TY - JOUR
T1 - Engineering of a chitosanase fused to a carbohydrate-binding module for continuous production of desirable chitooligosaccharides
AU - Zhou, Jianli
AU - Harindintwali, Jean Damascene
AU - Yang, Wenhua
AU - Han, Minghai
AU - Deng, Bin
AU - Luan, Hao
AU - Zhang, Wenshuai
AU - Liu, Xiaobo
AU - Yu, Xiaobin
N1 - Publisher Copyright:
© 2021 Elsevier Ltd
PY - 2021/12/1
Y1 - 2021/12/1
N2 - Chitooligosaccharides (CHOS) with multiple biological activities are usually produced through enzymatic hydrolysis of chitosan or chitin. However, purification and recycling of the enzyme have largely limited the advancement of CHOS bioproduction. Here, we engineered a novel enzyme by fusing the native chitosanase Csn75 with a carbohydrate-binding module (CBM) that can specifically bind to curdlan. The recombinase Csn75-CBM was successfully expressed by Pichia pastoris and allowed one-step purification and immobilization in the chitosanase immobilized curdlan packed-bed reactor (CICPR), where a maximum adsorption capacity of 39.59 mg enzyme/g curdlan was achieved. CHOS with degrees of polymerization of 2–5 (a hydrolysis yield of 97.75%), 3–6 (75.45%), and 3–7 (73.2%) were continuously produced by adjusting the ratio of enzyme and chitosan or the flow rate of chitosan. Moreover, the CICPR exhibited good stability and reusability after several cycles. The recombinase Csn75-CBM has greatly improved the efficiency of the bioproduction of CHOS.
AB - Chitooligosaccharides (CHOS) with multiple biological activities are usually produced through enzymatic hydrolysis of chitosan or chitin. However, purification and recycling of the enzyme have largely limited the advancement of CHOS bioproduction. Here, we engineered a novel enzyme by fusing the native chitosanase Csn75 with a carbohydrate-binding module (CBM) that can specifically bind to curdlan. The recombinase Csn75-CBM was successfully expressed by Pichia pastoris and allowed one-step purification and immobilization in the chitosanase immobilized curdlan packed-bed reactor (CICPR), where a maximum adsorption capacity of 39.59 mg enzyme/g curdlan was achieved. CHOS with degrees of polymerization of 2–5 (a hydrolysis yield of 97.75%), 3–6 (75.45%), and 3–7 (73.2%) were continuously produced by adjusting the ratio of enzyme and chitosan or the flow rate of chitosan. Moreover, the CICPR exhibited good stability and reusability after several cycles. The recombinase Csn75-CBM has greatly improved the efficiency of the bioproduction of CHOS.
KW - Bioproduction
KW - Carbohydrate-binding module
KW - Chitooligosaccharide
KW - Chitosanase
KW - One-step purification and immobilization
KW - Packed-bed reactor
UR - http://www.scopus.com/inward/record.url?scp=85114002415&partnerID=8YFLogxK
U2 - 10.1016/j.carbpol.2021.118609
DO - 10.1016/j.carbpol.2021.118609
M3 - 文章
C2 - 34561008
AN - SCOPUS:85114002415
SN - 0144-8617
VL - 273
JO - Carbohydrate Polymers
JF - Carbohydrate Polymers
M1 - 118609
ER -