PRODUCTION AND CHARACTERIZATION OF CRUDE INTRACELLULER PHYTASE FROM RECOMBINANT BACTERIA pEAS1AMP

Nuhriawangsa, A. M. P. and Bachruddin, Z. and SAJIDAN, and Wibowo, A. (2009) PRODUCTION AND CHARACTERIZATION OF CRUDE INTRACELLULER PHYTASE FROM RECOMBINANT BACTERIA pEAS1AMP. J.Indonesian Trop.Anim.Agric, 34 (4). pp. 265-271.

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    Abstract

    This research was aimed at producing a crude intracellular phytase characterized from recombinant bacteria. The recombinant bacteria (pEAS1AMP) was produced by way of transforming pET-22b(+) +pEAS1 into competent E. coli BL21 and E. coli BL21(DE3) cells. Crude intracellular phytase production was induced using 1,5 mM Isopropyl-β-D-thiogalactopyranosid (IPTG). Recombinant bacteria product and enzyme activity test followed the Sajidan method. E. coli BL21(+)pEAS1 and E. coli BL21 (DE3)(+)pEAS1 recombinant bacteria showed growth after 20 hours and 10 hours of transformation. Phytase activity of E. coli BL21 (DE3)(+)+pEAS1 showed higher than those of E. coli BL21(+)+pEAS1. Crude intracellular phytase of pEAS1AMP recombinant bacteria has an optimum activity at pH 5, 40o C, incubation period of 60 minutes, substrate concentration of 2%, molecular weight (MW) of 47.3 kDa, Km = 15.91 υM and Vm = 2.41 υM/second. Mg2+ acts as a cofactor but Fe3+ (10-4 M) acts as an inhibitor. Keywords: bacteria recombinant pEAS1AMP, competent cells, crude intracellular phytase

    Item Type: Article
    Subjects: S Agriculture > S Agriculture (General)
    Divisions: Fakultas Matematika dan Ilmu Pengetahuan Alam > Biologi
    Depositing User: mr azis r
    Date Deposited: 04 Jun 2013 18:00
    Last Modified: 04 Jun 2013 18:00
    URI: https://eprints.uns.ac.id/id/eprint/923

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