Isolation and Characterization Trypsin - Like Protease (PST) from Lactobacillus plantarum FNCC 0270

  • TRISMILAH TRISMILAH UNIVERSITAS INDONESIA
  • WAHONO SUMARYONO UNIVERSITAS PANCASILA
  • AMARILA MALIK UNIVERSITAS INDONESIA
  • MOHAMAD SADIKIN UNIVERSITAS INDONESIA

Abstract

Trypsin is an enzyme which is important for research as well as for the pharmaceutical industry, medicine and health, which is especially developed as a raw material for digestive enzymes. This study aimed to characterize PST from Lactobacillus plantarum FNCC 0270 that have not been studied as a producer of trypsin. Fermentation for producing PST by L . plantarum FNCC 0270 was performed in the LKB fermentor with working volume of 3.5 liters, at temperature of 37 oC, pH value of 8, agitation rate of 77 rpm and aeration of 0.5 vvm. The media composition were 3.64% baker’s yeast, 1.21% glucose, 0.13% skim milk. The fementation resulted in enzyme activities of 0.00129 U/mL and protein content of 0.49 mg/mL. Purification of the PST conducted by ultrafiltration Hollow Fiber Catridge 5 kD, presipitated with ammonium sulfate saturated (30-70%), 30 kD membrane polyesthersulfone, ion exchange column chromatography Q-XL and HiTrap affinity column chromatography. All of system increased purity of the crude PST. SDS-PAGE of PST indicated the molecular weights of 47.35; 38.42;21.40 and 12.96 kD. The PST was stable at pH value of 8 and temperature range of 25-35 oC. It is proved that the same half-life was 693.2 minutes. Km and Vmax values of the PST were 0.231 mM and 0.001 U/mL, respectively using N-α-benzoyl-DL-arginine-p-nitroanilide as the substrate. PST was inhibited by EDTA, Ca2+, Zn2+, Mg2+, Mn2+ and specific substrates. Based on the result of characterization of PST from L. plantarum FNCC 0270 can be concluded that the trypsin–like protease (PST) was trypsin.

References

1. Badan Pusat Statistik. Buletin Statistik Perdagangan Luar Negeri, Import, 2008. ISSN: 0216.3668, CV. Litama Printing. 9.
2. Anonim. Related topics: Financial & industry, cultures, enzymes, yeast. Diambil dari www.foodnavigator.com/Financial-Industry/World-enzyme-demand-to-hit-5.1bn-by-2009- 68k., diakses 19 Mei 2009.
3. McCann M. Pancreatic enzyme supplement for treatment of multiple food allergies. Ann Allergy. 1993. 71: 69.
4. Ambrus JL, Lassman HB, DeMarchi JJ. Absorption of exogenous and endogenous proteolytic enzymes. Clin Pharmacol Ther. 1967. 8: 362–8.
5. Khantaphan S, Benjakul S. Purification and characterization of trypsin from the Pyloric caeca of brownstripe red snapper (Lutjanus vita). Food Chemistry. 2010. 120: 658-64.
6. Malik A, Hermawati AK, Hestiningtyas M, Soemiati A, Radji M. Isolasi dan skrining molekuler bakteri asam laktat pembawa gen glukansukrase dari makanan dan minuman mengandung gula. Makara (Sains). 2012. 14: 57-62.
7. Wahyuntari B. Pemurnian dan karakterisasi protease ekstraseluler isolat prokariot termofilik ekstrim dari Tangkuban Perahu [disertasi]. Bogor: Jurusan Teknologi Pangan Institut Pertanian Bogor; 2001. 54.
8. Ignatius Sawabi. Makanan rakyat cegah kegemukan. Diambil dari http: //travel. kompas.Com /read/ 2010/08/16/11570266/Growol, diakses tanggal 11 Maret 2011.
9. Bradford M. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye bind. Anal. Biochem. 1976. 72: 248-54.
10. Desrosiers V, Le François NR, Blier PU. Trypsin-like enzyme from Atlantic wolffish (Anarhichas lupus) viscera: Purification and characterization. Journal of Aquatie Food Product Technology. 2008. 17(1).
11. Trismilah, Lutfi A. Membran polyethersulfone dan regenerated cellulose untuk ultrafiltrasi: Pengaruh pH terhadap proses ultrafiltrasi xilanase. Jurnal Sains Dan Teknologi Indonesia. 2009.11(2): 76-83.
12. Kishimura H, Tokuda Y, Klomklao S, Benjakul S, Ando S. Enzymatic characteristics of trypsin from Pyloric caeca of spotted mackerel (Scomber australisicus). Journal of Food Biochemistry. 2006. 30:466–77.
13. Whitcomb DC, Lowe ME. Human pancreatic digestive enzymes. Springer Science. 2006. 52:1-17.
14. Setyawati I, Wiratmini NI,Wiryatno J. Pertumbuhan, hispatologi ovarium dan fekunditas ikan nila merah (Oreochromis niloticus) setelah paparan pestisida organofosfat. Jurnal Biologi. 2011. XV(2): 44-8.
15. Baehaki A, Suhartono MT, Palupi NS, Nurhayati T. Purifikasi dan karakterisasi protease dari bakteri patogen Pseudomonas aeruginosa. Jurnal Teknologi dan Industri Pangan. 2008. XIX(1).
16. Lehninger AL. Dasar-dasar biokimia. Terjemahan Thenawijaya M. Jilid 1. Jakarta: Penerbit Erlangga; 1982.
17. Winarno FG. Enzim pangan. Jakarta: Penerbit Gramedia; 1988.
Published
2014-04-30
How to Cite
TRISMILAH, TRISMILAH et al. Isolation and Characterization Trypsin - Like Protease (PST) from Lactobacillus plantarum FNCC 0270. JURNAL ILMU KEFARMASIAN INDONESIA, [S.l.], v. 12, n. 1, p. 57-66, apr. 2014. ISSN 2614-6495. Available at: <http://jifi.farmasi.univpancasila.ac.id/index.php/jifi/article/view/187>. Date accessed: 08 feb. 2023.
Section
Articles