Thermostable aminopeptidase from <i>Pyrococcus horikoshii</i>

Ando, Susumu; Ishikawa, Kazuhiko; Ishida, Hiroyasu; Kawarabayasi, Yutaka; Kikuchi, Hisasi; Kosugi, Yoshitsugu

doi:10.1016/s0014-5793(99)00257-4

Cited by 37 publications

(43 citation statements)

References 14 publications

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“…4). Its aminopeptidase activity stops at the residue before the first proline in the peptide, which was also observed for the related deblocking aminopeptidase homolog in archaea (1). This may be due to the unique imide bond in proline, which restricts the overall N-terminal conformation of the oligopeptides and disrupts the contacts between residues and active sites of the enzyme.…”

Section: Fig 2 Genomic Region In Ementioning

confidence: 62%

“…YpdE in E. coli shows moderate similarity (ϳ24% identity) to the previously reported metal ion-dependent deblocking aminopeptidase (PH0519) in the archaeon Pyrococcus horikoshii (1,17) and to an aminopeptidase in Haloarcula marismortui (ϳ21% identity in an ϳ150-aa region) (8). It is known that PH0519 shows broad aminopeptidase activity on nonblocked peptides and blocked peptides by acyl group (1).…”

Section: Fig 2 Genomic Region In Ementioning

confidence: 87%

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Characterization of Two New Aminopeptidases in Escherichia coli

et al. 2005

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Two genes in the Escherichia coli genome, ypdE and ypdF, have been cloned and expressed, and their products have been purified. YpdF is shown to be a metalloenzyme with Xaa-Pro aminopeptidase activity and limited methionine aminopeptidase activity. Genes homologous to ypdF are widely distributed in bacterial species. The unique feature in the sequences of the products of these genes is a conserved C-terminal domain and a variable N-terminal domain. Full or partial deletion of the N terminus in YpdF leads to the loss of enzymatic activity. The conserved C-terminal domain is homologous to that of the methionyl aminopeptidase (encoded by map) in E. coli. However, YpdF and Map differ in their preference for the amino acid next to the initial methionine in the peptide substrates. The implication of this difference is discussed. ypdE is the immediate downstream gene of ypdF, and its start codon overlaps with the stop codon of ypdF by 1 base. YpdE is shown to be a metalloaminopeptidase and has a broad exoaminopeptidase activity.

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Section: Fig 2 Genomic Region In Ementioning

confidence: 62%

Section: Fig 2 Genomic Region In Ementioning

confidence: 87%

Characterization of Two New Aminopeptidases in Escherichia coli

et al. 2005

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“…Pyrococcus horikoshii OT3 is one of those organisms with the optimum growth temperature above 95°C (4). Based on its complete published genome sequence (8,9), we have expressed some of the enzymes from this organism and characterized them, most of which have proven to be highly thermostable (1,7). It is quite likely, therefore, that other useful enzymes that are active and stable at extremely high temperatures remain to be found.…”

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confidence: 99%

Hyperthermostable Endoglucanase from Pyrococcus horikoshii

Ando

Ishida

Kosugi

et al. 2002

Appl Environ Microbiol

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116

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An endoglucanase homolog from the hyperthermophilic archaeon Pyrococcus horikoshii was expressed in Escherichia coli, and its enzymatic characteristics were examined. The expressed protein was a hyperthermostable endoglucanase which hydrolyzes celluloses, including Avicel and carboxymethyl cellulose, as well as ␤-glucose oligomers. This enzyme is the first endoglucanase belonging to glycosidase family 5 found from Pyrococcus species and is also the first hyperthermostable endoglucanase to which celluloses are the best substrates. This enzyme is expected to be useful for industrial hydrolysis of cellulose at high temperatures, particularly in biopolishing of cotton products.In the textile industry, cellulases have been used in large quantity for biopolishing of cotton products. This process is essential for removing fuzz and giving a soft touch and clean appearance to the fabrics. The enzymes that are presently used for this purpose are mesophilic cellulases from fungi, and their optimum reaction temperatures are between 50 and 55°C. If they are replaced by a hyperthermostable enzyme with an optimum temperature close to 100°C, which will make it possible to treat cotton products in steam, the processing will be much more simple, quick, and efficient than in the presently employed method. Desizing, the step to remove starch from the fabrics, is performed at temperatures at least 70°C, and higher temperatures are preferred. Because amylases active at these temperatures are available, this process is performed at temperatures higher than 70°C. However, cellulase treatment, which usually follows desizing, is performed at lower temperatures, since a cellulase that is active and stable in this temperature range has not been available. If such a hyperthermostable cellulase is introduced, it will be possible to combine desizing and biopolishing in a single step.For the purpose of producing such a hyperthermostable cellulase, we investigated the possibility of utilizing the genetic resources of hyperthermophilic archaea. Pyrococcus horikoshii

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“…A new aminopeptidase from P. horikoshii has been identi®ed recently by Ando and coworkers (gene PH0519;Ando et al, 1999). They have shown that this enzyme has an aminopeptidase activity and cleaves the N-terminal amino acid from a variety of peptide substrates and that it also has a putative acylamino-acid-releasing (deblocking) activity for acyl (blocked) peptides.…”

Section: Introductionmentioning

confidence: 99%

Expression, purification, crystallization and preliminary crystallographic analysis of a deblocking aminopeptidase fromPyrococcus horikoshii

et al. 2005

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The deblocking aminopeptidase (DAP) of Pyrococcus horikoshii is a hyperthermophilic exoprotease that cleaves the N-terminal amino acid of peptide substrates with a putative deblocking activity for acylated peptides. DAP has been found to be homologous to a tetrahedral aminopeptidase from the halophilic Haloarcula marismortui. The latter enzyme is a dodecameric complex and has been revealed to be a self-compartmentalized protease whose central cavity harbouring the catalytic site is accessible through several channels of different size, unlike all other known proteolytic complexes. Three paralogues of DAP have been identi®ed in P. horikoshii, with about 40% identity between them. Each of them has been overexpressed in Escherichia coli, puri®ed and crystallized in the native and selenomethionine-substituted states. The results indicate that they form two kinds of assemblies, of 12 and of 24 subunits, with a molecular weight of $400 and $800 kDa, respectively. Crystals of the different variants of DAP and in their different oligomeric states diffract up to a resolution of 3 A Ê .

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Thermostable aminopeptidase from Pyrococcus horikoshii

Cited by 37 publications

References 14 publications

Characterization of Two New Aminopeptidases in Escherichia coli

Characterization of Two New Aminopeptidases in Escherichia coli

Hyperthermostable Endoglucanase from Pyrococcus horikoshii

Expression, purification, crystallization and preliminary crystallographic analysis of a deblocking aminopeptidase fromPyrococcus horikoshii

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