Characterization of thermostable native alkaline phosphatase from an aerobic hyperthermophilic archaeon, Aeropyrum pernix K1

Helianti, Is; Okubo, Takako; Morita, Yasutaka; Tamiya, Eiichi

doi:10.1007/s00253-006-0640-y

Cited by 12 publications

(10 citation statements)

References 24 publications

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“…The isolate NSW13-2 was provided as a potential ALP-producing strain and capable of growing in the pH range of 7-8 with maximum growth at pH 7.0 that will be useful for further study. Several ALPs have been investigated from the strains of bacteria and fungi such as the hyperthermophilic archaeon Pyrococcus abyssi (Zappa et al, 2001); the thermophile Thermotoga neapolitana and Thermus thermophilus (Pantazaki et al, 1998), the hyperthermophilic archaea, Aeropyrum pernix (Helianti et al, 2007), Bacillus sphaericus (Dhaked et al, 2005), Bacillus sp. (Mahesh et al, 2010) and E.coli EFRL 13 (Qureshi et al, 2010), Bacillus licheniformis MTCC1483 (Pandey & Banik, 2011), Rhizopus icrospores var.…”

Section: Isolation and Screening Of Alp-producing Halophillic Bacteriamentioning

confidence: 99%

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Characterization of Alkaline Phosphatase Producing Bacteria Isolated from Thai Fermented Fish Products

Sitdhipol¹,

Niwasabutra²,

Wannissorn³

et al. 2012

IJB

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Seventeen isolates of alkaline phosphatase (ALP) producing bacteria were screened and systematically studied. They were divided into 10 groups on the basis of their phenotypic characteristics and 16S rRNA gene sequence analyses. The Gram-positive coccal isolates in Group I (3 isolates), Group II (3 isolates) and Group III (1 isolate) showed 99.2%, 99.5-99.7% and 99.8% sequences similarity to Staphylococcus saprophyticus, S. napalensis and S. sciuri, respectively. Each Gram-positive rod-shaped isolates in Group IV and V belonged to the genus Bacillus and was closely related to B. vietnamensis and B. safensis with 99.2% and 99.4% sequences similarity, respectively. The Gram-positive, moderately halophilic rod-shaped bacteria in Group VI (3 isolates), Group VII (1 isolate), Group VIII (1 isolate) and Group IX (2 isolates) were closely related to Virgibacillus halodenitrificans (98.5-99.1%), Oceanobacillus iheyensis (99.3%), Halobacillus mangrove (97.9%) and H. dabanensis (98.5-98.6%), respectively. One Gram-negative isolate of moderately halophilic bacterium (Group X) was closely related to Idiomarina zobelli (98.0%). They possessed phosphatase activities ranged from 10.08-70.96 U ml-1 . The isolate NSW 13-2 (Group VI) showed the highest ALP.

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Section: Isolation and Screening Of Alp-producing Halophillic Bacteriamentioning

confidence: 99%

“…ALP activity assay was done by method described by Helianti et al (2007). Reaction mixture composing of 1.0 ml of 10 mM p-nitrophenylphosphate (pNPP) (Sigma) in 0.2 M Tris-HCl buffer pH 10.0 with 5 mM MgCl 2 , and 0.1 ml of the crude enzyme was incubated at 37 o C for 15 min.…”

Section: Screening Of Alkaline Phosphatase (Alp) Producing Bacteriamentioning

confidence: 99%

Characterization of Alkaline Phosphatase Producing Bacteria Isolated from Thai Fermented Fish Products

Sitdhipol¹,

Niwasabutra²,

Wannissorn³

et al. 2012

IJB

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“…3a). This clearly distinguished Gtd AP from other thermostable APs, which only retained about 8-20% of their maximal activity at the neutral pH [29,30]. As shown in Fig.…”

Section: Activity Of the G Thermodenitrificans T-2 Apmentioning

confidence: 91%

“…Further analysis showed that Gtd AP for total thermal inactivation was only 15 min by a heat treatment of 80°C and the half-life value at 70°C was 8 min (Fig. 3c), which is less stable than its counterparts in various thermophiles [9,22,29,30,[39][40][41][42]. For example, Thermus caldophilus retained 80% activity after incubation at 80°C for 12 h [39] and even E. coli AP exhibited a half-life value at 80°C more than 6 h [8].…”

Section: Activity Of the G Thermodenitrificans T-2 Apmentioning

confidence: 99%

A Moderately Thermostable Alkaline Phosphatase from Geobacillus thermodenitrificans T2: Cloning, Expression and Biochemical Characterization

Zhang

et al. 2008

Appl Biochem Biotechnol

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A gene-encoding alkaline phosphatase (AP) from thermophilic Geobacillus thermodenitrificans T2, termed Gtd AP, was cloned and sequenced. The deduced Gtd AP protein comprises 424 amino acids and shares a low homology with other known AP (<35% identity), while it exhibits the conservation of the active site and structure element of Escherichia coli AP. The Gtd AP protein, without a predicted signal peptide of 30 amino acids, was successfully overexpressed in E. coli and purified as a hexa-His-tagged fusion protein. The pH and temperature optima for purified enzyme are 9.0 and 65 degrees C, respectively. The enzyme retained a high activity at 45-60 degrees C, while it could be quickly inactivated by a heat treatment at 80 degrees C for 15 min, exhibiting a half-life of 8 min at 70 degrees C. The K(m) and V (max) for pNPP were determined to be 31.5 muM and 430 muM/min at optimal conditions. A divalent cation is essential, with a combination of Mg2+ and Co2+ or Zn2+ preferred. The enzyme was strongly inhibited by 10 mM ethylenediaminetetraacetic acid (EDTA) and vanadate but highly resistant to urea and dithiothreitol. The properties of Gtd AP make it suitable for application in molecular cloning or amplification.

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“…(Murakawa et al 2002) and Shewanella sp. SIB1 (Suzuki et al 2005), the aerobic hyperthermophilic archaeon Aeropyrum pernix (Helianti et al 2007) and the hyperthermophilic archaeon Pyrococcus abyssi (Zappa et al 2001) have been investigated. The activities of these enzymes are in the range 427-3,707 U/mg.…”

Section: Introductionmentioning

confidence: 99%

Cloning and expression of a highly active recombinant alkaline phosphatase from psychrotrophic Cobetia marina

Nasu

Ichiyanagi²,

Gomi³

2011

Biotechnol Lett

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Alkaline phosphatase catalyzes the hydrolysis of phosphomonoesters and is widely used in molecular biology techniques and clinical diagnostics. We expressed a recombinant alkaline phosphatase of the marine bacterium, Cobetia marina, in Escherichia coli BL21 (DE3). The recombinant protein was purified with a specific activity of 12,700 U/mg protein, which is the highest activity reported of any bacterial alkaline phosphatase studied to date. The molecular mass of the recombinant protein was 55-60 kDa, as determined by SDS-PAGE, and was observed to be a dimer by gel filtration analysis. The enzyme was optimally active at 45°C and the recombinant alkaline phosphatase efficiently hydrolyzed a phosphoric acid ester in luminescent and fluorescent substrates. Therefore, this enzyme can be considered to be extremely useful as a label conjugated to an antibody.

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Characterization of thermostable native alkaline phosphatase from an aerobic hyperthermophilic archaeon, Aeropyrum pernix K1

Cited by 12 publications

References 24 publications

Characterization of Alkaline Phosphatase Producing Bacteria Isolated from Thai Fermented Fish Products

Characterization of Alkaline Phosphatase Producing Bacteria Isolated from Thai Fermented Fish Products

A Moderately Thermostable Alkaline Phosphatase from Geobacillus thermodenitrificans T2: Cloning, Expression and Biochemical Characterization

Cloning and expression of a highly active recombinant alkaline phosphatase from psychrotrophic Cobetia marina

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