Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss].
Membrane-bound polyphenoloxidase (mPPO) an oxidative enzyme which is responsible for the undesirable browning reaction in Snake fruit (Salacca zalacca (Gaertn.) Voss) was investigated. The enzyme was extracted using a non-ionic detergent (Triton X-114), followed by temperature-induced phase partitio...
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| Format: | Article |
| Language: | English English |
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Elsevier
2013
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| Online Access: | http://psasir.upm.edu.my/id/eprint/30443/ http://psasir.upm.edu.my/id/eprint/30443/1/Purification%20and%20characterization%20of%20membrane.pdf |
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| author | Mohd Zaini, Nurul Aqilah Osman, Azizah Abdul Hamid, Azizah Ebrahimpour, Afshin Saari, Nazamid |
| author_facet | Mohd Zaini, Nurul Aqilah Osman, Azizah Abdul Hamid, Azizah Ebrahimpour, Afshin Saari, Nazamid |
| author_sort | Mohd Zaini, Nurul Aqilah |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Membrane-bound polyphenoloxidase (mPPO) an oxidative enzyme which is responsible for the undesirable browning reaction in Snake fruit (Salacca zalacca (Gaertn.) Voss) was investigated. The enzyme was extracted using a non-ionic detergent (Triton X-114), followed by temperature-induced phase partitioning technique which resulted in two separate layers (detergent-poor phase at the upper layer and detergent-rich phase at the lower layer). The upper detergent-poor phase extract was subsequently fractionated by 40–80% ammonium sulfate and chromatographed on HiTrap Phenyl Sepharose and Superdex 200 HR 10/30. The mPPO was purified to 14.1 folds with a recovery of 12.35%. A single prominent protein band appeared on native-PAGE and SDS–PAGE implying that the mPPO is a monomeric protein with estimated molecular weight of 38 kDa. Characterization study showed that mPPO from Snake fruit was optimally active at pH 6.5, temperature 30 °C and active towards diphenols as substrates. The Km and Vmax values were calculated to be 5.46 mM and 0.98 U/ml/min, respectively, when catechol was used as substrate. Among the chemical inhibitors tested, l-cysteine showed the best inhibitory effect, with an IC50 of 1.3 ± 0.002 mM followed by ascorbic acid (1.5 ± 0.06 mM), glutathione (1.5 ± 0.07 mM), EDTA (100 ± 0.02 mM) and citric acid (186 ± 0.16 mM). |
| first_indexed | 2025-11-15T09:06:32Z |
| format | Article |
| id | upm-30443 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T09:06:32Z |
| publishDate | 2013 |
| publisher | Elsevier |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-304432015-10-07T08:40:30Z http://psasir.upm.edu.my/id/eprint/30443/ Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss]. Mohd Zaini, Nurul Aqilah Osman, Azizah Abdul Hamid, Azizah Ebrahimpour, Afshin Saari, Nazamid Membrane-bound polyphenoloxidase (mPPO) an oxidative enzyme which is responsible for the undesirable browning reaction in Snake fruit (Salacca zalacca (Gaertn.) Voss) was investigated. The enzyme was extracted using a non-ionic detergent (Triton X-114), followed by temperature-induced phase partitioning technique which resulted in two separate layers (detergent-poor phase at the upper layer and detergent-rich phase at the lower layer). The upper detergent-poor phase extract was subsequently fractionated by 40–80% ammonium sulfate and chromatographed on HiTrap Phenyl Sepharose and Superdex 200 HR 10/30. The mPPO was purified to 14.1 folds with a recovery of 12.35%. A single prominent protein band appeared on native-PAGE and SDS–PAGE implying that the mPPO is a monomeric protein with estimated molecular weight of 38 kDa. Characterization study showed that mPPO from Snake fruit was optimally active at pH 6.5, temperature 30 °C and active towards diphenols as substrates. The Km and Vmax values were calculated to be 5.46 mM and 0.98 U/ml/min, respectively, when catechol was used as substrate. Among the chemical inhibitors tested, l-cysteine showed the best inhibitory effect, with an IC50 of 1.3 ± 0.002 mM followed by ascorbic acid (1.5 ± 0.06 mM), glutathione (1.5 ± 0.07 mM), EDTA (100 ± 0.02 mM) and citric acid (186 ± 0.16 mM). Elsevier 2013-01-15 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/30443/1/Purification%20and%20characterization%20of%20membrane.pdf Mohd Zaini, Nurul Aqilah and Osman, Azizah and Abdul Hamid, Azizah and Ebrahimpour, Afshin and Saari, Nazamid (2013) Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss]. Food Chemistry, 136 (2). pp. 407-414. ISSN 0308-8146 10.1016/j.foodchem.2012.08.034 English |
| spellingShingle | Mohd Zaini, Nurul Aqilah Osman, Azizah Abdul Hamid, Azizah Ebrahimpour, Afshin Saari, Nazamid Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss]. |
| title | Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss]. |
| title_full | Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss]. |
| title_fullStr | Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss]. |
| title_full_unstemmed | Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss]. |
| title_short | Purification and characterization of membrane-bound polyphenoloxidase (mPPO) from Snake fruit [Salacca zalacca (Gaertn.) Voss]. |
| title_sort | purification and characterization of membrane-bound polyphenoloxidase (mppo) from snake fruit [salacca zalacca (gaertn.) voss]. |
| url | http://psasir.upm.edu.my/id/eprint/30443/ http://psasir.upm.edu.my/id/eprint/30443/ http://psasir.upm.edu.my/id/eprint/30443/1/Purification%20and%20characterization%20of%20membrane.pdf |