Advanced search
2 files | 2.34 MB

Optimized expression of the Starmerella bombicola lactone esterase in Pichia pastoris through temperature adaptation, codon-optimization and co-expression with HAC1

Stijn De Waele (UGent) , Isabel Vandenberghe (UGent) , Bram Laukens (UGent) , Sören Planckaert (UGent) , Stijn Verweire (UGent) , Inge Van Bogaert (UGent) , Wim Soetaert (UGent) , Bart Devreese (UGent) and Katarzyna Ciesielska (UGent)
Author
Organization
Abstract
The Starmerella bombicola lactone esterase (SBLE) is a novel enzyme that, in vivo, catalyzes the intramolecular esterification (lactonization) of acidic sophorolipids in an aqueous environment. In fact, this is an unusual reaction given the unfavorable conditions for dehydration. This characteristic strongly contributes to the potential of SBLE to become a 'green' tool in industrial applications. Indeed, lactonization occurs normally in organic solvents, an application for which microbial lipases are increasingly used as biocatalysts. Previously, we described the production of recombinant SBLE (rSBLE) in Pichia pastoris (syn. Komagataella phaffii). However, expression was not optimal to delve deeper into the enzyme's potential for industrial application. In the current study, we explored codon-optimization of the SBLE gene and we optimized the rSBLE expression protocol. Temperature reduction had the biggest impact followed by codon-optimization and co-expression of the HAC1 transcription factor. Combining these approaches, we achieved a 32-fold improvement of the yield during rSBLE production (from 0.75 mg/l to 24 mg/L culture) accompanied with a strong reduction of contaminants after affinity purification.
Keywords
Lipase, Lactonase, Starmerella bombicola, Pichia pastoris, Protein purification, Green chemistry, UNFOLDED-PROTEIN RESPONSE, N-GLYCOSYLATION, LIPASE, SOPHOROLIPIDS, YEAST, GENE, DEGRADATION, LEVEL

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 709.20 KB
  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 1.63 MB

Citation

Please use this url to cite or link to this publication:

Chicago
De Waele, Stijn, Isabel Vandenberghe, Bram Laukens, Sören Planckaert, Stijn Verweire, Inge Van Bogaert, Wim Soetaert, Bart Devreese, and Katarzyna Ciesielska. 2018. “Optimized Expression of the Starmerella Bombicola Lactone Esterase in Pichia Pastoris Through Temperature Adaptation, Codon-optimization and Co-expression with  HAC1.” Protein Expression and Purification 143: 62–70.
APA
De Waele, Stijn, Vandenberghe, I., Laukens, B., Planckaert, S., Verweire, S., Van Bogaert, I., Soetaert, W., et al. (2018). Optimized expression of the Starmerella bombicola lactone esterase in Pichia pastoris through temperature adaptation, codon-optimization and co-expression with  HAC1. PROTEIN EXPRESSION AND PURIFICATION, 143, 62–70.
Vancouver
1.
De Waele S, Vandenberghe I, Laukens B, Planckaert S, Verweire S, Van Bogaert I, et al. Optimized expression of the Starmerella bombicola lactone esterase in Pichia pastoris through temperature adaptation, codon-optimization and co-expression with  HAC1. PROTEIN EXPRESSION AND PURIFICATION. 2018;143:62–70.
MLA
De Waele, Stijn, Isabel Vandenberghe, Bram Laukens, et al. “Optimized Expression of the Starmerella Bombicola Lactone Esterase in Pichia Pastoris Through Temperature Adaptation, Codon-optimization and Co-expression with  HAC1.” PROTEIN EXPRESSION AND PURIFICATION 143 (2018): 62–70. Print.
@article{8543724,
  abstract     = {The Starmerella bombicola lactone esterase (SBLE) is a novel enzyme that, in vivo, catalyzes the intramolecular esterification (lactonization) of acidic sophorolipids in an aqueous environment. In fact, this is an unusual reaction given the unfavorable conditions for dehydration. This characteristic strongly contributes to the potential of SBLE to become a 'green' tool in industrial applications. Indeed, lactonization occurs normally in organic solvents, an application for which microbial lipases are increasingly used as biocatalysts. Previously, we described the production of recombinant SBLE (rSBLE) in Pichia pastoris (syn. Komagataella phaffii). However, expression was not optimal to delve deeper into the enzyme's potential for industrial application. In the current study, we explored codon-optimization of the SBLE gene and we optimized the rSBLE expression protocol. Temperature reduction had the biggest impact followed by codon-optimization and co-expression of the HAC1 transcription factor. Combining these approaches, we achieved a 32-fold improvement of the yield during rSBLE production (from 0.75 mg/l to 24 mg/L culture) accompanied with a strong reduction of contaminants after affinity purification.},
  author       = {De Waele, Stijn and Vandenberghe, Isabel and Laukens, Bram and Planckaert, S{\"o}ren and Verweire, Stijn and Van Bogaert, Inge and Soetaert, Wim and Devreese, Bart and Ciesielska, Katarzyna},
  issn         = {1046-5928},
  journal      = {PROTEIN EXPRESSION AND PURIFICATION},
  keyword      = {Lipase,Lactonase,Starmerella bombicola,Pichia pastoris,Protein purification,Green chemistry,UNFOLDED-PROTEIN RESPONSE,N-GLYCOSYLATION,LIPASE,SOPHOROLIPIDS,YEAST,GENE,DEGRADATION,LEVEL},
  language     = {eng},
  pages        = {62--70},
  title        = {Optimized expression of the Starmerella bombicola lactone esterase in Pichia pastoris through temperature adaptation, codon-optimization and co-expression with  HAC1},
  url          = {http://dx.doi.org/10.1016/j.pep.2017.10.016},
  volume       = {143},
  year         = {2018},
}

Altmetric
View in Altmetric
Web of Science
Times cited: