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Updated: Jul 14, 2026

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology
Published on: September 16, 2013
[Biosynthesis of chitooligosaccharides by recombinant Escherichia coli]
Da-Wei Zhang1, Jie Chu, Yong-Ren Hao
1Shandong Key Laboratory of Microbial Technology, Shandong Academy of Sciences, Jinan 250014, China.
Abstract:
Acetyl-N-glucosaminyltransferase gene (nodC) was successfully cloned to Escherichia coli from Mesorhizobium loti. The recombinant E. coli harboring nodC gene was able to synthesize chitooligosaccharides (COs) in MMYNG medium. In optimized condition, a yield of 526 mg/L was obtained after 26 h cultivation in 10 L bioreactor. COs concentration reached up to 4.5% of the cell dry weight. The COs products were purified by charcoal adsorption and Bio-gel P4 chromatography, penta-N-acetylchitopentaose (m/z, 1034[M + H]+) and tetra-N-acetylchitopentaose (m/z, 831 [M + H]+) were identified as the dominating COs product using the method of liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS).
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