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Journal : Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology

PURIFICATION AND CHARACTERIZATION OF THE NEWLY THERMOSTABLE PROTEASE PRODUCED BY Brevibacillus thermoruber LII ISOLATED FROM PADANG CERMIN HOTSPRING, INDONESIA Zilda, Dewi Zeswita; Harmayani, Eni; Widada, Jaka; Asmara, Widya; Irianto, Hari Eko; Patantis, Gintung; Fawzya, Yusro Nuri
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 9, No 1 (2014): May 2014
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v9i1.91

Abstract

Thermo stability is among of the vital enzyme characteristics for industrial application. Brevibacillus thermoruber LII was obtained as a potential isolate from the previous researchwhich screened the potential thermostable protease producing bacteria from Indonesian hotspring.The newly thermostable protease produced by thermophilic Brevibacillus thermoruber LII hadbeen purified and characterized. It was predicted that the pure enzyme obtained from Brevibacillusthermoruber LII was homo hexameric, having molecular weight of 36 kDa unit protein and itsnative was 215 kDa. In addition, it was also a neutral metalo serine protease according tobiochemical tests that it was totaly inhibited by PMSF (Phenylmethanesulfonyl fluoride) and EDTA(Ethylenediaminetetraacetic acid). It showed optimum activity at pH of 8 and active in acidic buffer(up to pH of 4). All of metal ion in the form of chloride salt (2.5 mM) which were tested on theenzyme enhanced the enzyme activity but Li2+. Ca2+ion increased the activity and the stability ofenzyme against thermal. The enzyme also showed the stability against solvent. The protease LIIhad optimum temperature at 60oC without CaCl 2and 80 – 85oC with addition of 2.5 mM CaCl 2. TheK Mand V maxvalues for the purified protease LII were 27.2 mg/ml or 0.362 – 0.272 M for substrateHammersteinCasein (MM 75–100 kDa) and 261.1 µg/minute/ml, respectively.
SCREENING OF THERMOSTABLE PROTEASE PRODUCING MICROORGANISMS ISOLATED FROM INDONESIAN HOTSPRING Zilda, Dewi Seswita; Harmayani, Eni; Widada, Jaka; Asmara, Widya; Irianto, Hari Eko; Patantis, Gintung; Fawzya, Yusro Nuri
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 7, No 3 (2012): December 2012
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v7i3.5

Abstract

Although many proteases had been studied and characterized, only a few of them are commercially available.  Protease thermostability is one of the crucial properties for industrialapplication. This research aimed to isolate and to screen the potential isolate which produce thermostable protease. There were 6 isolates (BII-1, BII-2, BII-3, BII-4, BII-6 and LII), isolated using solid Minimal Synthetic Medium (MSM) supplemented with 1.5% skim milk, that have, protease activity. Based on the 16S-rRNA gene sequencing analysis, isolates BII-1, BII-2 and BII- 6 were identified as Bacillus licheniformis, isolates BII-3 and BII-4 were identified as Bacillus subtilis, while isolate LII was identified as Brevibacillus thermoruber. Three isolates (BII-6, BII-4 and LII) were then further investigated for the second screening step using liquid MSM supplemented with 1% skim milk. The isolates (BII-6, BII-4 and LII) optimally produced protease when they were cultivated at 35, 30 and 50o C respectively after 22 h of incubation. Protease produced by BII-6, BII-4 and LII had optimum temperature  of 65, 60 and 85o C, optimum pH at 7-8, 8 and 9 and stable up to 100 min at 55, 60 and 75o C respectively.
Co-Authors . Sismindari ., Wdjijono Aastuti, Wijayanti Dwi Abdul Salam M. Sofro, Abdul Salam Adi Heru Sutomo Aditya Krishar Karim AETH. Wahyuni, AETH. Afiono Agung Prasetyo Ag. Yuswanto Agnes Sri Harti Agnesia Endamg Tri Hastuti Wahyuni Agnesia Endang Tri Hastuti Wahyuni Agnesia Endang Tri Hastuti Wahyuni Agus Eko Srihanto, Agus Eko Agustinus Joko Nugroho, Agustinus Joko Akiyama, Koichi Al Supartinah Santoso, Al Supartinah Alimuddin . Alimuddin A, Alimuddin Alimuddin, A. Alma Linggar Jonarta, Alma Linggar Ambar Pertiwiningrum Anwar Rosyidi April H Wardhana Ardianata, Dana Arief Mulyono Arum Setiawan Asih Kurniawati B. Sardjono Bambang Hariono Bambang Sumiarto Bambang Sutrisno Banun Kusumawardani Boy M. Bachtiar Budi Mulyaningsih Budi Mulyono BUDI SETIADI DARYONO Budi Setiadi Daryono Charles Rangga Tabbu Charles Rangga Tabbu Chatarina Behar, Chatarina Dayono, Budi Setiadi Dewi Agustina Dewi Seswita Zilda Dito Anggoro, Dito Djaswadi Dasuki Doddi Yudhabuntara Dyah Haryuningtyas Dyah Irnawati Eko Agus Srihanto, Eko Agus Eni Harmayani Enny Yusuf Wachidah Yuniwarti Ety Aryati, Ety Gintung Patantis Gusti Ayu Yuniati Kencana Hardyanto Soebono Hardyanto Subono Hari Eko Irianto Heni Susilowati Heru Susetya Hidayanto, Nur Khusni Hidayati, Dewi Noor I Gusti Ketut Suarjana I Gusti Made Krisna Erawan, I Gusti Made Krisna I Gusti Ngurah Kade Mahardika I Wayan Suardana Ida Tjahajati Ignatius Mulyadi, Ignatius Ika Dewi Ana Ika Dyah Kusumawati, Ika Dyah Indwiani Astuti Istriyati ., Istriyati Istriyati Istriyati Istriyati, . Istriyati, . Istriyati, I. Iwan Dwiprahasto JAKA WIDADA Juni Handajani Karna Wijaya Khilyat Ulin Nur Zaini, Khilyat Ulin Nur Khrisdiana Putri, Khrisdiana LANGKAH SEMBIRING M. Haryadi Wibowo Mammed Sagi Mandojo Rukmo Marsetyawan HNE Soesatyo Marsetyawan HNE. Soesatyo, Marsetyawan HNE. Marsetyawan Soesatyo Masashi Kawaichi, Masashi Maxs Urias Ebenhaizar Sanam Maxs Urias Ebenhaizar Sanam, Maxs Urias Michael Haryadi Wibowo Michael Haryadi Wibowo MM.Firdiana Krisnaningsih Mustofa . Mustofa M, Mustofa Mustofa, M. Ning Rintiswati Nobuyuki Harada Nugroho, Dwi Aji Nugroho, Dwi Aji Nursyirwani . Nuryono ., Nuryono Nuryono, N. Osman Sianipar, Osman Pinandi Sri Pudyani, Pinandi Sri Purnama Edy Santosa Putri, Krisdiana Rahmat Setya Adji Regina TC Tandelilin, Regina TC Regina TC. Tandelilin, Regina TC. Reni Nurjasmi, Reni Rini Widayanti Risya Cilmiaty, Risya S Muharsini S Rahmah Umniyati, S Rahmah Sarwo Edy Wibowo Sebastian Margino Setiyono Setiyono Setyawan Budiharta Sidna Artanto, Sidna Sismindari . Sismindari Sismindari Sismindari, S. Siti Sunarintyas Soemitro Djojowidagdo Sri Darmawati Sri Lestari Sri Murwani Subronto Prodjoharjono Suhartono Taat Putra Surya Amanu Suryani Hutomo, Suryani Susi Iravati Syaiful Anwar Tita Ratya Utari Titik Purwati Widowati, Titik Purwati Tiyas Tono Taufiq, Tiyas Tono Tri Ratnaningsih Tri Untari Tri Wibawa Triyanto . Tsutomu Nohno W. Widodo Wajar, Dony wayan T Artama Wayan T. Artama Wayan T. Artama Wayan Tunas Artama Wayan Tunas Artama Widagdo Sri Nugroho Widagdo Sri Nugroho Widjijono Widjijono Widjijono, W. Widodo . Widya Hary Cahyati Wisnu Nurcahyo Yatri Drastini Yulita Kristanti, Yulita Yuni Wijayanti Yusro Nuri Fawzya ZAKI MUBARAK Zilda, Dewi Zeswita