Small cause, large effect: Structural characterization of cutinases from Thermobifida cellulosilytica

ABSTRACT We have investigated the structures of two native cutinases from Thermobifida cellulosilytica, namely Thc_Cut1 and Thc_Cut2 as well as of two variants, Thc_Cut2_DM (Thc_Cut2_ Arg29Asn_Ala30Val) and Thc_Cut2_TM (Thc_Cut2_Arg19Ser_Arg29Asn_Ala30Val). The four enzymes showed different activiti...

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Published in:Biotechnology and bioengineering 2017-11, Vol.114 (11), p.2481-2488
Main Authors: Ribitsch, Doris, Hromic, Altijana, Zitzenbacher, Sabine, Zartl, Barbara, Gamerith, Caroline, Pellis, Alessandro, Jungbauer, Alois, Łyskowski, Andrzej, Steinkellner, Georg, Gruber, Karl, Tscheliessnig, Rupert, Herrero Acero, Enrique, Guebitz, Georg M.
Format: Article
Language:English
Subjects:
Actinobacteria - enzymology
Aliphatic compounds
Carboxylic Ester Hydrolases - chemistry
Carboxylic Ester Hydrolases - ultrastructure
Crystal structure
cutinases
Enzyme Activation
Enzyme Stability
enzyme structure
Enzymes
Hydrophobicity
Molecular Docking Simulation - methods
poly(lactic acid)
polyester hydrolysis
Polyesters - chemistry
Polylactic acid
Protein Binding
Protein Conformation
Selectivity
Small angle X ray scattering
Static Electricity
Structural analysis
Structure-Activity Relationship
Surface charge
Surface properties
X-ray scattering
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Summary:ABSTRACT We have investigated the structures of two native cutinases from Thermobifida cellulosilytica, namely Thc_Cut1 and Thc_Cut2 as well as of two variants, Thc_Cut2_DM (Thc_Cut2_ Arg29Asn_Ala30Val) and Thc_Cut2_TM (Thc_Cut2_Arg19Ser_Arg29Asn_Ala30Val). The four enzymes showed different activities towards the aliphatic polyester poly(lactic acid) (PLLA). The crystal structures of the four enzymes were successfully solved and in combination with Small Angle X‐Ray Scattering (SAXS) the structural features responsible for the selectivity difference were elucidated. Analysis of the crystal structures did not indicate significant conformational differences among the different cutinases. However, the distinctive SAXS scattering data collected from the enzymes in solution indicated a remarkable surface charge difference. The difference in the electrostatic and hydrophobic surface properties could explain potential alternative binding modes of the four cutinases on PLLA explaining their distinct activities. Biotechnol. Bioeng. 2017;114: 2481–2488. © 2017 Wiley Periodicals, Inc. Differences in electrostatic and hydrophobic surface properties of cutinases from Thermobifida cellulosilytica explain distinct activities on poly(lactic acid).
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.26372