Purification and characterization of a hydrophobic amino acid—specific endopeptidase from Halobacterium halobium S9 with potential application in debittering of protein hydrolysates

Capiralla, Hemachander; Hiroi, Tetsuya; Hirokawa, Takahiko; Maeda, Shuichi

doi:10.1016/s0032-9592(02)00180-2

Cited by 49 publications

(24 citation statements)

References 15 publications

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“…Capiralla et al [18] reported that op$ mum pH value for extracellular serine protease enzyme of Halobacterium halobium S9 was 8.0. Gimenez et al [19] showed that the extracellular protease enzyme which isolated from Natrialba magadii exhibited an op$ mum ac$ vity between pH 8.0-10.0.…”

Section: Resultsmentioning

confidence: 99%

Investigation of extracellular protease activity of two different extremely halophilic archaea isolated from raw hide

Kesbic¹,

Yapıcı²

2017

LFJ

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ABSTRACT. In this study, it was aimed to inves% gate extracellular protease ac% vity of two diff erent extremely halophilic archaea and to iden% fy them by using phenotypic and molecular methods. The haloarchaea were isolated from salted raw hide. The isolates were screened for produc% on of protease enzyme and two isolates which have extracellular protease ac% vity were selected. To iden% fy the halophilic archaea, Gram staining procedure, an% bio% c sensi% vity tests and molecular methods were used. According to the 16S r-RNA molecular iden% fi ca% on results, isolate 1 iden% fi ed as Halococcus morrhuae JCM 8876 with % 99 sequence similari% es and isolate 2 iden% fi ed as Natrinema pallidum 153 with % 99 sequence similari% es. Azocasein was used as a substrate for measuring their proteases ac% vi% es daily and their op% mum ac% vi% es were determined. H. morrhuae and N. pallidum showed daily maximum proteases ac% vity on 6 and 4 days, respec% vely. According to the results of the study, it was found that proteases ac% vity was related with diff erent reac% on condi% ons. These condi% ons were detected as diff erent pH (4-11), temperature (4-70°C) and NaCl concentra% on (1-5 M) values. H. morrhuae showed the maximum ac% vity at pH 9.5, 2M NaCl concentra% on and 50°C temperature and N. pallidum displayed the op% mum ac% vity at pH 9.0, 2M NaCl concentra% on and 40°C temperature. KEY WORDS: salted hide, haloarchaea, extracellular protease enzyme, 16S r-RNA sequence analysis. INVESTIGAREA ACTIVITĂȚII PROTEAZICE EXTRACELULARE A DOUĂ ARCHAEA EXTREM HALOFILE DIFERITE IZOLATE DE PE PIELEA CRUDĂREZUMAT. În acest studiu, s-a urmărit cercetarea ac% vității proteazice extracelulare a două archaea extrem halofi le diferite și iden% fi carea acestora u% lizând metode feno% pice și moleculare. Haloarchaea au fost izolate de pe pielea crudă sărată. Izolatele au fost evaluate în funcție de capacitatea de producere a enzimelor de % p protează și s-au selectat două izolate care au ac% vitate proteazică extracelulară. Pentru a iden% fi ca archaea halofi le, s-au u% lizat proceduri de colorare Gram, teste de sensibilitate la an% bio% ce și metode moleculare. Conform rezultatelor iden% fi cării moleculare ARNr 16S, izolatul 1 a fost iden% fi cat ca Halococcus morrhuae JCM 8876 cu asemănări de secvență de 99%, iar izolatul 2 a fost iden% fi cat ca Natrinema pallidum 153 cu asemănări de secvență de 99%. S-a u% lizat azocaseina ca substrat pentru măsurarea zilnică a ac% vității proteazice și s-a determinat ac% vitatea op% mă a acestora. H. morrhuae și N. pallidum au prezentat ac% vitate proteazică maximă zilnică la 6, respec% v 4 zile. Conform rezultatelor studiului, s-a constatat că ac% vitatea proteazică a fost corelată cu diferite condiții de reacție. Aceste condiții au constat în valori diferite ale pH-ului (4-11), ale temperaturii (4-70°C) și ale concentrației de NaCl (1-5 M). H. morrhuae a prezentat ac% vitate maximă la pH 9,5, la concentrația de NaCI de 2M și la temperatura de 50°C, iar N. pallidum a prezentat ac% vitate op% m...

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Section: Resultsmentioning

confidence: 99%

Investigation of extracellular protease activity of two different extremely halophilic archaea isolated from raw hide

Kesbic¹,

Yapıcı²

2017

LFJ

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“…In general terms, haloarchaeal proteases show optimum activity at high salt concentration, although some of them can be stable and active at lower concentrations. The extracellular endopeptidase from Halobacterium halobium hydrolyzes polypeptides and oligopeptides with specificity for hydrophobic amino acids, especially proline [65]. The enzyme exhibits azocasein activity at low salt concentrations.…”

Section: Proteasesmentioning

confidence: 99%

“…The enzyme exhibits azocasein activity at low salt concentrations. This endopeptidase could be an important tool to be used in the food processing industry as well as in biomedical applications to produce peptides [65,66].…”

Section: Proteasesmentioning

confidence: 99%

Biocompounds from Haloarchaea and Their Uses in Biotechnology

Torregrosa-Crespo¹,

Galiana²,

MaríaMartínez-Espinosa³

2017

Archaea - New Biocatalysts, Novel Pharmaceuticals and Various Biotechnological Applications

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New advances in the understanding of haloarchaea physiology, metabolism, biochemistry, and molecular biology show that these kinds of microorganisms produce several compounds in response to the extreme conditions of their ecosystems. Thus, the complete metabolic and genetic machineries are fully adapted to nutrient starvation, high sun radiation, and high ionic strength. Due to these adaptations, some of the primary and secondary metabolites produced by haloarchaea are of high interest in terms of potential biotechnological uses. The principal goal of the chapter is to present a review about the main characteristics of these biocompounds and their potential uses in biomedicine, pharmacy, and industry.

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“…Furthermore, these enzymes are used in pharmaceuticals, medical diagnosis, and decomposition of gelatin on X-ray films as well as in textiles (Tunga, Shrivastava, & Banerjee, 2003). Although most available reports on proteases are derived from microbial origin (Tang et al, 2003;Soares, Castilho, Bon, & Freire, 2004;Ire, Okolo, & Moneke, 2011) but commercial use of proteases with different origins for hydrolysis of proteins was reported to be very promising due to the biological origin of enzymes (Capiralla, Hiroi, Hirokawa, & Maeda, 2002). Plant cysteine proteases suchas papain and stem bromelain are still extensively used for medicine, brewing wine, and food industry (Kaneda, Yonezawa, & Uchikoba, 1997).…”

Section: Introductionmentioning

confidence: 99%

Partial Characterization of Protease from the Leaves of Jatropha curcas

Ibrahim¹,

Olatominwa²,

Aliyu³

et al. 2012

IJB

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The crucial roles of proteases in food and other industries stimulate research to find additional sources of the enzyme especially from non-conventional sources. In this study, protease was partially characterized from Jatropha curcas leaves.The enzyme had pH and temperature optima of 4.0 and 45 o C respectively. A decline in residual activity of the enzyme was observed at above 45 o C and the activation energy (Ea) from Arrhenius plot was 0.57kJ/mol. The enzyme showed specificity in the order; casein>hemoglobin> albumin> ovalbumin. Initial velocity studies for the determination of kinetic parameters revealed a K M and V max of 0.48 mg/ml and 0.014µmol/min respectively with a computed index of physiological efficiency (K cat ) of 0.029 min -1 . Furthermore, Dixon-Webb's plot identified ionizable groups at the active site with pKa 1 and pKa 2 of 5.0 and 5.3 respectively as well as an enthalpy of ionization of 0.047 kcal/mol implicating aspartate as an important amino acid at the active site of the enzyme. The protease was highly sensitive to cysteine protease specific inhibitor iodoacetate while 1,10phenanthroline and ethylenediaminetetraacetate slightly inhibited the enzyme. Data from this study suggest that J. curcas protease possesses closely similar properties to other known industrial proteases.

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Purification and characterization of a hydrophobic amino acid—specific endopeptidase from Halobacterium halobium S9 with potential application in debittering of protein hydrolysates

Cited by 49 publications

References 15 publications

Investigation of extracellular protease activity of two different extremely halophilic archaea isolated from raw hide

Investigation of extracellular protease activity of two different extremely halophilic archaea isolated from raw hide

Biocompounds from Haloarchaea and Their Uses in Biotechnology

Partial Characterization of Protease from the Leaves of Jatropha curcas

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