Molecular and Cellular Biochemistry

, Volume 4, Issue 3, pp 217–220 | Cite as

Covalent coupling of alkaline bacillus subtilis protease to controlledpore silica with a new simplified coupling technique

  • Ralph A. Messing
  • Harold R. Stinson
Review and General Articles b. general articles


A controlled-pore silica with an average pore diameter of 510 A° was used as the carrier for the immobilization of alkaline Bacillus subtilis protease. The coupling of the enzyme was accomplished by a simple two-step procedure which involved the aqueous reaction of a bifunctional diazonium salt with the surface of the silica followed by the coupling of the enzyme through the remaining available azo functional group.

The stability of the coupled enzyme was compared to that of an adsorbed enzyme produced under similar conditions. Both immobilized enzymes were stored in water at room temperature and were repeatedly assayed at intervals up to 103 days. These protease preparations were assayed with 1% casein solutions at pH 7.8 at 37°C. Under these conditions, the adsorbed enzyme exhibited a half-life of approximately 48 days, while that of the coupled protease was somewhat greater than 80 days.


Bacillus Subtilis Immobilize Enzyme Average Pore Average Pore Diameter Diazonium 
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Copyright information

© Dr. W. Junk b.v. Publishers 1974

Authors and Affiliations

  • Ralph A. Messing
    • 1
  • Harold R. Stinson
    • 1
  1. 1.Research & Development Laboratories Corning Glass WorksCorningUSA

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