Immobilization and stability of the NAD-dependent hydrogenase from alcaligenes eutrophus and of whole cells

✍ Scribed by Peter Egerer; Helmut Simon; Atsuo Tanaka; Saburo Fukui

• Publisher: Springer Netherlands
• Year: 1982
• Tongue: English
• Weight: 412 KB
• Volume: 4
• Category: Article
• ISSN: 0141-5492
• DOI: 10.1007/bf00131570

✦ Synopsis

The purified soluble NAD-dependent hydrogenase from Alcaligenes eutrophus was immobilized to porous glass beads according to the glutaraldehyde method retaining about 80% of its original activity. :Entrapment of the purified hydrogenase in photo-crosslinkable prepolymers led to apparent activity yields of i0=80% dependent on the thickness of the gel film. The storage stability of entrapped hydrogenase (t/2 = 4 d) was considerably lower than that of glass-bound hydrogenase (t/2 = 150 d). During continuous production of NADH (turnover conditions), the half-life of entrapped hydrogenase was not longer than i0 h. Whole cells of A. eutrophus entrapped in a polyurethane matrix were used to produce NADH with hydrogen gas as electron donor. After. 18 runs for 4h each and storage periods overnight the residual activity was still about 50%.