Title
Synthesis and Characterization of Highly Stabilized Polymer–Trypsin Conjugates with Autolysis Resistance
Author
Yasushi Sasai, Hiroshi Kanno, Naoki Doi, Yukinori Yamauchi, Masayuki Kuzuya, Shin-ichi Kondo
Year
2017
Journal
Catalysts
Abstract
Protein digestion by trypsin has been widely used in many industrial and research applications. However, extensive use of trypsin is limited because of the rapid decrease in enzymatic activity caused by autolysis at optimal pH and temperature. To improve the enzymatic performance of trypsin, we synthesized highly stabilized polymer–trypsin conjugates using vinylmethylether-maleic acid copolymer (VEMAC) via multi-point attachment. The VEMAC modification significantly enhanced the thermal stability of trypsin, and the resulting conjugates showed a strong resistance to autolysis. VEMAC-modified trypsin (VEMAC-Tryp) showed maximum activity at 55 °C and at 1.4-fold higher levels than that of unmodified trypsin. Bovine serum albumin was effectively digested by VEMAC-Tryp, indicating that the modified trypsin can be used for digestion of high molecular weight substrates. VEMAC modification is a simple and cost-effective strategy to obtain fully active modified enzymes, and may be used to develop bioreactors.
Full Article
Instrument
J-820
Keywords
Circular dichroism, Secondary structure, Biochemistry