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HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS SUWANTO, ANTONIUS; MALIK, AMARILA; TJAHJONO, BUDI; HARLING, ROB
BIOTROPIA No. 20 (2003)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (405.968 KB) | DOI: 10.11598/btb.2003.0.20.182

Abstract

A transcriptional fusion for an Aeromonas caviae chitinase gene was constructed under the control of a constitutive promoter of the kanaraycin resistance gene (PKmR). The construct was inserted into a medium copy number broad host range plasmid vector to yield recombinant plasmid pAM340, which harbored transcriptional fusion PKmR- chi. Another transcriptional fusion, Ptac-chi, in a recombinant plasmid pAM630, was conducted as comparison. Triparental mating of E. coli  carrying the recombinant plasmids with Pseudomotws fluorescens  5100, a phyllosphere bacterium, was performed. Pseudomonas fluorescens  5100 exconjugants were examined for constitutive expression of chitinase employing a spectrophotometric assay; they showed stronger chitin degradation activity than Escherichia coli transformants. Using a fungal antagonism plate assay, this chitinolytic P. fluorescens, however, could not inhibit selected phytopathogenic fungi. Keywords:    Aeromonas   caviae/  chitinase   gene/transcriptional   fusion/PKm'V   Vtac-chilPseudomonas fluorescens
SOLATION AND CHARACTERIZATION OF A NOVEL BENZOATE- UTILIZING Serratia marcescens SURYANTO, DWI; SUWANTO, ANTONIUS
BIOTROPIA No. 21 (2003)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (424.953 KB) | DOI: 10.11598/btb.2003.0.21.184

Abstract

A new benzoate-utilizing strain, Serratia marcescens DS-8, isolated from the environment was characterized. The strain was enterobacilli, Gram negative, mesophilic, non halophilic, and aerobic bacterium that showed motile ovale-rod shaped cells. The isolate produced extracellular chitinase, protease, and prodigiosin (a red pigment produced by several Serratia strains yielding bright red or pink colonies). A physiological assay using Microbact* test showed that the strain was closely related to Klebsiella ozaenae  (49.85%) and Serratia liquefaciens  (24.42%), respectively. However, 16S rRNA sequence analysis indicated that the strain was closely related to  S. marcescens  DSM 30121 with similarity level of 98%. DS-8 strain was able to synthesize its own vitamins. Optimum growth in benzoate was obtained at pH between 7-8.5 and NaCl concentration of 1-1.5% (w/v). The isolate could grow in benzoate-containing medium up to 10 mM. Other carbon sources that could support the growth of DS-8 were casamino acid, glutamate, glucose, acetate, potato starch, and ethanol. Keywords: Serratia marcescens/aromatic degradation/168 rRNA sequence
ISOLATION AND CHARACTERIZATION OF MANNANOLYTIC THERMOPHILIC BACTERIA FROM PALM OIL SHELL AND THEIR MANNANASE ENZYME PRODUCTION PROPERTIES SUWANTO, ANTONIUS; SUMARDI; HENAWIDJAJA, MAGGY T; URWADARIA, T RESNAWATI P
BIOTROPIA No. 25 (2005)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2005.0.25.211

Abstract

A mannanolytic thermophilic bacterium (L-07) was isolated from palm oil shell after 2 days ofenrichment in liquid medium supplemented with 1% palm kernel meal as mannan source. Sequenceanalysis of 16S-rRNA indicated that L-07 was similar (98%) to  Geobacillus stearothermophilus, aspecies of thermophilic aerobic bacteria. We found that  G. stearothermophilus L-07 producedextracellular β-1,4-mannanases, but no β-manosidase and α-galactosidase activities. The growth of L-07reached its maximum (3.0 x 106 cell/ml) at 12-20 hours, while the highest  β-mannanase activity (0.52U/ml) was observed in culture medium after 36 hours of cultivation at 60oC. The medium containinglocust bean gum was the best for producing extracellular β-1,4-mannanases compared with kolang kaling,konjak, and palm kernel meal. SDS-PAGE and zymogram analysis demonstrated that crude mannanasecomplex of L-07 from locust bean gum containing medium comprised three active bands with molecularweight of 85, 73 and 50 kDa.   Keywords :  Extracellular enzyme/mannanase/Geobacillus stearothermophilus
Co-Authors . EFRIWATI . Melani . NURHAIMI-HARIS A A Hermosaningtyas A A Hermosaningtyas, A A ADI YULANDI Agus PURWANTARA Agus Purwito Agustin Wydia Gunawan Agustina, Delia Aini Nurul Alfred Michael ALINA AKHDIYA RUSMANA Amarila Malik Andi Khaeruni Andi Khaeruni Andreas Adhi Satya Andreas Adhi Satya Anja Meryandini ARI SUSILOWATI Arifudin Rafif , Muhammad Ghildan Arild Ranlym Arifin Arina Amalia Putri Aris Tjahjoleksono Aris Tri Wahyudi ARTINI PANGASTUTI Asmini Budiani Basil J NIKOLAU BIBIANA W LAY Budi Tjahjono Budinarta, Widyah C Hanny Wijaya CAHYA PRIHATNA Cahya Prihatna Cahya Prihatna CECILIA ANNA SEUMAHU CECILIA ANNA SEUMAHU DEBORA HADISUSANTO Delia Agustina Desniar - - DIAH ISKANDRIATI DIANA ELIZABETH WATURANGI Djoko Santoso Dondin Sajuthi DWI SURYANTO Dyah Kusuma Anggraini Edi Husen EDI HUSEN Edi Husen ERDELEN, WALTER ERNIN HIDAYATI ESTI PUSPITASARI Esti Puspitasari Esti Utarti Eunice Limantara EVELINE AYU Felicia Felicia Felicia Kartawidjajaputra Ferymon Mahulette Ferymon Mahulette Ferymon Mahulette, Ferymon G. A. Wattimena Gayuh Rahayu Griselda Griselda Griselda Herman Natadiputri Griselda, Griselda Hajrial ASWIDINNOO HAJRIAL ASWIDINNOOR Hariyatun Hariyatun Hariyatun, Hariyatun Hariyatun, Hariyatun HARLING, ROB HENAWIDJAJA, MAGGY T Hermosaningtyas, Anastasia Aliesa Iman Rusmana Inez H.S. Loeddin Suharsono It Jamilah IVAN HANJAYA JOANITA SADELI Jr., Muhammad Zairin KATHARINA JESSICA Kusharyoto, Wien Kusharyoto, Wien Lilis Nuraida Linda Wati LISTYA UTAMI KARMAWAN Ludovika Jessica Virginia Maggy T Suhartono Maggy T Suhartono MAGGY T. SUHARTONO Maggy T. Suhartono MAGGY THENAWIJAYA SUHARTONO Maria Dita Febriani Lumban Gaol Maria Indah Purnamasari Maria Indah Purnamasari Maria Indah Purnamasari Maria Sugiharti Meity S. Sinaga MEITY SURADJI SINAGA Muhamad Azwar Syah Muhammad Agus Muljanto Ni Nyoman Tri Puspaningsih NISA RACHMANIA MUBARIK NURITA TORUAN-MATHIUS PRIHASTO SETYANTO Prihasto Setyanto Prihatna , Cahya Puspitasari, Esti QURROTA A’YUN R. Sapto Hendri Boedi Soesatyo Raden Ajie Syahbarie RAHAYU WIDYASTUTI RASTI SARASWATI Rasti Saraswati RASTI SARASWATI Rasti Saraswati RATIH DEWI HASTUTI RATNA SETYANINGSIH RATNA SETYANINGSIH RIDWAN AFFANDI Rika Indri Astuti ROHANI CINTA BADIA GINTING Rustam, Yepy Hardi Satya, Andreas Adhi Sheila Sutanto Sheila Sutanto, Sheila Sumardi Suryo Wiyono SUSAN SOKA SUSILOWATI1 SUSILOWATI1 Tan Watumesa Agustina TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Tati Barus URWADARIA, T RESNAWATI P VICKY MEICY WIDANARNI WIDANARNI Widanarni Widanarni Wien Kusharyoto Yasinta Ratna Esti Wulandari Yepy Hardi Rustam Yogiara Yogiara YULIN LESTARI Yusminah Hala