磷酸糖与高浓度磷酸盐的高效纳滤分离

doi:10.11949/0438-1157.20250844

化工学报 ›› 2025, Vol. 76 ›› Issue (12): 6423-6438.DOI: 10.11949/0438-1157.20250844

磷酸糖与高浓度磷酸盐的高效纳滤分离

毛正鑫¹^,²(), 申佳昌¹^,², 刘梦鑫³, 季延洁³, 王钦宏⁴, 杨茂华¹(), 邢建民¹^,²()

^1.中国科学院过程工程研究所绿色过程与工程重点实验室，北京 100190
^2.中国科学院大学化学工程学院，北京 100049
^3.天津君诺生物技术有限公司，天津 300308
^4.中国科学院天津工业生物技术研究所低碳合成工程生物学全国重点实验室，天津 300308

收稿日期:2025-07-29 修回日期:2025-10-23 出版日期:2025-12-31 发布日期:2026-01-23
通讯作者: 杨茂华,邢建民
作者简介:毛正鑫(1995—), 男, 博士研究生, maozhengxin@ipe.ac.cn

Efficient separation of phosphorylated sugars and high-concentration phosphate by nanofiltration

Zhengxin MAO¹^,²(), Jiachang SHEN¹^,², Mengxin LIU³, Yanjie JI³, Qinhong WANG⁴, Maohua YANG¹(), Jianmin XING¹^,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:2025-07-29 Revised:2025-10-23 Online:2025-12-31 Published:2026-01-23
Contact: Maohua YANG, Jianmin XING

摘要/Abstract

摘要：

磷酸糖是重要的生化药物与糖类合成关键中间体。在多酶催化制备糖类物质时，伴随着磷酸糖的生成，磷酸盐（Pi）的浓度逐渐升高，高浓度Pi对酶活性具有抑制作用。因此，Pi的高效去除是实现糖类物质合成技术推广应用的关键。本研究针对磷酸糖/Pi模拟体系，考察了Pi总浓度、pH和溶液组成对分离效率的影响。对比分析了多种纳滤膜，结果表明，在高盐浓度下，纳滤膜NF4和NF7展现出良好的分离选择性能，可以实现磷酸糖与高浓度磷酸盐的高效分离。通过洗滤，Pi的去除率可达到80%以上。采用了DSPM-DE模型对分离机制进行解析，空间位阻和介电排斥是实现溶质分离的关键机制。本研究利用纳滤技术实现了高盐体系中磷酸糖与Pi的高效分离，同时解决了高盐环境对催化体系中酶活性的潜在抑制作用，确保了体系的长期稳定运行，为磷酸糖制备技术的工业应用奠定了基础。

关键词: 膜, 纳滤, 高盐溶液, 磷酸糖, 磷酸盐去除, 分离, 介电排斥

Abstract:

Phosphorylated sugars are important biopharmaceuticals and key intermediates in carbohydrate synthesis. In multi-enzyme catalytic preparation systems for sugars, the continuous generation of phosphorylated sugars will result in the accumulation of inorganic phosphate (Pi). Since high concentrations of Pi inhibit enzyme activity, the efficient removal of Pi is critical for the implementation of carbohydrate synthesis technology. However, the separation of phosphorylated sugars/Pi presents a significant difficulty due to their highly similar physicochemical properties. This study investigated the separation efficiency of phosphorylated sugars/Pi in a simulated solution, specifically examining the influence of total concentration, pH and solution composition. Comparative analysis of multiple nanofiltration membranes revealed that NF4 and NF7 exhibit superior separation selectivity under high-salinity conditions. Through rinsing, the removal rate of Pi reached over 80%. This performance robustly confirmed the feasibility of separating phosphorylated sugars from Pi in high-salt solutions. Furthermore, the separation mechanism was analyzed using the Donnan Steric Pore Model with Dielectric Exclusion (DSPM-DE). The analysis revealed that steric hindrance and dielectric exclusion are the key mechanisms underpinning the selective separation of these two solutes under high-salt conditions. This study successfully demonstrated efficient separation of phosphorylated sugars from Pi in high-salt systems by nanofiltration. The approach simultaneously mitigates potential enzyme inhibition in catalytic systems caused by high salinity, enabling long-term stable operation. These findings lay the groundwork for industrial implementation of phosphorylated sugars production technology.

Key words: membranes, nanofiltration, high salinity solution, phosphorylated sugar, phosphate removal, separation, dielectric exclusion

中图分类号:

TF 028.8

毛正鑫, 申佳昌, 刘梦鑫, 季延洁, 王钦宏, 杨茂华, 邢建民. 磷酸糖与高浓度磷酸盐的高效纳滤分离[J]. 化工学报, 2025, 76(12): 6423-6438.

Zhengxin MAO, Jiachang SHEN, Mengxin LIU, Yanjie JI, Qinhong WANG, Maohua YANG, Jianmin XING. Efficient separation of phosphorylated sugars and high-concentration phosphate by nanofiltration[J]. CIESC Journal, 2025, 76(12): 6423-6438.

图/表 20

表1 SOM的理化性质

Table 1 Physicochemical properties of SOM

SOM	MW/(g·mol^-1)	pK_a	Charge (pH6)	Stokes radius/nm	Diffusion coefficient/(10^-10 m²·s^-1)^③	Ref.
carbamazepine	236.27	15.96	0	0.36	6.81	[13]
dienestrol	266	10.50	—	0.47	5.21	[14]
trimethoprim	290.32	7.16	1	0.42	5.83	[6]
ethynyl estradiol	296	10.05	—	0.49	5.00	[14]
perfluoroheptanoic acid	364.02	-2.29	-1	0.35	7.00	[15]
FDP	340.12	—	-2^①	0.58^②	4.21	this work

表2 膜参数

Table 2 Membrane parameters

Product model	Material	MWCO/Da	Water flux/(L·m^-2·h^-1)
UE003	PES	3500	45^①
UE005	PES	5000	100^①
UX003	—	3000	60^①
NF2	PA	200—300	55^②
NF3	PA	300—400	70^②
NF4	PA	400—500	55^②
NF7	PA	400—700	60^②
NF8	PA	800—1000	65^②

图1 错流洗滤装置示意图a—恒温水浴;b—进料罐;c—隔膜泵;d—三通;e—压力表;f—螺旋缠绕卷式膜组件;g—透过液罐;h—计算机;i—电子天平;j—流量计;k—节流阀

Fig.1 Device diagram of cross-flow diafiltrationa—thermostatic water bath; b—feed tank; c—diaphragm pump; d—three-way; e—pressure gauge; f—spiral-wound membrane module; g—permeate tank; h—computer; i—electronic balance; j—flow meter; k—control valve

图2 不同pH的Pi组成

Fig.2 Distributions of phosphate species as a function of pH

图3 pH和浓度对Pi截留率和通量的影响

Fig.3 Effect of pH and concentration on phosphate rejection and flux

表3 离子物性参数

Table 3 Physical parameters of ions

Specie	Stokes radius/nm	Hydrated radius /nm	Hydration free energy/(kJ·mol^-1)	Diffusion coefficient/(10^-9 m²·s^-1)
Na⁺	0.184	0.358	-365	1.177
Cl^-	0.121	0.332	-340	2.028
$S O 4 2 -$	0.231	0.379	-1080	1.062
$H 2 P O 4 -$	0.256	0.302	-465	0.846
$H P O 4 -$	0.323	0.327	-1089	0.690
$P O 4 3 -$	—	0.339	-2765	0.612

表3 离子物性参数

Table 3 Physical parameters of ions

Specie	Stokes radius/nm	Hydrated radius /nm	Hydration free energy/(kJ·mol^-1)	Diffusion coefficient/(10^-9 m²·s^-1)
Na⁺	0.184	0.358	-365	1.177
Cl^-	0.121	0.332	-340	2.028
$S O 4 2 -$	0.231	0.379	-1080	1.062
$H 2 P O 4 -$	0.256	0.302	-465	0.846
$H P O 4 -$	0.323	0.327	-1089	0.690
$P O 4 3 -$	—	0.339	-2765	0.612

表4 模拟高盐体系的组成和浓度

Table 4 Compositions and concentrations of simulated high salinity solution

No.	Conc. of FDP/ (mmol·L^-1)	Conc. of Pi/(mmol·L^-1)	Ion strength/(mmol·L^-1)
1	2	18	24
2	4	36	48
3	8	72	96
4	15	135	180
5	30	270	360
6	8	216	240
7	16	192	240
8	24	168	240
9	32	144	240
10	48	96	240

图4 不同离子强度对磷酸糖/Pi截留率的影响（固定FDP的比例）

Fig.4 Effect of different ionic strengths on the rejection of phosphorylated sugars/phosphates (fixed the percentage of FDP)

图5 不同离子强度Pi离子通过NF4和NF7膜的跨膜传输活化能

Fig.5 Transport activation energy of Pi through the NF4 and NF7 membrane at different ionic strengths

图6 不同磷酸糖比例对磷酸糖/Pi截留率的影响（固定离子强度）

Fig.6 Effect of different percentage of FDP on the rejection of phosphorylated sugars/phosphates (fixed the ionic strengths)

图7 膜的纯水渗透系数（1 bar=105 Pa）

Fig.7 Pure water permeability coefficient of different membranes

图8 膜参数计算值

Fig.8 Calculated values for membrane parameters

图9 量化空间排斥、介电排斥和Donnan排斥（进料侧）对Pi截留的贡献

Fig.9 Distinction the contribution of steric, dielectric and donnan exclusion (feed side) on phosphates rejection

图10 DSPM-DE模型对离子截留行为的描述

Fig.10 Description of ion retention behavior by the DSPM-DE model

图11 NF4和NF7在不同pH下的Zeta电位值（a）及接触角（b）（L和R分别表示左右两侧）

Fig.11 Zeta potential values at different pH (a) and contact angles (b) of NF4 and NF7

图12 FDP转化液分离纯化流程示意图

Fig.12 Separation and purification process for FDP conversion solution

表5 实际体系纳滤测试结果

Table 5 Results of nanofiltration tests in the actual system

Batch		Volume/L	Concentration/(mmol·L^-1)		Rejection rate/%
Batch		Volume/L	Pi	Phosphorylated sugars	Pi	Phosphorylated sugars
1	Feed	0.77	89.50	28.04	66.85	100.00
1	Permeate	0.53	43.10	ND	66.85	100.00
2	Feed	1.00	104.16	40.05	63.15	100.00
2	Permeate	0.50	76.77	ND	63.15	100.00

图13 恒压洗滤分离磷酸糖/Pi

Fig.13 Separation of phosphorylated sugars/phosphates under constant-pressure diafiltration

图14 恒通量洗滤分离磷酸糖/Pi

Fig.14 Separation of phosphorylated sugars/phosphates under constant-flux diafiltration

图15 高浓度Pi (500 mmol·L-1)恒压洗滤

Fig.15 Constant pressure filtration with high concentration of Pi (500 mmol·L-1)

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磷酸糖与高浓度磷酸盐的高效纳滤分离

Efficient separation of phosphorylated sugars and high-concentration phosphate by nanofiltration

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No.	Conc. of FDP/ (mmol·L^-1)	Conc. of Pi/(mmol·L^-1)	Ion strength/(mmol·L^-1)
1	2	18	24
2	4	36	48
3	8	72	96
4	15	135	180
5	30	270	360
6	8	216	240
7	16	192	240
8	24	168	240
9	32	144	240
10	48	96	240

No.	Conc. of FDP/ (mmol·L^-1)	Conc. of Pi/(mmol·L^-1)	Ion strength/(mmol·L^-1)
1	2	18	24
2	4	36	48
3	8	72	96
4	15	135	180
5	30	270	360
6	8	216	240
7	16	192	240
8	24	168	240
9	32	144	240
10	48	96	240

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No.	Conc. of FDP/ (mmol·L^-1)	Conc. of Pi/(mmol·L^-1)	Ion strength/(mmol·L^-1)
1	2	18	24
2	4	36	48
3	8	72	96
4	15	135	180
5	30	270	360
6	8	216	240
7	16	192	240
8	24	168	240
9	32	144	240
10	48	96	240

No.	Conc. of FDP/ (mmol·L^-1)	Conc. of Pi/(mmol·L^-1)	Ion strength/(mmol·L^-1)
1	2	18	24
2	4	36	48
3	8	72	96
4	15	135	180
5	30	270	360
6	8	216	240
7	16	192	240
8	24	168	240
9	32	144	240
10	48	96	240

No.	Conc. of FDP/ (mmol·L^-1)	Conc. of Pi/(mmol·L^-1)	Ion strength/(mmol·L^-1)
1	2	18	24
2	4	36	48
3	8	72	96
4	15	135	180
5	30	270	360
6	8	216	240
7	16	192	240
8	24	168	240
9	32	144	240
10	48	96	240

No.	Conc. of FDP/ (mmol·L^-1)	Conc. of Pi/(mmol·L^-1)	Ion strength/(mmol·L^-1)
1	2	18	24
2	4	36	48
3	8	72	96
4	15	135	180
5	30	270	360
6	8	216	240
7	16	192	240
8	24	168	240
9	32	144	240
10	48	96	240