Magnetic Bead Cellulose as a Suitable Support for Immobilization of α-Chymotrypsin

• Published in: Applied biochemistry and biotechnology Vol. 168; no. 2; pp. 295 - 305
• Main Authors: Prikryl, Petr, Lenfeld, Jiri, Horak, Daniel, Ticha, Marie, Kucerova, Zdenka
• Format: Journal Article
• Language: English
• Published: New York Springer-Verlag 01.09.2012; Springer
• Subjects: Amino Acid Sequence; Animals; Biochemistry; Biological and medical sciences; Biotechnology; Cattle; Cellulose - chemistry; Chemistry; Chemistry and Materials Science; Chymotrypsin - chemistry; Chymotrypsin - metabolism; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Fundamental and applied biological sciences. Psychology; Gastric Juice - enzymology; General aspects; Humans; Immobilization techniques; Magnets - chemistry; Methods. Procedures. Technologies; Microspheres; Molecular Sequence Data; Pepsin A - chemistry; Pepsin A - metabolism; Proteolysis; Solubility; Sulfones - chemistry; Magnetic bead cellulose; Porcine pepsin A; Immobilized α-chymotrypsin; Enzyme immobilization; Human gastric juice; Human; Stomach; Serine endopeptidases; Support; Enzyme; Pepsin A; Cellulose; Immobilization; Pig; Peptidases; Chymotrypsin; Vertebrata; Mammalia; Hydrolases; Aspartic endopeptidases; Artiodactyla; Ungulata
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-012-9772-y

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 α-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. α-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.