Francisco M. Gírio

Production of oligosaccharides by autohydrolysis of brewery’s spent grain

Brewerys spent grain was treated with water in a process oriented towards the production of xylo-oligosaccharides (XOS). A wide range of temperatures and reaction times were tested and the effects of these operational variables on hemicellulose solubilization and reaction products were investigated. The maximal XOS yield (61% of the feedstock xylan) was obtained at 190 degrees C after 5 min of reaction. Several oligosaccharide mixtures with different molecular weight distributions were obtained depending on temperature and reaction time. Longer reaction times led to decreased oligosaccharide production and enhanced concentrations of monosaccharides, sugar decomposition products and acetic acid. With reaction times leading to the maximal yields of XOS, little decomposition into organic acids and aldehydes was found at all the temperatures assayed. From the composition of processed solids, it was calculated that 63-77% of the initial xylan was selectively solubilized in autohydrolysis treatments.

Wheat Straw Autohydrolysis: Process Optimization and Products Characterization

Wheat straw was subjected to autohydrolysis treatments in order to selectively hydrolyze the hemicellulose fraction. The effects of temperature (150–240°C) and non-isothermal reaction time on the composition of both liquid and solid phases were evaluated and interpreted using the severity factor (log R0). The operational conditions leading to the maximum recovery of hemicellulose-derived sugars were established for log R0 = 3.96 and correspond to 64% of the original (arabino)xylan with 80% of sugars as xylooligosaccharides. Under these conditions, a solubilization of 58% xylan, 83% arabinan, and 98% acetyl groups occurred. Glucan was mainly retained in the solid phase (maximum solubilization 16%), which enables an enrichment of the solid phase to contain up to 61% glucan. Delignification was not extensive, being utmost 15%. The yields of soluble products, including sugars, acetic acid, and degradation compounds, such as, furfural, 5-hydroxymethylfurfural furfural obtained suggest the fitness of liquid stream for fermentation purposes or to obtain xylooligosaccharides with potential applications in food, pharmaceutical, and cosmetic industries.

Assessment on the fermentability of xylooligosaccharides from rice husks by probiotic bacteria.

Liquors from rice husk autohydrolysis, containing xylooligosaccharides (XOS), other saccharides, and nonsaccharide compounds, were refined by membrane processing to increase the proportion of substituted XOS in refined liquors. XOS were assayed for composition and degree of polymerization (DP) distribution and hydrolyzed with commercial enzymes for obtaining XOS with DP in the range of 2-6. Nanofiltered, hydrolyzed liquors were subjected to ion exchange processing to yield a final product containing monosaccharides, XOS (accounting for 55.6% of the nonvolatile solutes), and other nonvolatile compounds. The solution obtained after enzymatic hydrolysis with commercial xylanases (in which 82.8% of XOS were in the DP range of 2-6) was examined as a medium for promoting the growth of Bifidobacterium adolescentis CECT 5781, B. longum CECT 4503, B. infantis CECT 4551, and B. breve CECT 4839. The growth rate of B. adolescentis (0.58 h(-1)) was higher than the ones determined for B. longum, B. infantis, and B. breve (0.37, 0.30, and 0.40 h(-1), respectively). The percentage of total XOS consumption by B. adolescentis was 77% after 24 h, the highest percentage of utilization corresponding to xylotriose (90%), followed by xylobiose (84%), xylotetraose (83%), and xylopentaose (71%).

Comparison of Two Posthydrolysis Processes of Brewery’s Spent Grain Autohydrolysis Liquor to Produce a Pentose-Containing Culture Medium

A readily fermentable pentose-containing hydrolysate was obtained from Brewerys spent grain by a two-step process consisting of an auto-hydrolysis (converting the hemicelluloses into oligosaccharides) followed by an enzymatic or sulfuric acid-catalyzed posthydrolysis (converting the oligosaccharides into monosaccharides). Enzymatic hydrolyses were performed with several commercial enzymes with xylanolytic and cellulolytic activities. Acid-catalyzed hydrolyses were carried out at 121 degrees C under various sulfuric acid concentrations and reaction times, and the effects of treatments were interpreted by means of a corrected combined severity factor (CS*), which varied in the range of 0.80-2.01. Under the tested conditions, chemical hydrolysis allowed higher pentose yields than enzymatic hydrolysis. Optimized conditions (defined by CS* = 1.10) allowed both complete monosaccharide recovery and low content of inhibitors. Liquors subjected to posthydrolysis under optimal conditions were easily fermented by Debaryomyces hansenii CCMI 941 in semiaerobic shake-flask experiments, leading to xylitol and arabitol as major fermentation products. The bioconversion process was improved by hydrolysate concentration and supplementation of fermentation media with casamino acids.

Kinetic modeling of brewery's spent grain autohydrolysis.

Isothermal autohydrolysis treatments of breweryapos;s spent grain were used as a method for hemicellulose solubilization and xylo‐oligosaccharides production. The time course of the concentrations of residual hemicelluloses (made up of xylan and arabinan) and reaction products were determined in experiments carried out at temperatures in the range from 150 to 190 °C using liquid‐to‐solid ratios of 8 and 10 g/g. To model the experimental findings concerning to breweryapos;s spent grain autohydrolysis several kinetic models based on sequential pseudo‐homogeneous first‐order reactions were tested. Xylan and arabinan were assumed to yield oligosaccharides, monosaccharides (xylose or arabinose), furfural, and other decomposition products in consecutive reaction steps. The models proposed provide a satisfactory interpretation of the hydrolytic conversion of xylan and arabinan. An additional model merging the two proposed models for xylan and arabinan degradation assuming that furfural was formed from both pentoses was developed and the results obtained are discussed. The dependence of the calculated kinetic coefficients on temperature was established using Arrhenius‐type equations.

Polyols production during single and mixed substrate fermentations in Debaryomyces hansenii

Abstract The kinetics of polyols production by Debaryomyces hansenii was studied both on single substrate and mixed substrate-containing media. From the single substrate experiments, polyols (xylitol and arabitol) and ethanol were produced from pentose sugars while ethanol was produced in significant amounts only from glucose-grown D. hansenii. The maximal xylitol volumetric productivity (Qxylitol), 0.28 g l−1 h−1 was obtained from D -xylose, whereas the maximal arabitol volumetric productivity (Qarabitol), 0.04 g l−1 h−1 was observed with arabinose. When D. hansenii was cultivated with mixed substrates, a simultaneous sugar consumption pattern occurred both for glucose/arabinose and xylose/arabinose mixtures. The addition of low amounts of xylose to an arabinose medium led to a fourfold increase in the arabitol volumetric productivity: 0.17 g l−1 h−1. Conversely, glucose addition had no effect on arabitol production. Xylitol was the main polyol produced for all tested cultivation conditions by D. hansenii. An enzymatic study of the first two xylose-catabolic enzymes in glucose and xylose-grown D. hansenii revealed that both enzymes were induced by D -xylose. Glucose caused total inhibition of xylitol dehydrogenase, whereas xylose reductase was only partially repressed.

Desulfurization of dibenzothiophene, benzothiophene, and other thiophene analogs by a newly isolated bacterium, Gordonia alkanivorans strain 1B

A novel bacterium, Gordonia alkanivorans strain 1B, was isolated from hydrocarbon-contaminated soil. Assessment of the biodegradation of distinct organic sulfur-compounds, such as dibenzothiophene (DBT), benzothiophene (BT), DBT sulfone, and alkylated tiophenic compounds, as the sole source of sulfure was investigated. G. alkanivorans strain 1B was able to remove selectively the sulfur from DBT while keeping intact the remaining carbon-carbon structure. Orthophenyl phenol (2-hydroxybiphenyl) was the only detected metabolic product. The bacterial desulfurization activity was repressed by sulfate. G. alkanivorans straini 1B consumed 310 μM DBT after 120 h of cultivation, corresponding to a specific desulfurization rate of 1.03 μmol/(g of dry cells·h). When an equimolar mixture of DBT/BT was used as a source of sulfur in the growth medium, G. alkanivorans strain 1B assimilated both compounds in a sequential manner, with BT as the preferred source of sulfur. Only when BT concentration was decreased to a very low level was DBT utilized as the source of sulfur for bacterial growth. Thespecific desulfurization overall rates of BT and DBT obtained were 0.954 and 0.813 μmol/(g of dry cells·h), respectively. The newly isolated G. alkanivorans strain 1B has good potential for application in the biodesulfurization of fossil fuels.

Effects of Aliphatic Acids, Furfural, and Phenolic Compounds on Debaryomyces hansenii CCMI 941

Debaryomyces hansenii is a polyol overproducing yeast that can have a potential use for upgrading lignocellulosic hydrolysates. Therefore, the establishment of its tolerance to metabolic inhibitors found in hydrolysates is of major interest. We studied the effects of selected aliphatic acids, phenolic compounds, and furfural. Acetic acid favored biomass production for concentrations <6.0 g/L. Formic acid was more toxic than acetic acid and induced xylitol accumulation (maximum yield of 0.21 g/g of xylose). All tested phenolics strongly decreased the specific growth rate. Increased toxicity was found for hydroquinone, syringaldehyde, and 4-methylcatechol and was correlated to the compounds hydrophobicity. Increasing the amount of furfural led to longer lag phases and had a detrimental effect on specific growth rate and biomass productivity.

Optimization of Brewery's spent grain dilute-acid hydrolysis for the production of pentose-rich culture media.

Dilute-acid hydrolysis of brewerys spent grain to obtain a pentose-rich fermentable hydrolysate was investigated. The influence of operational conditions on polysaccharide hydrolysis was assessed by the combined severity parameter (CS) in the range of 1.39–3.06. When the CS increased, the pentose sugars concentration increased to a maximum at a CS of 1.94, whereas the maximum glucose concentration was obtained for a CS of 2.65. The concentrations of furfural, hydroxymethylfurfural (HMF), as well as formic and levulinic acids and total phenolic compounds increased with severity. Optimum hydrolysis conditions were found at a CS of 1.94 with >95% of feedstock pentose sugars recovered in the monomeric form, together with a low content of furfural, HMF, acetic and formic acids, and total phenolic compounds. This hydrolysate containing glucose, xylose, and arabinose (ratio 10∶67∶32) was further supplemented with inorganic salts and vitamins and readily fermented by the yeast Debaryomyces hansenii CCMI 941 without any previous detoxification stage. The yeast was able to consume all sugars furfural, HMF, and acetic acid with high biomass yield, 0.68C-mol/C-mol, and productivity, 0.92 g/(L·h). Detoxification with activated charcoal resulted in a similar biomass yield and a slight increase in the volumetric productivity (11%).

The influence of hexoses addition on the fermentation of d-xylose in Debaryomyces hansenii under continuous cultivation

The effect of hexoses (glucose and galactose) addition to the feed xylose mineral medium of Debaryomyces hansenii chemostat cultures grown at a constant dilution rate of 0.055 h(-1) was studied. Xylitol was the major product detected amongst all tested conditions. The maximal values for xylitol yield and volumetric productivity (0.56 gg(-1) xylose and 0.21 gl(-1)h(-1), respectively) were obtained for a glucose/xylose feeding ratio of 10%, showing that the addition of small amounts of glucose, but not galactose, enhanced the xylitol production. A xylitol yield increase of 30%, compared with the sole xylose-containing feed medium, was observed. It was found that the oxygen requirement for D. hansenii growth is lower under glucose compared with xylose. Ethanol and glycerol were only produced for glucose/xylose feeding ratio above 30%. The byproducts accumulation was correlated with glucose metabolism, because a direct relationship between the increase of ethanol (and glycerol) concentration and the increase of glucose in the feed medium was found.