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Title:	Pengekstrakan dan penulenan protease daripada kulit biji melinjau (Gnetum gnemon, L) serta penggunaan dalam pelembutan daging
Authors:	Faisal Abdurrahman (P32145)
Supervisor:	Sahidan Senafi, Prof Madya Dr
Keywords:	Pengekstrakan protease Gnetum gnemon Pelembutan daging
Issue Date:	2-Dec-2015
Description:	Protease merupakan enzim yang memangkinkan penguraian ikatan peptida protein. Protease dihasilkan oleh haiwan, tumbuhan dan mikrob. Penggunaan protease dalam industri pemakanan adalah sangat meluas, antaranya seperti papain dan bromelin. Papain dihasilkan daripada getah betik (Carica papaya) dan bromelin daripada buah nenas (Ananas comosus). Kulit biji melinjau (Gnetum gnemon, L.) juga mengandungi protease. Dalam kajian ini, protease telah diekstrak daripada kulit biji melinjau, kemudian diseparatulenkan melalui kombinasi pemendakan amonium sulfat dan penurasan gel Sephadex G-75. Hasil kajian mendapati bahawa aktiviti protease melinjau dikesan pada puncak protein pada kromatografi penurasan gel dengan menggunakan azokasein sebagai substrat. Analisis gel SDS poliakrilamida bagi fraksi separatulen menghasilkan dua jalur yang berberat molekul 15,000 Dalton. Manakala analisis zimogram mengesahkan bahawa berat molekul protease melinjau adalah 15,000 Dalton. Aktiviti optimum bagi protease melinjau adalah pada pH 7.0 dan suhu optimum 60 oC. Pencirian protease melinjau menunjukkan bahawa protease melinjau stabil pada pH 6-8 dan pada suhu daripada 25-60 oC pengeraman selama 30 minit dan 60 minit. Kajian juga menunjukkan bahawa protease melinjau dapat direncatkan oleh HgCl2 dan Asid Iodol Asetik (IAA) serta trans-epoksisuksinil-L-lesilamida (4-guanidino)-butona (E-64) yang menunjukkan protease melinjau tergolong dalam kelas protease sistein. Kajian kinetik menunjukkan protease melinjau mempunyai nilai KM = 0.094 mM manakala Vmax = 7.483 mL/µmol. Protease kulit melinjau separatulin sesuai digunakan untuk melembutkan daging pejal di mana daging yang dieramkan dalam larutan protease kulit biji melinjau lebih lembut berbanding kawalan dan tidak berbeza secara signifikan (p>0.05) berbanding protease daripada papain komersial, betik dan nenas. Protease kulit biji melinjau berjaya menguraikan serat daging lembu pejal menjadi lebih halus secara signifikan (p<0.05) berbanding kawalan dan protease daripada getah betik. Protease kulit biji melinjau tidak menjejaskan kuliti daging seperti warna, kandungan lemak, kandungan protein serta keupayaan memegang air.,Protease is an enzyme that catalyses the peptide bond hydrolysis in the polypeptide chain of a protein. Source of proteases can be from animal, plant and microbial. Proteases have been use widely in food industry, for example papain and bromelin. Papain was extracted from papaya (Carica papaya) and bromelin was extracted from pineapple (Ananas comosus). Melinjau (Gnetum gnemon, L) seed was reported to contain protease which are effective in tenderise of meat. In this study, protease enzyme was extracted from Melinjau testa, and then purified through ammonium sulphate and Sephadex G-75 gel filtration. Activity of melinjau protease has been detected at the protein peak of gel filtration chromatography by using azocasein as the substrate. SDS polyacrylamide gel analysis for partially pure fraction showed one band with the molecular mass of 15,000 malton respectely. Likewise, zymogram analysis showed that protease molecular size from melinjau was 15,000 Dalton. Activity of the protease was optimum at pH 7.0 and at temperature 60oC. Characterisation also showed protease was stable at pH 6 to 8 and at temperature from 25oC to 60oC. The result also showed that Melinjau protease can be inhibited by HgCl2 and Iodic Acetic acid (IAA) with trans-epoxysuccinyl-L-leucilamide (4-guanidino)-butone (E-64), to show that protease of melinjau could be classified as cysteine protease. Kinetic studies showed that protease of Melinjau value has a KM = 0.094 mM and Vmax = 7.483 mL/µmol. Melinjau protease was successfully tenderized meat becomes finer meat significantly (p <0.05) compared to the control and papaya latex protease. Melinjau protease does not affect the meat quality like color, fat content, protein content and water holding capacity.,PhD
Pages:	194
Call Number:	QK494.5.G565F338 2015 tesis
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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