Continuous Production of L-Carnitine with NADH Regeneration by a Nanofiltration Membrane Reactor with Coimmobilized L-Carnitine Dehydrogenase and Glucose Dehydrogenase

概要

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L-Carnitine dehydrogenase(CDH)was partially purified from Pseudomonas putida IMA12014 for the stereospecific reduction of 3-dehydrocarnitine to L-carnitine. CDH and glucose dehydrogenase(GDH)were coimmobilized in a nanofiltration membrane bioreactor(NFMBR)for the continuous production of L-carnitine from 3-dehydrocarnitine with NADH regeneration. In the NFMBR, NAD was partially immobilized through rejection by the nanofiltration membrane and effectively regenerated by the conjugation reaction of CDH and GDH. Since 3-dehydrocarnitine was unstable at neutral pH, it was maintained under acidic conditions(pH0.7)and supplied to the NFMBR separately from the other substrates, glucose and coenzyme NAD. As 50mM 3-dehydrocarnitine in HCl solution, 0.05mM NAD, and 100mM glucose in 0.5M Tris buffer(pH8)were continuously supplied to the NFMBR with immobilized CDH(200U/ml)and GDH(200U/ml)at the retention time of 80min and temperature of 25°C, the maximum conversion, reactor productivity, and NAD regeneration number were 78%, 113g/l/d, and 780, respectively. The half-life of the NFMBR was longer than 500h.
社団法人日本生物工学会の論文
1999-03-25