No description available.

No description available.

The general biosynthetic pathway for tryptophan is known. However, little information has been gathered on how substrates and enzymes interact when phosphoribosylanthranilate isomerase (PRAI) and indole-3-glycerolphosphate synthase (IPGS) convert a substituted phenyl ring, PRA, into an indole moiety, IGP, via 1-(O-carboxyphenylamino)-1-deoxyribulose-5-phosphate (CdRP). There has been no serious synthetic approach to develop methodology to produce a plethora of substrate and product analogues of CdRP. The studies described in this thesis cover methodology focusing on secondary aryl amine formation, using reductive amination, nucleophilic substitution and epoxide ring opening, leading to CdRP analogues. Reductive aminations with D-ribose failed to produce any aryl glycosylamine precursor, possibly due to the low nucleophilicity of aryl amines such as aniline. Removing the aromaticity and using cyclohexylamine produced secondary amines in moderate yield in the presence of benzylpentanal, and NaBH3CN, at a pH of 5.5. This led to a successful reductive amination using anthranilate methyl ester. Secondary aryl amine synthesis via epoxide ring opening proved consistently reproducible. Using LiNTf2 and high equivalents of cyclohexylamine or aniline in neat conditions opened protected epoxides. This has led to the formation of advanced secondary aryl amine synthons and the development of methodology leading to target compounds with functionality at the 1,2 and 5 positions. Nucleophilic substitution using caesium base, high equivalents aniline at room temperature, gave a moderate yield of secondary aryl amines from sulfonyl and bromide good leaving groups. Raising the reaction temperature improved yields using low equivalents of aniline, with the optimal temperature being 50 °C. Ultimately using both the high equivalents of aniline or anthranilate methyl ester and warming the reaction in DMF gave the highest yields of secondary aryl amines. No overalkylated tertiary amine was isolated when a caesium base was used. Boc N-protection of 1-phenylamino-4-pentene and asymmetric dihydroxylation gave the corresponding diol, which was phosphorylated giving the protected target 1,4,5 compound. The methodology leading to the protected target 1,4,5 compound synthesis provides a means to the synthesis additional of CdRP analogues.

Thesis (Ph. D.)--Hong Kong University of Science and Technology, 2002. / On t.p. "Trp" is superscript. Includes bibliographical references (leaves 152-172). Also available in electronic version. Access restricted to campus users.

No description available.

No description available.

In helminths, including the free-living nematode, Caenorhabditis elegans, serotonin (5-HT) acts as an important neuroactive agent and is associated with carbohydrate metabolism, glucouse utilization, motility, feeding and reproductive behaviour. In mammals and other organisms, 5-HT is synthesized through the action of tryptophan hydroxylase (TPH). TPH is the rate limiting enzyme in the biosynthesis of 5-HT, and as such sets the pace for the formation of 5-HT. TPH is a member of a family of enzymes that hydroxylate aromatic amino acids and have an absolute requirement for the pterin cofactor, tetrahydrobiopterin (BH4). It is unknown if this same enzyme catalyzes the synthesis of 5-HT in C. elegans and other helminths. / Based on sequence information from the C. elegans Genome Data Base and RT-PCR, we have cloned a full-length C. elegans TPH cDNA (CeTPH) that shows high homology to mammalian TPH. The predicted coding sequence of CeTPH was subcloned into the prokaryotic expression vector, pET-15b, and the resulting construct was introduced into E. coli (BL21 DE3 pLys strain) for IPTG-inducible expression of CeTPH protein. Results show that CeTPH expressed in E. coli has TPH activity and also shows an absolute requirement for the cofactor, BH4, just as shown previously for the mammalian enzyme. It has been well established that 5-HT is present and is biologically active in the tissues of C. elegans. By way of characterizing furthur CeTPH, we examined the localization of TPH in whole mounts of C. elegans by immunofluoresence using a polyclonal antibody against TPH. / Taken together, the results of this thesis characterize at the structural, functional and in situ levels one of the most primitive forms of TPH enzyme ever cloned.

Since no effect on CP and diet type on the optimal Trp-to-Lys ratio (TLR) was found in two previous experiments, two extra experiments were planned. In the first one, piglets were fed a Lys-deficient basal diet supplemented with crystalline Lys (to create graded levels of SID Lys). GFR and PUN were measured every 5-d for 15-d; and were analyzed using the broken-line analysis to determine the Lys requirement. As SID Lys level increased, GFR increased linearly and PUN decreased linearly. The SID Lys requirement for was estimated to be 1.02%. To determine the optimal TLR, pigs were fed Trp-deficient basal diet supplemented with crystalline Trp (to create graded levels of SID TLR). Most of the parameters evaluated failed (P > 0.10) to yield an estimate. Based on the evidence we suggest that the optimal SID TLR for early-weaned pigs fed a wheat-barley diet lies below 17.75%.

Since no effect on CP and diet type on the optimal Trp-to-Lys ratio (TLR) was found in two previous experiments, two extra experiments were planned. In the first one, piglets were fed a Lys-deficient basal diet supplemented with crystalline Lys (to create graded levels of SID Lys). GFR and PUN were measured every 5-d for 15-d; and were analyzed using the broken-line analysis to determine the Lys requirement. As SID Lys level increased, GFR increased linearly and PUN decreased linearly. The SID Lys requirement for was estimated to be 1.02%. To determine the optimal TLR, pigs were fed Trp-deficient basal diet supplemented with crystalline Trp (to create graded levels of SID TLR). Most of the parameters evaluated failed (P > 0.10) to yield an estimate. Based on the evidence we suggest that the optimal SID TLR for early-weaned pigs fed a wheat-barley diet lies below 17.75%.

Universiẗat, Diss., 2006--Giessen.