Recombinant aldolase

Pinsach J, de Mas C, Lopez-Santm J (2006) A simple feedback control of Escherichia coli growth for recombinant aldolase production in fed-batch mode. Biochem Eng J 29 235-242 Pons R, Erra P, Solans C et al. (1993) Viscoelastic properties of gel-emulsions their relationship with structure and equilibrium properties. J Phys Chem 97 12320-12324 Princen HM (1979) Highly concentrated emulsions. 1. CyUndtical systems. J Colloid Interface Sd 71 55-66... 

Like many other antibodies, the activity of antibody 14D9 is sufficient for preparative application, yet it remains modest when compared to that of enzymes. The protein is relatively difficult to produce, although a recombinant format as a fusion vdth the NusA protein was found to provide the antibody in soluble form with good activity [20]. It should be mentioned that aldolase catalytic antibodies operating by an enamine mechanism, obtained by the principle of reactive immunization mentioned above [15], represent another example of enantioselective antibodies, which have proven to be preparatively useful in organic synthesis [21]. One such aldolase antibody, antibody 38C2, is commercially available and provides a useful alternative to natural aldolases to prepare a variety of enantiomerically pure aldol products, which are otherwise difficult to prepare, allovdng applications in natural product synthesis [22]. 

Von derOsten, C.H., Sinskey, A.J., Barbas El, C.F., Pederson, R.L., Wang, Y.F. and Wong, C.H., Use of a recombinant bacterial fructose-1,6-diphosphate aldolase in aldol reactions preparative syntheses of 1-deoxynojirimycin, 1-deoxymannojirimycin, l,4-dideoxy-l,4-imino-D-arahinitol, and fagomine. J. Am. Chem. Soc., 1989, 111, 3924. 

Most interestingly, two fructose 6-phosphate aldolase isozymes (transaldolase C (TALC) and D-fructose-6-phosphate aldolase (FSA)) were reported from the E. coli genome and characterized in recombinant form [41]. 

Figure 2 One-pot three-enzyme chemoenzymatic synthesis of sialosides containing sialic acid modifications. In this strategy, mannose or ManNAc derivatives are chemically or enzymatically synthesized. These compounds are then used by a recombinant coli K-12 sialic acid aldolase to obtain sialic acids and their derivatives followed by an N. meningitidis CMP-sialic acid synthetase for the formation of CMP-sialic acids. From which, sialic acids can be transferred to lactose, LacNAc, galactose, GalNAc, or their derivatives by a multifunctional P. muitocida sialyltransferase (PmSTl) or a P, damseia a2,6-sialyltransferase (Pd2,6ST) to form a2,3- or a2,6-linked sialosides in one pot without the isolation of intermediates.

NaBH4 reduction with the help of CeCl3 -7H20 to obtain threo derivatives 232 (O Scheme 61). An enzymatic route for the synthesis of L-fucose analogs modified at the non-reducing end is reported by Fessner et al. [86], Using 2-Hydroxy-2-methylpropanal 233 and dihydroxyacetone phosphate 234 as substrates, branched fucose derivative 237 has been prepared via recombinant L-fuculose 1-phosphate aldolase (FucA) and L-fucose ketol isomerase (Fuel) in E. coli (O Scheme 62). 

A procedure for large-scale production of 2-deoxy-5-thio-D-eryf/tro-pentose (O Scheme 15) has been developed. It uses a recombinant 2-deoxyribose-5-phosphate aldolase (DER A) from E. coli strain DH5a as the catalyst that combines acetaldehyde with racemic 3-thioglyceralde-hyde [98]. 

Glyceraldehyde may be converted to the glycolytic intermediate, glyceraldehyde 3-phosphate (GAP), by the action of the enzyme triokinase. This enzyme phosphorylates glyceraldehyde at the expense of another molecule of ATP. The GAP can then enter into the glycolytic pathway and be further converted to pyra-vate, or recombine with DHAP to form F16BP by the action of aldolase. 

C. H. von der Osten, A. Sinskey, C. F. Barbas, R. L. Pederson, Y.-F. Wang, and C.-H. Wong, Use of a recombinant bacterial fractose-l,6-diphospbate aldolase in aldol reactions Preparative syntheses of 1-deoxynojirimycin, 1-deoxymannoijirimycin, 1,4-imino-D-arabinitol, and fago-mine, J. Am. Chem. Soc. 777 3924 (1989). 

Vidal L, Ferrer P, Alvaro G et al. (2005) Influence of induction and operation mode on recombinant rhamnulose-l-phosphate aldolase production in Escherichia coli using the T5 promoter. J... 

This chapter describes recent developments in carbohydrate synthesis using microbial aldolases. Recombinant type II fructose-1,6> diphosphate aldolase and type I 2-deoxyribose-5-phosphate aldolase have been exploited for the synthesis of monosaccharides and analogs. A new procedure for the synthesis of dihydroxy acetone phosphate has been developed. Sialic acid aldolase has been used in conjunction with other enzyme-catalyzed modifications of monosaccharides for the synthesis of sialic acid-related sugars. 

Figure 7. Plasmid construction for the preparation of recombinant deoxyribose-5-phosphate aldolase (DERA). phosphopentomutase and nucleoside phosphorylase.

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