MOLECULAR WEIGHT PROFILE OF PROTEASE OF PINEAPPLE (Ananas comosus L.Merr) AND PAPAYA (Carica papaya L.) USING SDS-PAGE METHOD

Rizky Arcinthya Rachmania, Priyo Wahyudi, Aniza Mutia Wardani, Dini Rohmatul Insani

Abstract

A group of protease enzymes such as papain and bromelain is able to decipher the molecular structure of the protein into amino acids which will be very useful in many fields, especially in food and pharmaceutical industries. The objective of this study to determine the molecular weight profile of enzyme bromelain from pineapple bark (Ananas comosus L. Merr) and papain (Carica papaya L.) from papaya latex with different varieties using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) method. The precipitation was performed with ammonium sulfate ((NH4)2SO4)) 60 % addition and dialysized using a cellophane tubing with a pore size of 12,000 Dalton. The molecular weight of enzyme solution were determined using SDS-PAGE. The analysis results of molecular weight of enzyme bromelain of Subang and Bogor varieties were not different and were about 30.654 kDa, as well as the molecular weight of enzyme papain and Sukma California varieties were also not different and were about 23.485 kDa. It can be concluded that the different varieties of fruit of pineapple and papaya had no effect on the molecular weight of enzyme papain and bromelain.

Keywords

bromelain; papain; protease; SDS-PAGE

Full Text:

PDF

References

Asadayanti, D.D, Jenie, B.S. L., Kusumaningrumi, H. D dan Nurhidayat, N. 2010. Peningkatan Kadar Lovastatin Angkak oleh Monascus purpureus Ko-Kultur dengan Endomycopsis burtonii. Berita Biologi Jurnal Ilmu-ilmu Hayati 10 (3), 313-321.

Bachrudin, Z., Astuti, and Dewi, Y.S., 2000. Isolasi dan Seleksi Mikroba Penghasil Laktat dan Aplikasinya pada Fermentasi Limbah Industri Tahu. Prosiding Seminar Nasional Industri Enzim dan Bioteknologi. Mikrobiologi Enzim dan Bioteknologi.

Bintang, M., 2010. Biokimia Teknik Penelitian. Erlangga. Jakarta.

BioRad Laboratories, 2011. A Guide to Polyacrylamide Gel Electrophoresis and Detection. Life Science Group. US/EG.

Gautam, S., Mishra, S.K., Dash, V., Goyal, A.K., and Rath, G., 2010. Comperative Study of Extraction, Purification and Estimation of Bromelain from Stem and Fruit of Pineapple Plant. Thailand Journal Pharmacy. 34: 67-76.

Hadiati, S., and Luh, P.I.N., 2008. Budidaya Nanas. Balai Penelitian Tanaman Buah Tropika. Sumatera Barat.

Handayani, L., 2007. Uji Aktivitas Anti tuberculosis Ekstrak Getah Pepaya (Carica papaya L.) terhadap Bakteri Mycobacterium tuberculosis H37Rv. Skripsi. Fakultas Farmasi dan Sains Universitas Muhammadiyah Prof. Dr. Hamka. Jakarta.

Herdyastuti, N., 2006. Isolasi dan Karakterisasi Ekstrak Kasar Enzim Bromelin dari Batang Nanas (Ananas comosus (L.) Merr). Berkala Penelitian Hayati 12, 75-77.

Jisha, V.N., Smitha, R.B., Pradeep, S., Sreedevi, S., Unni, K.N., Sajith, S., Priji, P., Josh, M.S., and Benjamin, S., 2013. Versatility of Microbial Protease. Advances in Enzyme Research 1 (3), 39-51

Kalie, M.B., 2008. Bertanam Pepaya. Penebar Swadaya. Depok. 92-106.

Kuchel, P., and Ralston, G.B., 2006. Schaum’s Easy Outlines Biokimia, Terjemahan: Eva Laelasari, S.Si. Erlangga, Jakarta. 49-50.

Kusumadjaja, P.A., and Dewi, P.R., 2005. Penentuan Kondisi Optimum Enzim Papain dari Pepaya Burung Varietas Jawa. Indonesia Journal of Chemistry 5 (2), 147-148.

Masri, M., 2014. Isolasi dan Pengukuran Aktivitas Enzim Bromelin dari Ekstrak Kasar Bongol Nanas (Ananas comosus) pada Variasi Suhu dan pH. Jurnal Ilmiah Biologi 2 (2), 119-125.

Motyan, J.A., Toth, F., and Tozser, J., 2013. Research Applications of Proteolytic Enzyme in Molecular Biology. Biomolecules review 3 (1), 925 - 931.

Nitsawang, S., Kaul, R.H., and Kanasawud, P., 2006. Purification of Papain from Carica papaya Latex : Aqueous Two-Phase Extraction versus Two-Step Salt Precipitation. Enzyme and Microbial Technology 39 (6), 1103-1107.

Nurhidayati, T. 2003. Pengaruh Konsentrasi Enzim Papain dan Suhu Fermentasi terhadap Kualitas Keju Cottage. Kappa 4 (1), 13-17

Nurmala, L.W., 2012. Kajian Penggunaan Ammonium Sulfat Pada Pengendapan Enzim Bromelin dari Batang Nanas (Ananas comosus L. Merr) Sebagai Koagulan pada Pembuatan Keju Cottage. Skripsi. Universitas Pendidikan Indonesia. Bandung.

Pitpreecha, S., and Damrongsakkul, S., 2006. Hydrolysis of Raw Hide using Proteolytic Enzyme Extracted from Papaya Latex. Korean Journal of Chemical Engineering 23 (6), 972-976.

Putri R.A., Kusrijadi, A., and Suryatna, A., 2013. Kajian Penggunaan Ammonium Sulfat pada Pengendapan Enzim Protease (Papain) dari Buah Pepaya sebagai Koagulan dalam Produksi Keju Cottage. Jurnal Sains dan Teknologi Kimia 4 (2), 159-168.

Rao, M.B., Tanksale, M.A., Ghatge, M.S., and Deshpande, V.V., 1998. Molecular and Biotechnological Aspects of Microbial Proteases. Microbiology and Molecular Biology Reviews 62 (3), 599-601, 606-607.

Rath, A., Cunninghan, F., and Deber, C.M., 2013. Acrylamide Concentration Determines the Direction and Magnitude of Helical Membrane Protein Gel Shifts. Proceeding of the National Academy Science 110 (39) 15668-15673.

Sobir, 2009. Sukses Bertanam Pepaya Unggul Kualitas Supermarket. AgroMedia. Jakarta. 26-29, 146-147.

Wuryanti, 2004. Isolasi dan Penentuan Aktivitas Spesifik Enzim Bromelin dari Buah Nanas (Ananas comosus L.). Jurnal kimia Sains dan Aplikasi 3 (7), 83-87.

Refbacks

  • There are currently no refbacks.