One-step purification and characterization of an alkaline protease from haloalkaliphilic Bacillus sp.

In this study, protease-producing capacity of Bacillus pumilus D3, isolated from hydrocarbon contaminated soil, was evaluated and optimized. Optimum growing conditions for B. pumilus D3 in terms of protease production were determined as 1% optimum inoculum size, 35 C temperature, 11 pH and 48 h incubation time, respectively. Stability studies indicated that the mentioned protease was stable within the pH range of 7-10.5 and between 30 C and 40 C temperatures. Surprisingly, the activity of the enzyme increased in the presence of SDS with concentration up to 5 mM. The protease was concentrated 1.6-fold with ammonium sulfate precipitation and dialysis. At least six protein bands were obtained from dialysate by electrophoresis. Four clear protein bands with caseinolytic activity were detected by zymography. Dialysate was further purified by anion-exchange chromatography and the caseinolytic active fraction showed a single band between 29 and 36 kDa of reducing conditions.

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“…An optimum pH of 11 of alkaline protease was observed in the protease produced by Bacillus sp 41,42 .…”

Section: Optimum Reaction Temperature and Phmentioning

confidence: 94%

“…The optimum pH was found to be 11, indicating that this enzyme might be the alkaline protease 40,41 . An optimum pH of 11 of alkaline protease was observed in the protease produced by Bacillus sp 41,42 .…”

Section: Optimum Reaction Temperature and Phmentioning

confidence: 99%

Production of an alkaline protease using Bacillus pumilus D3 without inactivation by SDS, its characterization and purification

Özçelik

Aytar

Gedikli

et al. 2013

Journal of Enzyme Inhibition and Medicinal Chemistry

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“…The secreted protease was purified (10-fold) with 82% yield by a single step method on Phenyl Sepharose 6 Fast Flow column. The enzyme was active over the pH range of 8.5-12.0, the optimum being 10.0-11.0 [179]. A halotolerant B. clausii I-52 produced >50% of alkaline protease when incubated with 5% (w/v) NaCl.…”

Section: Alkaline Proteases From Halophilesαmentioning

confidence: 91%

Extreme Environments: Goldmine of Industrially Valuable Alkali-stable Proteases

Garg¹,

Singh²

2015

Enz Eng

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Since the advent and release of Bio 40 (the first detergent with bacterial alkaline protease) in 1959, exploration of the microbial alkaline proteases has been exploited beyond expectations. The inventory of microbial alkaline proteases is difficult to draft as it is increasing daily. Most of these proteases are the result of studies in which one or a few isolates were targeted for an alkali-stable enzyme. Although, the worldwide research on microbial alkaline proteases has been widely performed, we still need an improved enzyme capable of catalyzing reactions under extreme conditions (also called as extremozymes), e.g., high alkalinity and salinity, anhydrous environment, cold and hot environment, etc. As the search for extremozymes is in progress, it is imperative to explore the extreme environments to get some novel microbial strains (extremophiles) for alkali-stable proteases. This review article is an effort to compile the scattered information on some extremophiles and their alkali-stable proteases, which may have better specific industrial applications. It includes: (i) various extreme environments relevant to industrial biocatalysts, (ii) strategies of extremophilic microbes and enzymes which make them able to tolerate such conditions, and (iii) an overview of some important work done so far to explore industrially relevant extremophilic enzymes from extremophiles.

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“…The stability of haloalkaliphilic Bacillus sp. (Gupta et al, 2005) and Bacillus mojavensis A21 (Haddar et al, 2009) has been reported, thus showing their strong potential in detergent industry. Laundry detergents are also popular in household dishwashing detergents and in both industrial and institutional cleaning detergents (Godfrey and West, 1996).…”

Section: Author(s) Agree That This Article Remains Permanently Open Amentioning

confidence: 99%

Halophile isolation to produce halophilic protease, protease production and testing crude protease as a detergent ingredient

Sekar¹,

Packyam²,

Kim³

2016

Afr. J. Microbiol. Res.

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Halophilic enzymes are potentially useful in many industries, particularly in food fermentation, pharmaceutical, textile, and leather for the treatment of saline and hypersaline wastewaters. In this study, a halophilic bacterium was isolated from saltpan environment, and was identified as Bacillus sp. Mk22 through biochemical test and 16S rRNA gene sequencing. During protease screening, the isolates produced 24 mm clear zone around the bacterial colony. The maximum production of proteases was due to the following conditions: 45°C, pH 8, 12% NaCl, carbon source glucose, nitrogen source skim milk powder and 42-h culture time, respectively. The protein was purified 16.5 fold, having 24.01% recovery, in DEAE-cellulose chromatography and 64 kDa molecular weight. Ca and Zn enhanced protease activity, while Hg strongly inhibited it. The protease was used to destain blood, ink, coffee and was active and stable under more than one extreme condition of high salt, pH, and temperature.

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One-step purification and characterization of an alkaline protease from haloalkaliphilic Bacillus sp.

Cited by 121 publications

References 21 publications

Production of an alkaline protease using Bacillus pumilus D3 without inactivation by SDS, its characterization and purification

Production of an alkaline protease using Bacillus pumilus D3 without inactivation by SDS, its characterization and purification

Extreme Environments: Goldmine of Industrially Valuable Alkali-stable Proteases

Halophile isolation to produce halophilic protease, protease production and testing crude protease as a detergent ingredient

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