期刊
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
卷 168, 期 2, 页码 295-305出版社
HUMANA PRESS INC
DOI: 10.1007/s12010-012-9772-y
关键词
Enzyme immobilization; Human gastric juice; Immobilized alpha-chymotrypsin; Magnetic bead cellulose; Porcine pepsin A
资金
- Ministry of Education, Youth, and Sports of the Czech Republic [MSM 0021620806, CEH LC 06044]
- Czech Science Foundation [203/09/0857]
- [SVV-2011-262507]
Magnetic bead cellulose was prepared by a suspension method from the mixture of viscose and magnetite using thermal sol-gel transition and regeneration of cellulose. The prepared magnetic particles after their activation with divinyl sulfone were shown to be suitable magnetic carrier for immobilization of alpha-chymotrypsin and for its application in proteomic studies. The specific activity of the immobilized proteinase was high; its activity did not change in the course of storage. The following properties of the immobilized proteinase were compared with those of the soluble enzyme: pH and temperature dependence of the activity, self-cleavage activity, and possibility of repeated use. alpha-Chymotrypsin immobilized to magnetic bead cellulose was used for the proteolytic digestion of porcine pepsin A and human gastric juice and a possibility of direct use of enzyme reaction products for matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis was shown.
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