L-Lysine decarboxylase

The lysine sensor reported by Karube et al. was a micro-C02 sensor containing immobilized L-lysine decarboxylase (LDC, Fig. 3.18.C). L-lysine catalysed the following reaction... 

Most cultures from Collection IBSO produce lyases L-ornithine, L-arginine, and L-lysine decarboxylases. Neuraminidase (sialidase, or mucopolysaccharide - N-acetylneuraminilhydrolase) is the enzyme of the hydrolase group. As is usual neuraminidase activity is a property of pathogenic organisms. We found for the first time that luminous bacterial cultures of the species V. harveyi possess low neuraminidase activity. It may be probably one of the factors contributing to contamination of marine animals by luminous bacteria. 

L-Lysine Decarboxylase Origin Bacterium cadaveris Fluka... 

The absolute configuration of 21 was established by two methods. First, 21 was converted to 5-phthalimido[5-2H]valerate by the use of chemical and enzymic methods and shown to have the same optical rotatory properties as those of authentic (5/ )-5-phthalimido[5- H]valerate produced from L-glutamate by an established stereochemical route. Second, 21 was converted to [l- H]cadavarine with L-lysine decarboxylase, followed by treatment with diamine oxidase to form pelletierine. Retention of all of the deuterium in the pelletierine demonstrated that the deuterium must be in the pro-(R) position, since the oxidase reaction is known to labilize hydrogen at the pro-(S) position [Eq. (51)] ... 

There is an alternative way of viewing the above results, however. It could be that the biosynthetic pathway to the pyrrolidine ring of nicotine is similar (in part) to the route to the piperidine alkaloids. Part of the model suggested for the biosynthesis of the piperidine nucleus from lysine (see above) could be easily adapted to account for the C02 and nornicotine results, that is variable/in-complete equilibration of bound putrescine (arising by enzyme-mediated decarboxylation of ornithine) with unbound material. L-Ornithine decarboxylase (EC 4.1.1.17, L-ornithine carboxy-lyase) occurs widely in higher plants and like L-lysine decarboxylase requires pyridoxal phosphate as a co-factor. ... 

Decarboxylation of lysine 257b gives the important biosynthetic intermediate cadaverine 266 (Scheme 71). This reaction is catalyzed by L-lysine decarboxylase (EC 4.1.1.18), decarboxylation in or H20, giving... 

L-Lysine decarboxylase has been used extensively to prepare stereo-specifically labeled samples of cadaverine for use in biosynthetic studies (274-280). 

The conversion of lysine into piperidine alkaloids involves retention of hydrogen isotope at C-2/° The sequence is suggested to be that shown in Scheme 1, and catalysis of the reaction may be attributed to L-lysine decarboxylase. This enzyme, from the micro-organism Bacillus cadaveris, has been found to carry out the conversion of L-lysine into cadaverine with retention of configuration. Decarboxylation of L-[2- H]lysine by this enzyme then affords [15- H]-cadaverine. When this material is converted into alkaloids, e.g. iV-methyl-pelletierine (4 R = Me), the tritium attached to what becomes C-2 is lost cf. refs. 5 and 6. On the other hand, conversion of lysine into sedamine (27) in Sedum acre results in retention of the tritium originally present at C-2. The simplest explanation is that protonation of (26) in the micro-organism and plant proceeds with opposite stereochemistry. This is at variance, however, with current ideas on the stereochemistry of reactions that are catalysed by pyridoxal phosphate. ... 

This is disturbing, and so the stereochemistry of the overall reaction L-lysinecadaverineA -piperideine has been checked, using L-lysine decarboxylase from Escherichia coli and B. cadaveris, pea seedling diamine oxidase, and 5. acre plants, with confirmation of the above deductions the lysine decarboxylase from the two sources effected decarboxylation with the same stereochemical outcome. Solution of the problem must now await further experiments with the enzymes involved in alkaloid biosynthesis. 

The enzyme, i.e. lysine decarboxylase, that is required for the conversion of lysine into cadaverine, and thus the first step of alkaloid biosynthesis, has been isolated from chloroplasts of L. polyphyllus,28 Like the majority of amino-acid decarboxylases, this enzyme is dependent on pyridoxal 5 phosphate. Its activity was found not to be affected by the presence or absence of quinolizidine alkaloids. Control of the enzyme by simple product feedback inhibition therefore seems unlikely. The operational parameters of this enzyme resemble those of the 17-oxosparteine synthase. Co-operation between the two enzymes would explain why cadaverine is almost undetectable in vivo. 

L-Lysine and L-arginine are determined in a rapid fashion by using a bienzyme-immobilized system, decarboxylase-diamine oxidase ... 

Pecker, L.R, Hillebrandt, S., Rfigenhagen, C., Herminghaus, S., Landsmann, J. and Berlin, J. (1992) Metabolic effects of a bacterial lysine decarboxylase gene expressed in hairy root culture of Nicotiana glauca. Biotech. Lett., 14, 1035-40. 

Alkaloid metabolism in lupine was proved by Wink and Hartmann to be associated with chloroplasts (34). A series of enzymes involved in the biosynthesis of lupine alkaloids were localized in chloroplasts isolated from leaves of Lupinus polyphylls and seedlings of L. albus by differential centrifugation. They proposed a pathway for the biosynthesis of lupanine via conversion of exogenous 17-oxosparteine to lupanine with intact chloroplasts. The biosynthetic pathway of lupinine was also studied by Wink and Hartmann (35). Two enzymes involved in the biosynthesis of alkaloids, namely, lysine decarboxylase and 17-oxosparteine synthetase, were found in the chloroplast stoma. The activities of the two enzymes were as low as one-thousandth that of diaminopimelate decarboxylase, an enzyme involved in the biosynthetic pathway from lysine to diaminopimelate. It was suggested that these differences are not caused by substrate availability (e,g., lysine concentration) as a critical factor in the synthesis of alkaloids. Feedback inhibition would play a major role in the regulation of amino acid biosynthesis but not in the control of alkaloid formation. 

L-Lysine DL-o-Aminocaprolactam a,E-DiaminopimeIic acid Hydrolase + Racemase DAP decarboxylase... 

A new gene Idc which encodes lysine decarboxylase was found in addition to the formerly known cadA in Escherichia coli and the enzyme purified from the overexpression strain. The lysine decarboxylase encoded by Idc is constitutively produced by E. coli cells though the cadA encodes an inducible one [9]. It is interesting to know of the existence of this new lysine decarboxylase in lysine-producing Corynebacterium and to investigate the effects of the deletion of the gene on the amounts of L-lysine production. 

The decarboxylation by lysine decarboxylase of L-lysine (13) to give cadaverine (17) occurs with retention of configuration [protonation occurs on the a-face of the iraine (14)]. The oxidation of cadaverine (17) occurs with loss of the 1-pro-S proton, which is the proton originally sited at C-2 in L-lysine (13) c. Section 1.1). It follows that L-[2- H]lysine should... 

Lysine decarboxylase Bacillus cadaveris, E. coli L-Lysine Retention 249-251... 

Manometric deterrnination of L-lysine, L-arginine, L-leucine, L-ornithine, L-tyrosine, L-histidine, L-glutamic acid, and L-aspartic acid has been reviewed (136). This method depends on the measurement of the carbon dioxide released by the L-amino acid decarboxylase which is specific to each amino acid. 

Also known as beta-alanine because of its similarity to the canonical amino acid L-lysine, it has been overproduced in E. coli. A strain was prepared, which included an aspartate decarboxylase gene panD) from C. glutamicum, overexpression of aspartase (aspA) and phosphoenolpyruvate carboxylase (ppc), and acetyl-CoA synthase (acs) and it resulted in total titers of 32 g 1 after 39 h from rich media supplemented by glucose and ammonium sulfate [61]. 

Corynebacterium actively excretes amino acids through its cell wall membrane and does not degrade L-lysine due to the lack of lysine-decarboxylase. For 60 years all these characteristics have made this microbe the species of choice in L-lysine production. In addition it demonstrates the potential of natural biosynthesis pathways for commercial purposes. In contrast Escherichia coli entered the field of industrial amino acid fermentation not because of comparable advantages provided by nature but because of the availability of effective tools for genetic engineering. In the early 1980s such methods were state of the art for Escherichia coli but were only on an infant level for Corynebacterium. Developing industrial strains based on Escherichia coli, which at that time was not broadly covered by intellectual property (IP), provided room to build new IP in the field of amino acid fermentation. 

The biochemistry and molecular biology of quinolizidine alkaloid biosynthesis have not been fully characterized. Quinolizidine alkaloids are formed from lysine via lysine decarboxylase (LDC), where cadaverine is the first detectable intermediate (Scheme 6). Biosynthesis of the quinolizidine ring is thought to arise from the cyclization of cadaverine units via an enzyme-bound intermediate 176). LDC and the quinolizidine skeleton-forming enzyme have been detected in chlorop-lasts of L. polyphyllus 177). Once the quinolizidine skeleton has been formed, it is modified by dehydrogenation, hydroxylation, or esterification to generate the diverse array of alkaloid products. 

A -piperideine-2-carboxylic acid which is subsequently converted into pipecolic acid, the molecule that is entirely incorporated. The other lysine unit provides the C5N (N-8, C-9 to C-13) through cadaverine, which is generated by stereospecific decarboxylation of the amino acid catalyzed by lysine decarboxylase. Intramolecular cyclization of cadaverine affords 2,3,4,5-tetrahydropyridiniuni, which undergoes nucleophilic attach by pipecolic acid to generate l,2-biperidine-2-carboxylic acid. A further intramolecular cyclization generates imidazo-dipyridin-one which upon reduction, dehydration, and subsequent oxidation produces anosmine (Figure 5.88) [333, 334]. 

In general, alkaloids derive from the metabohsm of amino acids such as phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), omitine (Om), or lysine (Lys). Quinolizidine alkaloids derived from L-lysine. Its decarboxylation by means of the enzyme lysine decarboxylase gives cadaverine (Cad), the first detectable intermediate of this biosynthetic pathway (Scheme 14.1). 

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