Purification and characterization of a novel metalloprotease isolated from Aeromonas caviae

Nakasone, Noboru; Toma, Claudia; Song, Tianyan; Iwanaga, Masaaki

doi:10.1016/j.femsle.2004.06.025

Cited by 7 publications

(7 citation statements)

References 33 publications

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“…Other metalloproteases had been described in Aeromonas spp. For instance, Aeromonas caviae Ae6 metalloprotease was optimally active at pH 7.5 (Nakasone et al 2004), and a gelatinolytic metalloprotease from A. salmonicida showed optimal activity at 40°C and pH 7.5 (Arnesen et al 1995), which are different from that described here for protease K12. The enzyme characteristics of A. hydrophila strain K12 resembles that of some Bacillus species (Werlang and Brandelli 2005;Corrêa et al 2010) and is also similar to the Gram-negative Chryseobacterium sp.…”

Section: Resultscontrasting

confidence: 92%

Production and properties of keratinolytic proteases from three novel Gram-negative feather-degrading bacteria isolated from Brazilian soils

et al. 2011

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The keratinolytic potential and protease properties of three novel Gram-negative feather-degrading bacteria isolated from Brazilian soils was described. Aeromonas hydrophila K12, Chryseobacterium indologenes A22 and Serratia marcescens P3 were able to degrade feather meal, producing high amounts of soluble proteins and forming thiol groups. The proteases of strains K12, A22 and P3 had optimal pH of 8.0, 7.5 and 6.0, respectively; this last is an uncommon feature for bacterial keratinases. The optimal temperature was in the range 45-55°C. All three proteases were active towards azokeratin and were inhibited by EDTA, suggesting that they are keratinolytic metalloproteases. The proteolytic activity of K12 was stimulated by organic solvents and the detergent SDS, suggesting its potential application for detergent formulations and peptide synthesis. Strains A22, K12 and P3 have great potential for use in biotechnological processes involving hydrolysis of keratinous byproducts.

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Section: Resultscontrasting

confidence: 92%

Production and properties of keratinolytic proteases from three novel Gram-negative feather-degrading bacteria isolated from Brazilian soils

et al. 2011

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“…The size of the mature protein is in agreement with results from gel filtration chromatography and SDS-PAGE analysis (9) describing a protease of approximately 20 kDa and composed of a single polypeptide. The molecular mass of the mature AsaP1 is also in accordance with what has been described for the AP19 metallopeptidase of Aeromonas caviae (21) and the peptidyl-Lys metalloendopeptidases of Grifola frondosa and Pleurotus ostreatus, respectively (23). The AsaP1 sequence was found to be 91% identical to the sequence of A. hydrophila extracellular protease (Table 3), and Chang et al (3) reported that the A. hydrophila extracellular protease is detected as a 29-kDa protein in casein SDS-PAGE.…”

Section: Discussionsupporting

confidence: 85%

The AsaP1 Peptidase of Aeromonas salmonicida subsp. achromogenes Is a Highly Conserved Deuterolysin Metalloprotease (Family M35) and a Major Virulence Factor

et al. 2009

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Infections by the bacterium Aeromonas salmonicida subsp. achromogenes cause significant disease in a number of fish species. In this study, we showed that AsaP1, a toxic 19-kDa metallopeptidase produced by A. salmonicida subsp. achromogenes, belongs to the group of extracellular peptidases (Aeromonas type) (MEROPS ID M35.003) of the deuterolysin family of zinc-dependent aspzincin endopeptidases. The structural gene of AsaP1 was sequenced and found to be highly conserved among gram-negative bacteria. An isogenic ⌬asaP1 A. salmonicida subsp. achromogenes strain was constructed, and its ability to infect fish was compared with that of the wild-type (wt) strain. The ⌬asaP1 strain was found to infect Arctic charr, Atlantic salmon, and Atlantic cod, but its virulence was decreased relative to that of the wt strain. The 50% lethal dose of the AsaP1 mutant was 10-fold higher in charr and 5-fold higher in salmon than that of the wt strain. The pathology induced by the AsaP1-deficient strain was also different from that of the wt strain. Furthermore, the mutant established significant bacterial colonization in all observed organs without any signs of a host response in the infected tissue. AsaP1 is therefore the first member of the M35 family that has been shown to be a bacterial virulence factor.

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“…This property of the enzyme proved the role of Fe 2+ ions as cofactor of this enzyme. Fe‐metalloproteases were also reported for Aeromonas caviae and Chryseobacterium gleum . The inhibitory effect of some metal ions may be due to attachment with SH groups of Cys‐X breaking SS bridges thus changing the conformation of enzyme or may be due to the replacement with metals originally present in the enzyme structure …”

Section: Resultsmentioning

confidence: 88%

The biotechnological potential of subtilisin‐like fibrinolytic enzyme from a newly isolated Lactobacillus plantarum KSK‐II in blood destaining and antimicrobials

Kotb

2014

Biotechnology Progress

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An antimicrobial oxidative- and SDS-stable fibrinolytic alkaline protease designated as KSK-II was produced by Lactobacillus plantarum KSK-II isolated from kishk, a traditional Egyptian food. Maximum enzyme productivity was obtained in medium containing 1% lactose and 0.5% soybean flour as carbon and nitrogen sources, respectively. Purification of enzyme increased its specific activity to 1,140-fold with a recovery of 33% and molecular weight of 43.6 kDa. Enzyme activity was totally lost in the presence of ethylenediaminetetraacetic acid and was restored after addition of Fe(2+) suggesting that KSK-II is a metalloprotease and Fe(2+) acts as cofactor. Enzyme hydrolyzed not only the natural proteins but also synthetic substrates, particularly Suc-Ala-Ala-Pro-Phe-pNA. KSK-II can hydrolyze the Lys-X easier than Arg-X; thus, it was considered as a subtilisin-family protease. Its apparent Km , Vmax , and Kcat were 0.41 mM, 6.4 µmol mg(-1) min(-1) , and 28.0 s(-1) , respectively. KSK-II is industrially important from the perspectives of its maximal activity at 50°C (stable up to 70°C), ability to function at alkaline pH (10.0), stability at broad pH ranges (7.5-12.0) in addition to its stability toward SDS, H2 O2 , organic solvents, and detergents. We emphasize for the first time the potential of fibrinolytic activity for alkaline proteases used in detergents especially in blood destaining.

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Purification and characterization of a novel metalloprotease isolated from Aeromonas caviae

Cited by 7 publications

References 33 publications

Production and properties of keratinolytic proteases from three novel Gram-negative feather-degrading bacteria isolated from Brazilian soils

Production and properties of keratinolytic proteases from three novel Gram-negative feather-degrading bacteria isolated from Brazilian soils

The AsaP1 Peptidase of Aeromonas salmonicida subsp. achromogenes Is a Highly Conserved Deuterolysin Metalloprotease (Family M35) and a Major Virulence Factor

The biotechnological potential of subtilisin‐like fibrinolytic enzyme from a newly isolated Lactobacillus plantarum KSK‐II in blood destaining and antimicrobials

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