TY - JOUR
T1 - Increase of Candida antarctica lipase B production under PGK promoter in Pichia pastoris: effect of multicopies
AU - Robert, Julia Macedo
AU - Betancur, Maritza Ocampo
AU - Machado, Antonio Carlos Oliveira
AU - Arruda, Andrelisse
AU - Reis, Viviane Castelo Branco
AU - Almeida, Rodrigo Volcan
AU - Torres, Fernando Araripe Gonçalves
AU - Alegre, Pau Ferrer
AU - Valero, Francisco
AU - Freire, Denise Maria Guimarães
PY - 2019/4/1
Y1 - 2019/4/1
N2 - © 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.
AB - © 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.
KW - Candida antarctica, lipase B
KW - Constitutive expression
KW - Gene dosage
KW - Komagataella phaffii
KW - Multicopy vector integration
KW - Lipase/biosynthesis
KW - Saccharomycetales/genetics
KW - Pichia/genetics
KW - Promoter Regions, Genetic/genetics
KW - Candida/enzymology
KW - Gene Dosage/genetics
KW - Glycerol/metabolism
KW - Fungal Proteins/biosynthesis
KW - SYSTEM
KW - AMPLIFICATION
KW - BIODIESEL
KW - BIOCATALYST
KW - Candida antarctica
KW - GENE
KW - INTEGRATION
KW - COPY NUMBER
KW - PROTEIN EXPRESSION
KW - SURFACE-ANTIGEN
KW - lipase B
KW - SECRETION
UR - http://www.mendeley.com/research/increase-candida-antarctica-lipase-b-production-under-pgk-promoter-pichia-pastoris-effect-multicopie
U2 - 10.1007/s42770-019-00056-8
DO - 10.1007/s42770-019-00056-8
M3 - Article
C2 - 30827000
VL - 50
SP - 405
EP - 413
JO - Brazilian Journal of Microbiology
JF - Brazilian Journal of Microbiology
SN - 1517-8382
IS - 2
ER -