Characterization of the Aspergillus niger pelB gene: structure and regulation of expression

Margo Kusters-van Someren, Michel Flipphi, Leo de Graaff, Hetty van den Broeck, Harry Kester, Albert Hinnen, Jaap Visser

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61 Citations (Scopus)

Abstract

The nucleotide sequence of pelB, a member of the Aspergillus niger pectin lyase multigene family, has been determined. The pelB gene product, PLB, shares 65% amino acid identity with pectin lyase A (PLA) and 60% with pectin lyase D (PLD). Although growth of pelB multicopy transformants on pectin-containing media results in elevated pelB mRNA levels, pectin lyase B (PLB) is barely detectable. This is probably due to degradation of PLB by acid proteases, since multicopy transformants grown on pectin medium with a high concentration of phosphate, leading to a less rapid decline in pH, secrete detectable amounts of PLB. To produce PLB in high amounts under conditions where few other extracellular enzymes are present, we tried two strategies. Firstly, heterologous expression of the pelB gene in A. nidulans, and secondly, expression of the pelB gene under control of the constitutive A. niger pki promoter.

Original languageEnglish
Pages (from-to)113-120
Number of pages8
JournalMGG Molecular & General Genetics
Volume234
Issue number1
DOIs
Publication statusPublished - Jul 1 1992

Keywords

  • Aspergillus niger
  • Heterologous expression
  • Pectin lyase
  • Pyruvate kinase promoter

ASJC Scopus subject areas

  • Genetics

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    Kusters-van Someren, M., Flipphi, M., de Graaff, L., van den Broeck, H., Kester, H., Hinnen, A., & Visser, J. (1992). Characterization of the Aspergillus niger pelB gene: structure and regulation of expression. MGG Molecular & General Genetics, 234(1), 113-120. https://doi.org/10.1007/BF00272352