The Effect of Organic Nitrogen Sources on Recombinant Glucoamylase Production by Aspergillus niger in Chemostat Culture

Authors: Swift R.J.¹; Karandikar A.¹; Griffen A.M.¹; Punt P.J.²; van den Hondel C.A.M.J.J.²; Robson G.D.¹; Trinci A.P.J.¹; Wiebe M.G.¹

Source: Fungal Genetics and Biology, Volume 31, Number 2, November 2000 , pp. 125-133(9)

Publisher: Academic Press

Buy & download fulltext article:

Price: $52.63 plus tax (Refund Policy)

Abstract:

Aspergillus niger B1, a recombinant strain carrying 20 extra copies of the native glucoamylase gene, was grown in glucose-limited chemostat cultures supplemented with various organic nitrogen sources (dilution rate 0.12 ± 0.01 h^-1, pH 5.4). In cultures supplemented with l-alanine, l-methionine, casamino acids, or peptone, specific glucoamylase (GAM) production rapidly decreased to less than 20% of the initial level. Reducing the pH of the culture to 4.0 resulted in stable GAM production for up to 400 h. Morphological mutants (a light brown and a dark brown mutant) appeared in each fermentation and generally displaced B1. Light brown mutants had higher selection coefficients relative to B1 than dark brown mutants and became the dominant strain in all fermentations except those maintained at pH 4.0. Several mutants isolated from these cultures had reduced ability to produce GAM in batch culture, although few had lost copies of the glaA gene. Some mutants had methylated DNA. Copyright 2000 Academic Press.

Keywords: Aspergillus niger; chemostat culture; glucoamylase; recombinant protein; nitrogen source; genetic instability

Language: English

Document Type: Research article

Affiliations: 1: School of Biological Sciences, University of Manchester, 1.800 Stopford Building, Manchester, M13 9PL, United Kingdom 2: Department of Applied Microbiology and Gene Technology, TNO Nutrition and Food Research Institute, Zeist, 3700 AJ, The Netherlands

Publication date: 2000-11-01