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

• Published in: Brazilian Journal of Microbiology
• Main Authors: Robert, Julia Macedo, Betancur, Maritza Ocampo, Machado, Antonio Carlos Oliveira, Arruda, Andrelisse, Reis, Viviane Castelo Branco, Almeida, Rodrigo Volcan, Torres, Fernando Araripe Gonçalves, Alegre, Pau Ferrer, Valero, Francisco, Freire, Denise Maria Guimarães
• Format: Text
• Language: English
• Published: Springer International Publishing 2019
• Subjects: Biotechnology and Industrial Microbiology - Research Paper; Antarc*; Antarctica
• Online Access: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6863280/; http://www.ncbi.nlm.nih.gov/pubmed/30827000; https://doi.org/10.1007/s42770-019-00056-8

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 P(PGK1) 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s42770-019-00056-8) contains supplementary material, which is available to authorized users.