Induction of catalytic activity in proteins by lyophilization in the presence of a transition state analogue

Slade, Christopher J.; Vulfson, Evgeny N.

doi:10.1002/(sici)1097-0290(19980120)57:2<211::aid-bit9>3.0.co;2-q

Cited by 46 publications

(16 citation statements)

References 20 publications

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“…The crown ethers might bind to the active site during the dehydration step, thus keeping it nativelike, but not necessarily the remaining part of the molecule. That such local structural effects can occur in the presence of imprinting agents has been proven by the de novo introduction of catalytic activity to the protein bovine serum albumin (BSA) by colyophilization with a transition state analog (Slade and Vulfson, 1998). Similarly, imprinting with various agents has been reported to induce even larger structural alterations in various proteins upon lyophilization (Mishra et al, 1996).…”

Section: Molecular Imprintingmentioning

confidence: 99%

Effect of crown ethers on structure, stability, activity, and enantioselectivity of subtilisin Carlsberg in organic solvents

Santos

Vidal

Pacheco

et al. 2001

Biotech & Bioengineering

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Colyophilization or codrying of subtilisin Carlsberg with the crown ethers 18-crown-6, 15-crown-5, and 12-crown-4 substantially improved enzyme activity in THF, acetonitrile, and 1,4-dioxane in the transesterification reactions of N-acetyl-L-phenylalanine ethylester and 1-propanol and that of (±)-1-phenylethanol and vinylbutyrate. The acceleration of the initial rate, V 0 , ranged from less than 10-fold to more than 100-fold. All crown ethers activated subtilisin substantially, which excludes a specific macrocyclic effect from being responsible. The secondary structure of subtilisin was studied by Fourier-transform infrared (FTIR) spectroscopy. 18-Crown-6 and 15-crown-5 led to a more nativelike structure of subtilisin in the organic solvents employed when compared with that of the dehydrated enzyme obtained from buffer alone. However, the high level of activation with 12-crown-4 where this effect was not observed excluded overall structural preservation from being the primary cause of the observed enzyme activation. The conformational mobility of subtilisin was investigated by performing thermal denaturation experiments in 1,4-dioxane. Although only a small effect of temperature on subtilisin structure was observed for the samples prepared with or without 12-crown-4, both 18-crown-6 and 15-crown-5 caused the enzyme to denature at quite low temperatures (38°C and 56°C, respectively). No relationship between this property and V 0 was evident, but increased conformational mobility of the protein decreased its storage stability. The possibility of a "molecular imprinting" effect was also tested by removing 18-crown-6 from the subtilisin-18-crown-6 colyophilizate by washing. V 0 was only halved as a result of this procedure, an effect insignificant compared with the ca. 80-fold rate enhancement observed prior to washing in THF. This suggests that molecular imprinting is likely the primary cause of sub-tilisin activation by crown ethers, as recently suggested.

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Section: Molecular Imprintingmentioning

confidence: 99%

Effect of crown ethers on structure, stability, activity, and enantioselectivity of subtilisin Carlsberg in organic solvents

Santos

Vidal

Pacheco

et al. 2001

Biotech & Bioengineering

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“…In the past few years, polymers with catalytic properties have been synthesised via a technique called molecular imprinting [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. In this technique, a transition state analogue (TSA) of a selected reaction, either in its free or in an immobilised form, is added as a template to appropriate functional monomers which are co-polymerised with high amounts of cross-linker monomers.…”

Section: Introductionmentioning

confidence: 99%

Catalysis of an ester hydrolysis applying molecularly imprinted polymer shells based on an immobilised chiral template

Volkmann

Brüggemann

2006

Reactive and Functional Polymers

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“…Indeed, the 1990's provided a wealth of techniques to activate enzymes for use in organic media. Some of the prominent ones include the addition of polyols (Adlercreutz, 1993), crown ethers (Engbersen et al, 1996), transition-state analogs (Slade and Vulfson, 1998), and substrates and substrate mimics (Braco et al, 1990;Rich and Dordick, 1997). Beginning with a fundamental picture of enzymes in the nonaqueous milieu, the field of nonaqueous enzymology has recently seen cases where enzymes function in organic media as well as they do in water; again a striking realization that biocatalysis can be tailored by manipulating an enzyme's heterogeneous environment.…”

Section: Nonaqueous Enzymology-quantitative and Mechanistic Principlesmentioning

confidence: 99%

Engineering of molecular and cellular biocatalysts: Selected contributions by James E. Bailey

Dordick

Khosla

2002

Biotech & Bioengineering

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James (Jay) E. Bailey was a pioneer in biotechnology and biochemical engineering. During his 30 years in academia he made seminal contributions to many fields of chemical engineering science, including catalysis and reaction engineering, bioprocess engineering, mathematical modeling of cellular processes, recombinant DNA technology, enzyme engineering, and metabolic engineering. This article celebrates some of his contributions to the engineering of molecular and cellular biocatalysts, and identifies the influence he had on current and future research in biotechnology.

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Induction of catalytic activity in proteins by lyophilization in the presence of a transition state analogue

Cited by 46 publications

References 20 publications

Effect of crown ethers on structure, stability, activity, and enantioselectivity of subtilisin Carlsberg in organic solvents

Effect of crown ethers on structure, stability, activity, and enantioselectivity of subtilisin Carlsberg in organic solvents

Catalysis of an ester hydrolysis applying molecularly imprinted polymer shells based on an immobilised chiral template

Engineering of molecular and cellular biocatalysts: Selected contributions by James E. Bailey

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