Glycosylation influences activity, stability and immobilization of the feruloyl esterase 1a from Myceliophthora thermophila
| Autor: | Raymond M. D. Verhaert, Alexander Steinbüchel, Stefan Uthoff, Sun-Li Chong, Silvia Hüttner, Cyrielle Bonzom, Lisbeth Olsson, Ekaterina Mirgorodskaya |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
animal structures Glycosylation lcsh:Biotechnology Biophysics lcsh:QR1-502 macromolecular substances medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology lcsh:Microbiology Pichia pastoris 03 medical and health sciences chemistry.chemical_compound Feruloyl esterase 010608 biotechnology lcsh:TP248.13-248.65 medicine Escherichia coli Enzyme activity Heterologous production 030304 developmental biology 2. Zero hunger chemistry.chemical_classification Enzyme stability 0303 health sciences biology biology.organism_classification Enzyme assay carbohydrates (lipids) Mass spectrometry (MS) Enzyme Biochemistry chemistry biology.protein Original Article Specific activity lipids (amino acids peptides and proteins) Myceliophthora thermophila |
| Zdroj: | AMB Express, Vol 9, Iss 1, Pp 1-13 (2019) AMB Express |
| ISSN: | 2191-0855 |
| Popis: | Heterologous protein production is widely used in industrial biotechnology. However, using non-native production hosts can lead to enzymes with altered post-translational modifications, such as glycosylation. We have investigated how production in a non-native host affects the physicochemical properties and enzymatic activity of a feruloyl esterase from Myceliophthora thermophila, MtFae1a. The enzyme was produced in two microorganisms that introduce glycosylation (M. thermophila and Pichia pastoris) and in Escherichia coli (non-glycosylated). Mass spectrometric analysis confirmed the presence of glycosylation and revealed differences in the lengths of glycan chains between the enzymes produced in M. thermophila and P. pastoris. The melting temperature and the optimal temperature for activity of the non-glycosylated enzyme were considerably lower than those of the glycosylated enzymes. The three MtFae1a versions also exhibited differences in specific activity and specificity. The catalytic efficiency of the glycosylated enzymes were more than 10 times higher than that of the non-glycosylated one. In biotechnology, immobilization is often used to allow reusing enzyme and was investigated on mesoporous silica particles. We found the binding kinetics and immobilization yield differed between the enzyme versions. The largest differences were observed when comparing enzymes with and without glycosylation, but significant variations were also observed between the two differently glycosylated enzymes. We conclude that the biotechnological value of an enzyme can be optimized for a specific application by carefully selecting the production host. Electronic supplementary material The online version of this article (10.1186/s13568-019-0852-z) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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