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Information Journal Paper

Title

PERIPLASMIC EXPRESSION OF BACILLUS THERMOCATENULATUS LIPASE IN ESCHERICHIA COLI IN PRESENCE OF DIFFERENT SIGNAL SEQUENCES

Pages

  255-262

Abstract

 Efforts to express lipase in the periplasmic space of Escherichia colihave so far been unsuccessful and most of the expressed recombinant lipases accumulate in the insoluble cell fraction. To evaluate the role of native and heterologous signal peptides in translocation of the lipase across the inner membrane ofE. coli, the lipase gene (btl2) was cloned downstream of the native Bacillus signal peptide and also in fusion with thepel B, ans B and ans B/asp signal peptides. For this purpose, four recombinant expression vectors (pYRKP.P, pYRKP.N, pYRKP. A and pYRKP. AA) were constructed and expressed in E. coli. Osmotic shock analysis showed that recombinant lipase was overexpressed as INCLUSION BODIES in E. coli. The lipase INCLUSION BODIES were subsequently solublized, refolded and purified using single column ion-exchange chromatography.To evaluate localization of lipase in the cell, the purified lipases were subjected to CAPILLARY ISOELECTRIC FOCUSING and tandem mass spectrometryResults showed that all signal peptides were able to direct the lipase from the cytoplasm into the periplasmic space of E. coli, because the periplasmic space of E. coliis not suitable for lipase folding, the translocated lipase aggregates in this space as INCLUSION BODIES.

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    APA: Copy

    KARKHANE, ALI ASGHAR, YAKHCHALI, BAGHER, RASTGAR JAZII, FERDOUS, HEMMAT, JAFAR, SHARIATI, PARVIN, KHODABANDEH, MAHVASH, & ZOMORODIPOUR, ALIREZA. (2012). PERIPLASMIC EXPRESSION OF BACILLUS THERMOCATENULATUS LIPASE IN ESCHERICHIA COLI IN PRESENCE OF DIFFERENT SIGNAL SEQUENCES. IRANIAN JOURNAL OF BIOTECHNOLOGY, 10(4), 255-262. SID. https://sid.ir/paper/593047/en

    Vancouver: Copy

    KARKHANE ALI ASGHAR, YAKHCHALI BAGHER, RASTGAR JAZII FERDOUS, HEMMAT JAFAR, SHARIATI PARVIN, KHODABANDEH MAHVASH, ZOMORODIPOUR ALIREZA. PERIPLASMIC EXPRESSION OF BACILLUS THERMOCATENULATUS LIPASE IN ESCHERICHIA COLI IN PRESENCE OF DIFFERENT SIGNAL SEQUENCES. IRANIAN JOURNAL OF BIOTECHNOLOGY[Internet]. 2012;10(4):255-262. Available from: https://sid.ir/paper/593047/en

    IEEE: Copy

    ALI ASGHAR KARKHANE, BAGHER YAKHCHALI, FERDOUS RASTGAR JAZII, JAFAR HEMMAT, PARVIN SHARIATI, MAHVASH KHODABANDEH, and ALIREZA ZOMORODIPOUR, “PERIPLASMIC EXPRESSION OF BACILLUS THERMOCATENULATUS LIPASE IN ESCHERICHIA COLI IN PRESENCE OF DIFFERENT SIGNAL SEQUENCES,” IRANIAN JOURNAL OF BIOTECHNOLOGY, vol. 10, no. 4, pp. 255–262, 2012, [Online]. Available: https://sid.ir/paper/593047/en

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