Protein-coated Microcrystals for use in Organic Solvents: Application to Oxidoreductases

Abstract

Protein-coated microcrystals (PCMC), a biocatalyst preparation previously demonstrated to display substantially increased transesterification activity of proteases and lipases in organic solvents when compared to their as received counterparts [Kreiner M, Moore BD, Parker MC (2001) Chem. Commun. 12:1096--1097], was applied to oxidoreductases. Horse liver alcohol dehydrogenase (HLADH), catalase (CAT), soybean peroxidase and horseradish peroxidase were immobilised onto K(2)SO(4) crystals and dehydrated in a 1-step process, resulting in PCMC. These PCMC preparations showed enhanced activity in different organic solvents in most types of reactions tested. The highest activation was observed with HLADH (50-fold as active as enzyme as received) and CAT (25-fold).

Combi-protein coated microcrystals of lipases for production of biodiesel from oil from spent coffee grounds

“…Many other enzymes have also been tried by other workers and their PCMCs also showed marked enhancement in catalytic activity as compared to that of the corresponding forms as received from the vendor. Horse radish peroxidase (HRP), soybean peroxidase (SBP), horse liver alcohol dehydrogenase (HLADH) and catalase showed 18, 9, 50 and 25-fold increase respectively [16]. Earlier, PCMC of a lipase has been reported to work well for obtaining biodiesel from Madhuca indica oil [43].…”

“…This is due to the structural changes in the enzyme molecules which take place during the drying process [7,8]. However, crystallization and precipitation have been found out to be better option for "drying" enzymes for use in low water media [9][10][11][12][13][14][15][16]. One such approach is preparation of protein coated microcrystals [10,14].…”

“…In this biocatalyst design, enzyme gets precipitated over microcrystals of salts or amino acids or sugar molecules. Fairly high initial rates for catalysis in low water containing organic media have been obtained for PCMCs of horse radish peroxidase (HRP), soybean peroxidase (SBP), horse liver alcohol dehydrogenase (HLADH), catalase and lipases [16]. For example, PCMC of lipase from Candida antarctica (CAL B) and Rhizomucor miehei (Palatase) showed a 5 and 15 fold increase in catalytic rates as compared to "straight from the vendor" preparations in the kinetic resolution of phenyl ethanol [10].…”

Influence of Water Miscible Organic Solvents on α-chymotrypsin in Solution and Immobilized on Eupergit CM

“…The covalent attachment to, physical adsorption on, or entrapment in, various solid phase matrices has been used to improve the tolerance of these biomolecular structures towards physical and chemical stress (Wang et al 2001;Kreiner and Parker 2005) and can afford improved conditions for substrate and product mass transfer, and even provide enhanced stabilities (Kudryashova et al 2003).…”

Enzyme Technology of Peroxidases: Immobilization, Chemical and Genetic Modification

“…The protein-coated microcrystals preparations showed enhanced activity in different organic solvents when tested with substrates such as guaiacol, thioanisole, and indole [20] Immobilized CPO on aminopropylglass…”