A novel milk-clotting cysteine protease from Ficus johannis: Purification and characterization

Afsharnezhad, Moslem; Shahangian, S. Shirin; Sariri, Reyhaneh

doi:10.1016/j.ijbiomac.2018.10.006

Cited by 49 publications

(38 citation statements)

References 48 publications

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“…The protease relative activity was lower than 5% at 60 °C for incubation for 100 min. The exposure of hydrophobic groups caused by the thermal aggregation of protease under high temperature inactivates the protease (Afsharnezhad et al., 2019). The above results indicated that the protease exhibited considerable stability at 20 to 40 °C.…”

Section: Resultsmentioning

confidence: 99%

“…The results suggested that K m , V max , and k cat of the purified protease from the liner were 8.19 mg/ml, 49.50 U/ml·min, and 16.5 1/s, respectively (Table 2). K m represents the affinity of the enzyme toward a substrate; a higher K m value means a lower affinity of enzyme toward the substrate (Afsharnezhad et al., 2019). V max represents the maximum reaction rate, which is affected by the amount of protease involved in the reaction.…”

Section: Resultsmentioning

confidence: 99%

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Biochemical properties of extracellular protease from Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins

Wang

Xia

et al. 2021

Journal of Food Science

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The characteristics of the extracellular protease, produced by Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins were investigated. The protease was purified by ammonium sulfate, ion exchange, and gel filtration chromatography to obtain a 20.0 kDa extracellular protease. The protease reached maximal activity at pH 9.0 and 50 °C and was stable at pH 7.0 to 11.0 and 20 to 40 °C. Its protease activity was easily inhibited in the presence of Zn2+, Fe2+, and Fe3+. The enzymatic characterization of the protease revealed a Vmax 49.50 U/ml·min, Km 8.19 mg/ml, and the half‐life = 28.06 min, ΔH*d = 114.11 kJ/mol, ΔG*d = 89.24 kJ/mol, and ΔS*d = 77.00 J/mol·K at 50 °C. In addition, the protease hydrolyzed meat protein into small particles and produced soluble peptides. This study provides a basis for understanding the biochemical characteristics of the S. carnosus RT6 protease and its future application for fermented meat products.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Biochemical properties of extracellular protease from Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins

Wang

Xia

et al. 2021

Journal of Food Science

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Section: Discussionmentioning

confidence: 81%

“…The rEla cysteine protease exhibited maximum activity at 50 °C similar to the cysteine protease extracted from Cissus quadrangularis 39 . Cysteine proteases isolated from Zingiber montanum rhizome, Ficus johannis, Calotropis procera, Z. officinale, Ficus johannis, exhibited optimum temperature at 60 °C which are all in the optimum temperature range of rEla 4,32,39,41 . The enzyme activity was stable with temperature at the range of 10 to 70 °C and it displayed thermostable features and maintained up to 60% of its proteolytic activity after incubation at 50 °C for 3 h. In a study held by K. Jamir and K. Seshagirirao cysteine protease isolated from Zingiber montanum rhizome 50% inactivation was observed by incubating the protein for 45 min at 40 °C and the enzyme was completely inactivated at 70 °C 32 while, rEla retained about 40% of its maximum activity after 3 h incubation at 90 °C.…”

Section: Discussionmentioning

confidence: 81%

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Cohnella 1759 cysteine protease shows significant long term half-life and impressive increased activity in presence of some chemical reagents

Saghian

Mokhtari

Aminzadeh

2021

Sci Rep

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Thermostability and substrate specificity of proteases are major factors in their industrial applications. rEla is a novel recombinant cysteine protease obtained from a thermophilic bacterium, Cohnella sp.A01 (PTCC No: 1921). Herein, we were interested in recombinant production and characterization of the enzyme and finding the novel features in comparison with other well-studied cysteine proteases. The bioinformatics analysis showed that rEla is allosteric cysteine protease from DJ-1/ThiJ/PfpI superfamily. The enzyme was heterologously expressed and characterized and the recombinant enzyme molecular mass was 19.38 kD which seems to be smaller than most of the cysteine proteases. rEla exhibited acceptable activity in broad pH and temperature ranges. The optimum activity was observed at 50℃ and pH 8 and the enzyme showed remarkable stability by keeping 50% of residual activity after 100 days storage at room temperature. The enzyme Km and Vmax values were 21.93 mM, 8 U/ml, respectively. To the best of our knowledge, in comparison with the other characterized cysteine proteases, rEla is the only reported cysteine protease with collagen specificity. The enzymes activity increases up to 1.4 times in the presence of calcium ion (2 mM) suggesting it as the enzyme’s co-factor. When exposed to surfactants including Tween20, Tween80, Triton X-100 and SDS (1% and 4% v/v) the enzyme activity surprisingly increased up to 5 times.

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Thermostable trypsin‐like protease by Penicillium roqueforti secreted in cocoa shell fermentation: Production optimization, characterization, and application in milk clotting

Nogueira

Tavares

Santana

et al. 2021

Biotech and App Biochem

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The increased demand for cheese and the limited availability of calf rennet justifies the search for milk-clotting enzymes from alternative sources. Trypsin-like protease by Penicillium roqueforti was produced by solid-state fermentation using cocoa shell waste as substrate. The production of a crude enzyme extract that is rich in this enzyme was optimized using a Doehlert-type multivariate experimental design. The biochemical characterization showed that the enzyme has excellent activity and stability at alkaline pH (10-12) and an optimum temperature of 80 • C, being stable at temperatures above 60 • C. Enzymatic activity was maximized in the presence of Na + (192%), Co 2+ (187%), methanol (153%), ethanol (141%), and hexane (128%). Considering the biochemical characteristics obtained and the milk coagulation activity, trypsin-like protease can be applied in the food industry, such as in milk clotting and in the fabrication of cheeses.

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A novel milk-clotting cysteine protease from Ficus johannis: Purification and characterization

Cited by 49 publications

References 48 publications

Biochemical properties of extracellular protease from Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins

Biochemical properties of extracellular protease from Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins

Cohnella 1759 cysteine protease shows significant long term half-life and impressive increased activity in presence of some chemical reagents

Thermostable trypsin‐like protease by Penicillium roqueforti secreted in cocoa shell fermentation: Production optimization, characterization, and application in milk clotting

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