CAJ | 학술논문

采用反相悬浮法合成了温敏型聚合物聚N-异丙基丙烯酰胺(PNIPA)凝胶及其与丙烯酸钠(SA)的共聚凝胶P(NIPA-co-SA),对凝胶的形态、粒径分布、表面特性以及温敏性能进行了考察,共单体丙烯酸钠的引入使得凝胶的最大溶胀倍率和低临界溶解温度(LCST)都有明显的提高.将两种凝胶用于目标蛋白溶菌酶的体外复性过程,考察了其协助复性的效果.结果表明:当蛋白浓度为250 μg·mL~(-1)时,加入80 mg·mL~(-1) PNIPA凝胶可使溶菌酶的活性回收率由稀释复性的51.3%提高到72.9%;加入120 mg·mL~(-1) P(NIPA-co-SA)共聚凝胶可使活性回收率达到71.5%.溶菌酶的浓度越高,与稀释复性相比凝胶协助复性的效果就越好.复性后凝胶可方便地分离回收,PNIPA凝胶重复使用6次后,溶菌酶的活性回收率仍高于稀释复性15%以上.
채용반상현부법합성료온민형취합물취N-이병기병희선알(PNIPA)응효급기여병희산납(SA)적공취응효P(NIPA-co-SA),대응효적형태、립경분포、표면특성이급온민성능진행료고찰,공단체병희산납적인입사득응효적최대용창배솔화저림계용해온도(LCST)도유명현적제고.장량충응효용우목표단백용균매적체외복성과정,고찰료기협조복성적효과.결과표명:당단백농도위250 μg·mL~(-1)시,가입80 mg·mL~(-1) PNIPA응효가사용균매적활성회수솔유희석복성적51.3%제고도72.9%;가입120 mg·mL~(-1) P(NIPA-co-SA)공취응효가사활성회수솔체도71.5%.용균매적농도월고,여희석복성상비응효협조복성적효과취월호.복성후응효가방편지분리회수,PNIPA응효중복사용6차후,용균매적활성회수솔잉고우희석복성15%이상.
The thermo-sensitive polymers of PNIPA and P(NIPA-co-SA) gel particles were synthesized by inverse suspension polymerization. The morphology, size distribution and thermo-sensitive characteristics of the prepared gel particles were studied, and it was found that the maximal swelling ratio and lower critical solution temperature (LCST) of the prepared gel particles increase obviously with the addition of sodium acrylate (SA) monomer. The effects of the gels for assisting the refolding of denatured lysozyme were then examined; when the protein concentration is 250 μg·mL~(-1), the found optimum concentration of PNIPA and P(NIPA-co-SA) gel used for assisting the refolding process are 80 μg·mL~(-1) and 120 μg·mL~(-1), respectively, and using the above optimum concentrations, the activity recovery of lysozyme increases from 51.3% (when using simple dilution refolding) to 72.9% (refolding assisted by PNIPA gel particles) and 71.5% (refolding assisted by P(NIPA-co-SA gel particles), respectively. The further results indicate that the higher initial denatured lysozyme concentration, the assisting effect of the prepared gel particles for refolding is more efficient. After refolding, the gel particles could be removed easily; and after reused for 6 times, the activity recovery of lysozyme refolding by reused PNIPA gel particles can still be 15% higher than that refolding by simple dilute refolding.