Improved Production of Recombinant Myrosinase in Pichia pastoris
| Autor: | Zuzana Rosenbergová, Zuzana Hegyi, Natália Andelová, M Ferko, Martin Rebroš |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Signal peptide
Glycoside Hydrolases Arabidopsis thaliana QH301-705.5 Arabidopsis Protein Engineering Catalysis Pichia Article plant enzymes Pichia pastoris law.invention Inorganic Chemistry chemistry.chemical_compound law Physical and Theoretical Chemistry Biology (General) Molecular Biology QD1-999 Spectroscopy chemistry.chemical_classification biology Chemistry Myrosinase Arabidopsis Proteins Endoplasmic reticulum Organic Chemistry myrosinase General Medicine biology.organism_classification Ascorbic acid Recombinant Proteins Computer Science Applications Enzyme Biochemistry Sinigrin Recombinant DNA signal sequence |
| Zdroj: | International Journal of Molecular Sciences International Journal of Molecular Sciences, Vol 22, Iss 11889, p 11889 (2021) Volume 22 Issue 21 |
| ISSN: | 1422-0067 |
| Popis: | The effect of the deletion of a 57 bp native signal sequence, which transports the nascent protein through the endoplasmic reticulum membrane in plants, on improved AtTGG1 plant myrosinase production in Pichia pastoris was studied. Myrosinase was extracellularly produced in a 3-liter laboratory fermenter using α-mating factor as the secretion signal. After the deletion of the native signal sequence, both the specific productivity (164.8 U/L/h) and volumetric activity (27 U/mL) increased more than 40-fold compared to the expression of myrosinase containing its native signal sequence in combination with α-mating factor. The deletion of the native signal sequence resulted in slight changes in myrosinase properties: the optimum pH shifted from 6.5 to 7.0 and the maximal activating concentration of ascorbic acid increased from 1 mM to 1.5 mM. Kinetic parameters toward sinigrin were determined: 0.249 mM (Km) and 435.7 U/mg (Vmax). These results could be applied to the expression of other plant enzymes. |
| Databáze: | OpenAIRE |
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