A mutant l-asparaginase II signal peptide improves the secretion of recombinant cyclodextrin glucanotransferase and the viability of Escherichia coli

Noor Faizah Ismail, Salehhuddin Hamdan, Nor Muhammad Mahadi, Abdul Munir Abd. Murad, Amir Rabu, Farah Diba Abu Bakar, Peter Klappa, Rosli Md Illias

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

l-Asparaginase II signal peptide was used for the secretion of recombinant cyclodextrin glucanotransferase (CGTase) into the periplasmic space of E. coli. Despite its predominant localisation in the periplasm, CGTase activity was also detected in the extracellular medium, followed by cell lysis. Five mutant signal peptides were constructed to improve the periplasmic levels of CGTase. N1R3 is a mutated signal peptide with the number of positively charged amino acid residues in the n-region increased to a net charge of +5. This mutant peptide produced a 1. 7-fold enhancement of CGTase activity in the periplasm and significantly decreased cell lysis to 7. 8% of the wild-type level. The formation of intracellular inclusion bodies was also reduced when this mutated signal peptide was used as judged by SDS-PAGE. Therefore, these results provide evidence of a cost-effective means of expression of recombinant proteins in E. coli.

Original languageEnglish
Pages (from-to)999-1005
Number of pages7
JournalBiotechnology Letters
Volume33
Issue number5
DOIs
Publication statusPublished - May 2011

Fingerprint

Asparaginase
Cyclodextrins
Protein Sorting Signals
Periplasm
Escherichia coli
Recombinant proteins
Inclusion Bodies
Recombinant Proteins
Peptides
Amino acids
Polyacrylamide Gel Electrophoresis
Amino Acids
Costs and Cost Analysis
cyclomaltodextrin glucanotransferase
Costs

Keywords

  • Cell lysis
  • Cyclodextrin glucanotransferase
  • l-asparaginase II signal peptide
  • Mutation
  • Secretion

ASJC Scopus subject areas

  • Biotechnology

Cite this

A mutant l-asparaginase II signal peptide improves the secretion of recombinant cyclodextrin glucanotransferase and the viability of Escherichia coli. / Ismail, Noor Faizah; Hamdan, Salehhuddin; Mahadi, Nor Muhammad; Abd. Murad, Abdul Munir; Rabu, Amir; Abu Bakar, Farah Diba; Klappa, Peter; Illias, Rosli Md.

In: Biotechnology Letters, Vol. 33, No. 5, 05.2011, p. 999-1005.

Research output: Contribution to journalArticle

Ismail, Noor Faizah ; Hamdan, Salehhuddin ; Mahadi, Nor Muhammad ; Abd. Murad, Abdul Munir ; Rabu, Amir ; Abu Bakar, Farah Diba ; Klappa, Peter ; Illias, Rosli Md. / A mutant l-asparaginase II signal peptide improves the secretion of recombinant cyclodextrin glucanotransferase and the viability of Escherichia coli. In: Biotechnology Letters. 2011 ; Vol. 33, No. 5. pp. 999-1005.
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