Thesis (MScAgric)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: The aim of this study was to determine whether a lamb finishing ration that is balanced for essential amino acids (EAA), at a specific level of non-structural carbohydrates (NSC) would yield better feedlot performance when compared to a standard commercial finishing ration. Metionine (Met) and Lysine (Lys) were identified as the first limiting amino acids for growing lambs. These two amino acids, as well as Threonine (Thr), Arginine (Arg), Leucine (Leu), Isoleucine (Ile) and Phenylalanine (Phe) were included in an optimized protein (OP) feedlot ration at optimal levels. This ration was formulated to contain 157 g/kg crude protein (CP) and 477 g/kg NSC, with Met, Lys, Arg and Thr at 2.48, 7.45, 8.51 and 8.12 g/kg on dry matter (DM) basis respectively. The second treatment, FIN, was a standard commercial lamb finishing feed without optimized amino acids and contained similar total protein and NSC to OP of 152 g/kg CP and 468 g/kg NSC but with Met, Lys, Arg and Thr at 2.08, 5.49, 7.47 and 4.80 g/kg on DM basis respectively. A third treatment, a low protein (LP) diet served as a positive control and was formulated on lower specifications (139 g/kg CP, 455 g/kg NSC) and was also not optimized for amino acids. This treatment contained Met, Lys, Arg and Thr at 1.93, 4.99, 6.66 and 4.73 g/kg on DM basis respectively. Lambs grazing kikuyu pasture served as the negative control (CON) group. These lambs also received additional supplementary feed at 500 g/day as a production lick to be comparable to a scenario where lambs are finished on grazing. Forty cross-bred Merino x Döhne-Merino lambs with an average weaning weight of 24.35 ± 0.648 kg were finished in a feedlot for 57 days where after they were slaughtered at an average weight of 41.41 ± 1.259 kg. During the feedlot trial lamb performance was measured by monitoring daily growth rates and feed intake. Also, an in vivo digestibility study was carried out on the OP and FIN lambs. During the slaughter process the rumen was removed which was done to collect a sample of the rumen wall from next to the rumino-reticular fold. These rumen samples were mounted onto slides so that the development of the rumen could be examined. The M. longissimus dorsi from both sides of the carcass between the 2nd and 3rd last thoracic vertebra and the 4th and 5th lumbar vertebra were removed. There was no significant difference (P > 0.05) between any of the concentrate feed treatments with regards to the average daily gain (ADG), feed conversion ratio (FCR) or dressing percentage (DP). The CON lambs, as expected, had lower growth rates (P < 0.05) than the concentrate fed lamb and thus showed significant differences (P < 0.05) in terms of ADG. The DP of 45.96 % ± 0.711 for the CON lambs differed significantly (P < 0.05) from the OP (51.44 % ± 0.358), FIN (52.72 % ± 0.653) and LP (51.74 % ± 0.611) treatments. As expected the concentrate feeds were much more effective in maintaining higher growth rates when compared to the CON lambs while the optimizing of EAA in the OP diet did not lead to improved feedlot performance as the FIN and LP treatments were able to achieve similar (P > 0.05) growth rates. Within the feedlot treatments there was no significant difference (P > 0.05) with regard to the papillae length and rumen wall thickness. There was however a numerical increase in the papillae length as the NSC levels in the feed increased. The CON lambs differed from the OP lambs (P < 0.05) in terms of papillae length. This illustrated the importance of having increased levels of NSC in a feedlot diet as it is this fraction that is responsible for the initialisation and maintenance of rumen morphological development. The in vivo digestibility study therefore confirmed that the commercial finishing feed was just as effective as the optimized feed in terms of nitrogen retention as well as in maintaining suitable energy balance. Although the in vivo digestibility for Met and Lys in the OP feed was higher (P < 0.05) than the FIN feed, this did not lead to improved feedlot performance of the OP lambs. The increased digestibility of these amino acids is due to the fact that the OP diet was higher in levels of bypass amino acids than the FIN feed. The in vitro true digestibility (IVTD) of the OP feed was higher (P < 0.05) than that of the FIN feed. / AFRIKAANSE OPSOMMING: Die doel van hierdie studie was om te bepaal of ‘n lamafrond rantsoen wat gebalanseerd is vir beperkende essiensiële aminosure (EAA) teen ‘n spesifieke vlak van nie-strukturele koolhidrate (NSK) beter voerkraal prestasie teweeg sou bring wanneer gemeet word teen ‘n standaard kommersiële afrond rantsoen. Metionien (Met) en Lisien (Lis) is geïdentifiseer as die eerste beperkende aminosure vir groeiende lammers. Hierdie twee aminosure, asook Treonien (Tre), Leusien (Leu), Isoleusien (Ile) en Fenielalanien (Fen) is teen optimale vlakke ingesluit in ‘n geoptimeerde voerkraal rantsoen, OP. Hierdie rantsoen is geformuleer om 157 g/kg RP en 477 g/kg NSK te bevat asook Met, Lis, Arg en Tre teen 2.48, 7.45, 8.51 en 8.12 g/kg onderskeidelik. Die tweede behandeling, FIN, was ‘n kommersiële lamafrond voer waarin die aminosure nie geoptimeer is nie en het soortgelyke vlakke van proteïene en NSK bevat teen 152 g/kg RP en 468 g/kg NSK met Met, Lis, Arg en Tre teen 2.08, 5.49, 7.47, 4.80 g/kg onderskeidelik. ‘n Derde voer, LP, het gedien as ‘n positiewe kontrole en was ‘n lae proteïen voer met laer spesifikasies (139 g/kg RP, 455 g/kg NSK) waarin die aminosure ook nie geoptimeer is nie. Die LP voer het Met, Lis, Arg en Tre bevat teen 1.93, 4.99, 6.66 en 4.73 g/kg onderskeidelik. Die negatiewe kontrole behandeling, CON, is verteenwoordig deur lammers wat op kikuyu gewei het terwyl addisionele supplementêre voeding teen 500g/lam/dag voorsien is. Hierdie supplementêre voeding het gedien as ‘n produksie lek om sodoende vergelykbaar te wees met scenario waar lammers op weiding afgerond word. Veertig kruisgeteelde Merino x Döhne-Merino lammers met ‘n gemiddelde gewig van 24.35 ± 0.648 kg is vir 57 dae in ‘n voerkraal afgerond waarna hulle, teen ‘n gemiddelde gewig van 41.41 ± 1.259 kg, geslag is. Tydens die voerkraal proef is prestasie gemonitor deur die meet van daaglikse groei en voerinname. Hiertydens is daar ook ‘n in vivo verteringsproef op die OP en FIN lammers gedoen. Tydens die slagproses is die rumen verwyder waarna ‘n monster van die rumenwand langs die rumino retikulêre vou geneem is. Hierdie rumenmonsters is op skyfies geplaas sodat die ontwikkeling van die rumen ondersoek kan word. Die M. longissimus dorsi was aan beide kante van die karkas tussen die 2de en 3de laaste torakale werwels en die 4de en 5de lumbale werwels verwyder. Daar was geen betekenisvolle verskille (P > 0.05) tussen enige van die konsentraat behandelings ten opsigte van gemiddelde daaglikse toename (GDT), voeromset verhouding (VOV) of uitslag persentasie nie. Die CON lammers het egter, soos verwag, beduidend (P < 0.05) stadiger gegroei en het dus verskille getoon ten opsigte van GDT. Die uitslag persentasie van 45.96 % ± 0.711 vir die CON lammers het ook betekenisvol verskil (P < 0.05) van die OP (51.44 % ± 0.358), FIN (52.72 % ± 0.653) en LP (51.74 % ± 0.611) behandelings. Binne die voerkraal behandelings was daar geen betekenisvolle verskille (P > 0.05) ten opsigte van die papillae lengte en rumenwand dikte nie, alhoewel daar ‘n numeriese toename in papillae lengte was soos die NSK vlakke in die voer gestyg het. Die CON lammers het wel van die OP lammers verskil (P < 0.05) ten opsigte van papillae lengte. Hierdie bevinding het bevestig hoe belangrik NSK is in die inisiasie en instandhouding van die morfologiese ontwikkeling van die rumen. Die in vivo verterings studie het daarop gedui dat die kommersiële afrond voer net so effektief soos die geoptimeerde voer was in terme van stikstof retensie asook die handhawing van ‘n geskikte energie balans. Alhoewel die in vivo verteerbaarheid van Met en Lis in die OP hoër was (P < 0.05) as in die FIN voer, het hierdie verskille nie gelei tot beter groei in die OP lammers nie. Hierdie verskil in verteerbaarheid is toegeskryf aan die feit dat die aminosure in die OP voer meer rumen-beskermd was as dié in die FIN voer. Die in vitro verteringstudie het daarop gedui dat die OP voer beduidend beter (P < 0.05) verteer is as die FIN voer.

Thesis (PhD (Chemistry and Polymer Science)--University of Stellenbosch, 2006. / Synthetic peptides derived from the active core of a natural antimicrobial peptide were used as a template for the design of novel bolaamphiphilic peptides and hybrid molecules. The amphiphilic character of the original compounds was modified by using non-natural amino acids (AAs) – such as ω-AA – and varying the hydrophobic content. The outcomes of these modifications were studied focusing on structural and biological properties. Because of the bolaamphiphilic character, the alternation of polar and non-polar AAs and the use of hydrophobic AAs such as tyrosine and leucine, these novel molecules were designed to undergo self-assembly in response to certain stimuli (e.g. a pH increase). This significant property was investigated by means of different tools, such as fluorescence measurements, electron microscopy (EM), Fourier transform infrared spectroscopy (FT-IR) and circular dichroism (CD). By using fluorescence it was possible to determine the critical aggregation concentration (CAC) of the new compounds. Differences in amino acid composition, which were reflected into diverse secondary structures and hydrophobicity (H), resulted in different CAC values and aggregation profiles. The data were consistent with the literature and showed that (i) the aggregation of these basic compounds was triggered by a pH increase, (ii) the use of hydrophobic AA highly augmented the self-assembly tendency while (iii) the presence of proline strongly reduced it. EM revealed the morphology of the peptide assemblies: microtubes and microvesicles were identified and characterised by dimensions of 500 nm to 2 μm. The presence of 3-way junctions and vesicles budding out of the microtubes demonstrated that the self-assembly is a dynamic process. The aggregation was confirmed by FT-IR spectroscopy, by studying the dried peptide assemblies and the significant spectral signs the process left, especially in the amide II envelope. The relationship between hydrophobicity and self-assembly was expanded by experimentally and theoretically determining the hydrophobic content of the novel bolaamphiphiles. Data from liquid chromatography and computational calculations (two common ways used to determine the hydrophobicity of a given molecule) correlated well with the tendency to self-assemble, as expressed by CAC values. Importantly, some structural parameters (such as the presence of β-turn induced by proline) also showed significant influence on the aggregation, highly limiting the role of the peptides’ hydrophobicity. These novel peptide bolaamphiphiles displayed a very low haemolytic action and retained some antimicrobial activity at high concentrations against both Gram-positive and -negative bacteria. Unfortunately, the activity was greatly reduced at low concentrations, as clearly demonstrated by the use of two antimicrobial tests. The inability to provoke cell lysis was also evident when using liposomes mimicking a negative bacterial membrane. The loss of activity is possibly related to the modifications of the three-dimensional structure caused by the use of ω-AA and proline, which strongly alter the secondary structure. The results of this study were valuable in terms of understanding the relationships between self-assembly and structural parameters, such as AA compositions, hydrophobicity and secondary structure. Possible applications of the synthesised compounds were however limited as a result of the loss of the biological activity at low concentrations.

Thesis (PhD (Chemistry and Polymer Science))--University of Stellenbosch, 2008. / This dissertation can be broken into two parts comprising different strategies to synthesise novel poly-amino acid based polymers. The use of recently developed nickel(0) and cobalt(0) metal catalysts for the living polymerization of α-amino acid-N-carboxyanhydrides (NCAs) to synthesise novel poly-amino acid polymers, comprising a polar, hydrophilic block and a neutral hydrophobic block, were investigated in the first part of this study. The hydrophilic block was made up of a random sequence of arginine (Arg, R), glycine (Gly, G) and aspartic acid (Asp, D) - poly-RGD. This was followed by a polyleucine (Leu, L) hydrophobic block. Success was limited with this system due to polymer precipitation during the polymerization reaction. Because of this precipitation, the amino acid composition of the hydrophilic block was changed to a random sequence of glutamic acid (Glu, E), cystein (Cys, C) and aspartic acid – poly-ECD. Here also, the success was limited because of polymer precipitation. A novel approach to the synthesis of hybrid poly-amino acid – synthetic polymer materials constitutes the second part of this study. The final polymeric structure can be described as a carboxylic acid functionalized polyethylene glycol (PEG) sheathed polylysine polymer. The technology involves the synthesis of a lysine NCA functionalized at the ε-amino group with an α,ω-bis(carboxymethyl) ether PEG. The distal carboxylic acid group was protected as a benzyl ester during synthesis and subsequent polymerization of the PEG-lysine-NCA macro-monomer. The polymerization was successfully initiated using n-butyl amine to form short homopolymer strands. Copolymerization with lysine-NCA was also achieved as well as the successful initiation using a generation 1.0 dendritic amine initiator, N,N,N’,N’-tetrakis(3-aminopropyl)-1,4-butanediamine (DAB-Am-4). These polymers were characterized by 1H NMR.

Human alteration of the global nitrogen (N) cycle has had major impacts on naturally N-limited ecosystems worldwide. Peatlands, dominated by peat mosses, Sphagnum species, represent one such sensitive ecosystem. I have studied how this ecosystem is affected by increased N availability, using a small-scale N fertilization experiment in combination with a gradient study of three peatlands with varying N deposition. I found both in the experiment and in the gradient a similar pattern of Sphagnum decline accompanied by an increase of vascular plants associated with enhanced N supply. For one common Sphagnum species - both in the experiment and in the gradient study - I also found an identical, linear increase in soluble amino acid N (NAA) accumulation. As soluble amino acids function as N storage compounds among Sphagna, NAA is a suitable measure for Sphagnum N status, and indicates accumulation of excess N not used for growth. My results show that NAA can be used as a sensitive indicator to signal N pollution before the slow, and gradual, regime shift from Sphagnum to vascular plant dominance is visible. In an N-uptake experiment using Sphagnum specimens from the three peatlands varying in N deposition, I found a reduced N-uptake by both investigated Sphagnum species from a high N deposition site, in south-western Sweden. This potential of Sphagna to adjust to high N loads through N uptake regulation will, however, not prevent tissue N accumulation, and as a result a shift from Sphagnum to vascular plant dominance. In general I found similar patterns of N induced changes both in Sphagnum tissue chemistry and vegetation structure in the experiment and along the gradient study. Thus, I conclude that long-term, small-scale field experiments seem to offer reliable estimates of both the direction and strength of key vegetation responses in Sphagnum dominated peatlands. This is likely related to the key role of Sphagna as ecosystem engineers. In the experiment I found a marked time lag in vegetation response to N application treatments. The closed Sphagnum carpet did not collapse until after eight years of continuous treatments. Another result was that dwarf shrubs, e.g. cranberry Vaccinium oxycoccos, first increased, but later declined due to severe attacks by fungal diseases. One important conclusion is that long-term, manipulative field experiments are necessary for our ability to understand how ecosystems will respond to environmental change.

Thesis (MSc)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: In this study, the analysis of ammo acids usmg High Performance Liquid Chromatography (HPLC) with pre-column derivatisation was optimised. The derivatisation reagents include o-phthaldialdehyde (OPA), 9- fluorenylmethylchloroformate (FMOC) and iodoacetic acid (IDA). Detection was performed using UV and fluorescence in series. The developed method was utilised for the analysis and quantitation of amino acids in eighteen wines. The application of chemometric data evaluation was initiated. / AFRIKAANSE OPSOMMING: Hierdie ondersoek behels die optimisering van die aminosuuranalise deur gebruik te maak van Hoë Druk Vloeistof Chromatografie (HDVC) in kombinasie met pre-kolom derivatisering. Die derivativatiserings reagense sluit in o-phthaldialdehied (OPA), 9- fluorenielmetielchloroformaat (FMOC) en jodoasynsuur (IDA). Deteksie is gedoen deur gebruik gemaak van 'n ultraviolet (UV) en 'n fluorosensie detektor in serie. Die metode sodoende ontwikkel is gebruik vir die analise en kwantifisering van aminosure in agtien wyne. Die toepassing van chemometriese data evaluasie is ook geïnisieer.

The present study was divided between two different drug delivery goals, each involving naltrexone (NTX) or its active metabolite, 6-β-naltrexol (NTXOL). First, amino acid esters of NTX and NTXOL were prepared in order to test their candidacy for microneedle-enhanced transdermal delivery. Second, a 3-O-(-)-cytisine-naltrexone (CYT-NTX) codrug was prepared for screening as a potential oral delivery form of NTX and (-)-cytisine (CYT). The amino acid prodrugs were intended for the treatment of alcohol abuse, while the codrug was designed as a single agent for the treatment of alcoholism and tobacco-dependency co-morbidities. One hypothesis of this work was that prodrugs of NTX or NTXOL can be designed that possess superior skin transport properties through microneedle-treated skin compared to parent NTX or NTXOL. Nine amino acid ester prodrugs were prepared, and only three 6-O amino acid ester prodrugs of NTXOL were stable enough at skin pH (pH 5.0) to move forward to studies in 50% human plasma. 6-O-β-Ala-NTXOL, the lead compound, exhibited the most rapid bioconversion to NTXOL in human plasma (t1/2 = 2.2 ± 0.1 h); however, this in vitro stability value indicates that the prodrug may require hepatic enzyme-mediated hydrolysis for sufficiently rapid bioconversion to NTXOL in vivo. A second hypothesis of this work was that a CYT-NTX codrug could be designed with appropriate stability characteristics for oral delivery. CYT-NTX was found to be stable over the time course of 24 h in buffer systems of pH 1.5, 5.0, 7.4 and 9.0, and in 80% rat plasma, 80% human plasma, simulated gastric fluid and simulated intestinal fluid. Six (3 rats/group) Sprague-Dawley male rats were dosed i.v. with 1 mg/kg CYT-NTX codrug, or 10 mg/kg, p.o. Oral administration of a 10 mg/kg dose of CYT-NTX codrug resulted in rapid absorption and distribution (5 min) of CYT-NTX codrug, and NTX was released from codrug with a peak plasma concentration of 6.8 ± 0.9 nmol/L reached within 65 minutes. Plasma CYT was not detected; however, NTX delivery was achieved with a fraction absorbed value of 13%. Thus, CYT-NTX may hold promise as a potential oral codrug for further optimization and development.

Stainless steel (SS) is the material of choice in a number of process industries ranging from food and dairy to pharmaceuticals. Adsorption phenomena on SS surfaces are of paramount importance in these industries. For example, protein adsorption constitutes a major issue in process equipment, as the associated surface fouling decreases the efficiency of the overall process and leads to an increase in operational costs because of the need for regular cleaning. In addition, the adsorption of proteins at solid–liquid interfaces is an important research field with relevance in biosensor and biomaterial applications. The primary aim of this thesis was to understand the underlying adsorption properties of selected protein onto SS surfaces and to identify the influence of specific amino acids on bio-fouling. Protein adsorption experiments were carried out on 316 grade SS sensors using a quartz crystal microbalance with dissipation (QCM-D). The proteins consisted of milk proteins (α-lactalbumin, β-lactoglobulin, α-casein, β-casein, κ-casein and bovine serum albumin), blood proteins (cytochrome-c, haemoglobin and myoglobin) and proteins of industrial and medical relevance (α-chymotrypsinogen, human recombinant insulin, lysozyme and papain). The adsorption characteristics of the test proteins were studied and an empirical correlation relating the amount of protein adsorbed to their physical properties was proposed. Adsorption onto a SS surface was followed on the QCM-D in real time and the amounts adsorbed calculated using the Sauerbrey model. In addition, the binding kinetics was modelled using different theoretical models to describe the adsorption mechanism. In all the proteins tested, the conformational change model was found to fit considerably well the adsorption data. Finally, the data collected were used to identify the physical properties of proteins that induce surface binding, with hydrophobic and aromatic amino acids having the most effect on binding. A second aspect investigated in the present work was the determination of hydration water present in the adsorbed layer. In fact, water molecules, solvated ions and other small molecules in the vicinity of the surface all play an important role in protein adsorption and often constitute a large fraction of the total measured adsorbed mass. The fraction of water present on SS surfaces along with adsorbed proteins was determined using fluorescently labelled proteins through a comparative study that included QCM-D experiments as well as fluorescent light intensity measurements. The results were similar for all proteins tested, indicating that 32-45.8% of the total mass adsorbed composed of water. One last aspect considered in this thesis was the influence of the putative adhesive amino acid 3, 4-dihydroxyphenylalanine (DOPA). DOPA residues are present in high levels in the adhesive proteins from marine mussels, hence are thought to facilitate surface attachment. The role of DOPA residues in mediating protein adhesion on SS surfaces was studied using QCM-D. Two repetitive peptide motifs extracted from the sequence of the mussel foot protein mefp-5, KGYKYYGGSS and KGYKYY, were selected for this study. The two peptides contained unmodified tyrosine (Y) residues, which were chemo-enzymatically modified to DOPA using mushroom tyrosinase. Adsorption of the two sequences on SS surfaces was tested before and after modification of tyrosine residues to DOPA. Conversion was linearly related to the incubation time of the peptide fragments with mushroom tyrosinase, Amount of DOPA formed was 70-99% of the tyrosine content in the peptides. QCM-D adsorption experiments on the DOPA-modified sequences revealed four-fold greater adhesion than for unmodified mefp-5 motifs, indicating the paramount role that DOPA has on the adsorption of peptides on 316 grade stainless steel.

This thesis examines the synthesis of conformationally constrained β-amino acids and β- peptides, and the electron transfer properties of the latter when immobilised on gold. Additionally, cross metathesis on gold was investigated as a method for surface functionalisation. Chapter One introduces the concepts of electron transfer in nature, how it is facilitated by the secondary structure in α-peptides, and why β-peptides might be useful for studying electron transfer. This is followed by a discussion of the properties of β-peptides, including the enhanced stability and variety of helical secondary structures and the greater potential for functionalisation of the peptide backbone when compared to α-peptides. Finally, the conformational constraints of ring-systems on cyclic amino acids is discussed, with reference to the stabilising effect of these compounds on peptide secondary structures. Chapter Two describes the electrochemical analysis of β-hexapeptides immobilised on gold. The chapter is prefaced by a discussion of the important electron transfer mechanisms for peptides, the fabrication of peptide-gold self-assembled monolayer (SAM) interfaces, and the electron transfer in helical α-peptides. β-Peptides containing an electroactive ferrocene moeity were immobilised on gold and studied using cyclic voltammetry and chronoamperometry. The latter method was used to examine the dependence of the electron transfer rate on overpotential, thereby determining the likely mode of electron transfer through the β-peptides SSβ₆Fc, Fcβ₆SS and SC₁₅β₆Fc. These peptides exhibited very weak dependence on overpotential, characteristic of electron transfer behaviour of an electron hopping mechanism (which is also thought to occur in helical α-peptides). Both the dipole moment of the peptides and the structure of the sulfurlinker group were found to be important in determining the rate of electron transfer. Conversely, the equivalent α-peptide SSα₆Fc exhibited electron transfer behaviour characteristic of the less efficient tunnelling mechanism, which is thought to operate in strand-like peptides. Chapter Three examines the application of cross metathesis, using a Grubbs' second generation catalyst, as a means to functionalise olefin-terminated self-assembled monolayers on gold. Abstract iv Firstly, an introduction into the limited published research on cross metathesis on both planar surfaces and nanoparticles is given. Olefin-terminated thiol 3.18, suitable for immobilisation on gold, and solution phase olefin-terminated ferrocene 3.10 were synthesised as reactants for cross metathesis studies. An analytical methodology was developed involving the cross metathesis of surface-immobilised 3.18 with ferrocene 3.10 in dichloromethane, whereby the concentration of electroactive cross metathesis product 3.22 was monitored electrochemically as a function of time. The concentration of surface-immobilised product 3.22 was determined by integration of the oxidation peak area and found to be highly dependent on both the concentration of immobilised olefin reactant 3.18 and reaction time. Furthermore, the surface concentration of ferrocenyl model disulfide 3.21 and thiol 2.18 decayed markedly upon addition of Grubb's catalyst, as revealed by the decrease in the oxidation peak area, which suggested that catalystmediated desorption was occurring. Chapter Four details the solution-phase synthesis of ferrocene- and thiol-functionalised β- hexapeptides used in both the electron transfer studies described in chapter two, and in the determination of secondary structure using circular dichroism and NMR techniques. The synthesis of simple model compounds 4.14, 4.16 and 4.18 established the incompatibility of the deprotection of methyl and benzyl ester protecting groups with protected-thiol and disulfide linkers, leading to the use of N-hydroxysuccinmide-activated sulfur-linkers 4.20 and 4.22 in further synthesis. A number of β-hexapeptides were synthesised by amide coupling of β- tripeptides functionalised at the N- and C-termini. Structural studies of the methanol soluble β- hexapeptide 4.60 suggested that the covalent attachment of ferrocene moeity to the C-terminus of a β-peptide did not disrupt the formation of a 14-helix in solution. β-peptides containing functionality at both the C- and N-termini (such as SSβ₆Fc, SSβ₆Et and acetyl-protected SC₁₅β₆Fc) were not suitable for solution phase structural studies; however, molecular modelling suggested that helical conformations are the most stable these β-peptides in solution phase. Chapter Five outlines the synthesis of novel cyclic β-amino acids by two different general synthetic routes. The first uses an efficient conjugate addition/fluorination reaction of α,β- unsaturated esters with lithiated chiral secondary amines to prepare the novel cyclopentyl- and cyclohexyl-based fluorinated β-amino acids 2.43a and 2.43b. The high diastereoselectivity of this reaction, which introduces two stereocentres into the achiral unsaturated esters, is directed by the configuration of the attacking amine. The second methodology utilizes the versatile ringclosing metathesis reaction in the synthesis of novel cyclic β-amino acids. A stereoselective Abstract v trans-alkylation of olefinic β-amino acids gave the required β-dienes 5.62 and 5.77. Optimised cyclisation yields were achieved with a Grubb's 2nd generation catalyst for diene 5.62 and Grubb's 1st generation catalyst for diene 5.77, to give the trans-cycloheptyl- and cyclooctylbased β-amino acids 5.63 and 5.78, respectively. The attempted synthesis of cyclononyl-based β-amino acid 5.87 using both catalysts yielded only cyclic dimer products 5.88 and 5.89. The trans configuration of the 5.62 diene was confirmed by x-ray crystallography. Chapter Six is an experimental chapter and outlines the electrochemical setup and analysis, and the synthesis, purification and characterisation of compounds described in this thesis.

2007 metais ūkininkų E. Guževičiaus (Alytaus r.) ir V. Svoto (Vilkaviškio r.) ūkiuose atlikti gamybiniai lauko bandymai. Vilaviškio rajono dirvožemiai yra vidutiniškai humusingi ir humusingi, turintys didelį magnio ir mažą mineralinio azoto kiekį. Judriojo fosforo ir kalio čia esti vidutiniškai. Šie dirvožemiai priskiriami prie išplautžemių. Alytaus rajone vyrauja lengvo priemolio karbonatingi išplautžemiai ir karbonatingi giliau glėjiški rudžemiai, kuriuose gausu magnio, kiek mažiau kalio, vidutiniškai – humuso ir mineralinio azoto. Tyrimams pasirinkti skirtingų rajonų ūkiai su skirtingomis dirvožemio ir agroklimatinėmis sąlygomis, siekiant įvertinti skystųjų azoto trąšų su aminorūgštimis įtaką žieminių kviečių ir vasarinių miežių derliui ir derliaus kokybei. Nustatyta, kad skystosios amidinio azoto trąšos su aminorūgštimis nežymiai didino žieminių kviečių produktyvumo elementus, tačiau skirtingų koncentracijų poveikis buvo kitoks. Karbonatiniame išplautžemyje gausiausias grūdų derlius gautas miežius patręšus trąšomis su 1,5% aminorūgščių. Kituose laukeliuose gauti derliaus pokyčiai buvo neesminiai. Aminorūgštys įtakojo žaliųjų proteinų kaupimąsi grūduose. Nustatyta, kad skystosios amidinio azoto trąšos su aminorūgštimis didino žieminių kviečių grūduose baltymų kiekį, sedimentacijos ir šlapiojo gilitimo vertes. / In 2007, farm production field trials were carried out in the E. Guževičius (Alytus region) and V. Svotas (Vilkaviškis region) farms. In Vilkaviškis region the soils are average humous and humous, with high level of magnesium and low level of mineral nitrogen. The mobile phosphorus and potassium is found in the average quantities there. These soils belong to scour soil. In Alytus region light loam carbonative scour soil and carbonative deeper slimy brown soils predominate. These brown soils are rich in magnesium, potassium to a lesser extent, on average - humus, and mineral nitrogen. The farms from different regions not only with different soils, but even with variant agro-climate conditions were selected for the research, to evaluate the liquid nitrogen fertilizer with amino acids influence on winter wheat and spring barley harvest and crop quality and technological properties. Found that liquid amidive nitrogen fertilizer with amino acids slightly increased the productivity of winter wheat cells, but the effects of different concentrations were totally mixed. In the carbonative scour soil the largest grain yield was obtained when barley was feeded fertilizers with 1.5% amino acids. Other options to receive the crop change was minor. Amino acids affected the crude protein and fat accumulation in grain. Found that fertilization liquid amidive nitrogen fertilizer with amino acids increased the grain protein content, sedimentation, and wet gluten values.

The class of polycyclic hydrocarbons including adamantane, pentacyclo [5.4.0.02 ,6. 03,I0.05,9]undecane (PCU) and trishomocubane have proven to be an exciting investigation for synthetic chemists. Many derivatives have been shown to possess excellent antiviral and antibacterial properties, as well as potent anti-Parkinson agents. Some improve the lipophilic nature of biologically active drugs, while others affect the three-dimensional structure of peptides once incorporated as amino acid analogues. This investigation focussed on deriving routes to improve yields of racemic 4-amino-(D3)trishomocubane- 4-carboxylic acid (tris-amino acid), the synthesis of enantiomerically pure tris-amino acid, the incorporation of tris-amino acid into a short peptide, as well as the simulation of the rearrangement of PCU to trishomocubane by using computational tools. Research into developing a more efficient hydrolysis of trishomocubane-hydantoin (trishydantoin) to yield racemic tris-amino acid, led to the development of two novel compounds: the mono-Boc [Novel Compound 1, (NC1)] and bis-Boc [Novel Compound 2, (NC2)] protected hydantoin. Base hydrolysis of NC2 quantitatively yielded the racemic tris-amino acid, which was a significant improvement on previously documented synthetic routes. The first attempt to produce enantiomerically pure tris-amino acid was through the synthesis of diastereomeric derivatives of tris-hydantoin, chromatographic separation of the diastereomers, followed by base hydrolysis of the hydantoin ring to produce enantiomerically pure tris-amino acid. This research led to the development of two novel N-protected tris-hydantoin derivatives (NC3 and NC4). Failure to chromatographically separate the diastereomers resulted in the abandonment of this particular route. The use of enzymes was, therefore, attempted to produce enantiomerically pure tris-amino acid. A novel ester derivative of tris-amino acid (NC5) was synthesised, which was followed by the application of Pig Liver Esterase (PLE). PLE is an enzyme which cleaves ester functionalities. Some success was achieved but the extremely low yields of enantiomerically pure tris-amino acid did not warrant this enzyme as a viable route for production of the desired product. Solid phase techniques were employed for the production of a tripeptide consisting of alanine-glycine-tris-amino acid (ala-gly-tris). Some difficulty was encountered in extending the amino acid sequence due to suspected Schiff base interaction between the free amino group of tris-amino acid and the carbonyl functionality of glycine in the second position. A computational study, using ab initio methods, was performed on the rearrangement of the PCU diol to 7-fluoro-11-hydroxy-trishomocubane. Two mechanisms (Proposed Mechanism 1 and Proposed Mechanism 2) were explored and both showed that the stereochemistry of the hydroxyl groups has only a marginal influence on the transition state energies of the various isomers. Both mechanisms were also indicated to occur through an intramolecular SN2 mechanism. / Thesis (M.Sc.)-University of Natal, Durban, 2003.