Purification, biochemical characterization and structural modeling of a potential htrA-like serine protease from Bacillus subtilis DR8806

Farhadian, Shaghayegh; Asoodeh, Ahmad; Lagzian, Milad

doi:10.1016/j.molcatb.2015.02.001

Cited by 47 publications

(28 citation statements)

References 46 publications

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“…The activity of both enzymes was increased with temperature with 45 °C optimum temperature for the free enzyme 50 °C for the immobilized one. Our results are in line with other keratinase preparations from Bacillus subtilis [33] and Brevibacillus brevis [34]. As in the case of pH stability, in comparison with the free enzyme, the beads entrapped-keratinase displayed heat stability even in the absence of its substrate (Figure 3).…”

Section: Discussionsupporting

confidence: 89%

Optimization of azo-keratin hydrolysis by alginate-immobilized Keratinase produced from Bacillus licheniforms

El-Komy

Aldosary

Elnaghy

et al. 2019

Journal of advanced Biomedical and Pharmaceutical Sciences

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Recently, more attention to Keratinolytic materials which can be converted into feed-stuffs, biofertilizers, glues, and foils or used for production of amino acids. Here, we determined the optimum conditions for keratin hydrolysis by alginate-immobilization of crude keratinase isolated from Bacillus lichenoformis. The results of this study indicated that the optimum pH for keratin degradation by immobilized keratinase enzyme was pH 8. Moreover, the crude enzyme was stable for 2 hours at pH 4, 7 and 10 and retained 55.5 %, 60 % and 53 %, of its activity respectively for immobilized enzyme while free enzyme retained only 40 %, 40 % and 61 %, respectively. The optimum temperature for keratinase activity was 50 o C when immobilized in alginate. At temperatures 30, 45 and 60 o C, keratinase was stable for 2 hours and retained 57 %, 66 % and 57 %, of its activity respectively for immobilized enzyme while free enzyme retained only 51 %, 62 % and 51 %, respectively. Furthermore, the effects of activators and inhibitors on the enzyme activity have been investigated.

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Section: Discussionsupporting

confidence: 89%

Optimization of azo-keratin hydrolysis by alginate-immobilized Keratinase produced from Bacillus licheniforms

El-Komy

Aldosary

Elnaghy

et al. 2019

Journal of advanced Biomedical and Pharmaceutical Sciences

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“…The protease from B. subtilis VSG-4 reported by Giri et al ( 2011 ) had the maximum activity at pH 9 with a sharp decline in the activity in pH values lower than pH 9. An extracellular hrtA-like serine protease by B. subtilis DR8806 exhibited high activity in the range of pH from 5 to 10 with the maximum activity at pH 8 (Farhadian et al 2015 ). Similarly, a halophilic bacterium isolated from sea water catalyzed reactions in the pH range 8–11 and performed optimally at pH 10 (Raval et al 2014 ).…”

Section: Resultsmentioning

confidence: 99%

“…Maximum proteolytic activity of Bacillus strains HR-08 and KR-8102 isolated from soil of western and northern parts of Iran have been recorded at 65 and 50 °C, respectively (Moradian et al 2006 ). A serine protease by B. subtilis DR8806 showed highest activity at 45 °C and withstood at temperature up to 70 °C (Farhadian et al 2015 ). Cha et al ( 2005 ) reported that the protease from Bacillus sp.…”

Section: Resultsmentioning

confidence: 99%

“…A serine protease from Bacillus subtilis DR8806 was stimulated by K + , Ca 2+ , Mg 2+ and Fe 2+ at 10 mM concentration up to 134, 129, 128 and 112 %, respectively. Whereas, Na + ions had no significant effect on enzyme activity (Farhadian et al 2015 ). Metal-decreasing activity in the presence of Co 2+ , Ni 2+ was observed by Farhadian et al ( 2015 ), Shah et al ( 2010 ), Priya et al ( 2014 ), Ibrahim et al ( 2011 ) and Jain et al ( 2012 ).…”

Section: Resultsmentioning

confidence: 99%

“…They had shown the activity in the presence of wheat gluten was just 20 %, in comparison with casein. Furthermore, proteases produced by B. halodurans 373 CAS6 (Annamalai et al 2013 ), B. cereus TKU006 (Wang et al 2009 ) and B. subtilis DR8806 (Farhadian et al 2015 ) showed the most activity towards casein as a substrate.…”

Section: Resultsmentioning

confidence: 99%

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Production, purification and characterization of a thermotolerant alkaline serine protease from a novel species Bacillus caseinilyticus

Mothe

Sultanpuram

2016

3 Biotech

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Alkaline proteases are important enzymes in many industrial applications, especially as additives in laundry detergent industry. Though there are a number of Bacillus species which are reported to be producing proteases, the efficiency of a protease produced by a novel strain has to be studied in comparison to the others. Hence, in this study, an alkaline serine protease produced by a novel species Bacillus caseinilyticus was purified and characterized for its possible usage in detergent industry. Ammonium sulphate, dialysis and DEAE column chromatographic methods were used for purification of the isolated alkaline protease. The molecular weight of the protease was determined by SDS-PAGE and it was found to be 66 kDa. Peptide mass fingerprinting (PMF) was carried out using MALDI-TOF-TOF mass spectrometry and the peptides were found to be similar to that of subtilisin protease. Specific activity of purified protein was found to be 89.2 U/mg. Optimum pH and temperature for enzyme activity were at pH 8 and 60 °C, respectively, showing stability with 10 mM CaCl2. Phenyl methyl sulphonyl fluoride (PMSF) at both 5 and 10 mM concentrations completely inhibited the enzyme activity suggesting its serine nature. EDTA, metal ions Mg2+ and Ca2+ increased the enzyme activity. The one factor at a time optimisation of the protease production was carried to identify the important factors that affect its production. After optimisation, the protease was produced at lab scale, purified and characterised. This alkali, thermotolerant serine protease was found to be significantly stable in the presence of various surfactants and H2O2. Also, it was successfully able to remove blood stain when used as an additive along with commercial detergent suggesting its potential application in the laundry detergent industry.

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Purification, characterization and potential applications of a novel halotolerant metalloprotease from marine bacterium Vibrio sp. LA‐05

Zhang

Lang

et al. 2018

J of Chemical Tech & Biotech

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BACKGROUND: Many reported proteases showed low activity and stability under harsh operating conditions, e.g. high pH, salinity and surfactants, which hinders expansion of their applications in industry. Here, a novel halotolerant metalloprotease from marine bacterium Vibrio sp. LA-05 endowed with superior properties was purified and characterized, and its industrial application also evaluated. RESULTS:The protease was active and stable at 25-40 ∘ C and pH 6.0-10.0. Moreover, it was remarkably stable in high salt solution, tolerant to denaturing agents, organic solvents, surfactants and bleaching agents, and quite compatible and stable with many commercial detergents. Its residual activities were above 22% even after co-incubation with 20% NaCl for 480 h. Except Ariel and Tide, the residual activities were all above 55% after co-incubation with five other liquid detergents for 240 h. The corresponding theoretical analyses show that it is a metalloprotease containing 1 mol of zinc ion per mole of protease, the mature active protease predominantly in -helix form consists of 321 amino acids, and the central zinc ion along with 11 other conserved and crucial amino acid residues forms the catalytic domain. CONCLUSION: These excellent properties indicated that the protease has considerable potential as a bio-additive in the application of detergent formulations.

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Purification, biochemical characterization and structural modeling of a potential htrA-like serine protease from Bacillus subtilis DR8806

Cited by 47 publications

References 46 publications

Optimization of azo-keratin hydrolysis by alginate-immobilized Keratinase produced from Bacillus licheniforms

Optimization of azo-keratin hydrolysis by alginate-immobilized Keratinase produced from Bacillus licheniforms

Production, purification and characterization of a thermotolerant alkaline serine protease from a novel species Bacillus caseinilyticus

Purification, characterization and potential applications of a novel halotolerant metalloprotease from marine bacterium Vibrio sp. LA‐05

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