Glycosylation influences activity, stability and immobilization of the feruloyl esterase 1a from Myceliophthora thermophila
Journal article, 2019

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.

Enzyme activity

Escherichia coli

Pichia pastoris

Enzyme stability

Heterologous production

Mass spectrometry (MS)


Cyrielle Bonzom

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

Silvia Hüttner

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

Ekaterina Mirgorodskaya

University of Gothenburg

Sun-Li Chong

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

Zhejiang Agriculture and Forestry University

Stefan Uthoff

University of Münster

Alexander Steinbuechel

King Abdulaziz University

University of Münster

Raymond M. D. Verhaert

Proteonic Bv

Lisbeth Olsson

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

AMB Express

2191-0855 (ISSN)

Vol. 9 1 126

Subject Categories

Biochemistry and Molecular Biology

Analytical Chemistry

Biocatalysis and Enzyme Technology





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