Biocatalysis In Organic Media Research Articles

Cofactor Regeneration in Biocatalysis in Organic Media

Methods used for the regeneration of cofactors in organic media are reviewed. Substratedriven regeneration methods include the use of a second substrate of the same enzyme and the use of a second enzyme and its substrate. The use of mediators in oxidoreductions is described and examples of photochemical and electrochemical regeneration methods are presented. General problems and possibilities of cofactor regeneration in organic media are discussed.

‘Organic phase buffers’ control biocatalyst activity independent of initial aqueous pH

Combinations of triisooctylamine with its hydrochloride, or of triphenylacetic acid with its Na + salt, can function as buffers for use during biocatalysis in organic media. They can control the pH of an adjacent aqueous phase, even though both forms of each buffer remain in the organic phase. With 0.1 M aqueous NaCl, the mid-point pH values obtained with the two buffer systems are around pH 4.5 and 7.0, respectively. The activity of an immobilized subtilisin Carlsberg shows a strong dependence on the ratio of the two forms of the triphenylacetic acid buffer system. Without the buffer, the rate shows the normal dependence on the pH of the aqueous solution before drying; however, this is almost eliminated if the buffer is used. The amine buffer system can similarly affect the activity of an immobilized Rhizomucor miehei lipase.

Are Associated Ions Important for Biocatalysis in Organic Media?

The nature of the buffer species used in the drying process is important when lyophilized enzyme preparations are suspended in organic media. The activity of subtilisin Carlsberg in a transesterification reaction was found to vary depending on the nature of the buffer used. It was postulated that the large excess of salt present in the dried powder could be affecting enzymatic activity by alterations to the microscopic structure of the powder. To establish if this were true, microscopic changes were eliminated by covalently immobilising the enzyme onto a macroporous polymer support so that the counter-ions could be exchanged by washing with dilute salt solutions. It was found that in the immobilised samples no significant effects of salt ions were noted. This was the case even when salt ions were in considerable excess of that needed to balance protein charges. Hence the activity variations noted in freeze-dried powders are probably due to changes to the microscopic structure, rather than to molecular sca...

Continuous control of water activity during biocatalysis in organic media

This paper describes a newly developed technique to adjust and control the water activity in enzymatic reactions in organic media. A saturated salt solution of known water activity is circulated inside a silicone tube, submerged into the reaction medium. The circulating solution can both absorb and release water. Water activity control during lipase catalyzed esterification was demonstrated with diisopropyl ether as solvent.

Measurement of pH changes in an inaccessible aqueous phase during biocatalysis in organic media

The pH of the aqueous phase trapped within biocatalyst particles in organic media may be measured using very hydrophobic esters of fluorescein. These remain completely in a pentan-3-one phase, but they ionise there in response to the pH of an equilibrated aqueous phase. They show that the catalyst pH may be significantly shifted from the pre-adjusted value by partitioning of an acidic reactant (N-formyl-tyrosine during chymotrypsin-catalysed esterification) or product (acetic acid during lipase-catalysed ester hydrolysis).

Selective biotransformations. Patents and literature.

Stereo- and regioselective biotransformations is an area of intense research interest. The ability to take advantage of microorganisms and isolated enzyme systems to perform selective organic syntheses is well known. In recent years, several new methodologies have caused a rapid growth in the area of selective biotransformations. These novel techniques include genetic and protein engineering, biocatalysis in organic media, improved screening procedures, as well as an increased awareness of the value of biotechnology to solve important problems in organic synthesis. Recent US patents and scientific literature on stereo- and regioselective biotransformations are surveyed. Patent abstracts are summarized individually and a list of literature references are given.

Biocatalysis in organic media

Biocatalysis in organic media

Biocatalysis in organic media

The potentials of using organic reaction media in biotechnological conversions have already been demonstrated in several experimental studies. Examples of possible advantages are: possibility of higher substrate and/or product concentrations, favorable shift of reaction equilibria, reduced substrate and/or product inhibition, and facilitated product recovery. Especially water/organic solvent two-phase systems seem to possess several of these advantages. The solvent type will highly affect kinetics and stability of the (immobilized) biocatalyst, solubility and partitioning of reactants/products, and product recovery. Therefore the solvent choice can have a large influence on the economics of the two-liquid-phase biocatalytic process. Immobilization of the biocatalyst may be useful to provide protection against denaturating solvent effects. The polarity of the employed support material will also be decisive for the partitioning of substrates and products among the various phases. A classification of biphasic systems, which is based on the possible types of theoretical concentration profiles and aqueous phase configurations, is discussed. Reversed micelles and aqueous two-liquid-phase systems can be considered as special cases. The design of two-liquid-phase bioreactors is dependent on the state of the biocatalyst, free or immobilized, and on the necessity for emulsification of one of the two liquid phases in the other. Many mass-transfer resistances, e.g. across the liquid/liquid interface, in the aqueous phase, across the liquid/solid interface, and in the biocatalyst phase, can limit the overall reaction rate. The epoxidation of alkenes in water/solvent two-phase systems is discussed to give an example of the scope of biotechnological processes that is obtained by using organic media. Finally, a design calculation of a packed-bed organic-liquid-phasel immobilized-biocatalyst reactor for the epoxidation of propene is given to illustrate some of the above aspects.

Book review

Biocatalysis in Organic Media Edited by C. Laane, J. Tramper and M.D. Lilly Elsevier, 1987, 426 pages Large Scale Cell Culture Technology Lydersen, B.K. Hanser Publishers, Munich, 1987, 252 pgs., DM 98.00

Biocatalysis in organic media

Biocatalysis in organic media