Owing to the difficulty in isolating single molecules of chitosan oligosaccharides, it becomes challenging to study the functional activities of chitobiose with the same degree of polymerization.Hence, how to obtain oligosaccharides with the same degree of polymerization on a large scale is crucial for the research on the functional activities of chitobiose and its wide-scale popularization and application.The research of this subject aims to utilize relatively inexpensive N-acetylglucosamine as the substrate, with ATP as the cofactor, and synthesize chitobiose under the triculture-coupled whole-cell catalysis of polyphosphorylase (PPK), N-acetylhexosamine kinase (Nahk), and N,N′-diacetylated chitobiose phosphorylase (Chbp).The yield of chitobiose was 1 675 mg/L by three-bacteria coupling.After single-factor optimization, the maximum yield of chitobiose was 5 214 mg/L, an increase of 211%.To further reduce costs and simplify the process, chitosan disaccharide production strains were constructed and the expression of pathway genes was balanced through the combination of plasmids with different copy numbers.The engineered strain Escherichia coli BL21(DE3) (containing plasmids pRSFDuet1-Nahk-Chbp and pETDuet1-PPK) was adopted, achieving a chitosan disaccharide yield of 1 600 mg/L.This study provides references and guidance for the large-scale biological manufacturing of chitobiose.
MAO Yijie
,
LAI Xia
,
ZHANG Zimeng
,
WANG Zhijie
,
ZHANG Hongtao
. Biosynthesis of chitobiose via tri-microbial coupling strategy[J]. Food and Fermentation Industries, 2025
, 51(21)
: 35
-42
.
DOI: 10.13995/j.cnki.11-1802/ts.042624
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