Glycosylation influences activity, stability and immobilization of the feruloyl esterase 1a from Myceliophthora thermophila
Artikel i vetenskaplig tidskrift, 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.

Escherichia coli

Mass spectrometry (MS)

Enzyme activity

Pichia pastoris

Enzyme stability

Heterologous production


Cyrielle Bonzom

Chalmers, Biologi och bioteknik, Industriell bioteknik

Silvia Hüttner

Chalmers, Biologi och bioteknik, Industriell bioteknik

Ekaterina Mirgorodskaya

Göteborgs universitet

Sun-Li Chong

Chalmers, Biologi och bioteknik, Industriell bioteknik

Zhejiang A & F University

Stefan Uthoff

Universität Münster

Alexander Steinbuechel

Universität Münster

King Abdulaziz University

Raymond M. D. Verhaert

Proteonic Bv

Lisbeth Olsson

Chalmers, Biologi och bioteknik, Industriell bioteknik

AMB Express

21910855 (eISSN)

Vol. 9 1 126


Biokemi och molekylärbiologi

Analytisk kemi

Biokatalys och enzymteknik





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