Characterization of Thermo‐ and Detergent Stable Antigenic Glycosylated Cysteine Protease of <i>Euphorbia nivulia</i> Buch.‐Ham. and Evaluation of Its Ecofriendly Applications

Badgujar, Shamkant B.; Mahajan, Raghunath T.

doi:10.1155/2013/716545

Cited by 13 publications

(7 citation statements)

References 31 publications

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“…The influence of surfactants (SDS, Tween 20 & 80, and Triton X-100) on G. stearothermophilus protease stability was also examined. Specificity of our protease reduced remarkably after an addition of different concentrations of Triton X-100 and residual activity declined to 34% after addition of 1% Triton X-100 [59]. Sung et al [60] and Haddar et al [61] have reported similar results with decreased activity of proteases from B. subtilis and B. mojavensis A21 after an addition of 0.01% Tween 20 and Triton-X100, by 53 and 92%, respectively.…”

Section: Discussionsupporting

confidence: 71%

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Purification and characterization of cloned alkaline protease gene of Geobacillus stearothermophilus

Iqbal

Aftab

Afzal

et al. 2014

J. Basic Microbiol.

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Thermostable alkaline serine protease gene of Geobacillus stearothermophilus B-1172 was cloned and expressed in Escherichia coli BL21 (DE3) using pET-22b(+), as an expression vector. The growth conditions were optimized for maximal production of the protease using variable fermentation parameters, i.e., pH, temperature, and addition of an inducer. Protease, thus produced, was purified by ammonium sulfate precipitation followed by ion exchange chromatography with 13.7-fold purification, with specific activity of 97.5 U mg(-1) , and a recovery of 23.6%. Molecular weight of the purified protease, 39 kDa, was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable at 90 °C at pH 9. The enzyme activity was steady in the presence of EDTA indicating that the protease was not a metalloprotease. No significant change in the activity of protease after addition of various metal ions further strengthened this fact. However, an addition of 1% Triton X-100 or SDS surfactants constrained the enzyme specific activity to 34 and 19%, respectively. Among organic solvents, an addition of 1-butanol (20%) augmented the enzyme activity by 29% of the original activity. With casein as a substrate, the enzyme activity under optimized conditions was found to be 73.8 U mg(-1) . The effect of protease expression on the host cells growth was also studied and found to negatively affect E. coli cells to certain extent. Catalytic domains of serine proteases from eight important thermostable organisms were analyzed through WebLogo and found to be conserved in all serine protease sequences suggesting that protease of G. stearothermophilus could be beneficially used as a biocontrol agent and in many industries including detergent industry.

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Section: Discussionsupporting

confidence: 71%

“…The influence of surfactants (SDS, Tween 20 & 80, and Triton X‐100) on G. stearothermophilus protease stability was also examined. Specificity of our protease reduced remarkably after an addition of different concentrations of Triton X‐100 and residual activity declined to 34% after addition of 1% Triton X‐100 . Sung et al .…”

Section: Discussionmentioning

confidence: 71%

Purification and characterization of cloned alkaline protease gene of Geobacillus stearothermophilus

Iqbal

Aftab

Afzal

et al. 2014

J. Basic Microbiol.

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“…A number of studies have demonstrated the nematicidal effect of latex proteases from plants (Badgujar and Mahajan, 2013;Stepek et al, 2007;Sufiate et al, 2017b). However, this is the first report on nematicidal effect of S. grantii.…”

Section: Resultsmentioning

confidence: 82%

Role of Synadenium grantii latex proteases in nematicidal activity on Meloidogyne incognita and Panagrellus redivivus

et al. 2019

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Synadenium grantii is a Euphorbiaceae plant commonly found in Brazil, known as Janaúba or Leitosinha, whose latex is traditionally used for several purposes. However, it is not known whether the nematicidal action of this plant latex occurs due to the action of proteases. The present work aims to evaluate the nematicidal activity of proteases from Synadenium grantii latex on Meloidogyne incognita and Panagrellus redivivus. S. grantii latex used in the present study was collected from specimens found in Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil. The drained latex was collected in Eppendorf microtubes and immediately stored on ice at 4 °C. After this extraction, the latex was frozen (-20 °C) during 2 hours, thawed at room temperature (25 °C) and centrifuged at 10,000 g at 4 °C for 30 minutes to remove larger particles and concentrate the proteases. After the centrifugation, assays of enzymatic activity were performed in order to know in which of the phases the enzymes were found. S. grantii latex presented protease, but no chitinase activity. The results show that there was a significant difference (p <0.01) between the treated and control groups, with 100% mortality of Meloidogyne incognita and 72% average mortality of Panagrellus redivivus. In addition, it was demonstrated that the nematicidal action occurred due to the action of the proteases, since the control was only differentiated from the treatment by the presence of the enzymes with biological activity.

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“…DCM as a polar aprotic solvent decreased proteolytic activity of all lyophilized enzyme preparations at 2.5% and 5% (v/v) of concentration. Badgujar et al [25] reported that the proteolytic activity of cysteine protease was decreased with DCM. Ethanol as hydrophilic organic solvent and hexane and toluene as aprotic nonpolar organic solvents have increased proteolytic activity of lyophilized enzyme preparations with pumice and KCl.…”

Section: Organic Solvents Effect On Lyophilized Protease Preparationsmentioning

confidence: 99%

a PRODUCTION OF PROTEASE FROM BACILLUS SUBTILIS UNDER SSF AND EFFECT OF ORGANIC SOLVENTS ON LYOPHILIZED PROTEASE PREPARATIONS

2020

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Objective: In the present work, protease was produced from Bacillus subtilis under solid-state fermentation (SSF). The effect of lyophilization with different additives on the activity of protease in an organic solvent and kineteci properties was investigated. Methods: Production conditions of protease (fermentation time, moisture level, initial pH, temperature) were optimized. After production, it was partially purified and then, lyophilized with different additives from an aqueous buffer solution containing 98% (w/w) of different additives (pumice, KCl, without additive) for 72 h after freezing in liquid nitrogen. After that, the effect of organic solvents (2.5% and 5% of DCM, ethanol, hexane, toluene) on these lyophilized protease preparations was determined and their kinetic properties were determined. Results: Optimum protease production was obtained with 40% of moisture level, at pH 7.5, 37 °C after 24 h fermentation. It was partially purified by using ammonium sulphate precipitation (20-80%) with 5.8-fold and specific activity of 38 U/mg and then dialysed with 6.4-fold and a specific activity of 35 U/mg. Co-lyophilization of protease with pumice and KCl was increased activity of an enzyme in aqueous organic solvents when compared lyophilized protease without additive. Used solvents, except DCM, were increased activity of lyophilized protease with pumice/KCl. It was found that the lyophilization with pumice and KCl resulted in an increasing in the catalytic efficiency, while it was decreased in Km and Vmax values. Conclusion: The obtained findings demonstrated that protease from B. subtilis can effectively be produced under SSF by using wheat bran and used in industrial applications because of showing improved activity in an organic solvent by co-lyophilization with pumice/KCl.

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Characterization of Thermo‐ and Detergent Stable Antigenic Glycosylated Cysteine Protease of Euphorbia nivulia Buch.‐Ham. and Evaluation of Its Ecofriendly Applications

Cited by 13 publications

References 31 publications

Purification and characterization of cloned alkaline protease gene of Geobacillus stearothermophilus

Purification and characterization of cloned alkaline protease gene of Geobacillus stearothermophilus

Role of Synadenium grantii latex proteases in nematicidal activity on Meloidogyne incognita and Panagrellus redivivus

a PRODUCTION OF PROTEASE FROM BACILLUS SUBTILIS UNDER SSF AND EFFECT OF ORGANIC SOLVENTS ON LYOPHILIZED PROTEASE PREPARATIONS

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