Structural studies of Myceliophthora Thermophila Laccase in the presence of deep eutectic solvents
•Laccase activity were investigated in several DESs.•Laccase activity increase in lactic acid-betaine DES.•Kinetic studies with glycerol-choline chloride DES is a S-parabolic-I-parabolic mixed non-competitive inhibitor.•The crystal structures of laccase with lactic acid: choline chloride DES (LCDES)...
Saved in:
Published in | Enzyme and microbial technology Vol. 150; no. C; p. 109890 |
---|---|
Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
United Kingdom
Elsevier Inc
01.10.2021
Elsevier |
Subjects | |
Online Access | Get full text |
Cover
Loading…
Summary: | •Laccase activity were investigated in several DESs.•Laccase activity increase in lactic acid-betaine DES.•Kinetic studies with glycerol-choline chloride DES is a S-parabolic-I-parabolic mixed non-competitive inhibitor.•The crystal structures of laccase with lactic acid: choline chloride DES (LCDES) were obtained at 1.6 Å.•Presence of DES changes the local environment of the amino acids in laccase active sites.
In this work, we elucidated the interactions between Myceliophthora thermophila laccase and deep eutectic solvent (DES) by crystallographic and kinetics analyses. Four types of DESs with different hydrogen bond acceptor (HBA) and hydrogen bond donor (HBD), including lactic acid: betaine, glycerol: choline chloride, lactic acid: choline chloride and glycerol: betaine was used. The results revealed that different DES have different effects on laccase activity. Lactic acid-betaine (2:1) DES has shown to enhance laccase activity up to 300 % at a concentration ranged from 2% to 8% v/v, while glycerol: choline chloride and lactic acid: choline chloride DES choline chloride-based DES have found to possess inhibitory effects on laccase under the same concentration range. Detailed kinetic study showed that glycerol: choline chloride DES is a S-parabolic-I-parabolic mixed non-competitive inhibitor, where conformational changes can occur. The crystal structures of laccase with lactic acid: choline chloride DES (LCDES) were obtained at 1.6 Å. Crystallographic analysis suggested that the addition of LCDES causes changes in the laccase active site, but the increase in water molecules observed in the resulting crystal prevented laccase from experiencing drastic structural change. Fluorescence and circular dichroism spectroscopies were also applied to determine the effects of DES on the structural conformation of laccase. The results have confirmed that the presence of DES can trigger changes in the local environments of the amino acids in the active site of laccase which contributes to the changes in its activity and stability. |
---|---|
Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 USDOE |
ISSN: | 0141-0229 1879-0909 |
DOI: | 10.1016/j.enzmictec.2021.109890 |