Staphylococcus carnosus

The class I FruA isolated from rabbit muscle aldolase (RAMA) is the aldolase employed for preparative synthesis in the widest sense, owing to its commercial availability and useful specific activity of 20 U mg . Its operative stability in solution is limiting, but the more robust homologous enzyme from Staphylococcus carnosus has been cloned for overexpression [87], which offers unusual stability for synthetic purposes. Recently, it was shown that less polar substrates may be converted as highly concentrated water-in-oil emulsions [88]. 

Staphylococcus carnosus is genetically highly stable, a good secretor (Gram positive) able to translocate proteins containing several hydrophobic transmembrane regions [58], and it has no extracellular proteases, which makes it suitable for production of the secreted enzymes [58],... 

Hansson, M., Samuelson, P., Nguyen, T.N. and Stahl, S. (2002) General expression vectors for Staphylococcus carnosus enabled efficient production of the outer membrane protein A of Klebsiella pneumoniae. FEMS Microbiology Letters, 210 (2), 263—270. 

Kohlbrecher, D. Eisermann, R. Hengstenberg, W. Staphylococcal phos-phoenolpyruvate-dependent phosphotransferase system molecular cloning and nucleotide sequence of the Staphylococcus carnosus ptsi gene and expression and complementation studies of the gene product. J. Bacteriol., 174, 2208-2214 (1992)... 

Of the several aldolases that are now commercially available, the rabbit muscle enzyme with a useful specific activity of 20 Umg-1 still remains the most cost-efficient catalyst for preparative work, although the reported higher stability of the Staphylococcus carnosus enzyme is a challenge [252,285], Literally hundreds of aldehydes have so far been tested successfully by enzymatic... 

A particularly elegant example is shown in Scheme 11 5-deoxy-5-ethyl-D-xy-lulose (19) was prepared from glycerol by a four step, one-pot chemoenzymatic reaction sequence. The key step was the aldol reaction catalyzed by fructose-1,6-bisphosphate aldolase (FruA) from Staphylococcus carnosus [56]. 

Voit, H. Goetz, F. Mersmann, A.B. Overproduction of lipase with Staphylococcus carnosus (pLipPSl) under modified gravity in a centrifugal field bioreactor. Chem. Eng. Technol. 1989, 12, 364-373. 

Dilsen S, Paul W, Sandgathe A, Tippe D, Freudl R, Thommes J, Kula M R, Takers R, Wandrey C, Weuster-Botz D (2000). Fed-batch production of recombinant human calcitonin precursor fusion protein using Staphylococcus carnosus as an expression-secretion system. Appl. Microbiol. Biotechnol. 54 361-369. 

Groger et al. have explored xenon interacting with a mutant of the phospho-carrier protein from Staphylococcus carnosus using H- N HSQC experiments. Significant changes in the chemical shift values were observed for the mutant protein, especially those close to the engineered cavity. In contrast, the wild-type protein showed much smaller, nonspecific interactions when xenon was introduced into solution. 

Aldol reaction. More than 20 aldehydes are known to be substrates for the aldolases from Staphylococcus carnosus and S. aureus. Stereo-selectivity of the aldolases must be looked at for each acceptor substrate, because isomers are formed in different proportions. The oxidation and aldol reaction can be carried out simultaneously. 

Micrococcaceae. Staphylococcus spp. and to a lesser extent Micrococcus spp. are found in commercial starter cultures (Hammes et al., 1985). Three species are mentioned Staphylococcus carnosus. Staphylococcus xylosus and Micrococcus varians. 

The D-fractose 1,6-bisphosphate aldolase (FraA EC 4.1.2.13) catalyzes in vivo the glycolytic equilibrium addition of 22 to D-glyceraldehyde 3-phosphate (15) to give d-fructose 1,6-bisphosphate (23). The equilibrium constant of 10" strongly favors synthesis [43]. The class I FruA isolated from rabbit muscle has been the most extensively investigated aldolase for mechanistic, structural, and preparative purposes. Its stabiUty in solution is limiting, but the class I FruA (monomeric) from Staphylococcus carnosus [91] or the Zn -dependent, type II aldolase from E. coli [92] are much more stable with half-lives up to several weeks or months as compared to only a few days [93]. Typically, type I FruA enzymes are tetrameric proteins composed of subunits of approximately 40 kDa, while the type II FruA are dimers of approximately 39-kDa subunits. 

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