离子液体再生纤维素膜固定化木瓜蛋白酶的制备及在羊毛纤维修饰中的应用

doi:10.7503/cjcu20150498

摘要/Abstract

摘要：

利用离子液体1-丁基-3-甲基咪唑氯[Bmim]Cl溶解微晶纤维素Avicel, 得到再生纤维素膜, 作为载体固载木瓜蛋白酶, 将其用于羊毛纤维表面鳞片结构降解修饰. 固定化木瓜蛋白酶的优化条件: 以质量分数为6.5%的Avicel/离子液体混合溶液制备得到再生纤维素膜为载体; 改性液中(一定比例硅烷偶联剂与乙醇混合溶液)硅烷偶联剂NH₂(CH₂)₃Si(OC₂H₅)₃体积分数为30%, 改性温度50 ℃; 固定化体系pH=7.0, 吸附时间3 h, 交联时间4 h, 交联温度50 ℃, 戊二醛体积分数10%, 游离酶浓度300 mg/mL. 结果表明, 再生纤维素膜表面平滑规整, 木瓜蛋白酶被固定于再生纤维素膜2个表面, 固载密度高; 避免了游离酶浸入并侵蚀羊毛主体纤维内部而对鳞片结构降解不明显的缺点, 将酶的催化作用区域控制在羊毛纤维表面, 有效降解鳞片角质结构, 催化效能优良.

关键词: 木瓜蛋白酶, 固定化, 离子液体, 纤维素, 羊毛纤维鳞片结构

Abstract:

Ionic liquid(IL) 1-butyl-3-methylimidazolium chloride([Bmim]Cl) was used to dissolve microcrystalline cellulose Avicel to make IL-regenerated cellulose membrane for papain immobilization through silane modification method. It demonstrated that the immobilized papain could be obtained at the condition of 6.5%(mass fraction) Avicel/[BMIM]Cl for membrane as the solid support; 30% NH₂(CH₂)₃Si(OC₂H₅)₃ at 50 ℃ to functionalize the carrier; adsorbing for 3 h followed by cross-linking for 4 h at 50 ℃, pH=7.0 and enzyme concentration of 300 mg/mL, using 10%(volume fraction) glutaraldehyde as the cross-linker. After immobilization, papain was immobilized to the both sides of the regenerated cellulose membrane surfaces with high loading density. The immobilized papain could be limited on the wool fiber surface, only digesting the scale structure instead of native papain penetrating inside the fiber and generating the significant weight and strengthen loss.

Key words: Papain, Immobilization, Ionic liquid, Cellulose, Scale of wool

中图分类号:

O636.1⁺1

TrendMD:

李秋瑾, 赵芷芪, 袁亚梅, 巩继贤, 张健飞. 离子液体再生纤维素膜固定化木瓜蛋白酶的制备及在羊毛纤维修饰中的应用. 高等学校化学学报, 2015, 36(12): 2590.

LI Qiujin, ZHAO Zhiqi, YUAN Yamei, GONG Jixian, ZHANG Jianfei. Immobilization of Papain with Ionic Liquids-regenerated Cellulose Membrane for Wool Fabric Modification^†. Chem. J. Chinese Universities, 2015, 36(12): 2590.

图/表 5

Fig.1 TG(A) and DTG(B) analysis of Avicel(a) and [Bmim]Cl-regenerated cellulose membrane(b)

Fig.2 FTIR sprectra of original cellulose(a), [Bmim]Cl-regenerated cellulose membrane(b), and regenerated cellulose membrane after silane modification(c)

Fig.3 Effects of immobilization factors on activity of papain immobilized on [Bmim]Cl-regenerated cellulose membranes (A) Content of Avicel; (B) content of NH2(CH2)3Si(OC2H5)3; (C) temperature of silane modification; (D) pH; (E) adsorption time; (F) content of glutaraldehyde; (G) crosslinking time; (H) crosslinking temperature; (I) enzyme concentration.

Fig.4 SEM images of immobilized papain on Ils-regenerated cellulose membrane (A) Regenerated cellulose membrane; (B), (C) immobilized papain on regenerated cellulose membrane; (D)—(F) cross section of immobilized papain on regenerated cellulose membrane.

Fig.5 SEM images of wool fiber digested by immobilized papain (A) Original wool fiber; (B) wool fiber pretreated by H2O2; (C), (D) wool fiber hydrolyzed by free papain; (E), (F) wool fiber digested by immobilized papain.

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