Purification and characterization of a class I fructose 1,6-bisphosphate aldolase from Staphylococcus carnosus

Abstract

A fructose 1,6-bisphosphate aldolase (E.C.4.1.2.13) from Staphylococcus carnosus DSM 20501 was purified for the first time. The enzymatic activity was insensitive to high levels of E D T A indicating that the enzyme is a class I aldolase. This enzyme exhibits good stability at high temperatures and extreme stability over a wide p H range. The Km for fructose 1,6-bisphosphate as substrate was 0.022 mM. The S. carnosus aldolase is a monomeric enzyme with a molecular mass of about 33 kDa. It exhibits a relatively broad p H optimum between pH 6.5 and 9.0. Furthermore, the aldolase accepts other aldehydes in place of its natural substrate, glyceraldehyde 3-phosphate, allowing the synthesis of various sugar phosphates.

Purification and Characterization of Class-I and Class-II Fructose-1,6-bisphosphate Aldolases from the Cyanobacterium Synechocystis sp. PCC 6803

“…PCC 6803 CI-FBA. The CI-FBA of S. carnosus is active as a monomer in vivo, and it exhibits extreme stability at high temperatures and over a wide pH range (Brockamp and Kula 1990). Because these enzymes are highly homologous, it is thought they may also share similar characteristics.…”

Stereochemistry of nonnatural aldol reactions catalyzed by DHAP aldolases

“…This simple coupled enzymatic assay makes use of the known stereopreference (Brockamp and Kula, 1990a;Fessner et al, 1991) of the commercially available aldolases in retro-aldol reactions.…”

One-pot four-enzyme synthesis of ketoses with fructose 1,6-bisphosphate aldolases from Staphylococcus carnosus and rabbit muscle

“…5 In contrast, FruA from bacterial sources, such as FruA from Staphylococcus carnosus (FruA S.car ), is exceptionally temperature and pH stable. 5–6 In Sheldon’s work, the activities and stabilities of two bacterial FruA from S. carnosus and S. aureus were compared with RAMA to investigate their scope of synthetic application. 7 …”