Investigation of the Effect of Freezing on Protease-Catalyzed Peptide Synthesis Using Cryoprotectants and Frozen Organic Solvent
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Marion Haensler
Abstract
In order to investigate the effect of freezing on aqueous protease-catalyzed peptide synthesis systems, the influence of polyethylene glycols as cryoprotecting substances on α-chymotrypsin-catalyzed coupling of a N-protected acyl donor ester and various nucleophilic amino components was studied. Changes in S'-specificity of α-chymotrypsin in frozen aqueous systems were suppressed by polyethylene glycols even at concentrations below 1% (w/v). Furthermore, the influence of freeze-concentration in organic solvents on protease-catalyzed peptide synthesis was investigated for the first time. In frozen tert-butanol, α-chymotrypsin-catalyzed peptide synthesis took advantage from freeze-concentration, but in contrast to frozen aqueous systems, no changes in S'-specificity of the biocatalyst were observed. The results suggest that freeze-concentration is not the only cause of freezing-induced yield improvement in aqueous peptide synthesis systems, but interactions between enzyme and ice structures strongly contribute to the observed effects.
Copyright © 2000 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
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- Structural Analysis of Modified Forms of Recombinant IFN-β Produced under Stress-Simulating Conditions
- A Synthetic Peptide Derived from the Non-Structural Protein 3 of Hepatitis C Virus Serves as a Specific Substrate for PKC
- Neoglycolipids Derived from Phosphatidylethanolamine Serve as Probes in Cell Culture Studies on Glycolipid Metabolism
- Anomalous Binding of MgADP to Myosin of Skeletal Muscle
- Engineering, Purification and Applications of His-Tagged Recombinant Antibody Fragments with Specificity for the Major Birch Pollen Allergen, Bet v1
- Activation of Progelatinase B by Membranes of Human Polymorphonuclear Granulocytes
- Bait Region Cleavage and Complex Formation of Human α2M with a Porphyromonas gingivalisW50 Protease Is Not Accompanied by Enzyme Inhibition
- Adjuvant and Haemolytic Activities of 47 Saponins Derived from Medicinal and Food Plants
- Tissue-Specific Regulation of IRS-2/PI 3-Kinase Association in Aged Rats
- Investigation of the Effect of Freezing on Protease-Catalyzed Peptide Synthesis Using Cryoprotectants and Frozen Organic Solvent
- Cloning and Expression of Functional Equistatin
