Industrial Biotransformations

N-Acetyl-d-neuraminic acid aldolase
Escherichia coli
● Another process layout is realized by the Research Center Jülich, Germany, that already established
in 1991 the one-pot synthesis of Neu5Ac with the combined use of epimerase and aldolase.
Here a continuously operated membrane reactor is used, where the enzymes are retained by an
ultrafiltration membrane. By this technology kg quantities of N-acetylneuraminic acid were produced
(GlcNAc = 300 mM; pyruvate = 600 mM; pH = 7.5; T = 25 °C; conversion= 78 %; residence
time = 4 h; reactor volume = 0.44 L; space-time-yield = 655 g · L –1 ·d –1 ;enzymeconsumption=
10,000 U · kg –1 ). The advantage of this approach is a simplified downstream processing, since the
catalysts are already separated. The product is additionally pyrogen free.
3) Flow scheme
Not published.
4) Process parameters
conversion: 77 %
reactor type: fed batch
reactor volume: 200 L
capacity: multi kg
down stream processing: crystallization
enzyme supplier: Marukin Shoyu Co., Ltd., Japan
company: Marukin Shoyu Co., Ltd., Japan and Forschungszentrum Jülich
GmbH, Germany
5) Product application
● Please see page 338.
6) Literature
EC 4.1.3.3
● Ghisalba, O., Gygax, D., Kragl, U., Wandrey, C. (1991) Enzymatic method for N-acetylneuraminic
acid production, Novartis , EP 0428947
● Kragl, U., Gygax, D., Ghisalba, O., Wandrey, C. (1991) Enzymatic process for prepaing N-acetylneuraminic
acid, Angew. Chem. Int. Ed. Engl. 30, 827–828
● Kragl, U., Kittelmann, M., Ghisalba, O., Wandrey, C. (1995) N-Acetylneuraminic acid: from a
rare chemical isolated from natural sources to a multi-kilogram enzymatic synthesis for industrial
application, Ann. N. Y. Acad. Sci. 750, 300–305
● Maru, I., Ohnishi, J., Ohta, Y., Tsukada, Y. (1998) Simple and large-scale production of N-acetylneuraminic
acid from N-acetyl-d-glucosamine and pyruvate using N-acyl-d-glucosamine 2epimerase
and N-acetylneuraminate lyase, Carbohydrate Res. 306, 575–578
● Ohta, Y., Tsukada, Y., Sugimori, T., Murata, K., Kimura, A. (1989) Isolation of a constitutive Nacetylneuraminate
lyase-producing mutant of Escherichia coli and its use for NPL production,
Agric. Biol. Chem. 53, 477–481
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