Enhancement of the performance of the GH75 family chitosanases by fusing a carbohydrate binding module and insights into their substrate binding mechanisms

Jianli Zhou; Qiuya Gu; Yu Shen; Jean Damascene Harindintwali; Wenhua Yang; Shuliang Zou; Minghai Han; Chao Ma; Xiaobin Yu; Xiaobo Liu

doi:10.1016/j.lwt.2022.113390

Enhancement of the performance of the GH75 family chitosanases by fusing a carbohydrate binding module and insights into their substrate binding mechanisms

Jianli Zhou, Qiuya Gu, Yu Shen, Jean Damascene Harindintwali, Wenhua Yang, Shuliang Zou, Minghai Han, Chao Ma, Xiaobin Yu^*, Xiaobo Liu^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

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.

Original language	English
Article number	113390
Journal	LWT
Volume	163
DOIs	https://doi.org/10.1016/j.lwt.2022.113390
State	Published - 15 Jun 2022
Externally published	Yes

Keywords

Carbohydrate-binding module
Chitooligosaccharide
Chitosan
Chitosanase
Thermodynamic analysis

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10.1016/j.lwt.2022.113390

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@article{95d58a772e654f3e8e204d7110c936d3,

title = "Enhancement of the performance of the GH75 family chitosanases by fusing a carbohydrate binding module and insights into their substrate binding mechanisms",

abstract = "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.",

keywords = "Carbohydrate-binding module, Chitooligosaccharide, Chitosan, Chitosanase, Thermodynamic analysis",

author = "Jianli Zhou and Qiuya Gu and Yu Shen and Harindintwali, {Jean Damascene} and Wenhua Yang and Shuliang Zou and Minghai Han and Chao Ma and Xiaobin Yu and Xiaobo Liu",

note = "Publisher Copyright: {\textcopyright} 2022 The Authors",

year = "2022",

month = jun,

day = "15",

doi = "10.1016/j.lwt.2022.113390",

language = "英语",

volume = "163",

journal = "LWT",

issn = "0023-6438",

publisher = "Academic Press Inc.",

}

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

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 -

Enhancement of the performance of the GH75 family chitosanases by fusing a carbohydrate binding module and insights into their substrate binding mechanisms

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