Purification of a novel cysteine protease, procerain B, from Calotropis procera with distinct characteristics compared to procerain

Singh, Abhay Narain; Shukla, Anil Kumar; Jagannadham, Medicherla V.; Dubey, Vikash Kumar

doi:10.1016/j.procbio.2009.10.014

Cited by 45 publications

(34 citation statements)

References 28 publications

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“…MDA concentration was determined with the help of extinction coefficient (155 mM -1 cm -1 ). Protein content determination in all the above enzymatic preparation was done by Bradford method (1976) taking bovine serum albumin (BSA) as standard as followed in our earlier publication Singh et al (2010).…”

Section: Methodsmentioning

confidence: 99%

Effectivity of anti-oxidative enzymatic system on diminishing the oxidative stress induced by aluminium in chickpea (Cicer arietinum L.) seedlings

Singh

Verma

Dubey

2012

Braz. J. Plant Physiol.

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We explore physiological and biochemical response under increasing aluminium stress at different time interval in chickpea seedlings (Cicer arietinum L.). Germination percentage and root length were found to be highly reduced under increasing metal stress. Aluminium induced oxidative stress and led to fluctuations in antioxidative activity responses. Roots showed higher antioxidative activity compared to shoots. Low concentration of aluminium after a brief treatment induced higher superoxide dismutase (SOD; EC 1.15.1.1), ascorbate peroxidase (APX; EC 1.11.1.11) and guaiacol peroxidase (GPX; EC 1.11.1.7) activity whereas longer duration of treatment led to decrease in these activities. Malondialdehyde concentration indicated higher oxidative damage in roots compared to shoots. Taken together the data obtained indicated that high concentration and long exposition of aluminium increases oxidative stress and impairs antioxidative defense system, overall leading to poor growth and survival of seedlings.

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Section: Methodsmentioning

confidence: 99%

Effectivity of anti-oxidative enzymatic system on diminishing the oxidative stress induced by aluminium in chickpea (Cicer arietinum L.) seedlings

Singh

Verma

Dubey

2012

Braz. J. Plant Physiol.

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“…Relative activity (%) . 9 Photography of obtaining cheese at 60°C by using I puriWed protease enzyme from compositae (Scorzonera hispanica L.), II control…”

Section: Resultsmentioning

confidence: 99%

“…The ability of the compositain to retain its activity under conditions of varying pH (3)(4)(5)(6)(7)(8)(9)(10)(11) and temperature (40-80°C) was investigated. The enzyme was incubated under speciWed conditions of pH for 24 h, and the residual activity was determined.…”

Section: Dependence Of Enzyme Activity On Ph and Temperaturementioning

confidence: 99%

“…Despite being the largest class of proteases in plants, the functions and regulatory roles of plant serine proteases are poorly understood, probably due to a lack of identiWcation of their physiological substrates. Once thought to be rare in plants, in recent years, several serine proteases have been isolated and puriWed from diVerent plant parts of various plant species, including seeds, latex and fruits [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

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Purification and characterization of a novel serine protease compositain from compositae (Scorzonera hispanica L.)

Nadaroğlu

Demir

2012

Eur Food Res Technol

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In this research, a protease from the compositae (Scorzonera hispanica L.) was extracted and puriWed through (NH 4 ) 2 SO 4 precipitation, CM-Sephadex and Sephacryl S200. At the end of puriWcation by gel Wltration on a Sephacryl S-200 column, 87.11-fold puriWcation was achieved. It was shown that puriWed enzyme was homogeneous in terms of SDS-PAGE with molecular mass estimate of 30 kDa. The enzyme named compositain depicted an optimal pH of 8.0 and was stable at pH 7.0-9.0, and its optimal temperature was at 50°C. While Tween 80 (0.2 %) was activated to the puriWed protease enzyme, it was partially inhibited by 5 mM concentration of some metal salts and EDTA, PMSF, dithiothreitol, H 2 O 2 and -mercaptoethanol. The enzyme activity was stable even in the presence of detergents and organic solvents. In addition, it was investigated whether the puriWed and characterized protease enzyme would cause to congeal milk. As a result, it was determined that the compositain could congeal milk and it would be used for cheese production. The compositain had potential application in food processing.

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“…Unlike what happens with other peptidases when preparing electrophoresis samples (Priolo et al 2000;Obregón et al 2001) the presence of an inhibitor is not required to prevent autodigestion. Autolysis resistance proteases are very useful for industrial or therapeutic applications and frequently proteases are modified by protein engineering to avoid rapid autolysis (Singh et al 2010). Araujiain aII molecular mass measured by MALDI-TOF was 23.528 kDa (Fig.…”

Section: Resultsmentioning

confidence: 99%

Biochemical characterization, cDNA cloning, and molecular modeling of araujiain aII, a papain-like cysteine protease from Araujia angustifolia latex

Obregón

Lufrano

Liggieri

et al. 2011

Planta

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Araujiain aII, the protease with highest specific activity purified from latex of Araujia angustifolia (Apocynaceae), shows optimum proteolytic activity at alkaline pH, and it is completely inhibited by the irreversible inhibitor of cysteine proteases trans-epoxysucciny-L: -leucyl-amido(4-guanidino) butane. It exhibits esterolytic activity on several N-α-Cbz-amino acid p-nitrophenyl esters with a preference for Gln, Ala, and Gly derivatives. Kinetic enzymatic assays were performed with the thiol proteinase substrate p-Glu-Phe-Leu-p-nitroanilide (K (m) = 0.18 ± 0.03 mM, k (cat) = 1.078 ± 0.055 s(-1), k (cat)/K (m) = 5.99 ± 0.57 s(-1) mM(-l)). The enzyme has a pI value above 9.3 and a molecular mass of 23.528 kDa determined by mass spectrometry. cDNA of the peptidase was obtained by reverse transcription-PCR starting from total RNA isolated from latex. The deduced amino acid sequence was confirmed by peptide mass fingerprinting analysis. The N-terminus of the mature protein was determined by automated sequencing using Edman's degradation and compared with the sequence deduced from cDNA. The full araujiain aII sequence was thus obtained with a total of 213 amino acid residues. The peptidase, as well as other Apocynaceae latex peptidases, is a member of the subfamily C1A of cysteine proteases. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was suggested by molecular modeling.

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Purification of a novel cysteine protease, procerain B, from Calotropis procera with distinct characteristics compared to procerain

Cited by 45 publications

References 28 publications

Effectivity of anti-oxidative enzymatic system on diminishing the oxidative stress induced by aluminium in chickpea (Cicer arietinum L.) seedlings

Effectivity of anti-oxidative enzymatic system on diminishing the oxidative stress induced by aluminium in chickpea (Cicer arietinum L.) seedlings

Purification and characterization of a novel serine protease compositain from compositae (Scorzonera hispanica L.)

Biochemical characterization, cDNA cloning, and molecular modeling of araujiain aII, a papain-like cysteine protease from Araujia angustifolia latex

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