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Cheng Chin-Ho. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 93-100). / Abstracts in English and Chinese. / Table of content --- p.ii / Acknowledgements --- p.ix / Abstract (English) --- p.x / Abstract (Chinese) --- p.xi / Abbreviations --- p.xii / Chapter Chapter 1 - --- Introduction / Chapter 1-1 --- Definition of gels --- p.1 / Chapter 1-2 --- Organogels and organogelators --- p.3 / Chapter 1-3 --- Characterization of organogels and organogelators --- p.5 / Chapter 1-4 --- Classes of organogelators --- p.10 / Chapter 1-5 --- Applications --- p.17 / Chapter 1-6 --- Origin of research project --- p.20 / Chapter Chapter 2 - --- Synthesis and Characterization / Chapter 2-1 --- Structural modification of the Lead compound --- p.22 / Chapter 2-2 --- Retrosynthetic analysis --- p.23 / Chapter 2-3 --- Synthesis --- p.26 / Chapter 2-4 --- Characterization of the target compounds --- p.32 / Chapter 2-4-1 --- NMR spectrometry --- p.32 / Chapter 2-4-2 --- Mass spectrometry --- p.37 / Chapter 2-4-3 --- Elemental analysis --- p.38 / Chapter 2-4-4 --- Melting point determination --- p.39 / Chapter 2-4-5 --- Optical polarimetry --- p.40 / Chapter Chapter 3 - --- Investigation of Gelation Behaviors / Chapter 3-1 --- Gelation behaviors of bis-(urea valine) ethyl esters --- p.41 / Chapter 3-2 --- Gelation behaviors of bis-(urea valine) benzyl esters --- p.43 / Chapter 3-3 --- Effect of lengths of hydrocarbon chains on gelation behaviors --- p.46 / Chapter 3-4 --- Effect of ester protecting group on gelation behaviors --- p.47 / Chapter 3-5 --- Conclusions --- p.48 / Chapter Chapter 4 - --- Elucidation of Gelation Mechanisms / Chapter 4-1 --- FT-IR Spectroscopy --- p.49 / Chapter 4-2 --- Thermotropic behavior --- p.52 / Chapter 4-3 --- Morphological behavior --- p.54 / Chapter 4-4 --- Chiroptical behavior --- p.57 / Chapter 4-5 --- Conclusions --- p.58 / Chapter Chapter 5 - --- Summary --- p.59 / Chapter Chapter 6 - --- Experimental --- p.61 / References --- p.93 / Appendix NMR spectra / 1H NMR of (Boc-NH-V)2-Ar-C02Et 23 --- p.101 / 13C NMR of(Boc-NH-V)2-Ar-C02Et 23 --- p.102 / HNMR of (H2N-V)2-Ar-CO2Et 26 --- p.103 / 13C NMR of(H2N-V)2-Ar-CO2Et 26 --- p.104 / "1H NMR of 3,5-Di(tert-butylcarbonylamino)benzoic acid 36" --- p.105 / "13C NMR of 3,5-Di(tert-butylcarbonylamino)benzoic acid 36" --- p.106 / "1H NMR of Benzyl 3,5-di(tert-butylcarbonylamino)benzoate 37" --- p.107 / "13C NMR of Benzyl 3,5-di(tert-butylcarbonylamino)benzoate 37" --- p.108 / "1H NMR of Benzyl 3,5-diaminobenzoate 38" --- p.109 / "13C NMR of Benzyl 3,5-diaminobenzoate 38" --- p.110 / 1H NMR of(Boc-NH-V)2-Ar-CO2Bn 29 --- p.111 / 13C NMR of(Boc-NH-V)2-Ar-CO2Bn 29 --- p.112 / 1H NMR of (H2N-V)2-Ar-CO2Bn 30 --- p.113 / 13C NMR of (H2N-V)2-Ar-CO2Bn 30 --- p.114 / 1H NMR of O-Succinimidyl butylcarbamate 27a --- p.115 / 13C NMR of O-Succinimidyl butylcarbamate 27a --- p.116 / 1H NMR of O-Succinimidyl pentylcarbamate 27b --- p.117 / 13C NMR of O-Succinimidyl pentylcarbamate 27b --- p.118 / 1H NMR of O-Succinimidyj hexylcarbamate 27c --- p.119 / 13C NMR of O-Succinimidyl hexylcarbamate 27c --- p.120 / 1H NMR of O-Succinimidyl heptylcarbamate 27d --- p.121 / 13C NMR of O-Succinimidyl heptylcarbamate 27d --- p.122 / 1H NMR of O-Succinimidyl decylcarbamate 27e --- p.123 / 13C NMR of O-Succinimidyl decylcarbamate 27e --- p.124 / 1H NMR of O-Succinimidyl undecylcarbamate 27f --- p.125 / 13C NMR of O-Succinimidyl undecylcarbamate 27f --- p.126 / NMR of O-Succinimidyl tridecylcarbamate 27g --- p.127 / 13C NMR of O-Succinimidyl tridecylcarbamate 27g --- p.128 / 1H NMR of O-Succinimidyl hexadecylcarbamate 27h --- p.129 / 13C NMR of O-Succinimidyl hexadecylcarbamate 27h --- p.130 / 1H NMR of O-Succinimidyl nonadecylcarbamate 27i --- p.131 / 13C NMR of O-Succinimidyl nonadecylcarbamate 27i --- p.132 / 1H NMR of O-Succinimidyl heneicosylcarbamate 27j --- p.133 / 13C NMR of O-Succinimidyl heneicosylcarbamate 27j --- p.134 / 1H NMR of (n-C4H9-NHCONH-V)2-Ar-CO2Et 24a --- p.135 / 13C NMR of (n-C4H9-NHC0NH-V)2-Ar-CO2Et 24a --- p.136 / 1H NMR of (n-C5H11-NHCONH-V)2-Ar-C02Et 24b --- p.137 / 13C NMR of (N-C5H11-NHCONH-V)2-Ar-C02Et 24b --- p.138 / HNMR 0f(N-C6H13-NHC0NH-V)2-Ar-C02Et24c --- p.139 / 13C NMR of (n-C6H13-NHC0NH-V)2-Ar-C02Et 24c --- p.140 / HNMR of (n-C7H 15-NHCONH-V)2-Ar-C02Et 24d --- p.141 / 13C NMR of (n-C7H15-NHC0NH-V)2-Ar-C02Et 24d --- p.142 / 1H NMR of (n-C10H21-NHCONH-V)2-Ar-G02Et 24e --- p.143 / "13C NMR of(n-C10H21,-NHCONH-V)2-Ar-CO2Et 24e" --- p.144 / HNMR of (n-C11 H23-NHC0NH-V)2-Ar-C02Et 24f --- p.145 / 13C NMR 0f (n-C23H23-NHC0NH-V)2-Ar-C02Et 24f --- p.146 / HNMR of (n-Cl3H27-NHC0NH-V)2-Ar-C02Et 24g --- p.147 / 13C NMR of (n-C13H27-NHC0NH-V)2-Ar-C02Et 24g --- p.148 / 1H NMR of (n-C16H33-NHC0NH-V)2-Ar-C02Et 24h --- p.149 / 13C NMR of (n-C16H33-NHC0NH-V)2-Ar-C02Et 24h --- p.150 / 1H NMR of (n-C19H39-NHC0NH-V)2-Ar-C02Et 24i --- p.151 / 13C NMR of (n-C19H39-NHCONH-V)2-Ar-C02Et 24i --- p.152 / 1H NMR of (n-C21 H43-NHC0NH-V)2-Ar-C02Et 24j --- p.153 / 13C NMR of (^-C2iH43-NHC0NH-V)2-Ar-C02Et 24j --- p.154 / HNIVIR of (n-C4H9-NHC0NH-V)2-Ar-C02Bn 25a --- p.155 / 13C NMR of (n-C4H9-NHC0NH-V)2-Ar-C02Bn 25a --- p.156 / 1H NMR of(n-C5H11-NHC0NH-V)2-Ar-C02Bn 25b --- p.157 / 13C NMR of (n-C5H11-NHC0NH-V)2-Ar-C02Bn 25b --- p.158 / 1H NMR of(n--C6H13-NHC0NH-V)2-Ar-C02Bn 25c --- p.159 / 13C NMR of OC6Hl3-NHC0NH-V)2-Ar-C02Bn 25c --- p.160 / 1H NMR of(n-C7H15-NHC0NH-V)2-Ar-C02Bn 25d --- p.161 / 13C NMR of(n- C7H15-NHC0NH-V)2-Ar-C02Bn 25cl --- p.162 / 1H NMR of(n--C10H21-NHCONH-V)2-Ar-C02Bn 25e --- p.163 / 13C NMR of (n-C10H21NHCONH-V)2-Ar-C02Bn 25e --- p.164 / HNMR of(n-C11H23-NHC0NH-V)2-Ar-C02Bn 25f --- p.165 / 13C NMR of(n-C11H23-NHC0NH-V)2-Ar-C02Bn 25f --- p.166 / 1H NMR of(n-Cl3H27-NHC0NH-V)2-Ar-C02Bn 25g --- p.167 / 13C NMR of (n-C13H27-NHCONH-V)2-Ar-C02Bn 25g --- p.168 / 1H NMR of (n-Cl6H33-NHC0NH-V)2-Ar-C02Bn 25h --- p.169 / 13C NMR of (n-C16H33-NHCONH-V)2-Ar-C02Bn 25h --- p.170 / 1H NMR of (n-C19H39-NHC0NH-V)2-Ar-C02Bn 25i --- p.171 / 13C NMR of (n-Cl9H39-NHC0NH-V)2-Ar-C02Bn 25i --- p.172 / 1H NMR of (n-C21H43-NHC0NH-V)2-Ar-C02Bn 25j --- p.173 / 13C NMR of (n-C2lH43-NHC0NH-V)2-Ar-C02Bn 25j --- p.174 / "1H NMR of 1,3-didodecylurea 39" --- p.175 / "13C NMR of 1,3-didodecylurea 39" --- p.176 / "1H NMR of Ethyl 3,5-diaminobenzoate 32" --- p.177 / "13C NMR of Ethyl 3,5-diaminobenzoate 32" --- p.178

The coding properties of the 3 major valine tRNA isoacceptors of Drosophila melanogaster, the nucleotide sequences of tRNA[sub=Val, sub=3b] and tRNA[sub=Val, sub=4] and the nucleotide sequences of genes for these two tRNAs have been determined. Valyl-tRNA[sub=Val, sub=3a] binds strongly to ribosomes in response to the trinucleotide GUA and to a lesser extent with GUU and GUG. Valyl- tRNA[sub=Val, sub=3b] binds strongly in the presence of GUG and very weakly with the other 3 triplets whereas valyl- tRNA[sub=Val, sub=4] binds strongly in the presence of GUU, GUC, and GUA and weakly with GUG. The nucleotide sequences of tRNA[sub=Val, sub=3b] and tRNA[sub=Val, sub=4] were determined by a combination of techniques. For both tRNAs most of the sequence was determined by the method of Stanley and Vassilenko. The sequences at the 5' and 3'-ends of the molecules were determined by wandering-spot analysis. Regions of the molecules that could not be sequenced by these two techniques were determined by the gel read-off method. The use of tRNA modified with chloroacetaldehyde to overcome problems in sequencing RNA by the gel read-off method caused by secondary structure in the RNA is described. The nucleotide sequence of tRNA[sub=Val, sub=4] is: GUUU[sub=m]⁷CCGUm¹GGUG ѱAGCGGDU (acp³ U)AUCACA1ѱCUGCC[sub=m]UIACAm⁵CGCAGAAGm⁷GCCCCCGGѱC Gm¹ AUCCCGGGCGGAAACACCA. About 50% of the U residues at position 20 are modified to acp³U. One of the C residues at position 48 or 49 is probably modified to m5C. The nucleotide sequence of tRNA[sub=Val, sub=3b] is: GUUUCCGѱAGUGS1 AGCGGDacp³ UAUCACGѱGUGCUUC ACACGCACAAGm⁷- GDCCCCGGTѱCGm¹ AACCC GGGCGGGAACACCA. The C residue at position 48 is probably modified to m⁵C. The observed codon responses of the two tRNAs are discussed in relation to the anticodons found. Val The two tRNA[sub=Val, sub=4] genes of the recombinant plasmid pDt55 were sequenced by the Maxam and Gilbert method. This plasmid hybridizes to the 70BC site on the polytene chromosomes, a major site of tRNA[sub=Val, sub=4] hybridization. The two genes are of opposite polarity and are separated by 525 bp of DNA. The genes have identical sequences, which correspond to that expected from the sequence of tRNA[sub=Val, sub=4]. The nucleotide sequence of the tRNA[sub=Val, sub=3b] gene of recombinant plasmid pDt78R was also determined. This plasmid hybridizes to the 84D site, a major site of tRNA[sub=Val, sub=3b] hybridization. The sequence of the gene corresponds to that expected from the sequence of tRNA[sub=Val, sub=3b]. Comparison of the valine tRNA genes sequenced in this study and those determined by other workers shows that tRNA genes from major sites of tRNA[sub=Val, sub=3b] or tRNA[sub=Val, sub=4] hybridization to polytene chromosomes correspond exactly to the tRNA[sub=Val] sequences while tRNA tRNA[sub=Val] genes from minor sites of tRNA hybridization differ at 4 positions from the sequences expected on the basis of the tRNA sequences. The possible significance of this finding is discussed. / Medicine, Faculty of / Biochemistry and Molecular Biology, Department of / Graduate

Previous work in many laboratories has established that the addition of L-valine to a culture of Escherichia coli K-12 growing exponentially on minimal medium causes inhibition resulting in persistent linear growth. The inhibition can be removed by addition of isoleucine. Growth of other strains of E. coli e.g. E. coli B and two mutants of K-12 (E. coli AB1020 and AB1005) was not affected by addition of valine. E. coli LL5, another mutant of K-12, showed no inhibition of growth by valine added in concentrations up to 1 x 10ˉ⁴M. However, the addition of 1 x 10ˉ³M valine caused a decrease in the logarithmic growth rate, and 1 x 10ˉ²M valine caused persistent linear growth. The inhibition of growth by valine in LL5 was antagonized by L-isoleucine (as is the case with wild-type E. coli K-12), but not by α-ketobutyrate, threonine or pyruvate. Kinetic studies showed that the maximal inhibition of acetohydroxy acid synthetase (AHAS) by 1.5 x 10ˉ³ M valine was 84% for E. coli K-12, 55% for E. coli B, 75% for E. coli AB1005, 77% for E. coli AB1020 and only 20% for E. coli LL5. It is proposed that the persistence of linear growth of K-12 in valine-containing medium is the result of incomplete (84%) feed-back inhibition of AHAS. Analysis of the data for valine inhibition of AHAS was carried out by the method of Levitsky and Koshland (1969) using Hill Plots. E. coli strains K-12, AB1005 and AB10 20 showed "positive cooperativity" between inhibitor (valine) binding sites at low valine concentrations, and "negative cooperativity" between inhibitor binding sites at high valine concentrations. The AHAS of E. coli strains E and LL5, however, showed only negative cooperativity between binding sites for inhibitor, which could be the mechanism for incomplete inhibition of AHAS by valine. Preliminary kinetic analyses using Michaelis-Menten plots were carried out with strains K-12 and LL5. The levels of threonine deaminase (TD) and AHAS were also examined in four of the E. coli strains, K-12, B, AB1020 and LL5. AHAS was repressed and TD derepressed in E. coli K-12 grown (linearly) with 10ˉ³M valine. In E. coli strains B and AB1020, growth with 10ˉ³M valine had no effect on the levels of AHAS or (derepressed) TD. In E. coli strain LL5, growth with 10ˉ³M valine did not change the AHAS level but caused significant derepression of TD. Sensitivity of growth to valine has been correlated with three properties of the organism: 1) Feed-back sensitivity of acetohydroxy acid synthetase, to L-valine; 2) The level of acetohydroxy acid synthetase in the cell; 3) The level of biosynthetic threonine deaminase - the regulatory enzyme for isoleucine biosynthesis. / Medicine, Faculty of / Biochemistry and Molecular Biology, Department of / Graduate

Two different cloned human DNA segments encompassing transfer RNA gene and pseudogene clusters have been isolated from a human gene library harbored in bacteriophage lambda Charon 4-A. One clone (designated as λhVal7) encompassing a 20.5-kilobase (Kb) human DNA insert was found to contain a valine transfer RNA_AAC gene and several Alu-like elements by Southern blot hybridization analysis and DNA sequencing with the dideoxyribonucleotide chain-termination method in the bacteriophage M13mp19 vector. Another lambda clone (designated as λhLeu8) encompassing a 14.3-Kb segment of human DNA was found to contain a methionine elongator transfer RNA_CAT pseudogene and other as yet unidentified transfer RNA pseudogenes.

Master of Science / Department of Animal Sciences and Industry / Joel DeRouchey / Michael Tokach / Six experiments using a total of 2,974 nursery pigs were used to determine the effects of monosodium glutamate (MSG) and amino acids (AA) on nursery pig growth performance. Experiments 1 and 2 evaluated increasing dietary MSG for nursery pigs. Increasing dietary MSG up to 2% without balancing for sodium and chloride content decreased nursery pig performance, and feeding sodium levels equivalent to 1% MSG also decreased performance. When sodium and chloride were balanced, there were marginal effects of increasing dietary MSG on pig performance. Experiment 3 was conducted to determine the standardized ileal digestible (SID) lysine (Lys) requirement for pigs weighing 7- to 11- kg. The SID Lys requirement was estimated to be 1.45% and greater than 1.60% depending on the statistical model applied for both ADG and G:F. This experiment served to validate the SID Lys requirement for use in formulating diets for the subsequent experiments. Experiment 4 evaluated increasing SID valine (Val) to Lys ratio for nursery pigs weighing 7- to 10- kg. A SID Val:Lys ratio of 62.9% optimized ADG. Maximum feed efficiency (G:F) was captured using 71.7% SID Val:Lys ratio, however, 99% of maximum was achieved with SID Val at 64.4% of Lys. For ADFI, maximum performance was at 74% SID Val:Lys ratio, with 99% of maximum intake achieved at 68%. Experiments 5 and 6 investigated increasing SID isoleucine (Ile) to Lys ratio for 6- to 11- kg pigs. When ADG and ADFI were modeled, broken-line models reported maxima of 52.0% Ile:Lys ratio while quadratic models were as high as 64% of Lys.

O objetivo deste estudo foi avaliar o desempenho produtivo, o rendimento de carcaça e de cortes comerciais de frango alimentados com rações peletizadas contendo diferentes relações de valina/isoleucina durante as fases de crescimento e final (22 a 42 dias) desses animais. Foram utilizados 630 pintos machos de um dia de idade, da linhagem cobb 500. De 1 a 22 dias de idade, todas as aves receberam uma ração basal comum e aos 22 dias foram pesadas e distribuídas em um delineamento inteiramente casualizado, em esquema fatorial 3x3 (nove combinações de níveis de valina e isoleucina). Os frangos de corte foram submetidos aos seguintes tratamentos experimentais: T1 ração com relação valina/lisina 0,75% e isoleucina/lisina de 0,67%; T2 ração com relação valina/lisina 0,75% e isoleucina/lisina 0,71%; T3 ração com relação valina/lisina de 0,75% e isoleucina/lisina de 0,75%; T4 ração com relação valina/lisina de 0,79% e isoleucina/lisina 0,67%; T5 ração com relação valina/isoleucina de 0,79% e isoleucina/lisina 0,71%; T6 - ração com relação valina/isoleucina de 0,79% e isoleucina/lisina 0,75%; T7 - ração com relação valina/isoleucina de 0,83% e isoleucina/lisina 0,67%; T8 - ração com relação valina/isoleucina de 0,83% e isoleucina/lisina 0,71% e T9 - ração com relação valina/isoleucina de 0,83% e isoleucina/lisina 0,75%. As características avaliadas foram desempenho, o ganho de peso, o consumo de ração, a conversão alimentar, o rendimento de carcaça e cortes comerciais. Houve interação entre os fatores para todas as características avaliadas, exceto para peso corporal aos 28 dias. Os melhores resultados obtidos nas características avaliadas foram para as aves alimentadas com (0,83% de valina e 0,75% de isoleucina), exceto para peso corporal e ganho de peso aos 42 dias, em que os melhores resultados foram obtidos pelas aves que receberam (0,83% de valina e 0,75% de isoleucina) / The objective of this study was to evaluate the performance, carcass yield and commercial cuts fed pelleted diets containing different ratios of valine / isoleucine during the growing and final (22-42 days) phase of these animals. I was used 630 one day old male cobb 500 broiler chicks during 1-22 days of age, the birds were fed a common basal diet at 22 days the birds were weighed and distributed in a completely randomized design with factorial 3x3 (nine combinations of levels of valine and isoleucine). Broilers were submitted to the following experimental treatments: T1 - ration with respect valine / isoleucine and lysine 0.75% / 0.67% lysine; T2 - ration with respect valine / isoleucine and lysine 0.75% / 0.71% lysine; T3 - ration with respect valine / lysine and isoleucine 0.75% / 0.75% lysine; T4 - ration with respect valine / lysine and isoleucine 0.79% / 0.67% lysine; T5 - tion with respect valine / isoleucine isoleucine and 0.79% / 0.71% lysine; T6 - tion with respect valine / isoleucine isoleucine and 0.79% / 0.75% lysine; T7 - tion with respect valine / isoleucine isoleucine and 0.83% / 0.67% lysine; T8 - tion with respect valine / isoleucine isoleucine and 0.83% / 0.71% lysine and T9 - tion with respect valine / isoleucine isoleucine and 0.83% / 0.75% lysine. The characteristics evaluated were weight gain, feed intake, feed conversion and carcass yield and animal commercial cuts. There was a significant interaction between the factors for all characteristics evaluated, except for weight at 28 days. The best results in characteristics were evaluated for birds receiving (0.83% of valine and isoleucine 0.75%), except for body weight and weight gain at 42 days, in which the best results were obtained the animals receiving (0.83% and 0.75% of valine for isoleucine)

O objetivo deste estudo foi avaliar o desempenho produtivo, o rendimento de carcaça e de cortes comerciais de frango alimentados com rações peletizadas contendo diferentes relações de valina/isoleucina durante as fases de crescimento e final (22 a 42 dias) desses animais. Foram utilizados 630 pintos machos de um dia de idade, da linhagem cobb 500. De 1 a 22 dias de idade, todas as aves receberam uma ração basal comum e aos 22 dias foram pesadas e distribuídas em um delineamento inteiramente casualizado, em esquema fatorial 3x3 (nove combinações de níveis de valina e isoleucina). Os frangos de corte foram submetidos aos seguintes tratamentos experimentais: T1 ração com relação valina/lisina 0,75% e isoleucina/lisina de 0,67%; T2 ração com relação valina/lisina 0,75% e isoleucina/lisina 0,71%; T3 ração com relação valina/lisina de 0,75% e isoleucina/lisina de 0,75%; T4 ração com relação valina/lisina de 0,79% e isoleucina/lisina 0,67%; T5 ração com relação valina/isoleucina de 0,79% e isoleucina/lisina 0,71%; T6 - ração com relação valina/isoleucina de 0,79% e isoleucina/lisina 0,75%; T7 - ração com relação valina/isoleucina de 0,83% e isoleucina/lisina 0,67%; T8 - ração com relação valina/isoleucina de 0,83% e isoleucina/lisina 0,71% e T9 - ração com relação valina/isoleucina de 0,83% e isoleucina/lisina 0,75%. As características avaliadas foram desempenho, o ganho de peso, o consumo de ração, a conversão alimentar, o rendimento de carcaça e cortes comerciais. Houve interação entre os fatores para todas as características avaliadas, exceto para peso corporal aos 28 dias. Os melhores resultados obtidos nas características avaliadas foram para as aves alimentadas com (0,83% de valina e 0,75% de isoleucina), exceto para peso corporal e ganho de peso aos 42 dias, em que os melhores resultados foram obtidos pelas aves que receberam (0,83% de valina e 0,75% de isoleucina) / The objective of this study was to evaluate the performance, carcass yield and commercial cuts fed pelleted diets containing different ratios of valine / isoleucine during the growing and final (22-42 days) phase of these animals. I was used 630 one day old male cobb 500 broiler chicks during 1-22 days of age, the birds were fed a common basal diet at 22 days the birds were weighed and distributed in a completely randomized design with factorial 3x3 (nine combinations of levels of valine and isoleucine). Broilers were submitted to the following experimental treatments: T1 - ration with respect valine / isoleucine and lysine 0.75% / 0.67% lysine; T2 - ration with respect valine / isoleucine and lysine 0.75% / 0.71% lysine; T3 - ration with respect valine / lysine and isoleucine 0.75% / 0.75% lysine; T4 - ration with respect valine / lysine and isoleucine 0.79% / 0.67% lysine; T5 - tion with respect valine / isoleucine isoleucine and 0.79% / 0.71% lysine; T6 - tion with respect valine / isoleucine isoleucine and 0.79% / 0.75% lysine; T7 - tion with respect valine / isoleucine isoleucine and 0.83% / 0.67% lysine; T8 - tion with respect valine / isoleucine isoleucine and 0.83% / 0.71% lysine and T9 - tion with respect valine / isoleucine isoleucine and 0.83% / 0.75% lysine. The characteristics evaluated were weight gain, feed intake, feed conversion and carcass yield and animal commercial cuts. There was a significant interaction between the factors for all characteristics evaluated, except for weight at 28 days. The best results in characteristics were evaluated for birds receiving (0.83% of valine and isoleucine 0.75%), except for body weight and weight gain at 42 days, in which the best results were obtained the animals receiving (0.83% and 0.75% of valine for isoleucine)

Much of Nature is composed of highly modular and composable nested multicomponent living systems. Synthetic biology and bioengineering exploit this modularity to understand and engineer living things. This thesis explores two projects coupled by these principles, the first utilizing a synthetic genomics approach to probe the evolutionary history, flexibility, and modularity of core metabolism, and the second adapting and engineering components of a living material to generate living architecture and embed add program new behaviors into the living biocomposite. Chapter 1 details the synthetic resurrection of a core metabolic pathway lost from the metazoan lineage millions of years ago. All metazoans are auxotrophic for 9 of the 20 amino acids, the so-called “essential” amino acids. The pressures behind the loss of the 9 are a deep evolutionary puzzle. To investigate this event and probe the limits of core metabolic flexibility, we generated a synthetic valine prototrophic mammalian cell line, restoring valine self-sufficiency to the metazoan lineage. The restoration of this pathway implies the modern mammalian metabolism is still compatible with autogenous valine production, suggests profound modularity in core metabolism, and underscores the potential usefulness of large-scale synthetic genomics approaches in a answering deep evolutionary questions. Chapter 2 describes the engineering of a hybrid fungal-bacterial biocomposite by adapting and leveraging existing behaviors and microbial constituents of a living material. Fungal biocomposites are composed of a particulate lignocellulosic feedstock bound together into a bulk biocomposite by a network of dense fungal mycelium. Using a bioprospecting approach, we designed architectural and design strategies that relied on the natural substrate flexibility and growth patterns of the fungal component of the biocomposite to form origami-inspired human scale folding structures. Similarly, we isolated, characterized, and engineered a natural microbial component of the biocomposite’s own microbiome and used its pre-adapted ability to engraft in the growing biomaterial to embed new genetic functionalities in biocomposite objects. We believe that the strategy of bioprospecting useful components and behaviors holds promise for the development of future biomaterials adapted from living systems.

Chick feeding tests were conducted to study the effects of dietary imbalances among the three branched chain amino acids on growth rate. All diets fed in these tests contained about 18% protein. The indispensable amino acids (IAA) were found in two of the diets fed in the proportions found by Dobson et al. (1964) to be well balanced. One of the diets had all IAA at 85% of these balanced levels while the other had them all at 125% of these levels. The tests involved reducing the isoleucine, leucine and valine levels in the high IAA diet to the low levels in all combinations. Growth rates were similar with the diets containing all ten IAA at the low levels, all ten at the high levels, or seven at the high levels and isoleucine, leucine and valine at the low levels. Generally, when only one or two of these three were reduced to the low levels, growth rates were lower. Thus there appeared to be a three-way interaction among these amino acids with the reduction in weight gain being the result of an imbalance and not of a deficiency. The changes in growth rate noted when the leucine level was reduced indicated that the leucine level in Dobson's balanced diet was relatively high; the isoleucine level appeared to be relatively low. The interaction between leucine and valine appeared to be more significant than the other two two-way interactions. The effect of dietary level of these amino acids on the branched chain amino acid transaminase (BAT) activity in the liver and kidneys was determined. The differences noted were inconsistent. There tended to be a slightly higher activity in chicks fed the high levels of these amino acids, but the differences certainly were not as great as the changes in arginase activity reported by others when imbalanced diets were fed. Chicks fed the diet low in valine and high in the other nine IAA were selected for fast or slow growth on this diet. They were raised to maturity and produced eggs that were hatched for feeding tests with the diets containing different levels of the branched chain amino acids. Only a limited number of chicks from the two strains were hatched. Performance of the chicks was similar to that of their parents when fed the low valine diet.