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All Journal Livestock and Animal Research
Sajidan Sajidan
Department of Biology Faculty of Teaching and Education Sebelas Maret University Jl. Ir. Sutami 36A Surakarta, 57126
Agriculture, Biological Sciences & Forestry Veterinary

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In Vitro Stability of Phytase from Recombinant Bacteria E. Coli BL21 (DE3) EAS1-AMP Adi M.P. Nuhriawangsa; Sajidan Sajidan; Zaenal Bachruddin; Ali Wibowo
Sains Peternakan: Jurnal Penelitian Ilmu Peternakan Vol 9, No 1 (2011): Sains Peternakan
Publisher : Universitas Sebelas Maret (UNS)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20961/sainspet.v9i1.4742

Abstract

The objective of the research was to inquire the Km, Vm, activity, intracellular phytase stability exposed to pH variation, temperature variation and protease (pepsin and pancreas) in vitro. The phytase was produced from recombinant bacteria E. coli BL21(DE3) EAS1-AMP using 1.5 mM IPTG as inducer. Intracellular enzyme was extracted via freeze shock and centrifugation. Pure enzyme was acquired through NI-NTA agarose column. The enzyme was then tested for Km, Vm, phytase activity and stability against pH, temperature and protease. Treatment levels for stability against protease were P0: without protease, P1: addition of pepsin, P2: addition of pepsin and pancreas, and the data were statistically analyzed using analysis of variance of one-way Completely Randomized Design. Crude intracellular phytase had Vm 6.39 υM/sec, Km 34.82 υM, and 277 units activity. Intracellular phytas was stable at pH 4–6 and 0–550 C. Protease level influenced the activity of intracellular phytase (P<0.05). Intracellular phytase was stable against pepsin but not pancreas.
Co-Authors Adi M.P. Nuhriawangsa Ali Wibowo Zaenal Bachruddin