Progress 04/15/23 to 04/14/24
Outputs
Target Audience:Two PhD students who are supported by this project got regular training and education on related knowledge and lab techniques. The PIs also instructed the graduate students in their classes by presenting this project as an example of hybrid nanomaterials for nanobiotechnology and environmental remodation. The graduate student presented the results on MNP for resource recovery in ACS conferences. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?1. The project provided a training opportunity for two PhD students in two departments: environmental engineering and chemistry. 2. The project also provides training opportunities for a research associate who had a master degree. 3. The project involved one more undergraduate students in Environmental Engineering and provided training opportunities for the student to gain research experience and learn about P recovery. How have the results been disseminated to communities of interest?Three journal papers have been published, one manuscript is in preparation on P recovery from biorefinery wastestreams. The graduate students have presented the results in ACS meetings. The PI presented the project idea and results in invited seminars from peer universities. What do you plan to do during the next reporting period to accomplish the goals?We aim to develop phytase biocatalyst to convert organic P, i.e. phytate, to inorganic phosphate and to recover the coverted phosphate by the engineered biosorbent. We will tested the process by using the real world samples from corn ethanol biorefinery waste stream including steep water.
Impacts
What was accomplished under these goals?
This project aims to create a new class of hybrid bio-nanomaterial platform for resource recovery applications including selective capture and efficient recovery of phosphate ions from wastewater. Our research progresses to date are summarized below. We have successfully developed a modular MNP platform that can achieve easy separation under magnetic force and efficient conjugation of proteins onto the surface. Specifically, we developed an efficient and scalable synthesis method to construct the core-shell structured MNP platform that contained three domains, a Fe3O4 nanocluster core, a SiO2 middle shell, and a surface-grafted polymer corona with decorated SpyTag in high density. A model SpyCatcher-fused enhanced green fluorescent protein, EGFP-SpyCatcher, was successfully conjugated onto the SpyTag-functionalized MNP platform without adding enzymes or chemical cross-linkers. The conjugation of EGFP on MNP was highly specific and robust, which was not affected by the presence of other proteins and detergents. The MNP platform was demonstrated to be protective to the conjugated EGFP and significantly improved the shelf life of immobilized proteins. We characterized the MNP platform as a generic and modular water-dispersible protein immobilization approach that allows easy separation and reusability of proteins. Specifically, three different SpyCatcher-fused fluorescent proteins, including EGFP-SpyCatcher, RFP-SpyCatcher, and YFP-SpyCatcher, were successfully conjugated onto the SpyTag-functionalized MNP. SpyTag-functionalized MNP showed high specificity to conjugated EGFP-SpyCatcher or RFP-SpyCatcher from complex cell lysate. Modular assembly of multiple proteins on MNP was achieved either from the mixture of two different proteins or their cell lysate mixture. The MNP platform significantly improved the long-term stability of immobilized proteins and their robustness to extreme conditions, such as high temperature and freeze-thaw process. Furthermore, we investigated the application potential of our newly established MNP platform, First, we developed a new type of biosorbent by immobilizing Lanmodulin-SpyCatcher (LanM-Spycatcher) on the surface of SpyTag-functionalized magnetic nanoparticles (MNPs) for selective separation and recovery of REEs from waste streams. The biosorbent, referred to as MNP-LanM, had an adsorption activity of 6.01 ± 0.11 μmol-terbium/g-sorbent and fast adsorption kinetics. The adsorbed REEs could be desorbed with > 90% efficiency. The MNP-LanM selectively adsorbed REEs in the presence of a broad range of non-REEs. The protein storage stability of MNP-LanM increased by two-fold compared to free LanM-SpyCatcher. The MNP-LanM could be efficiently separated using a magnet and reused with high stability as it retained ~95% of the initial activity after eight adsorption-desorption cycles. Second, we have successfully constructed and produced SpyCatcher-fused PstS recombinant protein from E. coli by synthetic biology techniques. Specifically, we created expression plasmids for producing the recombinant fusion phosphate binding protein and express functional fusion protein in the E. coli host cell. Results demonstrated that the fusion protein can adsorb and desorb phosphate by controlling pH of the solution. We are developing phytase and PstS system for both P conversion and recovery.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Ye, Q., Jin, X., Zhu, B., Gao, H., & Wei, N. (2023). Lanmodulin-Functionalized Magnetic Nanoparticles as a Highly Selective Biosorbent for Recovery of Rare Earth Elements. Environmental Science & Technology, 57(10), 4276-4285.
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Progress 04/15/22 to 04/14/23
Outputs
Target Audience:Two PhD students who are supported by this project got regular training and education on related knowledge and lab techniques. The PIs also instructed the graduate students in their classes by presenting this project as an example of hybrid nanomaterials for nanobiotechnology and environmental remodation. The graduate student presented the results on MNP for resource recovery in ACS conferences. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?1. The project provided a training opportunity for two PhD students in two departments: environmental engineering and chemistry. 2. The project also provides training opportunities for a research associate who had a master degree. The research technician gained research experience through this project and has been admitted to the Department of Chemistry and Biochemistry to pursue PhD study in Fall 2020. 3. The project involved two undergraduate students in Environmental Engineering and provided training opportunities for the student to gain research experience and learn about P recovery. How have the results been disseminated to communities of interest?Three journal papers have been published. The graduate students have presented the results in ACS meetings. The PI presented the project idea and results in invited seminars from peer universities. What do you plan to do during the next reporting period to accomplish the goals?1. Conjugate the SpyCatcher-PstS protein onto the engineered magnetic nanoparticle with SpyTag, to create the proposed material MN-PBP 2. Characterize the structure-function relationships of the MN-PBP and optimize the material at the molecular level to enhance its performance in terms of P recovery efficiency and stability. 3. Assess the robustness of the MN-PBP under the environmentally relevant and application relevant conditions, which will provide a basis for further optimization.
Impacts
What was accomplished under these goals?
This project aims to create a new class of hybrid bio-nanomaterial platform for resource recovery applications including selective capture and efficient recovery of phosphate ions from wastewater. Our research progresses to date are summarized below. We have successfully developed a modular MNP platform that can achieve easy separation under magnetic force and efficient conjugation of proteins onto the surface. Specifically, we developed an efficient and scalable synthesis method to construct the core-shell structured MNP platform that contained three domains, a Fe3O4 nanocluster core, a SiO2 middle shell, and a surface-grafted polymer corona with decorated SpyTag in high density. A model SpyCatcher-fused enhanced green fluorescent protein, EGFP-SpyCatcher, was successfully conjugated onto the SpyTag-functionalized MNP platform without adding enzymes or chemical cross-linkers. The conjugation of EGFP on MNP was highly specific and robust, which was not affected by the presence of other proteins and detergents. The MNP platform was demonstrated to be protective to the conjugated EGFP and significantly improved the shelf life of immobilized proteins. We characterized the MNP platform as a generic and modular water-dispersible protein immobilization approach that allows easy separation and reusability of proteins. Specifically, three different SpyCatcher-fused fluorescent proteins, including EGFP-SpyCatcher, RFP-SpyCatcher, and YFP-SpyCatcher, were successfully conjugated onto the SpyTag-functionalized MNP. SpyTag-functionalized MNP showed high specificity to conjugated EGFP-SpyCatcher or RFP-SpyCatcher from complex cell lysate. Modular assembly of multiple proteins on MNP was achieved either from the mixture of two different proteins or their cell lysate mixture. The MNP platform significantly improved the long-term stability of immobilized proteins and their robustness to extreme conditions, such as high temperature and freeze-thaw process. Furthermore, we investigated the application potential of our newly established MNP platform, First, we developed a new type of biosorbent by immobilizing Lanmodulin-SpyCatcher (LanM-Spycatcher) on the surface of SpyTag-functionalized magnetic nanoparticles (MNPs) for selective separation and recovery of REEs from waste streams. The adsorbed REEs could be desorbed with > 90% efficiency. The MNP-LanM selectively adsorbed REEs in the presence of a broad range of non-REEs. The protein storage stability of MNP-LanM increased by two-fold compared to free LanM-SpyCatcher. The MNP-LanM could be efficiently separated using a magnet and reused with high stability as it retained ~95% of the initial activity after eight adsorption-desorption cycles. Second, we have successfully constructed and produced SpyCatcher-fused PstS recombinant protein from E. coli by synthetic biology techniques. Specifically, we created expression plasmids for producing the recombinant fusion phosphate binding protein and express functional fusion protein in the E. coli host cell. Results demonstrated that the fusion protein can adsorb and desorb phosphate by controlling pH of the solution. We are developing phytase and PstS system for both P conversion and recovery.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Ye, Q., Jin, X., Gao, H., & Wei, N. (2022). Site-specific and tunable co-immobilization of proteins onto magnetic nanoparticles via Spy chemistry. ACS Applied Bio Materials, 5(12), 5665-5674.
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Ye, Q., Jin, X., Zhu, B., Gao, H., & Wei, N. (2023). Lanmodulin-Functionalized Magnetic Nanoparticles as a Highly Selective Biosorbent for Recovery of Rare Earth Elements. Environmental Science & Technology, 57(10), 4276-4285.
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