Temperature Modulated Solubility-Activity Alterations for Poly(N-Isopropylacrylamide)-Lipase Conjugates

• Published in: Journal of biochemistry (Tokyo) 1994-09, Vol.116 (3), p.682-686
• Main Authors: Matsukata, Miki, Takei, Yoshiyuki, Aoki, Takashi, Sanui, Kohei, Ogata, Naoya, Sakurai, Yasuhisa, Okano, Teruo
• Format: Article
• Language: English
• Subjects: Acrylic Resins - chemistry; enzyme conjugate; lipase; Lipase - chemistry; Molecular Structure; poly(N-isopropylacrylamide)semitelechelic co-oligomer; Solubility; Temperature; temperature-induced phase transition; temperature-responsive polymer-temperature-modulated bio-separation system
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/oxfordjournals.jbchem.a124580

Chemical modification of proteins by use of functional polymers is expected to endow them with new properties without destroying their native functions, thus providing useful materials for application in different fields. We have synthesized poly(N-isopropylacryl-amide) [poly(IPAAm)] co-oligomer with N, N-dimethylacrylamide (DMAAm) and reactive end groups by telomerization of IPAAm. This co-oligomer exhibits a lower critical solution temperature (LCST) at 37°C. Using this temperature-responsive semitelechelic co-oligomer, we prepared polymer-enzyme conjugates of lipase by covalent coupling via carboxyl end-groups. This bioconjugate exhibits a LCST at 37°C, having rapid, reversible hydration-dehydration changes due to highly mobile free polymer end groups. The conjugate retained its native enzymatic activity below this critical temperature, above which it precipitated and its catalytic function was shut off. This conjugate can be readily separated from reaction mixtures as a precipitate by simple temperature changes after reaction and reused in cycles without denaturation. Such a modulated system is attractive for application as a novel bioreactor system.