Initial Xylose Research Articles

Activity of xylose reductase from Candida mogii grown in media containing different concentrations of rice straw hydrolysate.

Xylose reductase (XR) activity was evaluated in extracts of Candida mogii grown in media containing different concentrations of rice straw hydrolysate. Results of XR activity were compared to xylitol production and a similar behavior was observed for these parameters. Highest values of specific production and productivity were found for xylose reductase (35 U/g of cell and 0.97 U/[g of cell x h], respectively) and for xylitol (5.63 g/g of cell and 0.13 g/[g of cell x h]) in fermentation conducted in medium containing 49.2 g of xylose/L. The maximum value of XR:XD ratio (1.82) was also calculated under this initial xylose concentration with 60 h of fermentation.

The influence of aeration and hemicellulosic sugars on xylitol production by Candida tropicalis

The influence of other hemicellulosic sugars (arabinose, galactose, mannose and glucose), oxygen limitation, and initial xylose concentration on the fermentation of xylose to xylitol was investigated using experimental design methodology. Oxygen limitation and initial xylose concentration had considerable influences on xylitol production by Canadida tropicalis ATCC 96745. Under semi-aerobic conditions, the maximum xylitol yield was 0.62 g/g substrate, while under aerobic conditions, the maximum volumetric productivity was 0.90 g/l h. In the presence of glucose, xylose utilization was strongly repressed and sequential sugar utilization was observed. Ethanol produced from the glucose caused 50% reduction in xylitol yield when its concentration exceeded 30 g/l. When complex synthetic hemicellulosic sugars were fermented, glucose was initially consumed followed by a simultaneous uptake of the other sugars. The maximum xylitol yield (0.84 g/g) and volumetric productivity (0.49 g/l h) were obtained for substrates containing high arabinose and low glucose and mannose contents.

Application of a compatible xylose isomerase in simultaneous bioconversion of glucose and xylose to ethanol

Simultaneous isomerisation and fermentation (SIF) of xylose and simultaneous isomerisation and cofermentation (SICF) of glucose-xylose mixture was carried out by the yeastSaccharomyces cerevisiae in the presence of a compatible xylose isomerase. The enzyme converted xylose to xylulose andS. cerevisiae fermented xylulose, along with glucose, to ethanol at pH 5.0 and 30°C. This compatible xylose isomerase fromCandida boidinii, having an optimum pH and temperature range of 4.5–5.0 and 30–50°C respectively, was partially purified and immobilized on an inexpensive, inert and easily available support, hen egg shell. An immobilized xylose isomerase loading of 4.5 IU/(g initial xylose) was optimum for SIF of xylose as well as SICF of glucose-xylose mixture to ethanol byS. cerevisiae. The SICF of 30 g/L glucose and 70 g xylose/L gave an ethanol concentration of 22.3 g/L with yield of 0.36 g/(g sugar consumed) and xylose conversion efficiency of 42.8%.

Production of carotenoids by Phaffia rhodozyma growing on media made from hemicellulosic hydrolysates of Eucalyptus globulus wood

Phaffia rhodozyma NRRL Y-17268 cells were proliferated in xylose-containing media made from Eucalyptus wood. Wood samples were subjected to acid hydrolysis under mild operational conditions, and hydrolysates were neutralized with lime. Neutralized hydrolysates were treated with charcoal for removing inhibitors and then supplemented with nutrients to obtain culture media useful for proliferation of the red yeast P. rhodozyma. A set of experiments carried out in orbital shakers proved that hydrolysates containing 16.6 g xylose/L supplemented only with 3 g peptone/L performed well as fermentation media. At the end of experiments, xylose was depleted and 10.5 g cells/L were obtained. Biomass was highly pigmented and volumetric carotenoid concentrations up to 5.8 mg carotenoids/L (with 4.6 mg astaxanthin/L) were reached. Further experiments in batch fermentors using concentrated hydrolysates (initial xylose concentrations within 16.6 and 40.8 g/L) led to good biomass concentrations (up to 23.2 g cells/L) with increased pigment concentration (up to 12.9 mg total carotenoids/L, with 10.4 mg astaxanthin/L) and high volumetric rates of carotenoid production (up to 0.079 mg/L·h). © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59: 501–506, 1998.

Production of carotenoids by phaffia rhodozyma growing on media made from hemicellulosic hydrolysates of eucalyptus globulus wood

Phaffia rhodozyma NRRL Y-17268 cells were proliferated in xylose-containing media made from Eucalyptus wood. Wood samples were subjected to acid hydrolysis under mild operational conditions, and hydrolysates were neutralized with lime. Neutralized hydrolysates were treated with charcoal for removing inhibitors and then supplemented with nutrients to obtain culture media useful for proliferation of the red yeast P. rhodozyma. A set of experiments carried out in orbital shakers proved that hydrolysates containing 16.6 g xylose/L supplemented only with 3 g peptone/L performed well as fermentation media. At the end of experiments, xylose was depleted and 10.5 g cells/L were obtained. Biomass was highly pigmented and volumetric carotenoid concentrations up to 5.8 mg carotenoids/L (with 4.6 mg astaxanthin/L) were reached. Further experiments in batch fermentors using concentrated hydrolysates (initial xylose concentrations within 16.6 and 40.8 g/L) led to good biomass concentrations (up to 23.2 g cells/L) with increased pigment concentration (up to 12.9 mg total carotenoids/L, with 10.4 mg astaxanthin/L) and high volumetric rates of carotenoid production (up to 0.079 mg/L.h). Copyright 1998 John Wiley & Sons, Inc.

Fermentation of hemicellulosic sugars and sugar mixtures to xylitol by Candida parapsilosis

The ability of Candida parapsilosis to ferment xylose to xylitol in the presence of glucose and mannose was evaluated using a semidefined medium and aspenwood hemicellulose hydrolysate obtained by steam explosion acid hydrolysis. This natural growth substrate can be used with an appropriate dilution to give initial xylose, glucose, and mannose concentrations of 50, 27, and 6 g l −1, respectively. Above these values, inhibition of growth and metabolic activities appeared to be due to the level of by-products generated during pretreatment and hydrolysis of aspenwood. By supplementing the hemicellulose hydrolysate with yeast extract, xylitol yield was comparable to that measured in a semi-defined medium using a sugar mixture but the rate of xylitol production was twice as low. To improve the xylitol yield and productivity, an approach based on sugar assimilation is proposed.

Co-production of carotenoids and xylitol by Xanthophyllomyces dendrorhous (Phaffia rhodozyma)

Carotenoid production by Xanthophyllomyces dendrorhous (formerly Phaffia rhodozyma) when growing on xylose depended on the initial substrate concentration. Astaxanthin was the main carotenoid, others were echinenone, 3-hydroxyechinenone and canthaxanthin. Xylitol (an intermediate of xylose metabolism, with application as sweetener) accumulates in the fermentation media. Xylitol concentrations up to 22 g/l were obtained in experiments carried out with an initial xylose concentration of 42 g/l. Xylitol was assimilated after xylose depletion.

Fermentation of xylose and rice straw hydrolysate to ethanol by Candida shehatae NCL-3501.

Candida shehatae NCL-3501 utilized glucose and xylose efficiently in batch cultures. The specific rate of ethanol production was higher with mixtures of glucose and xylose (0.64-0.83 g g-1 cells d-1) compared to that with individual sugars (0.38-0.58 g g-1 cells d-1). Although the optimum temperature for growth was 30 degrees C, this strain grew and produced appreciable levels of ethanol at 45 degrees C. A stable ethanol yield (0.40-0.43 g g-1 substrate utilized) was obtained between 10 g L-1 and 80 g L-1 of initial xylose concentration. Conversion efficiency was further improved by immobilization of the cells in calcium alginate beads. Free or immobilized cells of C. shehatae NCL-3501 efficiently utilized sugars present in rice straw hemicellulose hydrolysate, prepared by two different methods, with 48 h. Ethanol yields of 0.45 g g-1 and 0.5 g g-1 from autohydrolysate, and 0.37 g g-1 from acid hydrolysate were produced by free and immobilized cells, respectively.

Pretreatment of sugar cane bagasse hemicellulose hydrolysate for xylitol production by yeast.

A total of six known xylitol-producing yeast strains were screened for production of xylitol from xylose. Candida sp. 11-2 proved to be the best producer. It was chosen to study its ability to produce xylitol from hemicellulose hydrolysate derived from sugar cane bagasse. The hydrolysate was prepared by dilute sulfuric acid (2-3% [w/v]) hydrolysis, with a high-solid, low-liquid ratio followed by leaching. Owing to the inhibitors present in the hydrolysate, different treatments were studied to overcome its effect. In order to reach higher xylitol productivity, treated hydrolysates were concentrated by vacuum evaporation in rotavapor to provide a higher initial xylose concentration. After treatment, Candida sp. 11-2 was able to ferment xylose in hemicellulose hydrolysate to produce xylitol.

FET-microbial sensor for xylose detection based on Gluconobacter oxydans cells

A potentiometric biosensor for xylose was devised utilizing Gluconobacter oxydans whole cells. Immobilization methods based on physical adsorption were used for G. oxydans cells and extracellular pH changes resulting from xylose dehydrogenation were monitored by a field effect transistor (FET). The G. oxydans, FET-based sensor detected xylose at a lower limit of 0.5 mM. From 5.0 to 30 mM xylose, the response of the sensor was linear. Expectedly, output signals were significantly suppressed by buffer (Tris-HCl). Responses were essentially stable for at least four weeks of storage and showed only a slight loss of initial xylose sensitivity. Xylitol exerted an insignificant influence on the sensor's response to xylose. However, the response to glucose was 5 times higher in relation to that of xylose at the same concentration (1 mM). For xylose determinations in the presence of glucose, a two-step assay is discussed.

Xylose reductase and xylitol dehydrogenase activities of D‐xylose‐xylitol‐fermenting Candida guilliermondii

AbstractThe effect of the inoculum age and initial xylose concentration on the activities of xylose reductase (XR) and xylitol dehydrogenase (XD) of Candida guilliermondii FTI 20037 were investigated. Under the experimental conditions used, the XR and XD activities were, respectively. NADPH and NAD or NADP dependent. The NADP/NADPH ratio for XR was not substantially influenced by the age of inoculum or the initial xylose concentration. The Km of XR (0.18 M) was about 4‐folds smaller than the Km of XD NAD‐dependent. A renewable xylose‐xylitol conversion was obtained with 40 h aged inoculum of Candida guilliermondii FTI 20037 grown in a medium containing an initial xylose concentration of 120g/l.

Fed-batch culture for xylitol production by Candida boidinii

Xylitol production by Candida boidinii NRRL Y-17213 was investigated in fed-batch fermentations with xylose (50, 100 g litre −1) and a mixture of glucose (25 g litre −1) and xylose (25 g litre −1). All fermentations were initially batch processes with high levels of aeration and rapid production of biomass. Faster growth occurred when a mixture of glucose and xylose, instead of xylose, was used as a substrate; glucose was assimilated first and maximal xylitol production was 39·41 g litre −1, compared with 46·5 and 59·3 g litre −1 in the processes with xylose alone. Fed-batch cultures were characterized with higher xylitol yields (0·57–0·68 g g −1) and production rates (0·32–0·46 g litre −1 h −1) compared with a batch process. Ethanol was accumulated in all processes, in smaller quantities, varying from 11 to 21% of the xylitol concentration. The fed-batch process with the highest initial xylose concentration (100 g litre −1) and the lowest level of aeration in the first phase, resulted in the highest yield of xylitol (75% of theoretical) and the highest productivity (approximately 35% higher than the productivities of the other two fed-batch experiments).

Xylitol production from xylose by two yeast strains: Sugar tolerance

The kinetics and enzymology ofd-xylose utilization are studied in micro-, semi-, and aerobic batch cultures during growth ofCandida guilliermondii andCandida parapsilosis in the presence of several initial xylose concentrations. The abilities of xylitol accumulation by these two yeast strains are high and similar, although observed under various growth conditions. WithCandida parapsilosis, optimal xylitol production yield (0.74 g/g) was obtained in microaerobiosis with 100 g/L of xylose, whereas optimal conditions to produce xylitol byCandida guilliermondii (0.69 g/g) arose from aerobiosis with 300 g/L of sugar. The different behavior of these yeasts is most probably explained by differences in the nature of the initial step of xylose metabolism: a NADPH-linked xylose reductase activity is measured with a weaker NADH-linked activity. These activities seem to be dependent on the degree of aerobiosis and on the initial xylose concentration and correlate with xylitol accumulation.

L-lactate production from xylose employingLactococcus lactis IO-1

The growth, substrate utilisation and L-lactate production ofLactococcus lactis IO-1 were examined on xylose, and glucose and xylose media. The yield of lactate on xylose was 0.47 g lactate/g xylose at an initial xylose concentration of 51.2 g/l and the μmax was 0.72 h−1. Xylose cultures were more susceptible to lactate inhibition than were glucose cultures but showed similar kinetic behaviour. The organism was capable of complete sugar utilisation when grown on a mixture of 20 g/l xylose and 20 g/l glucose and synthesised 0.66 g lactate/g sugar.

The effects of the oxygen transfer coefficient and substrate concentration on the xylose fermentation by Debaryomyces hansenii

Relevant production of xylitol by Debaryomyces hansenii requires semiaerobic conditions since in aerobic conditions the accumulated reduced adenine dinucleotide coenzyme is fully reoxidized leading to the conversion of xylitol into xylulose. For oxygen transfer coefficient values from 0.24 to 1.88 min-1, in shake flasks experiments, biomass formation increased proportionally to the aeration rate as shown in the oxygen transfer coefficient and xylose concentration isoresponse contours. The metabolic products under study, xylitol and ethanol were mainly growth associated. However, for oxygen transfer coefficient above 0.5 min-1 higher initial xylose concentration stimulated the rate of production of xylitol. This fact was less evident for ethanol production. The direct relationship between increased biomass and products formation rate, indicated that the experimental domain in respect to the aeration rate was below the threshold level before the decreasing in metabolic production rates reported in literature for xylose-fermenting yeasts. The fact that ethanol was produced, albeit in low levels, throughout the experimental design indicated that the semiaerobic conditions were always attained. Debaryomyces hansenii showed to be an important xylitol producer exhibiting a xylitol/ethanol ratio above four and a carbon conversion of 54% for xylitol.

Fumaric acid production from xylose by immobilized Rhizopus arrhizus cells

The production of fumaric acid by immobilized Rhizopus arrhizus TKK 204-1-1a mycelium was optimized in batch fermentations using statistical experimental design and empiric modelling. The maximum fumaric acid concentration was obtained at a xylose concentration of about 6% and a carbon:nitrogen ratio of about 160. In repeated batch fermentations with immobilized cells the highest volumetric productivity of fumaric acid reached was 87 mg/l per hour when the initial xylose concentration was 10%, the C:N ratio 160 and the residence time 1.75 days. The maximum product concentration was 16.4 g/l when the initial xylose concentration was 10%, the C:N ratio 160 and the residence time 10.25 days. The maximum yield from initial xylose (6.47%) was 23.7% with a product concentration of 15.3 g/l and volumetric productivity of 71 mg/l per hour at a residence time of 9 days and a C:N ratio of 188.3. Immobilization could increase the fumaric acid concentration to a level 3.4 times higher than that produced by free cells.

Alcoholic fermentation of D-xylose by immobilizedPichia stipitis yeast

Pichia stipitis NRRL Y-7124 yeast cells were for the first time immobilized both in agar gel beads and on fine nylon net for ethanol fermentation on D-xylose, in order to investigate the possibility of using the biocatalyst for improved utilization of the biomass pentose fraction. With free cells the initial xylose level affected little ethanol production, with a maximum of 22 g/l ethanol obtained in 5 days on 5% and of 40 g/l in 8 days on 10% xylose, and an average volumetric productivity of about 0.22 g/lh. The maximum ethanol concentration of 19.5% on 5% xylose with the nylon net attached cells in a continuous packed-bed column reactor was obtained with 35 h residence time. The volumetric productivities of 0.56 g/lh at 19.5 g/l ethanol and 1.0 g/lh at 15.0 g/l ethanol were markedly higher than those obtained with free cells. The stability of the immobilized biocatalyst was excellent. The same reactor could be used for at least 80 days without significant activity loss.

Red oak wood derived inhibitors in the ethanol fermentation of xylose byPichia stipitis CBS 5776

Xylose, the predominant sugar in red oak prehydrolysate, is fermented to ethanol byPichia stipitis CBS 5776. Toxic model compounds derived from red oak hemicelluloses, lignin, and extractives inhibited the fermentation. Treatment of the prehydrolysate with molecular sieve and mixed bed ion resins facilitated the ethanol fermentation giving about 10 g/l ethanol from 32 g/l initial xylose. Fermentation inhibitors derived from red oak lignin and extractives were identified.

Application of bioenergetics to modelling the microbial conversion of D‐xylose to 2,3‐butanediol

During the oxygen limiting growth of Klebsiella oxytoca, the xylose metabolism may be considered as consisting of three components: conversion to 2,3-butanediol by "fermentation," oxidation to carbon dioxide by respiration, and assimilation to cell mass. The amount of energy required for the assimilation of cell mass is assumed to determine the extent to which the two energy producing reactions occur. The activity of each energy producing pathway is also determined by the availability of oxygen and by the energy yield of each pathway. These relationships can be quantified by equating the ATP required for growth and maintenance to the ATP produced by the energy producing reactions. The resulting equation for butanediol production appears similar to the Luedeking and Piret model where the parameters alpha and beta are related to the maximum cell yield from ATP and the maintenance energy requirement. These parameters were estimated from 14 batch fermentations, and the resulting simulation was used to describe the effects of the oxygen transfer rate and the initial xylose concentration on the yields and rates of the 2,3-butanediol fermentation.