Comparison of polymer induced and solvent induced trypsin denaturation: the role of hydrophobicity.
Trypsin adsorption from aqueous buffer by various copolymers of allyl glycidyl ether-ethylene glycol dimethacrylate (AGE-EGDM) copolymer with varying crosslink density increases with increasing crosslink density and the effect slowly wears off after reaching a plateau at 50% crosslink density. The copolymer with 25% crosslink density was reacted with different amines with alkyl/aryl side chains to obtain a series of copolymers with 1,2-amino alcohol functional groups and varying hydrophobicity. Trypsin binding capacity again increases with hydrophobicity of the reacting amine and a good correlation between logPoctanol of the amine and protein binding is observed. The bound trypsin is denatured to the extent of 90% in spite of the presence of hydrophilic hydroxyl and amino groups. The behavior was comparable to that in mixtures of aqueous buffer and water-miscible organic co-solvents where the solvent concentration required to deactivate 50% of the enzyme (C50) is dependent on logPoctanol of the co-solvent.
Biotransformations Laboratory, Indian Institute of Chemical Technology, Uppal Road, Hyderabad 500007, India.
Lakshmi S
LS
Jasti
Biotransformations Laboratory, Indian Institute of Chemical Technology, Uppal Road, Hyderabad 500007, India. Electronic address: fadnavisnw@yahoo.com.
Nitin W
NW
Fadnavis
Centre for Biotechnology, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad 500085, India.
Uma
U
Addepally
Polymer Science & Engineering, Chemical Engineering Division, National Chemical Laboratory, Pashan Road, Pune 411008, India.
Siona
S
Daniels
Polymer Science & Engineering, Chemical Engineering Division, National Chemical Laboratory, Pashan Road, Pune 411008, India.
Sarika
S
Deokar
Polymer Science & Engineering, Chemical Engineering Division, National Chemical Laboratory, Pashan Road, Pune 411008, India.
Surendra
S
Ponrathnam
D000327:Adsorption; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D011108:Polymers / Q000737:chemistry*; D011489:Protein Denaturation; D012997:Solvents / Q000737:chemistry; D013499:Surface Properties; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism
D004800:Enzymes, Immobilized; D011108:Polymers; D012997:Solvents; D014357:Trypsin
Adsorption; Denaturation; Hydrophobicity; Polymer; Trypsin