Dual-cell co-catalytic system engineering enhances N-acetylneuraminic acid synthesis by Bacillus amyloliquefaciens

Xinyu Dong; Zhenying Han; Dian Zou; Ailing Guo; Changwen Ye; Xuetuan Wei

doi:10.1016/j.biortech.2025.132969

Dual-cell co-catalytic system engineering enhances N-acetylneuraminic acid synthesis by Bacillus amyloliquefaciens

Bioresour Technol. 2025 Jul 10:436:132969. doi: 10.1016/j.biortech.2025.132969. Online ahead of print.

Authors

Xinyu Dong¹, Zhenying Han², Dian Zou³, Ailing Guo⁴, Changwen Ye⁵, Xuetuan Wei⁶

Affiliations

¹ National Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: dongxinyu0401@163.com.
² National Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: hzy19991128@163.com.
³ National Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: zd498905497@163.com.
⁴ National Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: ailingguo234@163.com.
⁵ National Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: yectsrc@163.com.
⁶ National Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: weixuetuan@mail.hzau.edu.cn.

PMID: 40651703
DOI: 10.1016/j.biortech.2025.132969

Abstract

N-acetylneuraminic acid (NeuAc) is a bioactive nine-carbon sugar, and it is extensively applied in food, pharmaceutical, and cosmetic fields. However, feedback inhibition between key enzymes and metabolites adversely affects NeuAc synthesis. In this study, a novel Bacillus amyloliquefaciens dual-cell co-catalytic system was developed to relieve feedback inhibition effects, resulting in 62 % increase in NeuAc titer, and blocking consumption pathways of NeuAc precursors (N-acetylglucosamine (GlcNAc) and pyruvate) further increased NeuAc titer by 75 % (33.4 g/L). Subsequently, NeuAc-synthesizing enzyme promoter engineering increased NeuAc titer to 77.3 g/L. Finally, the maximum NeuAc titer (134.7 g/L) and GlcNAc conversion rate (77 %) were achieved under dual-cell co-catalytic optimized conditions (pH 7, cell concentration = 25 (OD₆₀₀), 0.8 mol/L GlcNAc, and xylose as carbon source). Overall, this study develops a dual-cell co-catalytic system of NeuAc efficient synthesis by Bacillus amyloliquefaciens, which exhibits great potential in the industrial production and application of NeuAc.

Keywords: Bioprocess optimization; Bioproduct synthesis; Feedback inhibition; Metabolic engineering; Promoter engineering.