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Perz, V; Zumstein, MT; Sander, M; Zitzenbacher, S; Ribitsch, D; Guebitz, GM.
Biomimetic Approach to Enhance Enzymatic Hydrolysis of the Synthetic Polyester Poly(1,4-butylene adipate): Fusing Binding Modules to Esterases.
Biomacromolecules. 2015; 16(12): 3889-3896. Doi: 10.1021/acs.biomac.5b01219
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Perz Veronika
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Abstract:
Mimicking a concept of nature for the hydrolysis of biopolymers, the Thermobifida cellulosilytica cutinase 1 (Thc_Cut1) was fused to a polymer binding module (PBM) to enhance the hydrolysis of the polyester poly(1,4-butylene adipate) (PBA). Namely, the binding module of a polyhydroxyalkanoate depolymerase from Alcaligenes faecalis (Thc_Cut1_PBM) was attached to the cutinase via two different linker sequences varying in length. In order to investigate the adsorption behavior, catalytically inactive mutants both of Thc_Cut1 and Thc_Cut1_PBM were successfully constructed by site-directed mutagenesis of serine 131 to alanine. Quartz crystal microbalance with dissipation monitoring (QCM-D) analysis revealed that the initial mass increase during enzyme adsorption was larger for the inactive enzymes linked with the PBM as compared to the enzyme without the PBM. The hydrolysis rates of PBA were significantly enhanced when incubated with the active, engineered Thc_Cut1_PBM as compared to the native Thc_Cut1. Thc_Cut1_PBM completely hydrolyzed PBA thin films on QCM-D sensors within approximately 40 min, whereas twice as much time was required for the complete hydrolysis by the native Thc_Cut1.
Find related publications in this database (using NLM MeSH Indexing)
Actinobacteria - chemistry
Actinobacteria - enzymology
Alanine - chemistry
Alanine - genetics
Alcaligenes faecalis - chemistry
Alcaligenes faecalis - enzymology
Amino Acid Substitution -
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Biomimetics - methods
Butylene Glycols - chemistry
Butylene Glycols - metabolism
Carboxylic Ester Hydrolases - chemistry
Carboxylic Ester Hydrolases - genetics
Carboxylic Ester Hydrolases - metabolism
Cloning, Molecular -
Escherichia coli - genetics
Escherichia coli - metabolism
Gene Expression -
Hydrolysis -
Kinetics -
Mutagenesis, Site-Directed -
Mutation -
Polymers - chemistry
Polymers - metabolism
Protein Engineering -
Protein Structure, Tertiary -
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Serine - chemistry
Serine - genetics
Structure-Activity Relationship -

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