One-step recovery of latex papain from Carica papaya using three phase partitioning and its use as milk-clotting and meat-tenderizing agent
| Autor: | Rubén Domínguez, José M. Lorenzo, Amira Leila Dib, Kahina Hafid, James John, Samira Becila, Mohammed Gagaoua, Melisa Lamri, Taha Mansour Sayah |
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| Rok vydání: | 2020 |
| Předmět: |
Latex
tert-Butyl Alcohol medicine.medical_treatment Metal ions in aqueous solution One-Step 02 engineering and technology Complex Mixtures Biochemistry law.invention 03 medical and health sciences chemistry.chemical_compound Magazine Structural Biology law Enzyme Stability Papain medicine Animals Molecular Biology 030304 developmental biology 0303 health sciences Protease Chromatography biology Carica Temperature Sodium Dietary General Medicine Hydrogen-Ion Concentration 021001 nanoscience & nanotechnology biology.organism_classification Enzyme assay Molecular Weight Dairying Drug Combinations Kinetics Milk chemistry Ammonium Sulfate Proteolysis biology.protein Separation method 0210 nano-technology |
| Zdroj: | International Journal of Biological Macromolecules. 146:798-810 |
| ISSN: | 0141-8130 |
| Popis: | Three Phase Partitioning (TPP) system as an elegant non-chromatographic and bulk separation method was successfully applied for the extraction and recovery of papain from the latex of Carica papaya. The optimized parameters of TPP allowed achieving a purification fold of 11.45 and activity recovery of 134% with 40% (NH4)2SO4, 1.0:0.75 ratio of crude extract: t-BuOH at pH and temperature of 6.0 and 25 °C, respectively. The recovered papain had a molecular weight of 23.2 kDa and revealed maximum activity at pH 6.0 and temperature of 50 °C. The maximum values of Km and Vmax parameters were 10.83 mg mL−1 and 33.33 U mL−1, respectively. The protease with 4 isoforms was stable at 40–80 °C and a pH range of 6.0–7.5 against numerous metal ions and none of them inactivated the recovered protease. Moreover, 10 mM Ca2+ improved 2-folds the activity and half-life of the protease at temperatures from 30 to 50 °C. The milk-clotting activity tests revealed high stability of latex papain at storage, namely at −20 °C compared to 4 °C and 25 °C for up than 5 weeks. As a meat tenderizing agent, it showed promising role under different treatments by improving the texture of tough meat. The findings indicated that one-step TPP system is a simple, quick, economical and very attractive process for fast recovery of latex papain compared to other proposed protocols. |
| Databáze: | OpenAIRE |
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