Protein rekombinan seperti vaksin, antibodi, hormon, dan obat-obatan, semakin dibutuhkan oleh ternak dan manusia. Hambatan utama untuk menghasilkan protein rekombinan pada Escherichia coli sebagai inang yang digunakan paling luas adalah degradasi oleh enzim proteolitik.
Hal ini disebabkan karena E. Coli memiliki sejumlah enzim proteolitik yang tersebar di dalam sitoplasmanya. Untuk itu, lebih dari 90% degradasi protein terjadi di dalam sitoplasmanya. Pada penelitian ini, peneliti telah menghasilkan mutant E. Coli BW25113 yang tidak memiliki gen penyandi enzim protease dengan menggunakan kombinasi metode pengerusakan kromosom dan metode transduksi phage P1.
Pembuatan mutan tersebut dimulai dengan pengerusakan gen penyandi enzim protease pada kromosom bakteri dengan produk PCR yang memiliki bagian yang homolog dengan gen target. Mutan-mutan yang dihasilkan kemudian digunakan untuk menghasilkan mutan ganda dengan metode Transduksi phage P1.
Analisis fenotif dan genotif menunjukkan bahwa kombinasi kedua metode tersebut sangat efektif untuk membuat lebih dari satu mutasi pada E. Untuk itu, mutan E. Coli yang telah diperoleh akan sangat bermanfaat untuk menghasilkan aneka protein rekombinan untuk ternak dan manusia. Cherepanov PP and Wackernagel W 1995. Gene disruption in Escherichia coli: TCR and Km R cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene, 158, 9-14.
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Enzim Proteolitik Adalah
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Protease is an enzyme that can hydrolyze protein into simpler compounds, i.e peptides and amino acids. Microbial Proteases have the potency to be applied in industries such as detergents, skins, silver recovery, dairy, baking, beverages and pharmaceutical industries. These hydrolytic enzyme are efficiently involved in the food industry to increase the nutritional value, digestibility, palatability, flavour and reducing allergenic compounds as well as in the management of domestic and industrial wastes. The purpose of this study was to investigate the ability of Stenotrophomonas sp.
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Isolated from Mount Bromo, East Java in producing protease. Protease activity of the bacterial isolate was qualitatively determined by formation of a clear zone surrounded their colonies on media containing skim milk (1%).
We analyzed its proteolic activity against some effects of the incubation period, pH, temperatures and addition of monovalent and divalent metal ionsquantitatively using a spectrophotometer? 280 nm.The results showed that the optimum activity after incubation for two days was 315.88 U/ mL. The enzyme has continued to its activity at pH 8 (419.68 U/mL) and maintained its stability at 398.22 U/mL with activities decreased to 94.87%, while its activity at 60°C was 519.86 U/mL and could maintain its stability at 419.58 U/ mL, the activity decreased to 74.75%. The addition of Ca2+ could activated its enzyme activity at the amount of 424.33U/mL, while without addition of the ion its activity was 400.29 U/mL. The addition with ion Mn²+, K+, Na+ and Cu 2+ could act as inhibitors that might reduced the activity of the enzyme. Published by: Research Center for Biology-LIPI Accredited by Panitia Penilai Majalah Ilmiah (P2MI)-LIPI Research Center For Biology - Cibinong Science Center (CSC) JL. Raya Jakarta - Bogor Km.46 Cibinong 16911 Bogor - Indonesia Phone: +62(0)1-8797636, Fax: +62(0)2 E-mail Portal: [email protected] url: e-journal.biologi.lipi.go.id E-mail Jurnal Reinwardtia: [email protected] url: E-mail Jurnal Treubia: [email protected] url: E-mail Jurnal Berita Biologi: [email protected] url: E-mail Jurnal Zoo Indonesia: [email protected] url: E-mail Jurnal Biologi Indonesia: [email protected]; [email protected] url: http://e-journal.biologi.lipi.go.id/index.php/jurnalbiologiindonesia.
ABSTRAK Helicobacter pylori berperan dalam infark miokard akut dan penyakit jantung koroner. Sitotoksin H.pylori dapat menginduksi sitokin inflamasi IL1-β dan TNFα yang mengaktivasi enzim proteolitik MMP-9 dan menyebabkan degradasi kolagen melalui plak aterosklerosis. Penelitian ini dilakukan untuk membuktikan kemampuan whole cell H.pylori dalam menginduksi degadrasi kolagen tipe IV melalui peningkatan aktifitas makrofag.
Aktivitas makrofag dilihat dari kadar TNFα dan IL1-β yang diukur dengan metode Elisa. Hasil uji ANOVA mengindikasikan perbedaan signifikan (p.
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Enzim Protease Pdf
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