Enzyme-polyelectrolyte coacervate complex and method of use

• US 5,834,271 A
• Filed: 06/21/1994
• Issued: 11/10/1998
• Est. Priority Date: 06/21/1994
• Status: Expired due to Term

• Current Assignee: Indiana University Research & Technology Corporation (Indiana University)
• Original Assignee: ADVANCED RESEARCH & TECHNOLOGY INSTITUTE
• Inventors: Xia, Jiulin, Muhoberac, Barry B., Dubin, Paul L.
• Primary Examiner(s): Weber, Jon P.
• Application Number:
  US08/263,041
• Time in Patent Office: 1,603 Days
• Field of Search: 435/174
• US Class Current: 435/174
• CPC Class Codes: C12N 11/02 Enzymes or microbial cells immobilised on or in an organic carrier
• CPC Class Codes: C12N 9/00 Enzymes; Proenzymes; Compositions thereof preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113; Processes for preparing, activating, inhibiting, separating or purifying enzymes preparation of malt C12C1/00
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)
• CPC Class Codes: C12P 1/00 Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
• CPC Class Codes: C12P 7/24 containing a carbonyl group
• CPC Class Codes: C12Q 1/00 Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: C12Q 1/37 involving peptidase or proteinase

Abstract

Described are unique preferred enzyme coacervate compositions and their use in catalyzing the reaction of substrates to which the enzyme is specific. Preferred coacervates of the invention are complex coacervates formed by complexing enzymes with polyelectrolytes.

Claims

1. An enzymatic process comprising the steps of:

first, forming a stable polyelectrolyte-enzyme coacervate dispersed as liquid droplets in a continuous liquid medium by mixing the polyelectrolyte with at least one enzyme so that the at least one enzyme is confined within the coacervate liquid droplets throughout the enzymatic process;

second, introducing a substrate into solution in the continuous liquid medium to react with the at least one enzyme to form a product, the substrate moving from the continuous liquid medium into the liquid droplets and the product moving from the liquid droplets into the continuous liquid medium; and

thereafter

harvesting the product from the continuous liquid medium; and

recovering the stable, liquid polyelectrolyte-enzyme coacervate for reuse.

2. An enzymatic process as defined in claim 1, wherein the continuous liquid medium comprises water.

3. A process as defined in claim 1, wherein the at least one enzyme comprises at least about 0.1 percent by weight of the coacervate.

4. A process as defined in claim 3, wherein the at least one enzyme comprises about 0.1 to abut 5 percent by weight of the coacervate.

5. A process as defined in claim 1, wherein the polyelectrolyte is a synthetic polymer.

6. A process as defined in claim 5, wherein the polyelectrolyte is selected from the group consisting of poly(acrylic acid) poly(alkyl acrylic acids), quaternized poly(vinylpyridines), poly(dialkyldiallylammoniumchlorides), and sodium poly(styrenesulfonate).

7. A process as defined in claim 1, wherein the coacervate comprises a plurality of enzymes.

8. An enzymatic process comprising the steps of:

forming a stable polyelectrolyte-enzyme liquid coacervate mixture by mixing the polyelectrolyte with at least one enzyme;

dispersing the polyelectrolyte-enzyme mixture as liquid coacervate droplets within a continuous liquid medium so that the at least one enzyme is confined within the liquid droplets throughout the enzymatic process;

introducing a substrate into solution in the continuous liquid medium to react with the at least one enzyme to form a product, the substrate moving from the continuous liquid medium into the liquid coacervate droplets and the product moving from the liquid coacervate droplets into the continuous liquid medium; and

thereafter

harvesting the product from the continuous liquid medium; and

recovering the stable, liquid polyelectrolyte-enzyme coacervate mixture for reuse.

9. A process as defined in claim 8, wherein the continuous liquid medium comprises water.

10. A process as defined in claim 8, wherein the at least one enzyme comprises at least about 0.1 percent by weight of the polyelectrolyte-enzyme coacervate mixture.

11. A process as defined in claim 10, wherein the at least one enzyme comprises about 0.1 to about 5 percent by weight of the polyelectrolyte-enzyme coacervate mixture.

12. A process as defined in claim 8, wherein the polyelectrolyte is a synthetic polymer.

13. A process as defined in claim 12, wherein the polyelectrolyte is selected from the group consisting of poly(acrylic acid), poly(alkyl acrylic acids), quaternized poly(vinylpyridines), poly(dialkyldiallylammoniumchlorides), and sodium poly(styrenesulfonate).

14. A process as defined in claim 8, wherein the coacervate comprises a plurality of enzymes.

15. An enzymatic process, comprising:

forming a reaction mass by charging to a reactor (i) a stable, polyelectrolyte-enzyme coacervate dispersed as liquid droplets in a continuous liquid medium by mixing the polyelectrolyte with at least one enzyme so that the at least one enzyme is confined within the liquid droplets throughout the enzymatic process, and (ii) a substrate, for which the at least one enzyme is specific, in solution in the continuous phase;

maintaining the reaction mass under conditions in which the at least one enzyme catalyzes reaction of the substrate to form a product while the at least one enzyme is incorporated in the coacervate liquid droplets; and

thereafter

harvesting the product from the continuous liquid medium; and

recovering the stable, liquid polyelectrolyte-enzyme coacervate for reuse.

16. A process as defined in claim 15, wherein the at least one enzyme comprises at least about 0.1 percent by weight of the coacervate.

17. A process as defined in claim 16, wherein the at least one enzyme comprises about 0.1 to about 5 percent by weight of the coacervate.

18. A process as defined in claim 15, wherein the polyelectrolyte is a synthetic polymer.

19. A process as defined in claim 18, wherein the polyelectrolyte is selected from the group consisting of poly(acrylic acid), poly(alkyl acrylic acids), quaternized poly(vinylpyridines), poly(dialkyldiallylammoniumchlorides), and sodium poly(styrenesulfonate).

20. A process as defined in claim 15, wherein the coacervate comprises a plurality of enzymes.

21. A process as defined in claim 15, wherein the coacervate is formed in situ in said reactor.

22. A composition comprising:

stable polyelectrolyte-enzyme coacervate dispersed as liquid droplets in a continuous liquid medium formed by mixing the polyelectrolyte with at least one enzyme so that the at least one enzyme is confined within the liquid droplets; and

a substrate which is soluble in the continuous phase, the coacervate being permeable to the substrate for which the at least one enzyme is specific.

23. A composition as defined in claim 22, wherein the at least one enzyme comprises at least about 0.1 percent by weight of the coacervate.

24. A composition as defined in claim 23, wherein the at least one enzyme comprises about 0.1 to about 5 percent by weight of the coacervate.

25. A composition as defined in claim 22, wherein the polyelectrolyte is a synthetic polymer.

26. A composition as defined in claim 25, wherein the polyelectrolyte is selected from the group consisting of poly(acrylic acid), poly(alkyl acrylic acids), quaternized poly(vinylpyridines), poly(dialkyldiallylammoniumchlorides), and sodium poly(styrenesulfonate).

27. A composition as defined in claim 26, wherein the at least one enzyme comprises at least about 0.1 percent by weight of the coacervate.

28. A composition as defined in claim 22, wherein the coacervate comprises a plurality of enzymes.