Structural and functional studies of Aspergillus oryzae cutinase: enhanced thermostability and hydrolytic activity of synthetic ester and polyester degradation.
Cutinases are responsible for hydrolysis of the protective cutin lipid polyester matrix in plants and thus have been exploited for hydrolysis of small molecule esters and polyesters. Here we explore the reactivity, stability, and structure of Aspergillus oryzae cutinase and compare it to the well-studied enzyme from Fusarium solani. Two critical differences are highlighted in the crystallographic analysis of the A. oryzae structure: (i) an additional disulfide bond and (ii) a topologically favored catalytic triad with a continuous and deep groove. These structural features of A. oryzae cutinase are proposed to result in an improved hydrolytic activity and altered substrate specificity profile, enhanced thermostability, and remarkable reactivity toward the degradation of the synthetic polyester polycaprolactone. The results presented here provide insight into engineering new cutinase-inspired biocatalysts with tailor-made properties.
Liu
Zhiqiang
Z
Department of Chemical and Biological Sciences, Polytechnic Institute of New York University, Brooklyn, New York 11201, USA.
Gosser
Yuying
Y
Baker
Peter James
PJ
Ravee
Yaniv
Y
Lu
Ziying
Z
Alemu
Girum
G
Li
Huiguang
H
Butterfoss
Glenn L
GL
Kong
Xiang-Peng
XP
Gross
Richard
R
Montclare
Jin Kim
JK
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology*; D002152:Calorimetry, Differential Scanning; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D003460:Crystallization; D004795:Enzyme Stability; D004952:Esters / Q000378:metabolism*; D006868:Hydrolysis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011091:Polyesters / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D013816:Thermodynamics
D004952:Esters; D011091:Polyesters; D002265:Carboxylic Ester Hydrolases; C019952:cutinase