No description available.

Plant biomass serves as a carbon and energy source for Streptomyces spp. which secrete degradative enzymes capable of breaking down the complex plant biomass into simple saccharides. Hemicellulose is a major component of plants and is composed of five and six carbon sugars, such as xylose and glucose. Enzymatic degradation of hemicellulose to obtain desired sugars has been a cornerstone of many industries, as well as the subject of worldwide research for additional sources of efficient enzymes for substrate conversion. In this study, environmentally-derived Streptomyces isolates were screened for their ability to hydrolyze oat-spelt and birchwood xylan in agar-based high throughput activity screens. Of the isolates tested, eight displayed high levels of substrate-degrading activity and were chosen for further characterization which included 16S rRNA gene analysis, microscopic analysis from both liquid and agar grown cultures, xylanase-specific activity, lignin peroxidase production and indole acetic acid production.Qualitative assessment of extracellular lactone signalling for all eight isolates was also performed. Putative lactone signalling was observed for Streptomyces isolates JLS1-C4, JLS1-A6, JLS2-D6 and KT1-B1 which exhibited xylanase-specific activities of 0.622 μmol/min/mg, 0.0243 μmol/min/mg, 0.721 μmol/min/mg, and 0.706 μmol/min/mg respectively. Streptomyces isolates JLS1-F12 and JLS1-C12 did not exhibit lactone signalling but did exhibit xylanase-specific activities of 0.125 μmol/min/mg and 0.0688 μmol/min/mg respectively. No xylanase-specific activity was detected for isolates JLS2- C7 and KT1-B8; however lactone signalling was observed for isolate KT1-B8. Streptomyces isolate JLS1-A6 degraded birchwood xylan optimally at pH 4 and 28°C with a maximal xylanase activity of 1.56 x10-3 μmol/min/mg. / UOIT

In the forest products industry the opportunity exists to extract currently under-utilised compounds from the process or waste streams and thereby derive more value from the wood entering the process. A big portion of the hemicellulose content of wood does not form part of the final product. Extracting the hemicelluloses from the waste streams or other locations in the process would allow them to be used more effectively. The predominant hardwood hemicellulose, xylan, is polymeric xylose. Xylose is an important platform sugar in bioconversion strategies and can be converted to fuels and other valuable chemicals. The xylan polymer can be hydrolysed to its xylose monomers by a number of conversion strategies; the most widely known being chemical and enzymatic digestion. Chemical conversion is usually done using acid at elevated temperatures, but high yields are often offset by degradation of the product. On the other hand, enzymatic hydrolysis can be better regulated to prevent unwanted degradation of the monomeric sugar products. Enzymatic hydrolysis has been pronounced the environmentally friendly choice of technology, although it is hampered by low conversions and high cost of enzymes. To date commercial enzymes for biomass conversion are not readily available most of which are still in development. In understanding how to best utilise a xylan, recovered from the pulping process, the potential to convert hardwood xylan to xylose with enzymes currently available on the market was studied. A hardwood xylan extracted from fully bleached Eucalyptus pulp with a chelating agent, Nitren, was used as substrate to evaluate the ability of some commercial enzymes to degrade the extracted xylan to xylose monomers. The enzymes used in this study were not dedicated biomass conversion enzymes, but rather chosen for their xylan degrading potential, i.e. xylanase content. By means of hydrolysis profiles on commercial Birchwood and Oat Spelts xylan as substrates and enzyme characterisation, Multifect xylanase was identified as most promising enzyme for xylan conversion. Multifect contained high levels of xylanase and xylosidase activity in the enzyme preparation. Commercial Birchwood xylan and the extracted Eucalyptus xylan were found to be chemically similar, both composed predominantly of xylose. The hydrolysis profiles obtained on Birchwood xylan could therefore serve as a benchmark against which the hy-drolysis of Eucaluptus xylan could be compared. Full conversion of the Eucalyptus xylan with Multifect could not be achieved, although Multifect completely degraded the Birchwood xylan. The maximum xylose yield that could be obtained on Eucalyptus xylan was 80 % and it was concluded that the remaining 20% was unhydrolysable by the enzyme, most likely due to the limitations in the employed extraction method. It was however noted that up to the point of 80 % conversion higher hydrolysis rates were observed on Eucalyptus xylan than Birchwood xylan with equal charges of Multifect. The differences in hydrolysis rates may have indicated that the Eucalyptus xylan is more accessible to enzyme attack than the Birchwood xylan, likely as a result of the extraction methods used to prepare the xylans. A simple economic evaluation illustrated the weight of various costs in process profitability. The most economic operation of a continuous steady state reactor is at a low enzyme charge, 17 IU/ℓ, and a long retention period, five days, due to the high cost of the enzyme compared to other factors. For a reduced retention time, an investigation into enzyme immobilisation and the use of a packed-bed type reactor is recommended. / Dissertation (MEng)--University of Pretoria, 2009. / Chemical Engineering / unrestricted

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Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Hemicellulose, a plant cell wall polysaccharide composed amongst other 6-carbon sugars of xylose, a 5-carbon fermentable sugar, has various side-chains which hinder breakdown by the hydrolytic enzyme xylanase. We executed a screen of a metagenomic library established from heat treated saw dust to isolate novel genes for plant cell wall esterases and isolated two clones that potentially could be utilised as acetyl xylan esterases: HEL12 and HEL37 from Citrobacter farmeri and Bacillus vallismortis respectively are confirmed short-chain acetyl xylan esterases (C2-C4) with an optimal temperature of 30°C and 35°C and pH 8.0. HEL12 (33kDa) and HEL37 (25kDa) are small, dextrous acetyl xylan esterases with HEL37: Km of 1.621mM for p-nitrophenyl acetate and Km of 3.571mM for the substrate p-nitrophenyl palmitate with a Vmax of 2.462 mMol/min/mg protein and Vmax of 0.4363 mMol/min/mg protein respectively. In addition HEL12 was active on other substrates with a Km of 1.321mM protein for p-nitrophenyl acetate and Km of 1.692mM for the substrate p-nitrophenyl butyrate with a Vmax of 3.812 and Vmax of 1.523 mMol/min/mg protein respectively. Both enzymes were assayed on various acetylated polymers including acetylated xylan to indicate their ability to hydrolyse plant lignocellulosic polymers. / AFRIKAANSE OPSOMMING: Met die herwinde interisering in die energie krisis, is “herwinbare energie” vinnig besig om die nuwe modewoord te raak tussen verskeie industriële bedrywighede. Hemicellulose, ‘n plant sel wand polisakkaride opgemaak uit xilose, ‘n 5-koolstof fermenteerbare suiker, het verskillende sykettings wat ‘n hindernis veroorsaak in die hidrolase van die komponent deur xilanase ensieme. HEL12 en HEL37 van bakterieë Citrobacter farmeri en Bacillus vallismortis afsonderlik, is deel van die hidrolitiese groep van ensieme wat die hidrolisasie van ester sy-bindings op die xilaan suikerstring kataliseer. Die ensieme was primêr geassesseer vir aktiwiteit deur middel van sintetiese substraat 4-nitrofeniel asetaat. HEL12 en HEL37 was gevind om kort-ketting asetiel xilaan esterase (C2-C4) te wees, met ‘n optimale temperatuur van 30°C en 35°C afsonderlik, beide met ‘n optimale pH van 8.0. Met ‘n geskatte gewig van omtrent +- 30kDa is beide die ensieme van die kleiner meer behendige asetiel xilaan esterase, met HEL37: Km = 1.621 op 4-nitrofeniel asetaat en Km = 3.571 op substraat 4-nitrofeniel palmetaat, en Vmax = 2.462 met ‘n Vmax = 0.4363 afsonderlik, en HEL12: Km = 1.321 op 4-nitrofeniel asetaat en Km = 1.692 op sustraat 4-nitrofeniel buteraat met ‘n Vmax = 3.812 en Vmax = 1.523 afsonderlik.

Interactions between polysaccharide derivatives and rod-like cellulose nanocrystals were studied by light scattering. Two replicates of cellulose nanocrystals (DOE-2-12A and DOE-2-12B) were probed with polarized and depolarized dynamic light scattering. X-ray photoelectron spectroscopy experiments showed sulfate groups on cellulose nanocrystals. Decay rates from polarized dynamic light scattering experiments exhibited a significantly smaller angular dependence for both samples. However, DOE-2-12B showed a smaller angular dependence than DOE-2-12A. Lengths and diameters of DOE-2-12A and DOE-2-12B obtained by Broersma's formula were 229 " 19 and 19 " 7 nm and 240 " 18 and 22 " 6 nm, respectively. The resultant length and diameter of DOE-2-12B were comparable to those for cellulose whiskers obtained from cotton. Adsorption of pullulan 4-chlorocinnamate (P4CC03) onto cellulose nanocrystals (DOE-2-12B) was also studied by UV-Vis spectroscopy, zeta-potential measurements, and polarized and depolarized dynamic light scattering. UV-Vis spectroscopy of the P4CC03/water binary system and in situ light scattering showed UV crosslinking of pullulan 4-chlorocinnamate occurred in binary and ternary systems but led to different aggregation behavior in the two ternary systems: PreX where P4CC03 crosslinking occurred prior to the addition of cellulose nanocrystals and Rxn where cellulose nanocrystals were present during UV exposure. These studies showed P4CC03 adsorbed onto cellulose nanocrystals and UV induced crosslinking occurred at the surface of the cellulose nanocrystals. Zeta-potential measurements also showed that P4CC03 adsorbed onto cellulose nanocrystals. Finally, adsorption of 2-hydroxypropyltrimethylammonium xylans (HPMAXs) of degree of molar substitution MS = 0.34 onto rod-like cellulose nanocrystals (DOE-2-12Bs) were probed with zeta-potential measurements and polarized and depolarized dynamic light scattering. Zeta-potential changes of HPMAX/water, HPMAX/DOE-2-12B/water, and DOE-2-12B/water systems showed HPMAX adsorption onto DOE-2-12Bs. Intensity autocorrelation functions from Hv and Vv mode exhibited partial heterodyning. Decay time distributions of the binary and ternary systems showed that aggregates existed in the binary system but disappeared in the ternary system. These observations revealed that HPMAX adsorbed onto a fraction of the cellulose nanocrystals in the ternary system at low concentrations of HPMAX. Decreasing translational and rotational diffusion coefficients with increasing HPMAX concentration indicated HPMAX adsorption onto cellulose nanocrystals. A significant HPMAX concentration dependence of the ratio of rotational diffusion coefficient to translational diffusion coefficient showed strong adsorptive interactions between HPMAX and DOE-2-12B. These studies showed there were interactions between polysaccharides and cellulose nanocrystals even in very dilute solutions. Also, it was shown that probe diffusion studies with rod-like cellulose nanocrystals is a promising strategy for probing complicated polymer solutions. / Ph. D.

The aim of this Master thesis was to investigate if the xylan content had any influence on the physical properties of softwood kraft pulps. To achieve pulps with different xylan content different kraft cooking conditions were used; two different temperatures and two different effective alkali levels. The cooking conditions used were 160°C with 30% effective alkali (EA) referred to as reference cook and 145°C with 17% effective alkali (EA) referred to as the mild cook. The first step in this study was to determine the cooking time needed for reaching a certain kappa number, i.e.30. It was also determined whether the difference in xylan content between the pulp samples was sufficient when these cooking conditions were used. When the correct cooking time and cooking conditions had been found new cooks were made using these conditions. All cooks were made at a liqour to wood ratio of 4:1.   The difference in xylan content between the corresponding pulp samples was found to be 3%. The physical testing showed no significant difference in the tensile strength between the two pulps after beating. There was a difference in tear index however and the pulp with the highest content of xylan had the lowest tear index. Zero-span index was the same for the two pulps when unbeaten. After beating the zero-span index decreased for the pulp with highest xylan content but stayed unchanged for the pulp with the lower amount of xylan. These results can be explained by the results from the fibre analysis which showed that the fibres with high xylan content were longer, thicker and had a higher coarseness. Thicker fibres are probably stiffer than thinner fibres which gives the paper fewer bonding points and a lower strength. The result from the zero-span test indicated that the fibres with higher xylan content are affected more by beating than fibres with the lower xylan content. / Målet med examensarbetet var att undersöka om och hur mycket xylaneti pappersmassan påverkar fiberns och därmed papperets fysikaliska egenskaper. Egenskaperna som undersöktes var drag- och rivstyrka samt zero-spanstyrka. Xylaninnehållet skulle varieras genom att kokförhållandena förändrades dels genom olika koktemperaturer dels olika satsningar av effektivt alkali vid given sulfiditet. Dessa var 160ºC med 30% effektivt alkali(EA) hädanefter benämnd referenskoket och 145ºC med 17% effektivt alkali(EA) som benämns det milda koket i fortsättningen.En bestämning av koktiden gjordes för att nå 30 i kappatal och två provkok, ett vid varje temperatur behövde göras.Skillnaden i xylanhalt mellan de slutliga massaproverna låg på c:a 3% enheter.   Styrkeproverna gav inga entydiga svar på om skillnaden i xylanhalt gav någon effekt på massastyrkan. Dragproverna visade att för omald massa var massan från referenskoket starkast men att massan från det mildare koket reagerade kraftigare på malningen. Redan vid 1000 varv hade den i princip samma dragindex som referensmassan vid samma malgrad. Zero-span mätningarna visade att fibrerna hade samma styrka när de var omalda.   Resultaten från fiberanalysenverifierade resultaten från styrketesterna eftersom en tjockare fiber bör ge en styvare fiber och därmed erhålls färre bindningspunkter.  Färre bindningspunkter ger en lägre dragstyrka och det krävs mindre energi för att bryta bindningarna. Efter malningen kan man se att zero-span styrkan har minskat betydligt för massan med högre xylanhalt medan referensmassan behöll styrkan. Dessutom har dragstyrkan ökat för båda massorna men mest för massan med högre xylanhalt. Det kan förklaras med att malningen ger små fibriller på ytan av fibern och en mjukare och böjligare fiber. Därmed ökar bindningsstyrkan då bindningsarean ökar och fibern blir mjukare och böjligare. Den ökade bindningsgraden samt den minskade fiberstyrkan kan förklara varför rivindex fortfarande var lägre för massan med högre xylanhalt trots att dragindex ökade med ökad malning.En annan förklaring kan vara att vid ett långt kok med låg temperatur är det troligt att lignin adsorberas på fibrerna.Ligninet på ytan ger en sämre bindningsförmåga vilket leder till att de är lättare att dra ur nätverket med lägre energiåtgång som följd.

Lösligheten av xylaner utvunna från trä undersöktes med Dynamisk Ljusspridning i syfte att observera eventuella förändringar i hydrodynamisk volym och ζ-potential. Xylaner extraherade från bok, björk och gran användes, där alkalisk extraherad bok erhölls kommersiellt och prover från björk och gran extraherades i förhand med subkritiskt vatten och alkalisk extraktion. Proverna analyserades vid koncentrationerna 1 g/L respektive 20 g/L. Förändringar i pH observerades stegvis med titrering av 1 g/L prover och förändringar över tid vid konstant pH observerades upp till en vecka efter upplösning. 20 g/L proverna undersöktes över tid vid bestämda pH enligt samma process för 1 g/L för både filtrerade och ofiltrerade prover. En undersökning av utvalda filter genomfördes där stora variationer observerades mellan det olika arterna men även mellan filtrens porstorlek. Med dynamisk ljusspridning observerades att populationer av högsta molekylvikt avlägsnades, där inverkan var störst för proven med lägst total koncentration och för högre koncentration efter en vecka. SEC visade att även populationer med mindre molekylvikt avlägsnades som medför att aggregering av proven kan ha förekommit, vilket kan ha förhindrat passingering genom filtren. Med några få undantag, samtliga provers hydrodynamiska volym visades vara heterogena, med endast små ökningar vid ökning i koncentration och icke-signifikanta förändringar över tid och med varierande pH. ζ-potentialvärdena visade markanta förändringar och tydliga trender med ändringar i pH och koncentration, där minskande pH och ökande koncentrationer resulterade i minskad stabilitet i lösning. Över tid ökades absolut ζ-potentialen för de prover med högre koncentration för alla pH miljöer. Filtrering av 20 g/L proverna resulterade i högre initial ζ-potential men efter en vecka ökade stabiliteten för de ofiltrerade till något högre slut ζ-potential för ofiltrerade. / The solubility of wood xylans was investigated using Dynamic Light Scattering to calculate number-average size and ζ-potential of samples in aqueous solution. The sources of xylan were a commercial alkaline extracted beech glucuronoxylan as well as pre-prepared samples of birch glucuronoxylan and spruce arabinoxylan, extracted using subcritical water extraction and alkaline extraction. The samples were analysed at concentrations of 1 g/L and 20 g/L, where the lower concentration samples were analysed at varying pH using titration and set pH over the span of hours and finally up to a week after preparation. The higher concentration samples were observed at set pH following the same procedure as the lower concentration for both filtered and unfiltered samples. An analysis of the filtration of 1 g/L samples was conducted by freeze-drying and weighing the samples before and after using 0.2 μm and 1.2 μm filters. These were then compared to unfiltered samples to evaluate the loss of total mass after filtration. The results of this investigation found significant variance between the samples and filters selected. Comparisons of filtered and unfiltered samples consistently showed that the filtration removed higher molecular weight portions of the samples, the most significant differences being observed at lower concentrations and higher concentration after 1 week. SEC analysis showed that not only larger molecular weight fraction but all the observed populations were removed during filtration, indicating that aggregates could be present, inhibiting the smaller molecular weight fractions from passing through. The size measurements showed that the vast majority of samples were heterogeneous in nature and with a slight increase in number-average size with increasing concentration and little change when varying both pH and over time. ζ-potential measurements showed substantial changes in stability when varying pH, coinciding well with predicted pKa values and generally decreased stability at higher concentrations. For higher concentrations lower initial absolute ζ-potential values were observed for both filtered and unfiltered however over time the unfiltered samples showed slightly greater increases for all measured pH values.

<p>The main goal for a packaging is to protect the product inside. Typical packaging nowadays is made of layers of paper and barriers consisting of plastics or aluminum foil. A problem with the barrier used today is the environmental thinking. Xylophane® is an environmental friendly and biodegradable alternative to the current barrier material used in packaging. It consists of the natural carbohydrate xylan and additives. Xylophane® is an efficient barrier to oxygen, grease and aroma and can prolong the shelf life of sensitive food.</p><p>The raw material xylan is water soluble and Xylophane® can be coated on paper, board and plastics without using other solvents. A problem with the drying process is the amount of energy needed and the consumption needs to be decreased. Also, the drying capacity of the equipment to be used is often limited and the amount of water to be dried off is critical for the success of the coating process. By increasing the dry content of Xylophane® without increasing the viscosity too much, the drying process can be more effective. In this thesis, studies were made of using a filler as an additive to increase the dry content without destroying the barrier properties.</p><p>With an experimental design, a suitable relationship between the ingredients xylan (X), plasticizer (P) and filler (F) was found. Xylan is the main component and is needed to get a good oxygen barrier. The plasticizer decreases the oxygen barrier properties but is needed to make the material more flexible. The filler is positive for the barrier properties. The chosen composition was X:P = 7:3 and X:F = 1:1. Some extra experiments were made to find a reasonable value of the dry content. Dry contents around 18 % work well with temperatures at and above 45°C, but to manage to perform coating at room temperature the dry content needs to be lower.</p>

The main goal for a packaging is to protect the product inside. Typical packaging nowadays is made of layers of paper and barriers consisting of plastics or aluminum foil. A problem with the barrier used today is the environmental thinking. Xylophane® is an environmental friendly and biodegradable alternative to the current barrier material used in packaging. It consists of the natural carbohydrate xylan and additives. Xylophane® is an efficient barrier to oxygen, grease and aroma and can prolong the shelf life of sensitive food. The raw material xylan is water soluble and Xylophane® can be coated on paper, board and plastics without using other solvents. A problem with the drying process is the amount of energy needed and the consumption needs to be decreased. Also, the drying capacity of the equipment to be used is often limited and the amount of water to be dried off is critical for the success of the coating process. By increasing the dry content of Xylophane® without increasing the viscosity too much, the drying process can be more effective. In this thesis, studies were made of using a filler as an additive to increase the dry content without destroying the barrier properties. With an experimental design, a suitable relationship between the ingredients xylan (X), plasticizer (P) and filler (F) was found. Xylan is the main component and is needed to get a good oxygen barrier. The plasticizer decreases the oxygen barrier properties but is needed to make the material more flexible. The filler is positive for the barrier properties. The chosen composition was X:P = 7:3 and X:F = 1:1. Some extra experiments were made to find a reasonable value of the dry content. Dry contents around 18 % work well with temperatures at and above 45°C, but to manage to perform coating at room temperature the dry content needs to be lower.