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LAPSE:2023.24363

Efficient Production of 3′-Sialyllactose by Single Whole-Cell in One-Pot Biosynthesis

Zhongkui Li, Xiangsong Chen, Zhijian Ni, Lixia Yuan, Lijie Sun, Yu Wang, Jinyong Wu, Jianming Yao

March 28, 2023

Abstract
Sialyllactose (SL) is one of the most important acidic oligosaccharides in human milk, which plays an important role in the health of infants. In this work, an efficient multi-enzyme cascade was developed in a single whole cell to produce 3′-SL. We constructed two compatible plasmids with double cloning sites to co-express four genes. Different combinations were assessed to verify the optimal catalytic ability. Then, the conversion temperature, pH, and stability under the optimal temperature and pH were investigated. Moreover, the optimal conversion conditions and surfactant concentration were determined. By using the optimal conditions (35 °C, pH 7.0, 20 mM polyphosphate, 10 mM cytidine monophosphate (CMP), 20 mM MgCl2), 25 mL and 4 L conversion systems were carried out to produce 3′-SL. Similar results were obtained between different volume conversion reactions, which led the maximum production of 3′-SL to reach 53 mM from 54.2 mM of sialic acid (SA) in the 25 mL system and 52.8 mM of 3′-SL from 53.8 mM of SA in the 4 L system. These encouraging results demonstrate that the developed single whole-cell multi-enzyme system exhibits great potential and economic competitiveness for the manufacture of 3′-SL.

Record ID

LAPSE:2023.24363

Keywords

3′-sialyllactose, CTP regeneration, human milk oligosaccharides, multi-enzyme, single whole cell

Subject

Biosystems

Suggested Citation

Li Z, Chen X, Ni Z, Yuan L, Sun L, Wang Y, Wu J, Yao J. Efficient Production of 3′-Sialyllactose by Single Whole-Cell in One-Pot Biosynthesis. (2023). LAPSE:2023.24363

Author Affiliations

Li Z: Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China; University of Science and Technology of China, Hefei 230026, China
Chen X: Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China; Huainan New Energy Research Center, Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Huaina
Ni Z: Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China; University of Science and Technology of China, Hefei 230026, China
Yuan L: Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China; Huainan New Energy Research Center, Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Huaina
Sun L: Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China; Huainan New Energy Research Center, Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Huaina
Wang Y: Wuhan Zhongke Optics Valley Green Biotechnology Co., Ltd., Wuhan 430200, China
Wu J: Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China; Huainan New Energy Research Center, Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Huaina
Yao J: Institute of Plasma Physics, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, China; University of Science and Technology of China, Hefei 230026, China; Huainan New Energy Research Center, Institute of Plasma Physics, Hefei

Journal Name

Processes

Volume

Issue

First Page

932

Year

2021

Publication Date

2021-05-26

DOI:

https://doi.org/10.3390/pr9060932