The Kluyveromyces lactis K1PMR1 gene is the functional homologue of Saccharomyces cerevisiae PMR1 which encodes a Ca2+-ATPase localized in the Golgi apparatus. We studied the effects of K1PMR1 inactivation on the glycosylation and secretion of native and heterologous proteins in K. lactis. We used acid phosphatase, recombinant human serum albumin and alpha-glucoamylase from Arxula adeninivorans as reporter proteins. The Klpmrl Delta strain showed enhanced secretion of the heterologous proteins analyzed; the improved rHSA production did not result from enhanced transcription but rather involved increased translation and/or secretion efficiency. The growth rate of mutant cells resulted slower as compared to that of wild-type strain. The addition of 10 mM calcium to the culture medium, however, not only completely relieved the growth defect of the mutant cells but also improved the rate of heterologous proteins production. Moreover, the addition of this ion in the culture medium of K. lactis did not suppress the glycosylation defects; this is an important difference with respect to S. cerevisiae where the glycosylation is partially restored by Ca2+ addition. The Klpmrl Delta strain as a host offers thus an additional advantage for those cases requiring that the produced recombinant protein would not result hyperglycosylated. (C) 2004 Elsevier B.V. All rights reserved.

KlPMR1 inactivation and calcium addition enhance secretion of non-hyperglycosylated heterologous proteins in Kluyveromyces lactis / Uccelletti, Daniela; F., Farina; Mancini, Patrizia; Palleschi, Claudio. - In: JOURNAL OF BIOTECHNOLOGY. - ISSN 0168-1656. - STAMPA. - 109:1-2(2004), pp. 93-101. (Intervento presentato al convegno 2nd Symposium on Recombinant Protein Production with Prokaryotic and Eukrayotic Cells tenutosi a Cernobbio, ITALY nel NOV, 2002) [10.1016/j.jbiotec.2003.10.037].

KlPMR1 inactivation and calcium addition enhance secretion of non-hyperglycosylated heterologous proteins in Kluyveromyces lactis

UCCELLETTI, Daniela;MANCINI, Patrizia;PALLESCHI, Claudio
2004

Abstract

The Kluyveromyces lactis K1PMR1 gene is the functional homologue of Saccharomyces cerevisiae PMR1 which encodes a Ca2+-ATPase localized in the Golgi apparatus. We studied the effects of K1PMR1 inactivation on the glycosylation and secretion of native and heterologous proteins in K. lactis. We used acid phosphatase, recombinant human serum albumin and alpha-glucoamylase from Arxula adeninivorans as reporter proteins. The Klpmrl Delta strain showed enhanced secretion of the heterologous proteins analyzed; the improved rHSA production did not result from enhanced transcription but rather involved increased translation and/or secretion efficiency. The growth rate of mutant cells resulted slower as compared to that of wild-type strain. The addition of 10 mM calcium to the culture medium, however, not only completely relieved the growth defect of the mutant cells but also improved the rate of heterologous proteins production. Moreover, the addition of this ion in the culture medium of K. lactis did not suppress the glycosylation defects; this is an important difference with respect to S. cerevisiae where the glycosylation is partially restored by Ca2+ addition. The Klpmrl Delta strain as a host offers thus an additional advantage for those cases requiring that the produced recombinant protein would not result hyperglycosylated. (C) 2004 Elsevier B.V. All rights reserved.
2004
glycosylation; heterologous proteins; kluyveromyces lactis; pmr1; secretion
01 Pubblicazione su rivista::01a Articolo in rivista
KlPMR1 inactivation and calcium addition enhance secretion of non-hyperglycosylated heterologous proteins in Kluyveromyces lactis / Uccelletti, Daniela; F., Farina; Mancini, Patrizia; Palleschi, Claudio. - In: JOURNAL OF BIOTECHNOLOGY. - ISSN 0168-1656. - STAMPA. - 109:1-2(2004), pp. 93-101. (Intervento presentato al convegno 2nd Symposium on Recombinant Protein Production with Prokaryotic and Eukrayotic Cells tenutosi a Cernobbio, ITALY nel NOV, 2002) [10.1016/j.jbiotec.2003.10.037].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/236973
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