evaluate the influences of pH, catalyst type (ZnO or TiO2), UV radiation (UVA or UVC), and oxidant agent (O2 or O3) on aromatic compounds concentration and fermentation performed. The results showed that treatment conditions which most contribute to reducing the toxicity of the hydrolysate in relation to ethanol production were pH 8.0 and the use of O3 as an oxidizing agent. Within the region evaluated, there was no difference between the use of TiO2 or ZnO and or UVA or UVC radiation. The heterogeneous AOPs were able to remove above 49 % of the furan, above 39 % of the total phenolic compounds, and 100 % of the low molecular weight phenolic compounds, without affecting the sugar concentrations in the hydrolysate. Furthermore, heterogeneous processes have provided relevant increases on sugars consumption (148 %) and maximum ethanol concentration (154 %) as compared with untreated hydrolysate. In addition, the AOPs treatment showed advantages such as it does not generate waste, does not degrade hydrolysate sugars, and does not lead to loss of hydrolysate volume due to the treatment. Based on the results, it can be concluded that AOPs are promising processes for application in hydrolysate treatment to reduce toxicity and consequently improve the fermentability of lignocellulosic hydrolysates.

Treatment of rice straw hemicellulosic hydrolysates with advanced oxidative processes: a new and promising detoxification method to improve the bioconversion process

BackgroundThe use of lignocellulosic constituents in biotechnological processes requires a selective separation of the main fractions (cellulose, hemicellulose and lignin). During diluted acid hydrolysis for hemicellulose extraction, several toxic compounds are formed by the degradation of sugars and lignin, which have ability to inhibit microbial metabolism. Thus, the use of a detoxification step represents an important aspect to be considered for the improvement of fermentation processes from hydrolysates. In this paper, we evaluated the application of Advanced Oxidative Processes (AOPs) for the detoxification of rice straw hemicellulosic hydrolysate with the goal of improving ethanol bioproduction by Pichia stipitis yeast. Aiming to reduce the toxicity of the hemicellulosic hydrolysate, different treatment conditions were analyzed. The treatments were carried out according to a Taguchi L16 orthogonal array to evaluate the influence of Fe+2, H2O2, UV, O3 and pH on the concentration of aromatic compounds and the fermentative process.ResultsThe results showed that the AOPs were able to remove aromatic compounds (furan and phenolic compounds derived from lignin) without affecting the sugar concentration in the hydrolysate. Ozonation in alkaline medium (pH 8) in the presence of H2O2 (treatment A3) or UV radiation (treatment A5) were the most effective for hydrolysate detoxification and had a positive effect on increasing the yeast fermentability of rice straw hemicellulose hydrolysate. Under these conditions, the higher removal of total phenols (above 40%), low molecular weight phenolic compounds (above 95%) and furans (above 52%) were observed. In addition, the ethanol volumetric productivity by P. stipitis was increased in approximately twice in relation the untreated hydrolysate.ConclusionThese results demonstrate that AOPs are a promising methods to reduce toxicity and improve the fermentability of lignocellulosic hydrolysates.

Ethanol Production Efficiency of an Anaerobic Hemicellulolytic Thermophilic Bacterium, Strain NTOU1, Isolated from a Marine Shallow Hydrothermal Vent in Taiwan

A new extremely thermophilic, anaerobic, gram-negative bacterium, strain NTOU1, was enriched and isolated from acidic marine hydrothermal fluids off Gueishandao island in Taiwan with 0.5% starch and 0.5% maltose as carbon sources. This strain was capable of growth utilizing various sugars found in lignocellulosic biomass as well as xylan and cellulose, and produced ethanol, lactate, acetate, and CO(2) as fermentation products. The results of a 16S rRNA gene sequence analysis (1,520 bp) revealed NTOU1 to belong to the genus Thermoanaerobacterium. When tested for the ability to grow and produce ethanol from xylose or rice straw hemicellulosic hydrolysate at 70°C, the strain showed the highest levels of ethanol production (1.65 mol ethanol mol xylose(-1)) in a medium containing 0.5% xylose plus 0.5% yeast extract. Maximum ethanol production from the rice straw hemicellulose was 0.509 g g(-1), equivalent to 98.8% theoretical conversion efficiency. Low concentrations of inhibitors (derived from dilute acid hydrolysis) in the rice straw hemicellulose hydrolysate did not affect the ethanol yield. Thus, Thermoanaerobacterium strain NTOU1 has the potential to be used for ethanol production from hemicellulose.

Comparison of cell growth and ethanol productivity on different pretreatment of rice straw hemicellulose hydrolysate by using Candida shehatae CICC 1766

In this work, the cell growth and ethanol productivity was tested forlignocellulosic biomass sugars of overliming-detoxified and NaOH-neutralized rice straw acid hydrolysate by using Candida shehatae CICC 1766. When the acetic acid reached 3 g/L in defined xylose medium, the final ethanol concentration was 33% of that in the medium without acetic acid addition. C. shehatae CICC 1766 could bear lower pH (ethanol yield 13.5 g/L at pH 4.0) in overliming-detoxified hydrolysate. At the optimal pH 5.0 the ethanol yield attended to 16.1 g/L. Ethanol yield in NaOH-neutralized hydrolysate was 13.7 g/L which is better than in overliming-detoxified hydrolysate (7.0 g/L). It suggested that NaOH-neutralized hydrolysate could be directly fermented by using C shehatae CICC 1766. Key words: Rice straw, Candida shehatae, hemicellulose hydrolysate, ethanol fermentation

The Influence of Initial Xylose Concentration, Agitation, and Aeration on Ethanol Production by Pichia stipitis from Rice Straw Hemicellulosic Hydrolysate

Rice straw hemicellulosic hydrolysate was used as fermentation medium for ethanol production by Pichia stipitis NRRL Y-7124. Shaking bath experiments were initially performed aiming to establish the best initial xylose concentration to be used in this bioconversion process. In the sequence, assays were carried out under different agitation (100 to 200 rpm) and aeration (V(flask)/V(medium) ratio varying from 2.5 to 5.0) conditions, and the influence of these variables on the fermentative parameters values (ethanol yield factor, Y(P/S); cell yield factor, Y(X/S); and ethanol volumetric productivity, Q(P)) was investigated through a 2(2) full-factorial design. Initial xylose concentration of about 50 g/l was the most suitable for the development of this process, since the yeast was able to convert substrate in product with high efficiency. The factorial design assays showed a strong influence of both process variables in all the evaluated responses. The agitation and aeration increase caused a deviation in the yeast metabolism from ethanol to biomass production. The best results (Y(P/S) = 0.37 g/g and Q(P) = 0.39 g/l.h) were found when the lowest aeration (2.5 V(flask)/V(medium) ratio) and highest agitation (200 rpm) levels were employed. Under this condition, a process efficiency of 72.5% was achieved. These results demonstrated that the establishment of adequate conditions of aeration is of great relevance to improve the ethanol production from xylose by Pichia stipitis, using rice straw hemicellulosic hydrolysate as fermentation medium.

The behavior of key enzymes of xylose metabolism on the xylitol production by Candida guilliermondii grown in hemicellulosic hydrolysate

A variety of raw materials have been used in fermentation process. This study shows the use of rice straw hemicellulosic hydrolysate, as the only source of nutrient, to produce high added-value products. In the present work, the activity of the enzymes xylose reductase (XR); xylitol dehydrogenase (XD); and glucose-6-phosphate dehydrogenase (G6PD) during cultivation of Candida guilliermondii on rice straw hemicellulosic hydrolysate was measured and correlated with xylitol production under different pH values (around 4.5 and 7.5) and initial xylose concentration (around 30 and 70 g l(-1)). Independent of the pH value and xylose concentration evaluated, the title of XD remained constant. On the other hand, the volumetric activity of G6PD increased whereas the level of XR decreased when the initial xylose concentration was increased from 30 to 70 g l(-1). The highest values of xylitol productivity (Q (P) approximately 0.40 g l(-1)) and yield factor (Y (P/S) approximately 0.60 g g(-1)) were reached at highest G6PD/XR ratio and lowest XR/XD ratio. These results suggest that NADPH concentrations influence the formation of xylitol more than the activity ratios of the enzymes XR and XD. Thus, an optimal rate between G6PD and XR must be reached in order to optimize the xylitol production.

Xylitol Production from Rice Straw Hemicellulose Hydrolyzate by Polyacrylic Hydrogel Thin Films with Immobilized Candida subtropicalis WF79

Xylose from rice straw hemicellulose hydrolysate was fermented for xylitol production using Candida subtropicalis WF79 cells immobilized in polyacrylic hydrogel thin films of 200 mum thickness. Cell immobilization was conducted by first suspending the yeast cells in a mixture of 2-hydroxyethyl methacrylate (HEMA, hydrophilic monomer), polyethylene glycol diacrylate (PEG-DA, crosslinking agent), and benzoin isopropyl ether (photoinitiator). The mixture was then allowed to form polyacrylic hydrogel thin films, between two pieces of glass sheets, by UV-initiated photopolymerization. The hemicellulose of rice straw was hydrolyzed using dilute sulfuric acid at 126 degrees C. The hydrolysate was neutralized with calcium hydroxide. After separating the solid residues and calcium sulfate precipitates by filtration, the hydrolysate was treated with charcoal to partially remove potential inhibitory substances, followed by vacuum concentration to obtain solutions of desired xylose concentrations for yeast fermentation. The thin films with immobilized yeast cells were submerged in the xylose solution from rice straw hydrolysate for fermentation in an Erlenmeyer flask. The maximum yield was 0.73 g of xylitol per gram of xylose consumed. In the 52.5-day long durability test, after 40 d of repeated batchwise operation, the fermentation activities of the cell immobilized in thin films began to decline to a yield of 0.57 g/g at the end.

Glucose-6-phosphate dehydrogenase and xylitol production by Candida guilliermondii FTI 20037 using statistical experimental design

The aim of this work was evaluate the G6PD and xylitol production by Candida guilliermondii grown in rice straw hemicellulosic hydrolysate. Fermentative assays were carried out to evaluate the effect of pH ( X 1), xylose concentration ( X 2), and inoculum level (X 3) on the production of G6PD and xylitol. For optimization of G6PD production it was employed the methodology of experimental design. The estimated model predicts a maximum specific activity of 0.64 U mg −1 being possible a variation of 0.55–0.72 U mg −1 for response specific activity of G6PD (As), in the confidence level of 95%. For the model validation, it was attained 0.60 U mg −1 with 70 g L −1 xylose, pH 7.5 and 1 g L −1 of inoculum level. Indeed, C. guilliermondii proved to be a potencial source of intracellular enzyme (G6PD) when grown in a low cost fermentation medium as well as efficient in xylose–xylitol bioconversion.

Evaluation of nutrient supplementation to charcoal-treated and untreated rice straw hydrolysate for xylitol production by Candida guilliermondii

Xylitol was produced by Candida guilliermondii from charcoal-treated and untreated rice straw hemicellulosic hydrolysate with or without nutrients (ammonium sulphate, calcium chloride, rice bran extract). Both, xylitol yield and volumetric productivity decreased significantly when the nutrients were added to treated and untreated hydrolysates. In the treated hydrolysate, the efficiency of xylose conversion to xylitol was 79% when the nutrients were omitted. The results demonstrated that rice straw hemicellulosic hydrolysate treated with activated charcoal was a cheap source of xylose and other nutrients for xylitol production by C. guilliermondii. The non-necessity of adding nutrients to the hydrolysate media would be very advantageous since the process becomes less costly.

Effect of pH and activated charcoal adsorption on hemicellulosic hydrolysate detoxification for xylitol production

AbstractBiotechnological conversion of xylose into xylitol using hydrolysates obtained from the hemicellulosic fraction of lignocellulosic materials is compromised by the presence of compounds released or formed during the hydrolysis process, some of them being toxic to microorganisms. In order to improve the bioconversion of these hydrolysates it is necessary to find methods to reduce their toxicity. In the present work, rice straw hemicellulosic hydrolysate was treated by six different procedures (all of them involving pH adjustment, with or without activated charcoal adsorption), before being used as a fermentation medium for xylitol production. The most effective method of treatment was to increase the initial pH (0.4) to 2.0 using solid NaOH, followed by the addition of activated charcoal (25 g kg−1) and increase in the pH to 6.5 using solid NaOH. Lignin degradation products were the most inhibitory compounds present in the hydrolysate; their removal was selective and strongly dependent on the pH employed in the treatment. The highest yield of xylitol was 0.72 g g−1 xylose, with a productivity of 0.55 g dm−3 h−1. Copyright © 2004 Society of Chemical Industry

Avaliação de diferentes tipos de carvão ativo na destoxificação de hidrolisado de palha de arroz para produção de xilitol

O hidrolisado hemicelulósico de palha de arroz foi tratado com cinco tipos de carvão ativo (pó e granulado) com o objetivo de remover, por adsorção, compostos tóxicos que podem agir como inibidores no processo de bioconversão de xilose em xilitol, por Candida guilliermondii. Os valores máximos de fator de rendimento em xilitol (Y P/S = 0,67g g-1) e produtividade volumétrica (Q P = 0,61g L-1 h-1) foram atingidos quando o hidrolisado foi tratado com carvão ativo em pó de partículas de tamanho pequeno (0,043mm), baixa granulometria (32% retidos em peneira de 325mesh) e grande área superficial (860m² g-1), características as quais favoreceram a adsorção dos compostos tóxicos.

Kinetic behavior of Candida guilliermondii yeast during xylitol production from highly concentrated hydrolysate

Rice straw hemicellulosic hydrolysate containing a high xylose concentration was used as fermentation medium to evaluate the kinetic behavior of Candida guilliermondii yeast (FTI 20037) during the bioconversion of xylose into xylitol. Assays were conducted first with detoxified and non-detoxified (raw) hydrolysates and semi-synthetic medium in agitated flasks, and second with detoxified hydrolysate in a stirred-tank bioreactor at a given oxygen transfer rate. The results for the agitated flasks showed that in detoxified hydrolysate the xylose-to-xylitol bioconversion by the yeast was as effective as in synthetic medium and 47% higher than in raw hydrolysate. In the stirred-tank bioreactor, the kinetic behavior of the yeast in detoxified hydrolysate was slower, resulting in smaller values of fermentative parameters, probably due to unsuitability of the oxygen transfer rate employed ( K L a=22 h −1).

Xylitol production from high xylose concentration: evaluation of the fermentation in bioreactor under different stirring rates.

To investigate the production of xylitol by the yeast Candida guilliermondii FTI 20037, in a bioreactor, from rice straw hemicellulosic hydrolysate with a high xylose concentration. Batch fermentation was carried out with rice straw hemicellulosic hydrolysate containing about 85 g xylose l(-1), in a stirred-tank bioreactor at 30 degrees C, under aeration of 1.3 vvm (volume of air per volume of medium per min) and different stirring rates (200, 300 and 500 rev min(-1)). The bioconversion of xylose into xylitol by the yeast depended on the stirring rate, the maximum xylitol yield (YP/S = 0.84 g g(-1)) being achieved at 300 rev min-1, with no need to pretreat the hydrolysate for purification. To determine the most adequate oxygen transfer rate is fundamental to improving the xylose-to-xylitol bioconversion by C. guilliermondii. For the microbial production of xylitol to be economically viable, the initial concentration of xylose in the lignocellulosic hydrolysate should be as high as possible, as with high substrate concentrations it is possible to increase the final product concentration. Nevertheless, there are few reports on the use of high xylose concentrations. Considering a process in bioreactor, from rice straw hemicellulosic hydrolysate, this is an innovator work.

Improvement of xylitol production by Candida guilliermondii FTI 20037 previously adapted to rice straw hemicellulosic hydrolysate.

To evaluate a simple and economical technique to improve xylitol production using concentrated xylose solutions prepared from rice straw hemicellulosic hydrolysate. Experiments were carried out with rice straw hemicellulosic hydrolysate containing 90 g l-1 xylose, with and without the addition of nutrients, using the yeast Candida guilliermondii previously grown on the hydrolysate (adapted cells) or on semi-defined medium (unadapted cells). By this method, the yield of xylitol increased from 17 g l-1 to 50 g l-1, and xylose consumption increased from 52% to 83%, after 120 h of fermentation. The xylitol production rates were very close to that (0.42 g l-1 h-1) attained in a medium simulating hydrolysate sugars. Yeast strain adaptation to the hydrolysate showed to be a suitable method to alleviate the inhibitory effects of the toxic compounds. Adapted cells of Candida guilliermondii can efficiently produce xylitol from hydrolysate with high xylose concentrations. Yeast adaptation helps the bioconversion process in hydrolysate made from lignocellulosic materials. This low-cost technique provides an alternative to the detoxification methods used for removal of inhibitory compounds. In addition, the use of adapted inocula makes it possible to schedule a series of batch cultures so that the whole plant can be operated almost continuously with a concomitant reduction in the overall operation time.

Hydrolysate detoxification with activated charcoal for xylitol production by Candida guilliermondii

A detoxification method using activated charcoal with concentrated rice straw hemicellulosic hydrolysate improved the conversion of xylose to xylitol by the yeast Candida guilliermondii by 22%. This was achieved when the hydrolysate:charcoal ratio was 40 g g−1, resulting in removal of 27% of phenolic compounds. Under this condition, the xylitol yield factor (0.72 g g−1) and volumetric productivity (0.61 g l−1 h−1) were close to those attained in a semi-defined medium simulating hydrolysate sugars.

Influence of k L a on bioconversion of rice straw hemicellulose hydrolysate to xylitol

The effect of overall oxygen mass transfer coefficient (k L a) on the conversion of xylose to xylitol by Candida guilliermondii FTI 20037 was investigated in batch experiments. Rice straw hemicellulose hydrolysate obtained by acid hydrolysis was employed as a xylose-rich medium. The results showed that this bioconversion strongly depended on the aeration rate. The maximum volumetric productivity (0.52 g/l h−1) and the highest xylitol yield (0.73 g/g) were achieved at an overall oxygen mass transfer coefficient of 15 h−1. Under these conditions 80% efficiency in relation to theoretical yield was attained.

Kinetics of Xylitol Fermentation by Candida guilliermondii Grown on Rice Straw Hemicellulosic Hydrolysate

The fermentation kinetics for the conversion of rice straw hemicellulosic hydrolysate to xylitol by the yeast Candida guilliermondii was evaluated under batch conditions. The fermentation was accomplished in a 1 L working volume stirred-tank reactor with aeration of 1.3 vvm and agitation of 300 rpm (kLa=15/h). The maximum specific rate of xylitol formation (0.12 g/g) was achieved when the specific growth rate was lowered to 1/5 of its highest value. From analysis of the fermentation kinetics, a linear correlation between specific growth rate (μx) and specific rate of xylitol formation (qp) was evident. Based on the Gaden model, this bioprocess was classified as growth-associated production and the relationship between μx and qp can be described by the equation qp=6.31μx.

Production of xylitol byCandida mogii from rice straw hydrolysate

The influence of aeration level, initial pH, initial cell concentration, and fermentation time on the xylitol production from rice straw hemicellulose hydrolysate by Candida mogii was studied. A multifactorial experimental design was adopted to evaluate this influence. A statistical analysis of the results showed that the aeration level and the initial pH had significant effects on yield factor, volumetric productivity, and xylose consumption. For the latter, fermentation time was also a significant variable. Based on the response surface methodology, models for the range investigated were proposed. The maximum values for the yield factor (Y p/s ) and volumetric productivity (Q p ) were, respectively, 0.71 g/g and 0.46 g(Lh).