Efficient separation of phosphorylated sugars from multi-enzyme system by ultrafiltration and membrane fouling mechanism

doi:10.1016/j.cjche.2025.08.003

中国化学工程学报 ›› 2025, Vol. 87 ›› Issue (11): 157-170.DOI: 10.1016/j.cjche.2025.08.003

Efficient separation of phosphorylated sugars from multi-enzyme system by ultrafiltration and membrane fouling mechanism

Zhengxin Mao^1,2, Jiachang Shen^1,2, Mengxin Liu³, Yanjie Ji³, Qinhong Wang⁴, Maohua Yang¹, Jianmin Xing^1,2

1. CAS Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China;
2. College of Chemical Engineering, University of Chinese Academy of Sciences, Beijing 100049, China;
3. Tianjin Juno Biotechnology Company, Tianjin 300308, China;
4. CAS, Key Laboratory of Low Carbon Synthetic Engineering Biology, Tianjin Institute of Industrial Biotechnology, Tianjin 300308, China

收稿日期:2025-06-21 修回日期:2025-07-31 接受日期:2025-08-03 出版日期:2025-11-28 发布日期:2025-08-20
通讯作者: Maohua Yang,E-mail:mhyang@ipe.ac.cn;Jianmin Xing,E-mail:jmxing@ipe.ac.cn
基金资助:
This work acknowledges the funding support provided by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDC0120402) and the National Key Research & Development Program of China (2022YFC2105103).

Efficient separation of phosphorylated sugars from multi-enzyme system by ultrafiltration and membrane fouling mechanism

Zhengxin Mao^1,2, Jiachang Shen^1,2, Mengxin Liu³, Yanjie Ji³, Qinhong Wang⁴, Maohua Yang¹, Jianmin Xing^1,2

1. CAS Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China;
2. College of Chemical Engineering, University of Chinese Academy of Sciences, Beijing 100049, China;
3. Tianjin Juno Biotechnology Company, Tianjin 300308, China;
4. CAS, Key Laboratory of Low Carbon Synthetic Engineering Biology, Tianjin Institute of Industrial Biotechnology, Tianjin 300308, China

Received:2025-06-21 Revised:2025-07-31 Accepted:2025-08-03 Online:2025-11-28 Published:2025-08-20
Contact: Maohua Yang,E-mail:mhyang@ipe.ac.cn;Jianmin Xing,E-mail:jmxing@ipe.ac.cn
Supported by:
This work acknowledges the funding support provided by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDC0120402) and the National Key Research & Development Program of China (2022YFC2105103).

摘要/Abstract

摘要： Phosphorylated sugars, recognized as central intermediates in carbohydrate metabolism and critical precursors for enzymatic synthesis of rare sugars, face significant technical barriers in their industrial-scale production. The multi-enzymatic preparation systems for these compounds inherently accumulate complex impurities, including protein-based catalysts, residual substrates, and oligosaccharide by-products, posing persistent challenges in product separation and biocatalyst recycling. To address this limitation, we conducted a systematic investigation of ultrafiltration-based separation strategies during the multi-enzyme-catalyzed synthesis of fructose-1,6-bisphosphate (FDP), with particular emphasis on membrane fouling mechanisms. By screening the ultrafiltration membranes, UE020 showed the best performance in the model system, achieving significant separation targets: 99.97% retention of bovine serum albumin, FDP/maltodextrin separation coefficient of 7.41, and FDP recovery of 93.63%. An analysis of the components of resistance revealed that concentration polarization induced by maltodextrin was the main factor constituting the resistance, irreversible resistance due to bovine serum albumin was a secondary effect, and the resistance constituted by FDP was negligible. A mitigation strategy employing powdered activated carbon for dynamic membrane formation significantly improved system performance, reducing irreversible resistance by 59.14% and enhancing flux recovery by 20.85%. In this study, ultrafiltration was strategically employed to achieve efficient separation of FDP and enzyme recovery. Significantly, we deciphered the synergistic fouling mechanisms arising from interactions within the multicomponent system containing phosphorylated sugars, oligosaccharides, and proteins. These findings provide a mechanistic framework for scaling up multi-enzymatic systems dedicated to phosphorylated sugar biosynthesis, effectively bridging the gap between laboratory-scale synthesis and industrial implementation.

关键词: Phosphorylated sugars, Multi-enzyme system, Ultrafiltration, Efficient separation, Membrane fouling

Abstract: Phosphorylated sugars, recognized as central intermediates in carbohydrate metabolism and critical precursors for enzymatic synthesis of rare sugars, face significant technical barriers in their industrial-scale production. The multi-enzymatic preparation systems for these compounds inherently accumulate complex impurities, including protein-based catalysts, residual substrates, and oligosaccharide by-products, posing persistent challenges in product separation and biocatalyst recycling. To address this limitation, we conducted a systematic investigation of ultrafiltration-based separation strategies during the multi-enzyme-catalyzed synthesis of fructose-1,6-bisphosphate (FDP), with particular emphasis on membrane fouling mechanisms. By screening the ultrafiltration membranes, UE020 showed the best performance in the model system, achieving significant separation targets: 99.97% retention of bovine serum albumin, FDP/maltodextrin separation coefficient of 7.41, and FDP recovery of 93.63%. An analysis of the components of resistance revealed that concentration polarization induced by maltodextrin was the main factor constituting the resistance, irreversible resistance due to bovine serum albumin was a secondary effect, and the resistance constituted by FDP was negligible. A mitigation strategy employing powdered activated carbon for dynamic membrane formation significantly improved system performance, reducing irreversible resistance by 59.14% and enhancing flux recovery by 20.85%. In this study, ultrafiltration was strategically employed to achieve efficient separation of FDP and enzyme recovery. Significantly, we deciphered the synergistic fouling mechanisms arising from interactions within the multicomponent system containing phosphorylated sugars, oligosaccharides, and proteins. These findings provide a mechanistic framework for scaling up multi-enzymatic systems dedicated to phosphorylated sugar biosynthesis, effectively bridging the gap between laboratory-scale synthesis and industrial implementation.

Key words: Phosphorylated sugars, Multi-enzyme system, Ultrafiltration, Efficient separation, Membrane fouling

Zhengxin Mao, Jiachang Shen, Mengxin Liu, Yanjie Ji, Qinhong Wang, Maohua Yang, Jianmin Xing. Efficient separation of phosphorylated sugars from multi-enzyme system by ultrafiltration and membrane fouling mechanism[J]. 中国化学工程学报, 2025, 87(11): 157-170.

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Efficient separation of phosphorylated sugars from multi-enzyme system by ultrafiltration and membrane fouling mechanism

Efficient separation of phosphorylated sugars from multi-enzyme system by ultrafiltration and membrane fouling mechanism

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