Purification and Properties of an Extracellular Aminopeptidase from Streptomyces Lividans 1326

Aphale, Jayant S.; Strohl, William R.

doi:10.1099/00221287-139-3-417

Cited by 34 publications

(16 citation statements)

References 26 publications

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“…Some other aminopeptidase inhibitors containing a 3-amino-2-hydroxy-carboxylic acid also inhibit bacterial aminopeptidases: Wilkes and Prescott (1985) have shown that bestatin and amastatin are slow-and tight-binding competitive inhibitors of leucine aminopeptidase from A. proteolytica. Leucine aminopeptidase from S. rimosus is also inhibited by bestatin and amastatin (Vitale et al 1986), and leucine aminopeptidase from S. lividans is inhibited by bestatin (Aphale and Strohl 1993). All these aminopeptidases belong to the PepA-type metallo-aminopeptidases that appear to be quite common in Streptomyces species (Gonzales and Robert-Baudouy 1996).…”

Section: Discussionmentioning

confidence: 97%

Lapstatin, a new aminopeptidase inhibitor produced by Streptomyces rimosus , inhibits autogenous aminopeptidases

Lampret¹,

Kidrič

Kralj³

et al. 1999

Archives of Microbiology

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Lapstatin, a low-molecular-weight aminopeptidase inhibitor, was purified to homogeneity from Streptomyces rimosus culture filtrates. The purification procedure included extraction with methanol, followed by chromatography on Dowex 50WX4, AG50WX4, and HPLC RP C18 columns. By amino acid analysis, mass spectrometry, and NMR spectroscopy, the structure of lapstatin was shown to be 3-amino-2-hydroxy-4-methylpentanoylvaline. Lapstatin inhibited the extracellular leucine aminopeptidases from Streptomyces rimosus, Streptomyces griseus, and Aeromonas proteolytica with an IC50 in the range of 0.3-2.4 microM. IC50 values for other enzymes tested were at least tenfold higher. Leucine aminopeptidase from Streptomyces griseus was inhibited in a competitive manner, with an inhibition constant of 5 x 10(-7) M. Lapstatin is the first low-molecular-weight compound isolated from streptomycetes shown to inhibit an autogenous aminopeptidase.

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Section: Discussionmentioning

confidence: 97%

Lapstatin, a new aminopeptidase inhibitor produced by Streptomyces rimosus , inhibits autogenous aminopeptidases

Lampret¹,

Kidrič

Kralj³

et al. 1999

Archives of Microbiology

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“…A variety of LAP from many tissues and organisms has been isolated and characterized (Van Wart 1981;Taylor 1993). The enzymes were obtained from Escherichia coli (Vogt 1970), Aeromonas proteolytica (Pre-scott and Wilkes 1976), marine Pseudomonas (Merkel et al 1981;Qua et al 1981) and Streptomyces lividans (Aphale and Strohl 1993). Several genes encoding for bacterial LAP have been also isolated (Garcia-Alvarez et al 1991;Glaser et al 1991;Guenet et al 1992).…”

Section: Introductionmentioning

confidence: 99%

Purification and properties of an extracellular leucine aminopeptidase from the Bacillus sp. N2

Lee

Chun

Kho

et al. 1998

Journal of Applied Microbiology

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N. 1998. A halophilic bacterium was isolated from fermented anchovy sauce and identified as Bacillus species. An extracellular leucine aminopeptidase from Bacillus sp. N2 was purified to homogeneity using four successive purification steps. The enzyme has a native molecular mass of about 57 000 Da using FPLC gel filtration analysis and a molecular mass of 58 000 Da using SDS-polyacrylamide gel electrophoresis. This monomeric leucine aminopeptidase showed maximum enzyme activity at pH 9·5. The optimum temperature was 50°C when L-Leu-p-nitroanilide was the substrate. The leucine aminopeptidase was inactivated by 1,10-phenanthroline, dithiothreitol and sodium dodecyl sulphate. Enzyme activity was increased by addition of Co 2¦ . It is likely that Co 2¦ plays an important role in the catalysis or stability of the Bacillus sp. N2 leucine aminopeptidase. Sodium chloride (0-4·5 mol l −1 ) increased the hydrolytic activity towards L-Leu-p-nitroanilide. The N-terminal amino acid sequence was GluArg-Glu-Leu-Pro-Phe-Lys-Ala-Lys-His-Ala-Tyr-Ser-Thr-Ile. The purified enzyme had a high specificity for L-Leu-p-nitroanilide.

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“…Using a purification method consisting of seven steps, we homogeneously purified LAP from G. thermoleovorans 47b. Microbial LAP has been purified and characterized from A. sojae [4], A. oryzae [18], Aeromonas proteolytica [6], Alteromonas B-207 [16], B. stearothermophilus [11], Pseudomonas putida [7], Streptomyces griseus [2], S. peptidofaciens [24], S. rimosus [25] and S. lividans [1]. Gonzales and RobertBaudouty [5] summarized that almost half (47%) of the 102 bacterial aminopeptidases are monomers and the remainder (53%) display a multimeric structure.…”

Section: Discussionmentioning

confidence: 99%

Purification and characterization of hyperthermotolerant leucine aminopeptidase from Geobacillus thermoleovorans 47b

Deejing¹,

Yoshimune²,

Lumyong³

et al. 2005

J IND MICROBIOL BIOTECHNOL

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A thermophilic bacterium, which we designated as Geobacillus thermoleovorans 47b was isolated from a hot spring in Beppu, Oita Prefecture, Japan, on the basis of its ability to grow on bitter peptides as a sole carbon and nitrogen source. The cell-free extract from G. thermoleovorans 47b contained leucine aminopeptidase (LAP; EC 3.4.11.10), which was purified 164-fold to homogeneity in seven steps, using ammonium sulfate fractionation followed by the column chromatography using DEAE-Toyopearl, hydroxyapatite, MonoQ and Superdex 200 PC gel filtration, followed again by MonoQ and hydroxyapatite. The enzyme was a single polypeptide with a molecular mass of 42,977.2 Da, as determined by matrix-assisted laser desorption ionization and time-of-flight mass spectrometry, and was found to be thermostable at 90 degrees C for up to 1 h. Its optimal pH and temperature were observed to be 7.6-7.8 and 60 degrees C, respectively, and it had high activity towards the substrates Leu-p-nitroanilide (p-NA)(100%), Arg-p-NA (56.3%) and LeuGlyGly (486%). The K(m) and V(max) values for Leu-p-NA and LeuGlyGly were 0.658 mM and 25.0 mM and 236.2 micromol min(-1) mg(-1) protein and 1,149 micromol min(-1) mg(-1) protein, respectively. The turnover rate (k(cat)) and catalytic efficiency (k(cat)/ K(m)) for Leu-p-NA and LeuGlyGly were 10,179 s(-1) and 49,543 s(-1) and 15,470 mM(-1 ) s(-1) and 1981.7 mM(-1 ) s(-1), respectively. The enzyme was strongly inhibited by EDTA, 1,10-phenanthroline, dithiothreitol, beta-mercaptoethanol, iodoacetate and bestatin; and its apoenzyme was found to be reactivated by Co(2+) .

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Purification and Properties of an Extracellular Aminopeptidase from Streptomyces Lividans 1326

Cited by 34 publications

References 26 publications

Lapstatin, a new aminopeptidase inhibitor produced by Streptomyces rimosus , inhibits autogenous aminopeptidases

Lapstatin, a new aminopeptidase inhibitor produced by Streptomyces rimosus , inhibits autogenous aminopeptidases

Purification and properties of an extracellular leucine aminopeptidase from the Bacillus sp. N2

Purification and characterization of hyperthermotolerant leucine aminopeptidase from Geobacillus thermoleovorans 47b

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