AKTIVITAS PROTEOLITIK ENZIM PAPAIN DARI GETAH BUAH PEPAYA (Carica papaya L.) VARIETAS CALINA DAN PERANANNYA SEBAGAI ANTIBAKTERI; PROTEOLYTIC ACTIVITY OF PAPAIN ENZYME FROM PAPAYA FRUIT LATEX (Carica papaya L.) CALINA VARIETY AND ITS ROLE AS ANTIBACTERIAL AGENT

Jannah, Roudotul, Astuti, Endang

2015 | Skripsi | FMIPA UGM

Isolation of crude papain with both oven drying and freeze drying methods, and partial purification of crystal papain by Kimmel and Smith method was performed, followed by the determination of proteolytic activity and antibacterial activity of papain enzyme from papaya fruit latex (Carica papaya L.) Calina variety. The purpose of this research is to determine the effect of crude papain drying method for its proteolytic activity, the comparison of proteolytic activity of crude papain and crystal papain from partial purification and the determination of antibacterial activity of papain enzyme. Crude papain was extracted by drying papaya latex that has been mixed with NaCl solution. Partial purification was carried out by Kimmel and Smith method until the third fraction. The proteolytic activity was determined by conducting Sigma SSCASE01.001 (1999) protocol with casein used as the substrate. Casein hydrolysis was analyzed with addition of Folin Ciocalteu reagent that produced blue complex whose absorbance measured by using UV-Vis spectrophotometer. Antibacterial activity assay of papain enzyme was performed by agar diffusion method for Escherichia coli, while Most Probable Number (MPN) method was used for fecal coliform and total coliform obtained from Code River water. The result shows that drying method affects the quality of papain. Proteolytic activity of crude papain with freeze drying method is higher than oven drying. Proteolytic activity of crude papain is higher than crystal papain from partial purification. Antibacterial assay shows that crude papain from oven drying forms inhibition zone whereas the third fraction shows no inhibition zone. MPN test shows that crude papain from freeze drying method could inhibit the growth of fecal coliform but not to total coliform.

Kata Kunci : Papain enzyme; Kimmel and Smith; proteolytic activity; agar diffusion; MPN