Characterization and stability of extracellular alkaline proteases from halophilic and alkaliphilic bacteria isolated from saline habitat of coastal Gujarat, India

Dodia, M. S.; Joshi, Rupal H.; Patel, Rajesh; Singh, Satya P.

doi:10.1590/s1517-83822006000300015

Cited by 50 publications

(24 citation statements)

References 28 publications

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“…The optimum production (190 U/mL) was evident during stationary phase at 56-60 h, suggesting the prominent role of this enzyme in both, primary metabolisms and ecological sustenance. Alkaline proteases from other haloalkaliphilic bacteria studied in our group, reflected the optimum production during the late exponential phase and beginning of the stationary phase [22][23][24]. Interestingly, the enzyme production from S-20-9 was rapidly declined after reaching to optimal level; nevertheless, the production was quite stable up to 104 h (Fig.…”

Section: Growth Kinetics and Protease Productionmentioning

confidence: 76%

“…So far, alkaline proteases from halophiles and haloalkaliphiles have been relatively less explored compared to alkaliphiles. However, as revealed from the recent literature [14][15][16][17][18][19][20] and our own studies [21][22][23][24][25], the proteases are wide spread among the haloalkaliphiles and many of them possess the potential that can be novel for biotechnological applications. In order to obtain commercially viable yields, it is essential to optimize fermentation media for the growth and protease production.…”

Section: Introductionmentioning

confidence: 99%

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Production and optimization of a commercially viable alkaline protease from a haloalkaliphilic bacterium

Joshi

Dodia

Singh

2008

Biotechnol Bioproc E

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Twenty five haloalkaliphilic bacterial strains were isolated from sea water along the Coastal Gujarat (India) and screened for their ability to secret alkaline proteases. Among them, a potent strain S-20-9 (GenBank accession number EU118360), resembling to Halophilic Bacterium MBIC3303 on the basis of 16S rRNA gene sequencing, was selected for the optimization of enzyme production. S-20-9 produced protease optimally, under aerobic conditions during mid-stationary phase over a broad range of salt (5~25%, w/v) and pH (7~10). The optimum production was at pH 9 and 15% (w/v) NaCl. The production was suppressed by lactose, maltose, sucrose, and inorganic nitrogen sources, especially ammonium ions. Further, the production was significantly stimulated by KH 2 PO 4 and suppressed by glucose. Similarly, the production was also suppressed at higher concentrations of gelatin, yeast extract, peptone, and casamino acids, indicating towards a threshold value for nitrogen requirement. The growth and protease production were enhanced by mono-valent cation (KCl), while the divalent cations acted as inhibitors. The study holds significance as only few reports are available on the alkaline proteases from haloalkaliphilic bacteria, particularly those from moderate saline habitats. © KSBB hÉóïçêÇëW=Ü~äç~äâ~äáéÜáäáÅ=Ä~ÅíÉêá~I=~äâ~äáåÉ=éêçíÉ~ëÉI=éêçÇìÅíáçåI=çéíáãáò~íáçåI=ëÉ~=ï~íÉê= = = = =

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Section: Growth Kinetics and Protease Productionmentioning

confidence: 76%

Section: Introductionmentioning

confidence: 99%

Production and optimization of a commercially viable alkaline protease from a haloalkaliphilic bacterium

Joshi

Dodia

Singh

2008

Biotechnol Bioproc E

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“…2). Alkaline proteases from some other organisms have also displayed the similar pH range; 9-11 [2,9,16,40]. However, the studies on pH stability of AH-6 protease demonstrated a relatively different pattern than the activity profile.…”

Section: Enzyme Purification By Hydrophobic Interaction Chromatographymentioning

confidence: 99%

“…While proteolytic enzymes are reported from halo-neutrophilic bacteria, alkaline proteases from haloalkaliphiles have rarely been investigated [12,17,22,41,51]. We have been involved with the screening and characterization of alkaline proteases from haloalkaliphiles [9,19,27,39,40]. Enzymes derived from these bacteria are potential biocatalysts due to their optimal activities under the conditions of high salt, surfactants, high temperature and alkaline pH [34], whereas the mesophilic counter parts fail to act under similar conditions [55].…”

Section: Introductionmentioning

confidence: 99%

Purification and stability characteristics of an alkaline serine protease from a newly isolated Haloalkaliphilic bacterium sp. AH-6

Dodia¹,

Rawal²,

Bhimani³

et al. 2007

J Ind Microbiol Biotechnol

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An alkaline protease secreting Haloalkaliphilic bacterium (Gene bank accession number EU118361) was isolated from the Saurashtra Coast in Western India. The alkaline protease was purified by a single step chromatography on phenyl sepharose 6 FF with 28% yield. The molecular mass was 40 kDa as judged by SDS-PAGE. The enzyme displayed catalysis and stability over pH 8-13, optimally at 9-11. It was stable with 0-4 M NaCl and required 150 mM NaCl for optimum catalysis at 37 degrees C; however, the salt requirement for optimal catalysis increased with temperature. While crude enzyme was active at 25-80 degrees C (optimum at 50 degrees C), the purified enzyme had temperature optimum at 37 degrees C, which shifted to 80 degrees C in the presence of 2 M NaCl. The NaCl not only shifted the temperature profile but also enhanced the substrate affinity of the enzyme as reflected by the increase in the catalytic constant (K(cat)). The enzyme was also calcium dependent and with 2 mM Ca(+2), the activity reached to maximum at 50 degrees C. The crude enzyme was highly thermostable (37-90 degrees C); however, the purified enzyme lost its stability above 50 degrees C and its half life was enhanced by 30 and sevenfold at 60 degrees C with 1 M NaCl and 50 mM Ca(+2), respectively. The activity of the enzyme was inhibited by PMSF, indicating its serine type. While the activity was slightly enhanced by Tween-80 (0.2%) and Triton X-100 (0.05%), it marginally decreased with SDS. In addition, the enzyme was highly stable with oxidizing-reducing agents and commercial detergents and was affected by metal ions to varying extent. The study assumes significance due to the enzyme stability under the dual extremities of pH and salt coupled with moderate thermal tolerance. Besides, the facts emerged on the enzyme stability would add to the limited information on this enzyme from Haloalkaliphilic bacteria.

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“…Waktu produksi optimum protease terjadi pada fase eksponensial akhir dari kurva pertumbuhan isolat bakteri Salmonella sp. Menurut Dodia et al (2006) waktu produksi optimum protease alkalin dari isolat bakteri S 5 yang diisolasi dari Coastal Gujarat, India juga terjadi pada akhir fase eksponensial.…”

Section: Penentuan Waktu Produksi Optimum Enzim Protease Dan Kurva Peunclassified

Isolasi Dan Karakterisasi Protease Alkalin Dari Isolat Bakteri Limbah Ternak Di Exfarm Fakultas Peternakan Unsoed

Zusfahair

Lestari

Asnani

2011

Molekul

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Protease is one of the widely used enzymes for the industry. The potential resource of microorganism that produced protease is milk cow waste. In this research, isolation and characterization has been done toward isolated protease from milk cow waste of the Exfarm's Animal Husbandry Faculty at University of Jenderal Soedirman, Purwokerto. The research used experiment method and the parameters observed were the genus of bacteria which produce protease and the activity of protease. The characterizations of protease were determination of optimum pH and temperature, the influence of metal ions, EDTA, surfactant, and commercial detergent toward enzyme activity, and also the study of enzyme stability. The results from the research showed that the isolated bacteria from the Exfarm's of Animal Husbandry Faculty of UNSOED, which produced protease was Salmonella sp. Characterization of isolated Salmonella sp. from 45% ammonium sulphate fraction indicated that the optimum temperature was 50 ºC, optimum pH was 8, the enzyme was activated by Ca 2+ dan Mg 2+ ion, whereas it was inhibited by Zn 2+ , Cu 2+ ions and EDTA. The addition of Tween-80 with the concentration of 0.2% and 0.4% increased protease activity, however the addition of Tween-80 with concentration higher than 0.6% decreased the protease activity. Enzyme protease from isolated Salmonella sp. was relatively stable with the addition of commercial detergent such as Attack, Surf, and Bukrim.

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Characterization and stability of extracellular alkaline proteases from halophilic and alkaliphilic bacteria isolated from saline habitat of coastal Gujarat, India

Cited by 50 publications

References 28 publications

Production and optimization of a commercially viable alkaline protease from a haloalkaliphilic bacterium

Production and optimization of a commercially viable alkaline protease from a haloalkaliphilic bacterium

Purification and stability characteristics of an alkaline serine protease from a newly isolated Haloalkaliphilic bacterium sp. AH-6

Isolasi Dan Karakterisasi Protease Alkalin Dari Isolat Bakteri Limbah Ternak Di Exfarm Fakultas Peternakan Unsoed

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