Purification and characterization of a serine protease secreted by Brevibacillus sp. KH3 for reducing waste activated sludge and biofilm formation

Maeda, Takeyasu; Yoshimura, Takayuki; García‐Contreras, Rodolfo; Ogawa, Hiroaki

doi:10.1016/j.biortech.2011.08.098

Cited by 41 publications

(8 citation statements)

References 32 publications

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“…Earlier researchers reported that proteases isolated from Bacillus sp. were of the serine type [7,22,[33][34][35]. However, protease from Bacillus sp.…”

Section: Resultsmentioning

confidence: 99%

Purification and Characterization of Halophilic Organic Solvent Tolerant Protease from Marine Bacillus sp. APCMST-RS7 and Its Antioxidant Potentials

Maruthiah

Immanuel

Palavesam

2015

Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci.

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Halophilic organic solvent tolerant proteolytic bacterium was isolated from the marine sediment of Rajakkamankalam estuary and identified as Bacillus sp. APCMST-RS7. The alkaline protease synthesized by Bacillus sp. APCMST-RS7 was purified by using ammonium sulphate precipitation, dialysis and DEAE-Sepharose Fast Flow column. The purified protease showed 6.60-fold purity, 26.02 U/mg specific activity with 24.30 % yield. The molecular weight of the purified alkaline protease was 32 kDa. This purified protease registered maximum activity at pH 8 and it was stable between pH 8-9 after 1.30 h of incubation. The optimum temperature registered was 50°C and it was stable between 30 and 40°C even after 1.30 h of incubation. This enzyme also showed maximum activity at 1.5 M NaCl concentration. The Km and V max values registered were, 0.0002 g/l and 1428.57 U/ml, respectively. Further, magnesium chloride, barium chloride and copper sulphate influenced this enzyme activity remarkably and it was also found to be enhanced by many of the tested surfactants and solvents. Here, the serine protease inhibitor totally inhibited the enzyme activity; hence it is of serine protease family. The candidate bacterium effectively deproteinized the shrimp shell waste with maximum protease activity. The shrimp shell hydrolysate also exhibited maximum antioxidant activity.

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“…Earlier researchers reported that proteases isolated from Bacillus sp. were of the serine type [7,22,[33][34][35]. However, protease from Bacillus sp.…”

Section: Resultsmentioning

confidence: 99%

Purification and Characterization of Halophilic Organic Solvent Tolerant Protease from Marine Bacillus sp. APCMST-RS7 and Its Antioxidant Potentials

Maruthiah

Immanuel

Palavesam

2015

Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci.

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“…Determination of protease activity was performed using the standard assay condition as described above. Temperature stability of the protease was investigated by incubating the enzyme in 50 mM Tris-HCL (pH 8.0) within temperature range of 10 to 100°C for 1 h. The residual enzyme activity was determined by azocasein at pH 9.0 and 70°C for 1 h [ 12 ].…”

Section: Methodsmentioning

confidence: 99%

Purification and Characterization of Alkaline-Thermostable Protease Enzyme from Pitaya (Hylocereus polyrhizus) Waste: A Potential Low Cost of the Enzyme

Amid

Manap

Zohdi

2014

BioMed Research International

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The thermoalkaline protease enzyme from pitaya (Hylocereus polyrhizus) waste was purified by a factor of 221.2 with 71.3% recovery using ammonium sulphate precipitation, gel filtration, and cation exchange chromatography. Gel filtration chromatography together with sodium dodecyl sulphate gel electrophoresis (SDS-PAGE) revealed that the enzyme is monomeric with a molecular weight of 26.7 kDa. The apparent K m and V max of the protease were 2.8 mg/mL and 31.20 u/min, respectively. The optimum pH and temperature were 8.0 and 70°C. The enzyme was highly active and stable over a wide pH range (from pH 3.0 to pH 11.0 with the optimum activity at pH 8.0). The protease has broad specificity toward azocasein, casein, hemoglobin, and gelatine. Activity of the enzyme was inhibited by Fe2+ and Zn2+, while protease activity was increased in the presence of Ca2+ and Mg2+ and Cu2+ by factors of 125%, 110%, and 105%, respectively. The alkaline protease showed extreme stability toward surfactants and oxidizing agent. The purified protease exhibited extreme stability in the presence of organic solvents and inhibitors. In addition, the enzyme was relativity stable toward organic solvents and chelating agents, such as ethylenediaminetetraacetic acid (EDTA). The enzyme, derived from pitaya peel, possesses unique characteristics and could be used in various industrial and biotechnological applications.

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“…The main industrial producers of serine proteases are strains belonging to the genus Bacillus ( 60 , 61 ). This genus has an important role in white biotechnology for several reasons – it is very well studied, majority of species are non-pathogenic, generally recognized as safe (for example B. cereus , B. stearothermophilus , B. licheniformis , B. subtilis and so on) and many functional extracellular proteins are secreted directly into the culture medium ( 60 , 62 ).…”

Section: Industrially Important Producers Of Keratinasesmentioning

confidence: 99%

Microbial keratinases: enzymes with promising biotechnological 4 applications

Vidmar

Vodovnik

2018

Food Technol. Biotechnol.

110

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SUMMARYKeratin is a complex and structurally stable protein found in human and animal hard tissues, such as feathers, wool, hair, hoof and nails. Some of these, like feathers and wool, represent one of the main sources of protein-rich waste with significant potential to be transformed into value-added products such as feed, fertilizers or bioenergy. A major limitation impeding valorization of keratinous substrates is their recalcitrant structure and resistance to hydrolysis by common proteases. However, specialized keratinolytic enzymes produced by some microorganisms can efficiently degrade these substrates. Keratinases have already found a purpose in pharmaceutical, textile and leather industries. However, their wider implementation in other processes, such as cost-effective (pre)treatment of poultry waste, still requires optimization of production and performance of the available enzymes. Here we present a comprehensive review covering molecular properties and characteristics of keratinases, their classification, traditional and novel approaches in discovery of novel enzymes, production, characterization, improvement and biotechnological applications.

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Purification and characterization of a serine protease secreted by Brevibacillus sp. KH3 for reducing waste activated sludge and biofilm formation

Cited by 41 publications

References 32 publications

Purification and Characterization of Halophilic Organic Solvent Tolerant Protease from Marine Bacillus sp. APCMST-RS7 and Its Antioxidant Potentials

Purification and Characterization of Halophilic Organic Solvent Tolerant Protease from Marine Bacillus sp. APCMST-RS7 and Its Antioxidant Potentials

Purification and Characterization of Alkaline-Thermostable Protease Enzyme from Pitaya (Hylocereus polyrhizus) Waste: A Potential Low Cost of the Enzyme

Microbial keratinases: enzymes with promising biotechnological 4 applications

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