Increase of Candida antarctica lipase B production under PGK promoter in Pichia pastoris: effect of multicopies

Julia Macedo Robert, Maritza Ocampo Betancur, Antonio Carlos Oliveira Machado, Andrelisse Arruda, Viviane Castelo Branco Reis, Rodrigo Volcan Almeida, Fernando Araripe Gonçalves Torres, Pau Ferrer Alegre, Francisco Valero, Denise Maria Guimarães Freire

Research output: Contribution to journalArticleResearch

1 Citation (Scopus)

Abstract

© 2019, Sociedade Brasileira de Microbiologia. The effect of gene dosage on the production of Candida antarctica lipase B (CalB) in the methylotrophic yeast Komagataella phaffii, at high densities in a simple medium containing crude glycerin as the sole carbon source, is described. The use of crude glycerin, the main by-product of biodiesel production from vegetable oils, will reduce the production cost of the bioprocess. Two K. phaffii strains were constructed with one or three copies of LipB, an optimized version of the gene encoding CalB under the control of the constitutive PPGK1 promoter. These two constructs were tested and compared on batches using minimal-salts medium with crude glycerin. The strain with three copies achieved a higher enzyme yield (48,760 U/L, 2.3-fold higher than the one-copy strain), with 42 g/L biomass, with no effects on growth.
Original languageEnglish
Pages (from-to)405-413
Number of pages9
JournalBrazilian Journal of Microbiology
Volume50
Issue number2
DOIs
Publication statusPublished - 1 Apr 2019

Keywords

  • Candida antarctica, lipase B
  • Constitutive expression
  • Gene dosage
  • Komagataella phaffii
  • Multicopy vector integration
  • Lipase/biosynthesis
  • Saccharomycetales/genetics
  • Pichia/genetics
  • Promoter Regions, Genetic/genetics
  • Candida/enzymology
  • Gene Dosage/genetics
  • Glycerol/metabolism
  • Fungal Proteins/biosynthesis
  • SYSTEM
  • AMPLIFICATION
  • BIODIESEL
  • BIOCATALYST
  • Candida antarctica
  • GENE
  • INTEGRATION
  • COPY NUMBER
  • PROTEIN EXPRESSION
  • SURFACE-ANTIGEN
  • lipase B
  • SECRETION

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