College of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China.
College of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang 212013, China.
Colloids Surf B Biointerfaces. 2022 Sep;217:112694. doi: 10.1016/j.colsurfb.2022.112694. Epub 2022 Jul 6.
In this study, the upper critical solution temperature (UCST)-responsive polymers poly (ethylene oxide) monomethyl ether-block-poly(acrylamide-co-acrylonitrile) (PEG-b-p(AAM-co-AN) were synthesized and successfully utilized to immobilize β-glucosidase in crude enzyme solution. These UCST-responsive β-glucosidase biocatalysts (PEG-b-p(AAM-co-AN@LytA-Glu) have specific UCST with tunable transition temperature, which could be tuned the separation temperature to the desired temperature range. The P2 @ LytA-Glu with an UCST of about 42.9 ℃ was exploited by one-step covalent immobilization of β-glucosidase in crude enzyme solution. The prepared P2 @ LytA-Glu exhibited significantly improved temperature, pH, storage, and operation stabilities compared with that of free enzyme. The catalytic rate of P2 @ Glu-LytA was 14.5% higher than that of P2-Glu (immobilized pure β-glucosidase), which indicated that one-step immobilization of crude enzyme directly from crude enzyme solution was feasible, and it can greatly save the purification step and reduce the experimental cost. The engineered UCST-responsive immobilized enzymes are potentially useful for the practical green biocatalysis.
在这项研究中，合成了上临界溶液温度（UCST）响应聚合物聚（氧化乙烯）单甲醚嵌段-聚（丙烯酰胺-co-丙烯腈）（PEG-b-p（AAM-co-AN），并成功地将其用于固定粗酶溶液中的β-葡萄糖苷酶。这些 UCST 响应型β-葡萄糖苷酶生物催化剂（PEG-b-p（AAM-co-AN@LytA-Glu）具有特定的 UCST 和可调谐的转变温度，可以将分离温度调谐到所需的温度范围。通过一步共价固定化在粗酶溶液中，将具有约 42.9℃ UCST 的 P2@LytA-Glu 用于β-葡萄糖苷酶的一步固定化。与游离酶相比，制备的 P2@LytA-Glu 表现出显著提高的温度、pH、储存和操作稳定性。P2@Glu-LytA 的催化速率比 P2-Glu（固定化的纯β-葡萄糖苷酶）高 14.5%，这表明直接从粗酶溶液一步固定化粗酶是可行的，它可以大大节省纯化步骤并降低实验成本。工程化的 UCST 响应型固定化酶在实际绿色生物催化中具有潜在的应用价值。
