title: New method for the immobilization of pullulanase onto hybrid magnetic (Fe3O4-κ-carrageenan) nanoparticles by electrostatic coupling with pullulanase/chitosan complex.
abstract: We present a simple method to immobilize pullulanase onto hybrid magnetic (Fe3O4-κ-carrageenan) nanoparticles, involving the in situ synthesis of magnetic carrageenan nanoparticles and the formation of pullulanase/chitosan complex. The complex behavior of pullulanase with chitosan as a function of pH and protein-polysaccharide ratio was studied by turbidimetric titration. Then, the as-prepared immobilized enzymes were characterized by vibrating-sample magnetometer, transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray diffractometer, and thermogravimetric analysis. It was found that the activity retention of immobilized pullulanase and amount of enzyme loaded reached 95.5% and 96.3 mg/g, respectively, under optimal conditions. The immobilized enzyme exhibited great operational stability (retaining approximately 61% residual activity after ten consecutive reuses), demonstrating that enzyme leakage during the catalysis reaction was efficiently reduced. Furthermore, the activity of immobilized pullulanase was significantly (p < 0.01) higher than that of free pullulanase in a low pH range (pH < 3.0) and temperature over 60 °C, and the immobilized enzymes retained 45% of their initial activity after 5 h at 60 °C, compared to 21% for the free enzyme. These results indicated that immobilized pullulanase was efficient in terms of catalytic activity and can be applied to continuous starch processing applications in the food industry.
authors: †The State Key Laboratory of Food Science and Technology, School of Food Science and Technology and ‡Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
authors: Jie
authors: J
authors: Long
authors: †The State Key Laboratory of Food Science and Technology, School of Food Science and Technology and ‡Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
authors: Zhengzong
authors: Z
authors: Wu
authors: †The State Key Laboratory of Food Science and Technology, School of Food Science and Technology and ‡Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
authors: Xingfei
authors: X
authors: Li
authors: †The State Key Laboratory of Food Science and Technology, School of Food Science and Technology and ‡Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
authors: Enbo
authors: E
authors: Xu
authors: †The State Key Laboratory of Food Science and Technology, School of Food Science and Technology and ‡Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
authors: Xueming
authors: X
authors: Xu
authors: †The State Key Laboratory of Food Science and Technology, School of Food Science and Technology and ‡Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
authors: Zhengyu
authors: Z
authors: Jin
authors: †The State Key Laboratory of Food Science and Technology, School of Food Science and Technology and ‡Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
authors: Aiquan
authors: A
authors: Jiao
mesh_terms: D002351:Carrageenan / Q000737:chemistry*; D002384:Catalysis; D048271:Chitosan / Q000737:chemistry*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D005290:Ferric Compounds / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D060328:Magnetic Phenomena; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles; D017550:Spectroscopy, Fourier Transform Infrared; D055672:Static Electricity; D013696:Temperature
chemical_list: D004800:Enzymes, Immobilized; D005290:Ferric Compounds; C000499:ferric oxide; D002351:Carrageenan; D048271:Chitosan; D006026:Glycoside Hydrolases; C100181:pullulanase
keywords: immobilization; magnetic Fe3O4−κ-carrageenan nanoparticle; pullulanase; pullulanase/chitosan complex; stability