Efficient separation of HEPES and CO2-derived fructose by nanofiltration: Optimizing pH for improved separation and proposing a potential mechanism
Utilization of an in vitro multi-enzyme catalytic system for the synthesis of fructose from carbon dioxide is a promising approach towards achieving carbon neutrality. However, the lack of effective separation methods for the buffer HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) and the product fructose within the system has hindered the industrialization of in vitro multi-enzyme catalytic synthesis systems. This study aimed to separate and recover HEPES and fructose using nanofiltration technology. We investigated the influence of various parameters, including total solution concentration, solute concentration ratio, pH, and temperature, on the nanofiltration separation performance. Experimental results revealed that increasing the pH to 9.0 significantly decreased the fructose retention rate while maintaining a nearly complete HEPES retention rate. The HEPES-fructose mixed solution was further subjected to diafiltration using a spiral-wound membrane, ultimately yielding fructose and HEPES solutions with purities of 90.22 % and 92.93 %, respectively. Combining separation test results with models such as DSPM-DE, and using characterization methods such as ATR-FTIR and pore size distribution analysis, we observed pore swelling during nanofiltration of the HEPES-fructose solution at pH 9.0. A mechanism for this pore swelling, induced by the dissociation of HEPES under the influence of pH, was proposed: Dissociated HEPES in the solution interacts with the membrane surface at the liquid-membrane interface, causing reversible swelling of the membrane pores at the membrane surface, thereby reducing the retention rate of fructose. The nanofiltration process developed in this study effectively separates HEPES and fructose, enabling resource recycling and cost reduction. Furthermore, this study provides theoretical foundations and practical guidance for the design of nanofiltration separation strategies for similar systems using HEPES as a buffer, holding significant scientific and practical value.
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