| Functional polymer materials science is the fastest developed areas of polymer materials, which integrated the disciplines of chemistry, physics, and biology and so on. According to the nature and functions, functional polymer materials could be divided into the following branches, namely, separation polymer materials, polymer membranes, the active polymer materials (photoactive, electro-active and bioactive polymer), polymer liquid crystal materials, and smart polymer materials. The present work focuses on the separation polymer materials, and four types of functional polymer materials were prepared and applied in plant phosphorylation proteomics, protein crystallization and environmental microbiology respectively. The main findings were as follows:(1) A novel immobilized titanium ion affinity chromatography material (Ti4+-IMAC) was prepared in the aqueous environment using polyvinyl alcohol as raw material. SEM and IR spectroscopy were used to monitor the synthesis process. The selectivity for phosphopeptide enrichment from sample was improved by optimizing the pH and components of the loading buffer, and the optimal buffer system was found to be potassium hydrogen phthalate/HCl (pH2.5) solution. The final optimized protocol was adapted to salt-stressed maize leaves of Nongda108for phosphoproteome analysis, and numerous phosphorylated proteins/peptides related with salt stress were found for the first time. Thses research would provide solid evidence for comprehensive understanding of the mechanism of plant response to external salinity stress.(2) Molecularly imprinted polymer (MIPs) for tyrosine-phosphorylated peptides was synthesized by precipitation polymerization method using phenyl phosphate (PPA) as a template, Ti4+-immobilized ethylene glycol methacrylate phosphate as functional monomer, poly(ethylene glycol) diacrylate and N,N-methylene bisacrylamide as crosslinker, deionized water as porogen. The performance of MIPs was firstly evaluated by rebinding PPA, and then a competition experiment was conducted to assess the selectivity and specificity for PPA when mixed with benzoic acid, a structural analogue. The imprinting factor of the MIPs was up to2.04, compared with just0.24for benzoic acid, suggesting this kind of MIPs held superior selectivity and specificity. The selectivity and capacity of the MIPs for pTyr peptides from a mixture of peptides were considerably higher than that of commercial TiO2. Finally, MIPs were used for pTyr enrichment from a complex sample containing pTyr peptide and tryptic digestion of β-casein, where it demonstrated a clear preference for the pTyr peptide over ones containing phosphorylated serine.(3) Molecularly imprinted film based on polyurethane (PU-MIF) was obtained by the reaction among polyethylene glycol, castor oil and hexamethylene diisocyanate using methanotroph bacteria as template. PU-MIF was firstly characterized by biocompatibility test, selectivity and water retention capacity. Finally, PU-MIF was applied in the cell sorting of methanotroph bacteria.(4) Precipitant-immobilized molecularly imprinted polymers (piMIPs) were obtained by the combination of traditional molecularly imprinted technique and precipitating agent. For five model proteins, piMIPs facilitated better and faster crystals formation compared to conventional nucleants. piMIPs also can grow crystals which were missed using other nucleants, such as catalase. And results demonstrated if the type of immobilized precipitant was consistent with free one in solution, cognate piMIPs performed best in producing larger crystals and higher diffraction resolution. By this method, high-quality single crystal of a highly flexible and unsolved segment of Fragile X Mental Retardation Protein, whose absence causes the most common inherited mental retardation in human, was obtained without changing its structure. A novel KH domain-KHO and an intermolecular disulfide bond were identified for the first time. |
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