TY - JOUR
T1 - Enhancement of the performance of the GH75 family chitosanases by fusing a carbohydrate binding module and insights into their substrate binding mechanisms
AU - Zhou, Jianli
AU - Gu, Qiuya
AU - Shen, Yu
AU - Harindintwali, Jean Damascene
AU - Yang, Wenhua
AU - Zou, Shuliang
AU - Han, Minghai
AU - Ma, Chao
AU - Yu, Xiaobin
AU - Liu, Xiaobo
N1 - Publisher Copyright:
© 2022 The Authors
PY - 2022/6/15
Y1 - 2022/6/15
N2 - Chitooligosaccharides (COSs) with relatively higher degrees of polymerization (DP) exhibited better biological properties than COSs with lower DP. Here, we demonstrated a potential strategy to enhance the affinity, activity, and thermostability of the chitosanase Csn75 from Aspergillus fumigatus CJ22-326 through fusion of a carbohydrate binding module (CBM) that can specifically bind to chitosan. Compared with the Csn75, the specific enzyme activity of the Csn75-CBM32 and the Csn75-2CBM32 had increased by 59.18% and 14.29%, respectively. The free energy (ΔG), enthalpy (ΔH), and entropy (-TΔS) of the three enzymes (Csn75, Csn75-CBM32, and Csn75-2CBM32) toward (GlcN)6 was (−7.19, −7.54, and −8.08 kcal/mol), (−5.37, 1.75, and 0.86 kcal/mol), and (1.45, 9.27, and 8.03 kcal/mol), respectively. The substrate binding of the Csn75 and the fusion enzymes (Csn75-CBM32 and Csn75-2CBM32) were mainly driven by the hydrophobic (entropy effect) and favorable intermolecular forces (enthalpy contribution), respectively. Moreover, the DP of COSs in the final hydrolysate of Csn75 was 2–4, but that of Csn75-CBM32 and Csn75-2CBM32 was 3–5. This strategy provides a potential alternative for the bioproduction of desirable COSs to meet the requirement of application in food systems.
AB - Chitooligosaccharides (COSs) with relatively higher degrees of polymerization (DP) exhibited better biological properties than COSs with lower DP. Here, we demonstrated a potential strategy to enhance the affinity, activity, and thermostability of the chitosanase Csn75 from Aspergillus fumigatus CJ22-326 through fusion of a carbohydrate binding module (CBM) that can specifically bind to chitosan. Compared with the Csn75, the specific enzyme activity of the Csn75-CBM32 and the Csn75-2CBM32 had increased by 59.18% and 14.29%, respectively. The free energy (ΔG), enthalpy (ΔH), and entropy (-TΔS) of the three enzymes (Csn75, Csn75-CBM32, and Csn75-2CBM32) toward (GlcN)6 was (−7.19, −7.54, and −8.08 kcal/mol), (−5.37, 1.75, and 0.86 kcal/mol), and (1.45, 9.27, and 8.03 kcal/mol), respectively. The substrate binding of the Csn75 and the fusion enzymes (Csn75-CBM32 and Csn75-2CBM32) were mainly driven by the hydrophobic (entropy effect) and favorable intermolecular forces (enthalpy contribution), respectively. Moreover, the DP of COSs in the final hydrolysate of Csn75 was 2–4, but that of Csn75-CBM32 and Csn75-2CBM32 was 3–5. This strategy provides a potential alternative for the bioproduction of desirable COSs to meet the requirement of application in food systems.
KW - Carbohydrate-binding module
KW - Chitooligosaccharide
KW - Chitosan
KW - Chitosanase
KW - Thermodynamic analysis
UR - http://www.scopus.com/inward/record.url?scp=85130581335&partnerID=8YFLogxK
U2 - 10.1016/j.lwt.2022.113390
DO - 10.1016/j.lwt.2022.113390
M3 - 文章
AN - SCOPUS:85130581335
SN - 0023-6438
VL - 163
JO - LWT
JF - LWT
M1 - 113390
ER -