期刊
CARBOHYDRATE POLYMERS
卷 273, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.carbpol.2021.118609
关键词
Bioproduction; Chitooligosaccharide; Chitosanase; Carbohydrate-binding module; One-step purification and immobilization; Packed-bed reactor
资金
- National First-class Discipline Program of the Light Industry Technology and Engineering [LITE2018-11]
- Open Project of Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of Education [KLCCB-KF202007]
- Guizhou Science and Technology Plan Project [[2020]1Y144]
- Natural Science Research Project of Education Department of Guizhou Province [[2015]368]
The research engineered a novel enzyme, recombinase Csn75-CBM, for the bioproduction of CHOS, which successfully achieved a maximum adsorption capacity of 39.59 mg enzyme/g curdlan in a packed-bed reactor. Continuous production of CHOS with good stability and reusability was demonstrated by adjusting the enzyme-chitosan ratio or flow rate.
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 Csn75CBM 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.
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