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Pencirian Produksi Amilase oleh Saccaromyces cerevisiae W303A Rekombinan Thontowi, Ahmad; Puspaningsih, Ni Nyoman Tri; Hadi, Sofjan; Purkan, Purkan; Irawan, Bambang
JURNAL BIOLOGI INDONESIA Vol 3, No 3 (2002): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (398.112 KB) | DOI: 10.14203/jbi.v3i3.3464

Abstract

ABSTRACTCharacterization of Amylase Production by Saccharomyces cerevisiae W303A Recombinants. Cloning of amylase gene from Endomycopsis fibuligera ITB.R.cc.64 into S. cerevisiae W303a can effectively increase the yeast function to digest starch directly into ethanol. Production of amylase by S. cerevisiae W303a recombinants (I and P) were done by growing in yeast peptone starch (YPS) medium. The result showed that the recombinants could be produced of amylase by gave clear zone after staining by iodium vapor. The optimum condition of production of amylase by S. cerevisiae W303a recombinants were pH 7.0, 40?C temperature incubation, and gave maximum activity after 36 hours incubation. Amylase activity of I was higher than P recombinant for these condition respectively.Key words: Characterization, amylase, S. cerevisiae W303a
Construction of pY-Af Vector for Expression of Thermostable α-L-Arabinofuranosidase in Saccharomyces cerevisiae Wirajana, I Nengah; Puspaningsih, Ni Nyoman Tri; Wasito, Eddy Bagus; Kusuma, Soekry Erfan; Kimura, Tetsuya; Sakka, Kazuo
ANNALES BOGORIENSES Vol 14, No 2 (2010): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/51

Abstract

In this research,  construction  of expression  vector  for thermostable α -L-arabinofuranosidase  in Saccharomyces cerevisiae was conducted. BJ1824 was conducted The  Escherichia coli/S. cerevisiae  shuttle vector, pYES2 was  used  as  parental  vector  in  construction.  The  abfA  gene  encoding  α-L- arabinofuranosidase  from Geobacillus  thermoleovorans  IT-08  was  amplified  by  PCR,  in  which  the  plasmid  pTP510 was  used  as  a template. The amplification product was treated with  SacI and XhoI and then subcloned to the pYES2 vector, which was previously digested with  SacI and  XhoI. The recombinant plasmid was designated as pY-Af and propagated  first  in  E.  coli  Top 10,  and  then  transformed  into  S.  cerevisiae  BJ1824.  For  α- Larabinofuranosidase (AbfA) production, the yeast transformants were grown in YNBG selective medium and YPG rich medium, using galactose as an inducer. The AbfA activity was assayed by measuring the amount of p-nitrophenol (pNP) released  from  p-nitrophenyl-α-L-arabinofuranoside  (pNPA) substrate at pH 6.0 and 70 C  for  30  min.  The  recombinant  AbfA  activity  was  detected  in  either  of  culture  medium  (0.98%),  cellassociated (14.17%) and intracellular (84.85%) when recombinant yeast was grown in YPG rich medium.Key words: α-L-arabinofuranosidase; Saccharomyces cerevisiae; expression vector
PENCIRIAN PRODUKSI AMILASE OLEH SACCAROMYCES CEREVISIAE W303A REKOMBINAN Thontowi, Ahmad; Puspaningsih, Ni Nyoman Tri; Hadi, Sofjan; Purkan, Purkan; Ni'mahtuzahroh, Ni'mahtuzahroh; Irawan, Bambang
JURNAL BIOLOGI INDONESIA Vol 3, No 3 (2002): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v3i3.3464

Abstract

ABSTRACTCharacterization of Amylase Production by Saccharomyces cerevisiae W303A Recombinants. Cloning of amylase gene from Endomycopsis fibuligera ITB.R.cc.64 into S. cerevisiae W303a can effectively increase the yeast function to digest starch directly into ethanol. Production of amylase by S. cerevisiae W303a recombinants (I and P) were done by growing in yeast peptone starch (YPS) medium. The result showed that the recombinants could be produced of amylase by gave clear zone after staining by iodium vapor. The optimum condition of production of amylase by S. cerevisiae W303a recombinants were pH 7.0, 40?C temperature incubation, and gave maximum activity after 36 hours incubation. Amylase activity of I was higher than P recombinant for these condition respectively.Key words: Characterization, amylase, S. cerevisiae W303a