Highly Stable
            <scp>l</scp>
            -Lysine 6-Dehydrogenase from the Thermophile
            <i>Geobacillus stearothermophilus</i>
            Isolated from a Japanese Hot Spring: Characterization, Gene Cloning and Sequencing, and Expression

Heydari, Mojgan; Ohshima, Toshihisa; Nunoura-Kominato, Naoki; Sakuraba, Haruhiko

doi:10.1128/aem.70.2.937-942.2004

Cited by 21 publications

(29 citation statements)

References 27 publications

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“…In the case of saccharopine reductase, the substrates are glutamate and L-2-aminoadipate-6-semialdehyde, which form a Schiff base intermediate that is reduced by NADPH to saccharopine (76). Lysine 6-dehydrogenase catalyzes the oxidative deamination of the ⑀-amino group of lysine in the presence of NAD ϩ to form L-2-aminoadipate-6-semialdehyde (77).…”

Section: Resultsmentioning

confidence: 99%

Evolution and Multifarious Horizontal Transfer of an Alternative Biosynthetic Pathway for the Alternative Polyamine sym-Homospermidine

Shaw

Elliott

Kinch

et al. 2010

Journal of Biological Chemistry

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Polyamines are small flexible organic polycations found in almost all cells. They likely existed in the last universal common ancestor of all extant life, and yet relatively little is understood about their biological function, especially in bacteria and archaea. Unlike eukaryotes, where the predominant polyamine is spermidine, bacteria may contain instead an alternative polyamine, sym-homospermidine. We demonstrate that homospermidine synthase (HSS) has evolved vertically, primarily in the ␣-Proteobacteria, but enzymatically active, diverse HSS orthologues have spread by horizontal gene transfer to other bacteria, bacteriophage, archaea, eukaryotes, and viruses. By expressing diverse HSS orthologues in Escherichia coli, we demonstrate in vivo the production of co-products diaminopropane and N 1 -aminobutylcadaverine, in addition to sym-homospermidine. We show that sym-homospermidine is required for normal growth of the ␣-proteobacterium Rhizobium leguminosarum. However, sym-homospermidine can be replaced, for growth restoration, by the structural analogues spermidine and sym-norspermidine, suggesting that the symmetrical or unsymmetrical form and carbon backbone length are not critical for polyamine function in growth. We found that the HSS enzyme evolved from the alternative spermidine biosynthetic pathway enzyme carboxyspermidine dehydrogenase. The structure of HSS is related to lysine metabolic enzymes, and HSS and carboxyspermidine dehydrogenase evolved from the aspartate family of pathways. Finally, we show that other bacterial phyla such as Cyanobacteria and some ␣-Proteobacteria synthesize sym-homospermidine by an HSS-independent pathway, very probably based on deoxyhypusine synthase orthologues, similar to the alternative homospermidine synthase found in some plants. Thus, bacteria can contain alternative biosynthetic pathways for both spermidine and sym-norspermidine and distinct alternative pathways for sym-homospermidine.Polyamines are primordial, small flexible organic polycations found in almost all cells of bacteria, archaea, and eukaryotes (1). In bacteria and archaea, the key polyamines (see Fig. 1A) are the triamines spermidine, sym-norspermidine, and sym-homospermidine (referred to herein as norspermidine and homospermidine), and occasionally more than one triamine can be found in the same cell. In eukaryotes, which contain spermidine (and in some plants, yeasts, and animals, the tetraamine spermine), polyamines are required for growth, cell proliferation, and normal cellular physiology. Polyamine biosynthesis is essential in the fungi Saccharomyces cerevisiae (2), Schizosaccharomyces pombe (3), Aspergillus nidulans (4), and Ustilago maydis (5), the kinetoplastid parasites Trypanosoma brucei (6) and Leishmania donovani (7), and the diplomonad parasite Giardia lamblia (8). In mouse, polyamines are essential for early embryo development (9, 10), and they are also essential for seed development in the flowering plant Arabidopsis thaliana (11).The universal distribution of polyamines suggests that...

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Section: Resultsmentioning

confidence: 99%

Evolution and Multifarious Horizontal Transfer of an Alternative Biosynthetic Pathway for the Alternative Polyamine sym-Homospermidine

Shaw

Elliott

Kinch

et al. 2010

Journal of Biological Chemistry

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“…These results indicated that the enzyme was a dimer, similar to the A. tumefaciens enzyme (78 kDa) (7). This was in contrast to the hexameric enzyme (260 kDa) isolated from G. stearothermophilus (14). However, the A. denitrificans enzyme eluted as hexamer (240 kDa) when a high concentration of L-lysine (10 mM) was supplemented to both the enzyme solution and the elution buffer.…”

Section: Molecular Mass Subunit Structure and Amino Acid Sequencesmentioning

confidence: 47%

“…Our observations suggest that the dimeric enzyme itself exhibits catalytic activity and it assembled into a hexamer in the presence of L-lysine. Previous studies indicated that the A. tumefaciens enzyme associated as a tetramer (160 kDa) in the presence of L-lysine (7) and the G. stearothermophilus enzyme was a hexamer in the absence of L-lysine (14). Thus, the A. denitrificans enzyme is different from the A. tumefaciens enzyme and the G. stearothermophilus enzyme.…”

Section: Molecular Mass Subunit Structure and Amino Acid Sequencesmentioning

confidence: 88%

“…Moreover, the stability of the A. denitrificans enzyme incubated with L-lysine is near that of the G. stearothermophilus enzyme. The enzyme showed the maximum activity at approximately pH 9.7 and 50 o C. The optimum pH of the A. denitrificans enzyme (pH 9.7) is similar to that of the enzyme from A. tumefaciens (pH 9.7) (7) and G. stearothermophilus (pH 10.0) (14). http://bmbreports.org …”

Section: Effects Of Ph and Temperature On Stability And Activity Of Tmentioning

confidence: 94%

“…Among microbial Lys 6-DH enzymes, the A. tumefaciens protein has been extensively studied (4,6,(7)(8)(9)(10) and is presently used for L-lysine determination (11,12) and P-6-C preparation (13). Recently, Geobacillus stearothermophilus was characterized as a lysine dehydrogenase producer (14). However, it is not clear which amino group of L-lysine the enzyme cleaves because the reaction product from L-lysine was only examined via thin layer chromatography.…”

Section: Introductionmentioning

confidence: 99%

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