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A new type of immobilized halophilic nuclease H, Ca-enzyme complex, from a moderate halophile, Micrococcus varians subsp. halophilus was demonstrated. The 4-day cultivation of the bacterium in 3 m NaCl CM medium at 25°C was favorable for our purpose, producing less 5′-nucleotidase, but flocculation of the cells grown at 25°C was poorer than that of the cells grown at 30°C. On the other hand, almost all of the nuclease H was precipitated to form Ca-enzyme complex by addition of 70 mm Ca2+ to the supernatant of the broth. The nuclease H activity of the Ca-enzyme complex was increased by preferential adsorption of exogenous nuclease H. A column bioreactor packed with this enhanced Ca-enzyme complex was used for 5′ -mononucleotide production by continuous RNA degradation.

Ca-enzyme complex of halophilic nuclease H of halophilic Micrococcus varians subsp. halophilus for 5'-nucleotide production by RNA degradation.