Isolation of Proteolytic Enzyme from Pineapple Crown

The pineapple waste from the industry has contributed to an increase in Malaysia and worldwide every year. A major type of endopeptidase enzymes found is fruit bromelain, stem ananain, comasain. This study aims extract purify protease crown MD2 pineapple. Protease was extracted purified using anion exchange chromatography, gel filtration, desalting before being identified liquid chromatography-mass spectrometry (LC-MS). Proteolytic activity determined well diffusion method Casein Digestion Unit. In present study, proteolytic assay showed that 1 kg cultivar produced 126.0 ± 3.86 U/ml, a specific 3937.50 U/mg. U/mL, U/mg total 3.94 × 109 U. molecular weight enzyme range 25 35 kDa under optimum condition pH 7 37°C. Purification yielded band at 20–25 3 9 60°C. From LC-MS analysis, similar ananain accession number A0A199VSS3 (according Uniprot). It had five unique peptides covered 97/356 amino acids (44.9% coverage). (EC 3.4.22.31) classified subfamilies cysteine C1A (clan CA, family C1), peptidase related papain. conclusion, as ananain-like crown.