Dramatic enhancement of enzymatic activity in organic solvents by lyoprotectants

Journal Article · Fri Mar 05 04:00:00 EST 1993 · Biotechnology and Bioengineering; (United States)

DOI:https://doi.org/10.1002/bit.260410509· OSTI ID:6191576

Dabulis, K; Klibanov, A M ^[1]

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1. Massachusetts Inst. of Tech., Cambridge (United States)

When seven different hydrolytic enzymes (four proteases and three lipases) were lyophilized from aqueous solution containing a ligand, N-Ac-L-Phe-NH[sub 2], their catalytic activity in anhydrous solvents was far greater (one to two orders of magnitude) than that of the enzymes lyophilized without the ligand. This ligand-induced activation was expressed regardless of whether the substrate employed in organic solvents structurally resembled the ligand. Furthermore, nonligand lyoprotectants [sorbitol, other sugars, and poly(ethylene glycol)] also dramatically enhanced enzymatic activity in anhydrous solvents when present in enzyme aqueous solution prior to lyophilization. The effects of the ligand and of the lyoprotectants were nonadditive, suggesting the same mechanism of action. Excipient-activated and nonactivated enzymes exhibited identical activities in water. Also, addition of the excipients directly to suspensions of nonactivated enzymes in organic solvents had no appreciable effect on catalytic activity. These observations indicate that the mechanism of the excipient-induced activation is based on the ability of the excipients to alleviate reversible denaturation of enzymes upon lyophilization. Activity enhancement induced by the excipients is displayed even after their removal by washing enzymes with anhydrous solvents. Subtilisin Carlsberg, lyophilized with sorbitol, was found to be a much more efficient practical catalyst than its regular' counterpart.

• OSTI ID: 6191576
• Journal Information: Biotechnology and Bioengineering; (United States), Journal Name: Biotechnology and Bioengineering; (United States) Vol. 41:5; ISSN BIBIAU; ISSN 0006-3592
• Country of Publication: United States
• Language: English

Related Subjects

560300* -- Chemicals Metabolism & Toxicology
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALCOHOLS
BIODEGRADATION
BIOLOGICAL EFFECTS
CARBOHYDRATES
CARBOXYLESTERASES
CHEMICAL REACTIONS
DECOMPOSITION
DIURETICS
DRUGS
ENZYME ACTIVITY
ENZYMES
ESTERASES
GLYCOLS
HYDROLASES
HYDROXY COMPOUNDS
LIGANDS
LIPASES
LYOPHILIZATION
MATHEMATICAL MODELS
METABOLISM
MONOSACCHARIDES
ORGANIC COMPOUNDS
ORGANIC POLYMERS
ORGANIC SOLVENTS
PEPTIDE HYDROLASES
POLYETHYLENE GLYCOLS
POLYMERS
PROTEINS
RESPONSE MODIFYING FACTORS
SACCHARIDES
SOLVENTS
SORBITOL