Bacterial extracellular lignin peroxidase

• US 5,200,338 A
• Filed: 11/30/1988
• Issued: 04/06/1993
• Est. Priority Date: 11/30/1988
• Status: Expired due to Fees

• Current Assignee: Idaho Research Foundation, Inc.
• Original Assignee: Idaho Research Foundation, Inc.
• Inventors: Ramachandra, Muralidhara, Crawford, Donald L.
• Primary Examiner(s): Brown, Johnnie R.
• Assistant Examiner(s): Webber, Pamela S.
• Application Number:
  US07/277,802
• Time in Patent Office: 1,588 Days
• Field of Search: 435/200, 435/209, 435/192, 435/190, 435/28, 435/886, 435/253.5, 435/277
• US Class Current: 435/200
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12N 15/76 for Actinomyces; for Streptomyces
• CPC Class Codes: C12N 9/0065 acting on hydrogen peroxide as acceptor (1.11)
• CPC Class Codes: C12P 1/06 by using actinomycetales
• CPC Class Codes: C12R 2001/465 Streptomyces
• CPC Class Codes: Y10S 435/886 Streptomyces

Abstract

A newly discovered lignin peroxidase enzyme is provided. The enzyme is obtained from a bacterial source and is capable of degrading the lignin portion of lignocellulose in the presence of hydrogen peroxide. The enzyme is extracellular, oxidative, inducible by lignin, larch wood xylan, or related substrates and capable of attacking certain lignin substructure chemical bonds that are not degradable by fungal lignin peroxidases.

Claims

1. A lignin peroxidase enzyme having a molecular weight of about 18 kD, enzyme activity that produces Cα

-oxidation and Cα

-Cβ

cleavage of lignin and lignin degradation products, and a specific lignin peroxidase activity of greater than 0.30 enzyme U/mg.

2. The enzyme, according to claim 1, purified by at least one of ultrafiltration, ammonium sulphate precipitation, and Sepharose gel permeation chromatography.

3. The enzyme, according to claim 1, wherein said enzyme is inducible by lignin or larchwood xylan;

4. The enzyme, according to claim 1, wherein said enzyme is isolated from a member of Actinomycetales.

5. The enzyme, according to claim 4, wherein said member is from Streptomyces.

6. The enzyme, according to claim 5, wherein said member is from Streptomyces viridosporus.

7. A composition comprising a partially purified bacterial lignin peroxidase enzyme capable of degrading the lignin portion of lignocellulose in the presence of hydrogen peroxide and a carrier.

8. The composition, according to claim 7, wherein said enzyme has a molecular weight of about 18 KD, is inducible by lignin and larch wood xylan, and is capable of Cα

-Cβ

cleavage of lignin and lignin degradation products.

9. The composition according to claim 7, wherein said composition exhibits, a specific lignin peroxidase activity of greater than 0.30 enzyme U/mg.

15. The composition according to claim 7, wherein said composition exhibits a specific activity of greater than 1.0 enzyme U/mg.

10. A method for the degradation of lignocellulose comprising contacting said lignocellulose with a bacterial enzyme preparation of lignin peroxidase in the presence of hydrogen peroxide for a sufficient time for said degradation.

11. A method, according to claim 10, wherein said lignin substrate is selected from hardwood, softwood, and grasses.

12. A method, according to claim 10, further comprising the addition of, glucose and glucose oxidase to provide said hydrogen peroxide.

13. A method, according to claim 12, further comprising the addition of cellulase.

16. The method of claim 10, wherein said lignin peroxidase has a molecular weight of about 18 kD, enzyme activity that produces Cα

-oxidation and Cα

-Cβ

cleavage of lignin and lignin degradation products, and a specific lignin peroxidase activity of greater than 0.30 enzyme U/mg.

14. Actinomycete lignin peroxidase P3.