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Characterization of a Novel N -Acetylneuraminate Lyase from Staphylococcus carnosus TM300 and Its Application to N -Acetylneuraminic Acid Production

2012; American Chemical Society; Volume: 60; Issue: 30 Linguagem: Inglês

10.1021/jf3014102

1520-5118

María Inmaculada García‐García, Agustín Sola-Carvajal, Francisco Garcı́a-Carmona, Álvaro Sánchez‐Ferrer,

Amino Acid Enzymes and Metabolism

The possibility of incorporating N-acetylneuraminic acid (Neu5Ac) in infant formulas and other functional foods has opened up the need to synthesize N-acetylneuraminic acid using N-acetylneuraminate lyases (NALs) by reversible aldol condensation of pyruvate and N-acetyl-d-mannosamine. Until now, NALs have been cloned from pathogenic microorganisms; however, this Article describes the expression and characterization of an N-acetylneuraminate lyase from the Staphylococcus carnosus TM300, a GRAS microorganism used in fermented meat. ScNAL showed a high level of expression in E. coli (403 mg L–1 culture). This, combined with its simple two-step purification procedure, the highest recovery described to date (86%), its kinetic parameters, which are in the same order of magnitude as best reported NALs, and its optimum pH and temperature, make ScNAL a promising and cheap biocatalyst. To confirm its biotechnological potential, the Neu5Ac was synthesized in 3 h in simple industrial working conditions with a high degree of conversion (94%).