Engineering of a chitosanase fused to a carbohydrate-binding module for continuous production of desirable chitooligosaccharides

Jianli Zhou, Jean Damascene Harindintwali, Wenhua Yang, Minghai Han, Bin Deng, Hao Luan, Wenshuai Zhang, Xiaobo Liu*, Xiaobin Yu

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Article number118609
JournalCarbohydrate Polymers
Volume273
DOIs
StatePublished - 1 Dec 2021

Keywords

  • Bioproduction
  • Carbohydrate-binding module
  • Chitooligosaccharide
  • Chitosanase
  • One-step purification and immobilization
  • Packed-bed reactor

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