Sri Budiarti Poerwanto
Departemen Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Institut Pertanian Bogor

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PEMURNIAN DAN KARAKTERISASI INHIBITOR PROTEASE DARI Chromohalobacter sp. 6A3, BAKTERI YANG BERASOSIASI DENGAN SPONS Xetospongia testudinaria [Purification and Characterization of Protease Inhibitor from Chromohalobacter sp. 6A3, Bacteria-associated with S Nurhayati, Tati; Suhartono, Maggy T.; Nuraida, Lilis; Poerwanto, Sri Budiarti
Jurnal Teknologi Dan Industri Pangan Vol 21, No 2 (2010): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (464.732 KB) | DOI: 10.6066/3411

Abstract

PEMURNIAN DAN KARAKTERISASI INHIBITOR PROTEASE DARI Chromohalobacter sp. 6A3, BAKTERI YANG BERASOSIASI DENGAN SPONS Xetospongia testudinaria [Purification and Characterization of Protease Inhibitor from Chromohalobacter sp. 6A3, Bacteria-associated with Sponge Xetospongia testudinaria]           Tati Nurhayati1)*, Maggy T. Suhartono2), Lilis Nuraida2), Sri Budiarti Poerwanto3) 1) Departemen Teknologi Hasil Perairan, Fakultas Perikanan dan Ilmu Kelautan, Institut Pertanian Bogor 2) Departemen Ilmu dan Teknologi Pangan, Fakultas Teknologi Pertanian, Institut Pertanian Bogor 3) Departemen Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Institut Pertanian Bogor   Diterima 26 November 2009/ Disetujui  15 Desember 2010 ABSTRACT   Various sponges has been reported to produce protease inhibitor which could inhibit protease activity of pathogenic bacteria. The previous research showed that bacteria-associated with sponge could produce bioactive compound similar to their host, including protease inhibitor. The purposes of this research were to purify protease inhibitor from Chromohalobacter sp. 6A3 and to study the characteristics of the protease inhibitor. The result showed that the protein can be extracted by 30 % (v/v) acetone, purified by gel filtration (Sephadex G-75) and finally, purified by anion exchanger (Sephadex A-50). The molecular weight of the purified protease inhibitor after gel filtration was estimated as 21,31 kDa and 17,05 kDa, but anion exchanger gave protein with estimated molecular weight of 21,31 kDa The optimum temperature and pH were 30 oC and 5 respectively. The protease inhibitor could resist heating at 30 oC for 10 minutes. Incubation of the inhibitor at 30 oC, pH 5, still retained its activity until 3 hours. The purified enzyme inhibitor was also still active after incubated at pH from 5 to 6 for 1 hour. The most susceptible substrate (enzyme) for the inhibitor was protease from P. aeruginosa. The protease inhibitor was inhibited by metal ions except Na+ 1mM. Activity of the inhibitor increased twofold by SDS 5 mM. IC 50 of the protease inhibitor was 3.48 nM. The protease inhibitor inhibited the enzyme uncompetitively.   Key words: chromohalobacter, protease inhibitor, sponge  
Influence Of Glucose And Yeast Extract Toward Production Of Pseudomonas Aeruginosa-Protease Inhibitor From Chromohalobacter Sp. 6a3 (Bacteria Associated With Sponge Xetospongia Testudinaria) Nurhayati, Tati; Thenawidjaja, Maggy; Nuraida, Lilis; Poerwanto, Sri Budiarti
Jurnal Teknologi Industri Pertanian Vol 19, No 2 (2009): Jurnal Teknologi Industri Pertanian
Publisher : Jurnal Teknologi Industri Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (219.286 KB)

Abstract

One way to inhibit protease activity is search is compound which can inhibit the enzyme known as protease inhibitor. The bacteria associated with sponge Xetospongia testudinaria, Chromohalobacter sp. 6A3, as producer Pseudomonas aeruginosa-protease inhibitor. Because the compound is important, determination medium composition for producing is very important to be conducted.  The purpose of this research was to determinate the glucose and yeast extract consentration accurately so protease inhibitor would be produced in a short time.  The accurate medium composition for producing the protease inhibitor were 0.1%(w/v) yeast extract; 0.05% (w/v) glucose;  0.5%(w/v) special peptone; 0.2%(v/v) trace element; and 2%(w/v) NaCl at pH 7.Keywords: Chromohalobacter sp., protease inhibitor, sponges.
Preliminary Characterization of Protease Inhibitor from Bacteria-Associated with Sponge from Panggang Island, Seribu Islands NURHAYATI, TATI; SUHARTONO, MAGGY THENAWIDJAJA; NURAIDA, LILIS; POERWANTO, SRI BUDIARTI
HAYATI Journal of Biosciences Vol 13, No 2 (2006): June 2006
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (157.733 KB) | DOI: 10.4308/hjb.13.2.58

Abstract

Pathogenic bacteria produced protease that involved in molecular mechanism of foodborne disease. Produced protease involved in molecular mechanisms of foodborne diseases. The purpose of this research was to screen, identify and characterize the potential microorganisms associated with sponge as producer of protease inhibitor. Among 96 isolates examined, four isolates i.e 10A6, 6A3, 9A51, and 1A12 yielded protease inhibitors which were potential to inhibit protease substrates (40-90%). One of the most potential protease inhibitor producer, the bacteria isolate 6A3, was identified as Chromohalobacter sp. Chromohalobacter sp.6A3 produced protease inhibitor with optimum temperature and pH 300 C and 5, respectively. The inhibitor activity was stable when incubated at 400 C for ten minutes or at 300 C for 8 hours. Key words: Bacteria, Chromohalobacter sp., protease inhibitor, screen, sponge
PEMURNIAN DAN KARAKTERISASI INHIBITOR PROTEASE DARI Chromohalobacter sp. 6A3, BAKTERI YANG BERASOSIASI DENGAN SPONS Xetospongia testudinaria [Purification and Characterization of Protease Inhibitor from Chromohalobacter sp. 6A3, Bacteria-associated with S Nurhayati, Tati; Suhartono, Maggy T.; Nuraida, Lilis; Poerwanto, Sri Budiarti
Jurnal Teknologi dan Industri Pangan Vol 21, No 2 (2010): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (464.732 KB)

Abstract

 Various sponges has been reported to produce protease inhibitor which could inhibit protease activity of pathogenic bacteria. The previous research showed that bacteria-associated with sponge could produce bioactive compound similar to their host, including protease inhibitor. The purposes of this research were to purify protease inhibitor from Chromohalobacter sp. 6A3 and to study the characteristics of the protease inhibitor. The result showed that the protein can be extracted by 30 % (v/v) acetone, purified by gel filtration (Sephadex G-75) and finally, purified by anion exchanger (Sephadex A-50). The molecular weight of the purified protease inhibitor after gel filtration was estimated as 21,31 kDa and 17,05 kDa, but anion exchanger gave protein with estimated molecular weight of 21,31 kDa The optimum temperature and pH were 30 oC and 5 respectively. The protease inhibitor could resist heating at 30 oC for 10 minutes. Incubation of the inhibitor at 30 oC, pH 5, still retained its activity until 3 hours. The purified enzyme inhibitor was also still active after incubated at pH from 5 to 6 for 1 hour. The most susceptible substrate (enzyme) for the inhibitor was protease from P. aeruginosa. The protease inhibitor was inhibited by metal ions except Na+ 1mM. Activity of the inhibitor increased twofold by SDS 5 mM. IC 50 of the protease inhibitor was 3.48 nM. The protease inhibitor inhibited the enzyme uncompetitively.
Preliminary Characterization of Protease Inhibitor from Bacteria-Associated with Sponge from Panggang Island, Seribu Islands NURHAYATI, TATI; SUHARTONO, MAGGY THENAWIDJAJA; NURAIDA, LILIS; POERWANTO, SRI BUDIARTI
HAYATI Journal of Biosciences Vol 13, No 2 (2006): June 2006
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.13.2.58

Abstract

Pathogenic bacteria produced protease that involved in molecular mechanism of foodborne disease. Produced protease involved in molecular mechanisms of foodborne diseases. The purpose of this research was to screen, identify and characterize the potential microorganisms associated with sponge as producer of protease inhibitor. Among 96 isolates examined, four isolates i.e 10A6, 6A3, 9A51, and 1A12 yielded protease inhibitors which were potential to inhibit protease substrates (40-90%). One of the most potential protease inhibitor producer, the bacteria isolate 6A3, was identified as Chromohalobacter sp. Chromohalobacter sp.6A3 produced protease inhibitor with optimum temperature and pH 300 C and 5, respectively. The inhibitor activity was stable when incubated at 400 C for ten minutes or at 300 C for 8 hours. Key words: Bacteria, Chromohalobacter sp., protease inhibitor, screen, sponge
Influence Of Glucose And Yeast Extract Toward Production Of Pseudomonas Aeruginosa-Protease Inhibitor From Chromohalobacter Sp. 6a3 (Bacteria Associated With Sponge Xetospongia Testudinaria) Nurhayati, Tati; Thenawidjaja, Maggy; Nuraida, Lilis; Poerwanto, Sri Budiarti
Journal of Agroindustrial Technology Vol 19, No 2 (2009): Jurnal Teknologi Industri Pertanian
Publisher : Department of Agroindustrial Technology, Bogor Agricultural University

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

One way to inhibit protease activity is search is compound which can inhibit the enzyme known as protease inhibitor. The bacteria associated with sponge Xetospongia testudinaria, Chromohalobacter sp. 6A3, as producer Pseudomonas aeruginosa-protease inhibitor. Because the compound is important, determination medium composition for producing is very important to be conducted.  The purpose of this research was to determinate the glucose and yeast extract consentration accurately so protease inhibitor would be produced in a short time.  The accurate medium composition for producing the protease inhibitor were 0.1%(w/v) yeast extract; 0.05% (w/v) glucose;  0.5%(w/v) special peptone; 0.2%(v/v) trace element; and 2%(w/v) NaCl at pH 7.Keywords: Chromohalobacter sp., protease inhibitor, sponges.