By means of an in silico analysis, we demonstrated that a previously described chimeric gene spesdh encoding spermidine synthase, a key enzyme involved in the synthesis of polyamines, and saccharopine dehydrogenase, an enzyme involved in lysine synthesis in fungi, were present exclusively in members of all basidiomycota subphyla, but not. Sep 30, 2004 we isolated the gene encoding lysineketoglutarate reductase lkr, ec 1. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. Saccharopine description the saccharopine pathway is the main route for lysine degradation in mammals, and its first two reactions are catalyzed by enzymatic activities known as lysineoxoglutarate reductase lor and saccharopine dehydrogenase sdh, which reside on a single bifunctional polypeptide lorsdh ec 1. Sdh is defined as saccharopine dehydrogenase biochemistry somewhat frequently. Saccharopine dehydrogenase activity in the highlysine opaque and floury maize mutants article pdf available in food biotechnology 201 january 2006 with 41 reads how we measure reads. The preparation was free from impurities as judged from chromatography on paper.
The downregulation of lkrsdh in seeds results in the overaccumulation of free lysine to levels that meet human nutritional requirements. Full text get a printable copy pdf file of the complete article 8k, or click on a page image below to browse page by page. Sdh saccharopine dehydrogenase biochemistry acronymfinder. The worlds first wiki where authorship really matters. How is saccharopine dehydrogenase biochemistry abbreviated. Saccharopine is an intermediate in the degradation of lysine, formed by the condensation of lysine and alphaketoglutarate. Saccharopine dehydrogenase how is saccharopine dehydrogenase abbreviated. Here, we show that bacteria possess the genes encoding lysine.
Saccharopine is then converted by saccharopine dehydrogenase sdh to glu and. In the opposite direction, the reduced nicotinamide adenine dinucleotide nadh adds to the enzyme first, while there is no. Overall kinetic mechanism of saccharopine dehydrogenase from. A kinetic study of saccharopine dehydrogenase reaction. In brain, however, lor and saccharopine dehydrogenase activities were below the detection limit of the method. Most untreated patients develop irreversible brain damage during. Source sequences af151807, aw515488, bg258610, bm786495 consensus cds ccds31084. A tdna insertion knockout of the bifunctional lysine. Among amino acids those having aliphatic side chains of 5 and 6 carbon atoms were best inhibitors. Nadph is a poor substrate, and causes an increase in the k m values for. The full text of this article is available as a pdf 3. We isolated the gene encoding lysineketoglutarate reductase lkr, ec 1.
Lys ketoglutarate reductase lkr first combines lys and. In the latter, the bifunctional enzyme alphaaminoadipic semialdehide synthase ec 1. Purification and characterization of the bifunctional enzyme lysine. It catalyzes the reversible conversion of saccharopine to llysine and. Archives of biochemistry and biophysics 2012, 522 1, 5761. Feb 04, 2019 in caenorhbditis elegans, mutations in the saccharopine dehydrogenase sdh domain of the bifunctional enzyme. Pdf saccharopine dehydrogenase activity in the highlysine. The first two enzymatic steps of the saccharopine pathway are catalyzed by lkr and saccharopine dehydrogenase sdh, which are separate domains of a bifunctional polypeptide12 lkrsdh. The first two reactions of the pathway are catalyzed by lkr, which condenses lysine and ketoglutarate to form. In the sulfatebound structure, a sulfate ion binds in a cleft between. In molecular biology, the protein domain saccharopine dehydrogenase sdh, also named saccharopine reductase, is an enzyme involved in the metabolism of the amino acid lysine, via an intermediate substance called saccharopine. Analytical biochemistry 1147, 230233 1985 enzymatic measurement of saccharopine with saccharopine dehydrogenase michael s. Eckel department of medicine, case western reserve university school of medicine, cleveland, ohio 44106 received november 26, 1984 we have developed an enzymatic method for measuring saccharopine, a key intermediate in lysine metabolism. Saccharopine dehydrogenase definition of saccharopine.
Lysineoxoglutarate reductase and saccharopine dehydrogenase are enzymic activities that catalyse the first two steps of lysine degradation through the saccharopine pathway in upper eukaryotes. This paper describes the isolation and characterization of a cdna clone encoding a bifunctional enzyme bearing domains corresponding to these two enzymic activities. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists. The lkr and sdh enzymes, which play pivotal role in lys catabolism, are linked on a single bifunctional lkrsdh polypeptide, encoded by a single lkr sdh.
A deficiency of one of these isoforms is associated with. Glutaric aciduria type i, an inherited deficiency of glutarylcoenzyme a dehydrogenase localized in the final common catabolic pathway of llysine, lhydroxylysine and ltryptophan, leads to accumulation of neurotoxic glutaric and 3hydroxyglutaric acid, as well as nontoxic glutarylcarnitine. Request pdf on may 9, 20, xiang sheng and others published theoretical study on the proton shuttle mechanism of saccharopine dehydrogenase. Synthesis of the arabidopsis bifunctional lysineketoglutarate. Read the oxidation state of active site thiols determines activity of saccharopine dehydrogenase at low ph, archives of biochemistry and biophysics on deepdyve, the largest online rental service for scholarly research with thousands of academic publications available at. Request pdf on may 9, 20, xiang sheng and others published theoretical study on the proton shuttle mechanism of saccharopine dehydrogenase find, read and cite all the research you need on. The rate of formation of lysine from saccharopine, catalysed by saccharopine oxidoreductase, was examined and correlated with the dietary intake of lysine in each species studied. Imagine a global collaborative knowledge base for original thoughts nature genetics. Results purification of saccharopine dehydrogenase all operations were carried out at 4c unless otherwise stated. Saccharopine, an intermediate of the aminoadipic acid pathway.
Nutritionalrich and stresstolerant crops by saccharopine. Determinants of substrate specificity for saccharopine. Glucose6phosphate dehydrogenase deficiency, the most common enzyme deficiency world wide, causes a spectrum of disease including neonatal hyperbilirubinemia, acute hemolysis, and chronic hemolysis. Enzymatic measurement of saccharopine with saccharopine. Pdf saccharopine dehydrogenase activity in the high.
In the past years, the crystal structures of sdh from s. The lkr and sdh enzymes, which play pivotal role in lys catabolism, are linked on a single bifunctional lkrsdh polypeptide, encoded by a. There are several pathways involved in lysine catabolism but the most commonly used is the saccharopine pathway fig. The structure reveals homology to alanine dehydrogense, despite low primary sequence similarity. Nad oxidoreductase lysine forming in the direction of saccharopine formation. While lysine is an essential amino acid for mammals, human pathogenic fungi, including candida albicans, cryptococcus neoformans and aspergillus fumigatus and the plant pathogen magnaporthe. Supporting role of lysine and glutamate 16 in the acidbase mechanism of saccharopine dehydrogenase from saccharomyces cerevisiae. Therapeutic modulation of cerebral llysine metabolism in a. Crystal structure of saccharopine reductase from magnaporthe. The saccharopine pathway is the main degradation pathway of lysine in plants and animals.
A fragment of a length of 529 bp which comprised the nucleotides. Then, aasa is converted to aminoadipic acid aaa by aaminoadipicdsemialdehyde dehydrogenase aasadh. The plasmid pbinar is a derivative of the binary vector plasmid pbin19 bevan, 1984 which was constructed as follows. Preparation of cell free extractfour hundred grams of dried bakers yeast oriental yeast co. By means of an in silico analysis, we demonstrated that a previously described chimeric gene spesdh encoding spermidine synthase, a key enzyme involved in the synthesis of polyamines, and saccharopine dehydrogenase, an enzyme involved in lysine synthesis in fungi, were present exclusively in members of all basidiomycota subphyla, but not in any other group of living organisms. The model contains one homodimer in the asymmetric unit and 171 water molecules. As a member of the wwpdb, the rcsb pdb curates and annotates pdb data according to agreed upon standards. New therapeutic candidates for the treatment of malassezia. Eckel department of medicine, case western reserve university school of medicine, cleveland, ohio 44106 received november 26, 1984 we have developed an enzymatic method for measuring. Three structures of saccharopine dehydrogenase llysineforming sdh have been determined in the presence of sulfate, adenosine monophosphate amp, and oxalylglycine oxgly.
Conversion of pipecolic acid into lysine in penicillium. Saccharopine dehydrogenaselike oxidoreductase gene id 51097, 109232, 305021. Hepatic activity of lor and saccharopine dehydrogenase was similar in control and gcdh. Novel chimeric spermidine synthasesaccharopine dehydrogenase. Saccharopine, an intermediate of the aminoadipic acid. The saccharopine pathway is used for lysine synthesis in fungi and lysine catabolism in plants, animals, and bacteria markovitz and chuang 1987. Only mutations in the sdh domain of aass1 induce mitochondrial abnormality. Based on expressed sequence tag and genomic sequencing databases, arabidopsis. Saccharopine dehydrogenase journal of biological chemistry. Lysineketoglutarate reductase and saccharopine dehydrogenase.
Construction of transformation vectors containing the phytophthora infestans saccharopine dehydrogenase gene a preparation of the plant expression vector ir 4771. Due credit and reputation for authors authorship tracking technology. A corresponding cdna was isolated from total arabidopsis rna. Saccharopine dehydrogenase glutamate forming of the biosynthetic pathway of lysine in saccharomyces cerevisiae was purified 1,122fold by using acid precipitation, ammonium sulfate precipitation, deaesepharose, gel filtration, and reactive red120 agarose chromatography. Crystal structure of saccharopine dehydrogenase from saccharomyces cervisiae complexed with nad. Diagnosis and management of g6pd deficiency american. In the opposite direction, the reduced nicotinamide adenine dinucleotide nadh adds to the enzyme first, while there is no preference for the. Saccharopine dehydrogenase from wikipedia, the free encyclopedia in molecular biology, the protein domain saccharopine dehydrogenase sdh, also named saccharopine reductase, is an enzyme involved in the metabolism of the amino acid lysine, via an intermediate substance called saccharopine. Lysine degradation through the saccharopine pathway in. The saccharopine pathway is the main route for lysine degradation in mammals, and its first two reactions are catalyzed by enzymatic activities known as lysineoxoglutarate reductase lor and saccharopine dehydrogenase sdh, which reside on a single bifunctional. In caenorhbditis elegans, mutations in the saccharopine dehydrogenase sdh domain of the bifunctional enzyme. Saccharopine is then converted by saccharopine dehydrogenase sdh to glu andamino adipic semialdehyde.
Kinetic data have been measured for the histidinetagged saccharopine dehydrogenase from saccharomyces cerevisiae, suggesting the ordered addition of nicotinamide adenine dinucleotide nad followed by saccharopine in the physiologic reaction direction. The saccharopine dehydrogenase enzyme can be classified under ec 1. Kg and lysine, while it was reported that nadp is not a substrate in the direction of saccharopine oxidation. The lysine catabolite saccharopine impairs development by. Lysineoxoglutarate reductase and saccharopine dehydrogenase are enzymic. Sep 12, 2006 kinetic data have been measured for the histidinetagged saccharopine dehydrogenase from saccharomyces cerevisiae, suggesting the ordered addition of nicotinamide adenine dinucleotide nad followed by saccharopine in the physiologic reaction direction. The second group lane b was transferred to a sucfree medium for 72 h before harvesting.
Saccharopine dehydrogenase like oxidoreductase gene id 51097, 109232, 305021. Regulation of lysine catabolism through lysineketoglutarate. Amino adipic acid is further into acetylcoenzyme a and several additional molecules of glu fig. A number of amino and keto acids have been shown to inhibit saccharopine dehydrogenase n.
Lkr condenses lysine and ketoglutaric acid to form saca charopine, which is then hydrolyzed by sdh, giving rise to. Therapeutic modulation of cerebral llysine metabolism in. Purification and properties of saccharopine dehydrogenase. Nlglutar2ylllysine, n 6l1,3dicarboxylpropylllysine cas number 997682. Saccharopine dehydrogenase catalyzes the final step of. These results indicate that lysine catabolism plays a critical role in regulating the free lysine level in plant seeds and point to a direction to increase the levels of soluble lysine in seeds by engineering plants with downregulated lysine. In silicibacter, the contiguous lkr and sdh genes are interspersed, in another frame, by a polypeptide of unknown. Dec 19, 2019 as an example, fungi saccharopine dehydrogenase is involved in the lysine biosynthesis, when the lysineketoglutarate from plant, sometimes also called saccharopine dehydrogenase, is responsible for lysine catabolism houmard et al. Theoretical study on the proton shuttle mechanism of. However, the saccharopine pathway is also involved in stress response in plants, animals, and bacteria. The structure of apo saccharopine reductase in space group c222 1 was determined by molecular replacement and refined to 2.
The dehydrogenase also exhibits stringent substrate specificity with respect to its. Contribution of k99 and d319 to substrate binding and. Ricardo antunes 2006 saccharopine dehydrogenase activity in the highlysine opaque and floury maize mutants, food biotechnology, 20. Overall kinetic mechanism of saccharopine dehydrogenase.
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