Method of producing a product with crosslinked crystals of thermolysin
US5801022A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jun 7, 1995 |
| Grant date | Sep 1, 1998 |
| Priority date | — |
| Expiry date | Jun 7, 2015 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10S530/81
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent such as glutaraldehyde, and if desired lyophilizing the crosslinked crystals for storage. Crosslinking of the protein crystals provides stabilization for use under harsh conditions and for lyophilizing. The crystals crosslinked may be microcrystals having a cross-section of 10.sub.-1 mm or less. Crosslinked thermolysin, esterase, elastase, asparaginase and lysozyme crystals and crosslinked crystals of lipase from Geotrichum candidum and Candida cylindracea and of porcine origin can be used to convert a substrate to a product. Crosslinked thermolysin crystals are prepared that retain at least 96% of their initial activity after incubation for 4 days in the presence of a concentration of Pronase.TM. such as a thermolysin:Pronase.TM. ratio of 1:40 that causes the soluble uncrosslinked form of thermolysin that is crystallized to form the crystals that are crosslinked to lose at least 99% of its initial activity after incubation for 90 minutes under the same conditions. Crosslinked thermolysin crystals can be used to produce aspartame by combining the …
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.