Academic literature on the topic 'Cellobiosio'

2,3-Epoxypropyl and 3,4-epoxybutyl 6′-deoxy-6′-iodo-β-cellobioside, together with 3,4-epoxybutyl 6′- deoxy-6′-fluoro-β-cellobioside, were prepared as putative inhibitors and reporter groups for events occurring at the active site of some β-glucan hydrolases. As well, related syntheses gave the previously unknown 6-deoxy-6-fluoro- and 6′-deoxy-6′-fluoro-cellobioses.

The 2,3-epoxypropyl, 3,4-epoxybutyl and 4,5-epoxypentyl β-glycosides of D-glucose, cellobiose and laminaribiose have been prepared. As well, the 4,5-epoxypentyl β-glycosides of cellotriose, laminaritriose and two other trisaccharides have been synthesized. 3,4-Epoxybutyl β-cellobioside has also been prepared with a 14C-label in the cellobiose residue.

We have measured the hydrolyses of α- and β-cellobiosyl fluorides by the Cel6A [cellobiohydrolase II (CBHII)] enzymes of Humicola insolens and Trichoderma reesei, which have essentially identical crystal structures [Varrot, Hastrup, Schülein and Davies (1999) Biochem. J. 337, 297-304]. The β-fluoride is hydrolysed according to Michaelis-Menten kinetics by both enzymes. When the ~ 2.0% of β-fluoride which is an inevitable contaminant in all preparations of the α-fluoride is hydrolysed by Cel7A (CBHI) of T. reesei before initial-rate measurements are made, both Cel6A enzymes show a sigmoidal dependence of rate on substrate concentration, as well as activation by cellobiose. These kinetics are consistent with the classic Hehre resynthesis-hydrolysis mechanism for glycosidase-catalysed hydrolysis of the ‘wrong’ glycosyl fluoride for both enzymes. The Michaelis-Menten kinetics of α-cellobiosyl fluoride hydrolysis by the T. reesei enzyme, and its inhibition by cellobiose, previously reported [Konstantinidis, Marsden and Sinnott (1993) Biochem. J. 291, 883-888] are withdrawn. 1H NMR monitoring of the hydrolysis of α-cellobiosyl fluoride by both enzymes reveals that in neither case is α-cellobiosyl fluoride released into solution in detectable quantities, but instead it appears to be hydrolysed in the enzyme active site as soon as it is formed.

Cellobiohydrolase II hydrolyses alpha- and beta-D-cellobiosyl fluorides to alpha-cellobiose at comparable rates, according to Michaelis-Menten kinetics. The stereochemistry, absence of transfer products and strict hyperbolic kinetics of the hydrolysis of alpha-cellobiosyl fluoride suggest that the mechanism for the alpha-fluoride may be the enzymic counterpart of the SNi reaction observed in the trifluoroethanolysis of alpha-glucopyranosyl fluoride [Sinnott and Jencks (1980) J. Am. Chem. Soc. 102, 2026-2032]. The absolute factors by which this enzyme accelerates fluoride ion release are small and greater for the alpha-fluoride than for the beta, suggesting that its biological function may not be just glycoside hydrolysis. Cellobiohydrolase I hydrolyses only beta-cellobiosyl fluoride, which is, however, an approx. 1-3% contaminant in alpha-cellobiosyl fluoride as prepared and purified by conventional methods. Instrumental assays for the various components of the cellulase complex are discussed.

The catalytic domain of the CeIF processive endocellulase, a family 48 glycosyl hydrolase from Clostridium cellulolyticum has been crystallized in the presence of a newly synthesized inhibitor (methyl 4-S-β-cellobiosyl-4-thio-β-cellobioside), by vapour diffusion, using PEG as a precipitant. The protein crystallizes in the orthorhombic P212121 space group and diffracts to a resolution of 2.0 Å. The unit-cell parameters are a = 61.4, b = 84.5, c = 121.9 Å.

ABSTRACT Two thermostable endocellulases, CelA and CelB, were purified fromThermotoga neapolitana. CelA (molecular mass, 29 kDa; pI 4.6) is optimally active at pH 6.0 at 95°C, while CelB (molecular mass, 30 kDa; pI 4.1) has a broader optimal pH range (pH 6.0 to 6.6) at 106°C. Both enzymes are characterized by a high level of activity (high V max value and low apparentKm value) with carboxymethyl cellulose; the specific activities of CelA and CelB are 1,219 and 1,536 U/mg, respectively. With p-nitrophenyl cellobioside theV max values of CelA and CelB are 69.2 and 18.4 U/mg, respectively, while the Km values are 0.97 and 0.3 mM, respectively. The major end products of cellulose hydrolysis, glucose and cellobiose, competitively inhibit CelA, and CelB. The Ki values for CelA are 0.44 M for glucose and 2.5 mM for cellobiose; the Ki values for CelB are 0.2 M for glucose and 1.16 mM for cellobiose. CelB preferentially cleaves larger cellooligomers, producing cellobiose as the end product; it also exhibits significant transglycosylation activity. This enzyme is highly thermostable and has half-lives of 130 min at 106°C and 26 min at 110°C. A single clone encoding thecelA and celB genes was identified by screening a T. neapolitana genomic library in Escherichia coli. The celA gene encodes a 257-amino-acid protein, while celB encodes a 274-amino-acid protein. Both proteins belong to family 12 of the glycosyl hydrolases, and the two proteins are 60% similar to each other. Northern blots of T. neapolitana mRNA revealed that celA andcelB are monocistronic messages, and both genes are inducible by cellobiose and are repressed by glucose.

Cellulosic insulation paper is usually used in oil-immersed transformer insulation systems. In this study, the molecular dynamics method based on reaction force field (ReaxFF) was used to simulate the pyrolysis process of a cellobiose molecular model. Through a series of ReaxFF- Molecular Dynamics (MD) simulations, the generation path of ethanol at the atomic level was studied. Because the molecular system has hydrogen bonding, force-bias Monte Carlo (fbMC) is mixed into ReaxFF to reduce the cost of calculation by reducing the sampled data. In order to ensure the reliability of the simulation, a model composed of 20 cellobioses and a model composed of 40 cellobioses were respectively established for repeated simulation in the range of 500–3000 K. The results show that insulating paper produced ethanol at extreme thermal fault, and the intermediate product of vinyl alcohol is the key to the aging process. It is also basically consistent with others’ previous experiment results. So it can provide an effective reference for the use of ethanol as an indicator to evaluate the aging condition of transformers.

Streptomyces lividans CelB is a family-12 endoglucanase that hydrolyses cellulose with retention of anomeric configuration. A recent X-ray structure of the catalytic domain at 1.75 Å resolution has led to the preliminary assignment of Glu-120 and Glu-203 as the catalytic nucleophile and general acid–base respectively [Sulzenbacher, Shareck, Morosoli, Dupont and Davies (1997) Biochemistry 36, 16032–16039]. The present study confirms the identity of the nucleophile by trapping the glycosyl-enzyme intermediate with the mechanism-based inactivator 2´,4´-dinitrophenyl 2-deoxy-2-fluoro-β-d-cellobioside (2FDNPC). The kinetics of inactivation proceeded in a saturable fashion, yielding the parameters kinact = 0.29±0.02 min-1 and Kinact = 0.72±0.08 mM. Uncompetitive inhibition was observed at high concentrations of 2FDNPC (Ki = 9±1 mM), a behaviour that was also observed with the substrate 2´,4´-dinitrophenyl β-d-cellobioside (kcat = 40±1 s-1, Km = 0.35±0.03 mM, Ki = 24±4 mM). Protection against inactivation was afforded by the competitive inhibitor cellobiose. The electrospray ionization (ESI) mass spectrum of the intact labelled CelB indicated that the inactivator had labelled the enzyme stoichiometrically. Reactivation of the trapped intermediate occurred spontaneously (kH2O = 0.0022 min-1) or via transglycosylation, with cellobiose acting as an acceptor ligand (kreact = 0.024 min-1, Kreact = 54 mM). Digestion of the labelled enzyme by pepsin followed by LC–ESI–tandem MS (MS–MS) operating in neutral loss mode identified a labelled, singly charged peptide of m/z 947.5 Da. Isolation of this peptide by HPLC and subsequent collision-induced fragmentation by ESI–MS–MS produced a daughter-ion spectrum that corresponded to a sequence (QTEIM) containing Glu-120. The nucleophile Glu-120 and the putative acid–base catalyst Glu-203 are conserved in all known family-12 sequences.

Bioethanol produced from lignocellulosic biomass represents a promising alternative among biofuels. To date a cost-effective method for the industrial production of bioethanol from vegetal biomass has not been developed. One of the most attractive strategies is the construction of a CBP (Consolidated BioProcessing) microbe able both to hydrolyze the complex polymers of lignocellulosic biomass and to convert these into ethanol. In this context, the present study focused on the development of an industrial CBP microbe for the conversion of cellobiose into ethanol. To this purpose, it was necessary to define a new screening method for the selection of a yeast strain, suitable for the industrial bioethanol production having high fermentative abilities and considerable tolerance to inhibitors commonly present in lignocellulosic hydrolysates. The selection started from a collection of oenological yeasts. These strains, although showing interesting fermentative abilities, did not exhibit a good tolerance to inhibitors such as furfural, acetic acid, formic acid and lactic acid. Therefore, a new isolation programme was necessarily conducted in order to select efficient fermenting yeast strains able to tolerate high concentrations of inhibitory compounds. The isolation procedure, conducted in the presence of an inhibitors cocktail, allowed to obtain a wide collection of yeasts with interesting features for their future applications in the field of second generation bioethanol. Among them, few S. cerevisiae yeasts exhibited remarkable fermenting vigour at high temperature and promising inhibitors tolerance. In particular, S. cerevisiae T2 was selected as host for the development of a recombinant strain able to produce the BglI β-glucosidase of Saccharomycopsis fibuligera, one of the most efficient cellobiose hydrolyzing yeast species. For the first time, in this study, an industrial yeast strain secreting β-glucosidase BglI was described. However, the hydrolytic activity of the recombinant strain must be necessarily increased in order to produce an efficient cellulolytic CBP microbe. On the basis of the preliminary results obtained, this multi-disciplinary work represents a first step towards the development of microbes for the single-step conversion of lignocellulosic biomass to ethanol.
Il bioetanolo di origine lignocellulosica rappresenta una delle alternative più promettenti tra i biocarburanti. Dal punto di vista industriale, la produzione di bioetanolo da biomassa vegetale non è ancora sostenibile. Una delle strategie più interessanti proposte è la costruzione di un microganismo CBP (Consolidated BioProcessing) capace di idrolizzare i polimeri complessi della biomassa cellulosica e di convertirli efficacemente in etanolo. In questa prospettiva, questo lavoro di tesi si è focalizzato sullo sviluppo di un microbo CBP di tipo industriale per la conversione di cellobiosio in alcol etilico. A tal scopo, è stato necessario mettere a punto un nuovo metodo per la selezione di un ceppo di lievito idoneo alla produzione di bioetanolo su scala industriale caratterizzato da elevate performance fermentative e da una notevole capacità di tollerare gli inibitori normalmente presenti negli idrolizzati lignocellulosici. La selezione di tale microrganismo è partita da una collezione di ceppi di lievito di origine enologica. I ceppi enologici saggiati, pur dimostrando elevate capacità fermentative, non si sono purtroppo rivelati tolleranti nei confronti di inibitori quali furfurale, acido acetico, acido formico ed acido lattico. È stato quindi necessario eseguire un programma di isolamento mirato ad ottenere ceppi di lievito altamente fermentanti e capaci di tollerare elevate concentrazioni di inibitori. L’isolamento, eseguito in condizioni selettive per la presenza di un cocktail di inibitori, ha consentito di ottenere una ampia ceppoteca di lieviti con caratteristiche promettenti per la loro futura applicazione nel campo del bioetanolo di seconda generazione. Tra di essi, alcuni lieviti S. cerevisiae si sono distinti per vigore fermentativo ad elevata temperatura e per una consistente tolleranza agli inibitori. In particolare, il ceppo S. cerevisiae T2 è stato selezionato come host strain per lo sviluppo di un ceppo ricombinante capace di secernere la betaglucosidasi BglI di Saccharomycopsis fibuligera, specie di lievito tra le più efficienti per l’idrolisi del cellobiosio. Per la prima volta in questo lavoro di tesi è stato descritto un ceppo di lievito industriale betaglucosidasico. In ogni caso, l’attività idrolitica del ceppo ricombinante dovrà essere necessariamente incrementata al fine di ottenere un efficiente microrganismo CBP cellulosolitico. In base ai risultati ottenuti, questo studio rappresenta un primo passo verso lo sviluppo di microrganismi idonei alla conversione one-step di biomassa lignocellulosica in etanolo.

Upgrade of biomass to valuable chemicals is a central topic in modern research due to the high availability and low price of this feedstock. For the difficulties in biomass treatment, different pathways are still under investigation. A promising way is in the photodegradation, because it can lead to greener transformation processes with the use of solar light as a renewable resource. The aim of my work was the research of a photocatalyst for the hydrolysis of cellobiose under visible irradiation. Cellobiose was selected because it is a model molecule for biomass depolymerisation studies. Different titania crystalline structures were studied to find the most active phase. Furthermore, to enhance the absorption of this semiconductor in the visible range, noble metal nanoparticles were immobilized on titania. Gold and silver were chosen because they present a Surface Plasmon Resonance band and they are active metals in several photocatalytic reactions. The immobilized catalysts were synthesized following different methods to optimize the synthetic steps and to achieve better performances. For the same purpose the alloying effect between gold and silver nanoparticles was examined.

Dissertation (PhD)--Stellenbosch University, 2007.
PCT patent registered: https://www.google.com/patents/WO2009034414A1?cl=en&dq=pct/ib2007/004098&hl=en&sa=X&ei=b7AxUsSZK4jB0gWi14HgCQ&ved=0CEkQ6AEwAg USA: https://www.google.com/patents/US20110129888?dq=pct/ib2007/004098&ei=b7AxUsSZK4jB0gWi14HgCQ&cl=en
USA patent registered: https://www.google.com/patents/US20110129888?dq=pct/ib2007/004098&ei=b7AxUsSZK4jB0gWi14HgCQ&cl=en
ENGLISH ABSTRACT: The conversion of cellulosic biomass into fuels and chemicals has the potential to positively impact the South African economy, but is reliant on the development of low-cost conversion technology. Perhaps the most important progress to be made is the development of “consolidated bioprocessing” (CBP). CBP refers to the conversion of pretreated biomass into desired product(s) in a single process step with either a single organism or consortium of organisms and without the addition of cellulase enzymes. Among the microbial hosts considered for CBP development, Saccharomyces cerevisiae has received significant interest from the biotechnology community as the yeast preferred for ethanol production. The major advantages of S. cerevisiae include high ethanol productivity and tolerance, as well as a well-developed gene expression system. Since S. cerevisiae is non-cellulolytic, the functional expression of at least three groups of enzymes, namely endoglucanases (EC 3.2.1.4); exoglucanases (EC 3.2.1.91) and β-glucosidases (EC 3.2.1.21) is a prerequisite for cellulose conversion via CBP. The endo- and exoglucanases act synergistically to efficiently degrade cellulose to soluble cellodextrins and cellobiose, whereas the β-glucosidases catalyze the conversion of the soluble cellulose hydrolysis products to glucose. This study focuses on the efficient utilization of cellobiose by recombinant S. cerevisiae strains that can either hydrolyse cellobiose extracellularly or transport and utilize cellobiose intracellularly. Since it is generally accepted that S. cerevisiae do not produce a dedicated cellobiose permease/transporter, the obvious strategy was to produce a secretable β-glucosidase that will catalyze the hydrolysis of cellobiose to glucose extracellularly. β-Glucosidase genes of various fungal origins were isolated and heterologously expressed in S. cerevisiae. The mature peptide sequence of the respective β-glucosidases were fused to the secretion signal of the Trichoderma reesei xyn2 gene and expressed constitutively from a multi-copy yeast expression vector under transcriptional control of the S. cerevisiae PGK1 promoter and terminator. The resulting recombinant enzymes were characterized with respect to pH and temperature optimum, as well as kinetic properties. The maximum specific growth rates (μmax) of the recombinant strains were compared during batch cultivation in high-performance bioreactors. S. cerevisiae secreting the recombinant Saccharomycopsis fibuligera BGL1 enzyme was identified as the best strain and grew at 0.23 h-1 on cellobiose (compared to 0.29 h-1 on glucose). More significantly, was the ability of this strain to anaerobically ferment cellobiose at 0.18 h-1 (compared to 0.25 h-1 on glucose). However, extracellular cellobiose hydrolysis has two major disadvantages, namely glucose’s inhibitory effect on the activity of cellulase enzymes as well as the increased risk of contamination associated with external glucose release. In an alternative approach, the secretion signal from the S. fibuligera β-glucosidase (BGL1) was removed and expressed constitutively from the above-mentioned multi-copy yeast expression vector. Consequently, the BGL1 enzyme was functionally produced within the intracellular space of the recombinant S. cerevisiae strain. A strategy employing continuous selection pressure was used to adapt the native S. cerevisiae disaccharide transport system(s) for cellobiose uptake and subsequent intracellular utilization. RNA Bio-Dot results revealed the induction of the native α-glucoside (AGT1) and maltose (MAL) transporters in the adapted strain, capable of transporting and utilizing cellobiose intracellularly. Aerobic batch cultivation of the strain resulted in a μmax of 0.17 h-1 and 0.30 h-1 when grown in cellobiose- and cellobiose/maltose-medium, respectively. The addition of maltose significantly improved the uptake of cellobiose, suggesting that cellobiose transport (via the combined action of the maltose permease and α-glucosidase transporter) is the rate-limiting step when the adapted strain is grown on cellobiose as sole carbon source. In agreement with the increased μmax value, the substrate consumption rate also improved significantly from 0.25 g.g DW-1.h-1 when grown on cellobiose to 0.37 g.g DW-1.h-1 upon addition of maltose to the medium. The adapted strain also displayed several interesting phenotypical characteristics, for example, flocculation, pseudohyphal growth and biofilm-formation. These features resemble some of the properties associated with the highly efficient cellulase enzyme systems of cellulosome-producing anaerobes. Recombinant S. cerevisiae strains that can either hydrolyse cellobiose extracellularly or transport and utilize cellobiose intracellularly. Both recombinant strains are of particular interest when the final goal of industrial-scale ethanol production from cellulosic waste is considered. However, the latter strain’s ability to efficiently remove cellobiose from the extracellular space together with its flocculating, pseudohyphae- and biofilm-forming properties can be an additional advantage when the recombinant S. cerevisiae strain is considered as a potential host for future CBP technology.
AFRIKAANSE OPSOMMING: Die omskakeling van sellulose-bevattende biomassa na brandstof en chemikalieë beskik oor die potensiaal om die Suid-Afrikaanse ekonomie positief te beïnvloed, indien bekostigbare tegnologie ontwikkel word. Die merkwaardigste vordering tot dusvêr kon in die ontwikkeling van “gekonsolideerde bioprosessering” (CBP) wees. CBP verwys na die eenstap-omskakeling van voorafbehandelde biomassa na gewenste produkte met behulp van ‘n enkele organisme of ‘n konsortium van organismes sonder die byvoeging van sellulase ensieme. Onder die mikrobiese gashere wat oorweeg word vir CBP-ontwikkeling, het Saccharomyces cerevisiae as die voorkeur gis vir etanolproduksie troot belangstelling by die biotegnologie-gemeenskap ontlok. Die voordele van S. cerevisiae sluit in hoë etanol-produktiwiteit en toleransie, tesame met ‘n goed ontwikkelde geen-uitdrukkingsisteem. Aangesien S. cerevisiae nie sellulose kan benut nie, is die funksionele uitdrukking van ten minste drie groepe ensieme, naamlik endoglukanases (EC 3.2.1.4); eksoglukanases (EC 3.2.1.91) en β-glukosidases (EC 3.2.1.21), ‘n voorvereiste vir die omskakeling van sellulose via CBP. Die sinergistiese werking van endo- en eksoglukanases word benodig vir die effektiewe afbraak van sellulose tot oplosbare sello-oligosakkariede en sellobiose, waarna β-glukosidases die finale omskakeling van die oplosbare sellulose-afbraak produkte na glukose kataliseer. Hierdie studie fokus op die effektiewe benutting van sellobiose m.b.v. rekombinante S. cerevisiae-rasse met die vermoeë om sellobiose ekstrasellulêr af te breek of dit op te neem en intrasellulêr te benut. Aangesien dit algemeen aanvaar word dat S. cerevisiae nie ‘n toegewyde sellobiosepermease/ transporter produseer nie, was die mees voor-die-hand-liggende strategie die produksie van ‘n β-glukosidase wat uitgeskei word om sodoende die ekstrasellulêre hidroliese van sellobiose na glukose te kataliseer. β-Glukosidase gene is vanaf verskeie fungi geïsoleer en daaropvolgend in S. cerevisiae uitgedruk. Die geprosesseerde peptiedvolgorde van die onderskeie β-glukosidases is met die sekresiesein van die Trichoderma reesei xyn2-geen verenig en konstitutief vanaf ‘n multikopie-gisuitdrukkingsvektor onder transkripsionele beheer van die S. cerevisiae PGK1 promotor en termineerder uitgedruk. Die gevolglike rekombinante ensieme is op grond van hul pH en temperatuur optima, asook kinetiese eienskappe, gekarakteriseer. Die maksimum spesifieke groeitempos (μmax) van die rekombinante rasse is gedurende aankweking in hoë-verrigting bioreaktors vergelyk. Die S. cerevisiae ras wat die rekombinante Saccharomycopsis fibuligera BGL1 ensiem uitskei, was as the beste ras geïdentifiseer en kon teen 0.23 h-1 op sellobiose (vergeleke met 0.29 h-1 op glukose) groei. Meer noemenswaardig is the ras se vermoë om sellobiose anaërobies teen 0.18 h-1 (vergeleke met 0.25 h-1 op glukose) te fermenteer. Ekstrasellulêre sellobiose-hidroliese het twee groot nadele, naamlik glukose se onderdrukkende effek op die aktiwiteit van sellulase ensieme, asook die verhoogde risiko van kontaminasie wat gepaard gaan met die glukose wat ekstern vrygestel word. ’n Alternatiewe benadering waarin die sekresiesein van die S. fibuligera β-glucosidase (BGL1) verwyder en konstitutief uitgedruk is vanaf die bogenoemde multi-kopie gisuitrukkingsvektor, is gevolg. Die funksionele BGL1 ensiem is gevolglik binne-in die intrasellulêre ruimte van die rekombinante S. cerevisiae ras geproduseer. Kontinûe selektiewe druk is gebruik om die oorspronklike S. cerevisiae disakkaried-transportsisteme vir sellobiose-opname and daaropvolgende intrasellulêre benutting aan te pas. RNA Bio-Dot resultate het gewys dat die oorspronklike α-glukosied (AGT1) en maltose (MAL) transporters in die aangepaste ras, wat in staat is om sellobiose op te neem en intrasellulêr te benut, geïnduseer is. Aërobiese kweking van die geselekteerde ras het gedui dat die ras teen 0.17 h-1 en 0.30 h-1 groei in onderskeidelik sellobiose en sellobiose/maltose-medium. Die byvoeging van maltose het die opname van sellobiose betekenisvol verbeter, waarna aangeneem is dat sellobiose transport (via die gekombineerde werking van die maltose permease en α-glukosidase transporter) die beperkende stap gedurende groei van die geselekteerde ras op sellobiose as enigste koolstofbron is. In ooreenstemming hiermee, het die substraatbenuttingstempo ook betekenisvol toegeneem van 0.25 g.g DW-1.h-1, gedurende groei op sellobiose, tot 0.37 g.g DW-1.h-1 wanneer maltose by die medium gevoeg word. Die geselekteerde ras het ook verskeie interessante fenotipiese kenmerke getoon, byvoorbeeld flokkulasie, pseudohife- en biofilm-vorming. Hierdie eienskappe kom ooreen met sommige van die kenmerke wat met die hoogs effektiewe sellulase ensiem-sisteme van sellulosomeproduserende anaerobe geassosieer word. Hierdie studie beskryf die suksesvolle konstruksie van ‘n rekombinante S. cerevisiae ras met die vermoë om sellobiose ekstrasellulêr af te breek of om dit op te neem en intrasellulêr te benut. Beide rekombinante rasse is van wesenlike belang indien die einddoel van industriële-skaal etanolproduksie vanaf selluloseafval oorweeg word. Die laasgenoemde ras se vermoë om sellobiose effektief uit die ekstrasellulêre ruimte te verwyder tesame met die flokkulasie, pseudohife- en biofilm-vormings eienskappe kan ‘n addisionele voordeel inhou, indien die rekombinante S. cerevisiae ras as ‘n potensiële gasheer vir toekomstige CBP-tegnologie oorweeg word.

The solubility of cellobiose in 18 organic liquids and water was measured at 20°C. Hydrogen bond acceptors were the most effective solvents. Three models were analyzed to evaluate their accuracy and to understand factors that affect cellobiose solubility: Hansen solubility parameters (HSP), linear free energy relationship (LFER), and UNIQUAC functional-group activity coefficients (UNIFAC). The HSP of cellobiose were determined and the model was able to distinguish between most good and poor solvents, however, proved to be occasionally unreliable due to a false negative. The LFER model produced an empirical equation involving contributions from solvent molar refraction, polarizability, acidity, basicity, and molar volume, which predicted cellobiose solubilities to within ±2 log units. LFER indicated that good solvents were highly polarizable and had low molar volume, which was consistent with the good solvents found for cellobiose. A modified version of UNIFAC that includes an association term (A-UNIFAC) predicted the solubility of cellobiose in water and alcohols to within ±0.6 log units, indicating that A-UNIFAC can be used to predict the solubility of cellobiose and other carbohydrates provided additional data to extend the model to solvents other than water and alcohols.

This thesis reports some insight into fundamental chemistry of cellobiose decomposition in hot-compressed water (HCW). This includes the effect of mild temperature, initial cellobiose concentration, weakly acidic condition and AAEM chlorides on decomposition behaviour of cellobiose. The new knowledge provides fundamental understanding on decomposition mechanisms of sugar oligomers into monomers and other products. Such knowledge is also essential to understanding the decomposition behaviour of more complex cellulose and biomass conversion in HCW.

Pennycress is an annual cover crop in temperate North America and its seeds contain around 30% of oil and 20% of crude protein. Pennycress oil can be used for biodiesel production, while the seed meal has limited use in animal feed due to its relative high content of phenolic compounds and crude fiber. The nutritional value of pennycress meal (PM) can be improved by processing with GRAS fungal strains. In this study, three fungal strains, Rhizopus oryzae (RO), Mucor indicus (MI), and Aspergillus oryzae (AO), were used to ferment PM that contains 21% of total amino acids and 17% of structure carbohydrates. The fermentation was performed by inoculating each strain to the sterilized PM with initial moisture of 60% and incubated statically at 28 °C for 6 days. Amino acids profile, structure carbohydrates, soluble sugar, phytate, and mycotoxins including total aflatoxins, zearalenone (ZEN), and deoxynivalenol (DON) were monitored on the samples after fermentation. As compared to control without fermentation, the total amino acids were improved by 4.0% with RO and 5.9% with AO. Threonine, arginine, alanine, and lysine were significantly enriched in RO and AO treated meal. RO and MI degraded the fiber component into cellobiose, which was increased by 3 and 5.8-fold, respectively. Phytate was reduced by 46.6% with RO, 37.3% with AO, and 33.3% with MI. Compared with the control, ZEN was reduced by 39.3%, 32%, and 50% in AO, MI and RO treated meal, respectively. Total aflatoxin content was low in PM, and MI and RO treatments further reduced its content after fermentation. No significant change of DON was observed in the PM fermented by each strain. This study demonstrated the potential of using fungi to improve the feeding value of PM, which could potentially promote the plantation of oilseed crops in the region.

Abstract Cellulose, the one of the most abundant biomaterials available in nature, is a polymer with cellobiose as the smallest repeating unit, with a degree of polymerization that can go up to 1000 for wood cellulose. The strength-to-weight ratio of nanocellulose is eight times greater than steel (Patchiya Phanthong et al). Nanocellulose in suspension (NCS) at a varied concentration helps increase properties of cement without changing the density of the cement slurry. Being mindful of challenges in oil and gas wells, efforts were made to enhance cement properties using nanocellulose within conventional and water-extended cement systems. Samples of 15.8-ppg conventional and 12 ppg water-extended cements were prepared by varying the proportion of nanocellulose within an aqueous suspension. Rheology, sedimentation, compressive strength and mechanical properties were analyzed for a conventional 15.8-ppg cement system with varying NCS proportions of 0, 2, 4, and 5% by weight of cement (BWOC). Similar work was performed for a 12 ppg water-extended cement system by varying NCS differently in proportions of 0, 5, 10, and 20% BWOC. Two-inch cubes were set at 170°F for 24 hours for each sample. They were crushed using hydraulic crush compressive strength equipment, and the force used to break the sample was recorded. Compressive strength for this cement system was measured to be 2450, 3250, 3450, and 3875 psi, respectively, for samples with 0, 2, 4, and 5% BWOC concentrations of NCS. An increase in the strength of cement with an increase in NCS percentage was observed for the 15.8-ppg slurry design, which may be attributed to the size and shape of the NCS. However, similar study carried out with 12 ppg water extended slurries showed decrease in overall compressive strength. Nano-sized particles fill the pores within the sample, impacting structural network development. Additionally, cellulose, having a fiber-like structure, may provide inter-particulate reinforcement. Based on the results of the 15.8-ppg cement system and the high tensile strength of nanocellulose, it can be determined that NCS has a positive effect for increasing mechanical properties. By applying nanocellulose, a tailored cement system (dependable barrier) can be designed to minimize risk and maximize production from oil and gas wells. Nanocellulose is of increasing interest for a range of applications relevant to the fields of material science and biomedical engineering because of its renewable nature, anisotropic shape, excellent mechanical properties, good biocompatibility, tailorable surface chemistry, and interesting optical properties. Low-volume NCS additions can alter the structure of the cured cement system and increase its mechanical properties. This reinforcing mechanism may provide a new opportunity for achieving higher strength cementitious materials.