PROCESS FOR RAPIDLY SCREENING ENZYME AND ENZYME MIXTURES FOR THE HYDROLYSIS OF LIGNO-CELLULOSIC BIOMASS

• US 20140315230A1
• Filed: 11/23/2012
• Published: 10/23/2014
• Est. Priority Date: 11/23/2011
• Status: Abandoned Application

• Current Assignee: C5-6 Italy SRL
• Original Assignee: C5-6 Italy SRL
• Inventors: Volpati, Laura, Paravisi, Stefano, Rivas Torres, Beatriz
• Application Number:
  US14/359,349
• Publication Number:
  US 20140315230A1
• Time in Patent Office: Days
• US Class Current: 435/18
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: G01N 2333/924 acting on glycosyl compounds (3.2)
• CPC Class Codes: G01N 2333/938 acting on beta-galactose-glycoside bonds, e.g. beta-galactosidase
• CPC Class Codes: G01N 2333/942 acting on beta-1, 4-glucosidic bonds, e.g. cellulase

Abstract

The invention is to a novel method for evaluating the hydrolysis conversion of a enzyme or enzyme mixture relative to a specific ligno-cellulosic biomass. The method evaluates the hydrolysis conversion of a enzyme or enzyme mixture relative to a specific ligno-cellulosic biomass by preparing a pretreated ligno-cellulosic biomass suspension, a buffer solution and an enzyme or enzyme mixture solution; dispensing a portion of one or more of the three components into a plurality of test vials, subjecting one or more of the plurality of test vials to varying incubation periods, reducing the rate of incubation at the end of each incubation period, and analyzing the resulting samples for dissolved sugars to determine the rate of reaction.

Claims

1-9. -9. (canceled)

10. A process for evaluating the hydrolysis conversion of a enzyme or enzyme mixture relative to a specific ligno-cellulosic biomass, the process comprising the steps of:

a. Preparing a pre-treated ligno-cellulosic biomass suspension, a buffer solution and an enzyme or enzyme mixture solution,

b. Dispensing a portion of the pre-treated ligno-cellulosic suspension and buffer solution into at least two test vials of a plurality of test vials,

c. Adding a portion of the enzyme or enzyme mixture solution to at least one test vial containing the pre-treated ligno-cellulosic biomass suspension, leaving at least one test vial void of enzyme or enzyme mixture solution,

d. Dispensing the buffer solution into at least one test vial which is void and will remain void of the enzyme or enzyme mixture solution and the pretreated ligno-cellulosic biomass suspension,

e. Analyzing the at least one test vial which is void of enzyme or enzyme mixture solution but contains pre-treated ligno-cellulosic biomass suspension and buffer solution for soluble sugars prior to incubating the test vials,

f. incubating the test vials for a first time at a first temperature and analyzing at least one of the test vials containing the enzyme or enzyme mixture solution for soluble sugars at the first time,

g. incubating the remaining test vials for a second time and analyzing at least one of the remaining test vials containing the enzyme or enzyme mixture solution for soluble sugars,

h. incubating the remaining test vials and analyzing at least one test vial at each of the several pre-determined times of incubation for dissolved sugars to determine the rate of reaction.

11. The process of claim 10, wherein the volume of the portion of the biomass suspension in each test vials is less than a value selected from the group consisting of 2.0, 1.0, 0.5, 0.3, 0.2, and 0.1 mL.

12. The process of claim 10, wherein the dry matter percent content by weight of the ligno-cellulosic suspension in the test vials is higher than a value selected from the group consisting of 1, 2, 4, 10, 12, 15, 17, and 20.

13. The process of claim 11, wherein the dry matter percent content by weight of the ligno-cellulosic suspension in the test vials is higher than a value selected from the group consisting of 1, 2, 4, 10, 12, 15, 17, and 20.

14. The process of claim 10, wherein the at least one of the test vials of steps e, f, g, and h are analyzed by means of High-Performance Liquid Chromatography.

15. The process of claim 10, wherein the at least one of the test vials of steps e, f, g, and h are analyzed by means of optical absorbance.

16. The process of claim 15, wherein in step e, f, g, and h a reagent is added to each vial of the plurality of test vials before analyzing the test vials.

17. The process of claim 16, wherein the reagent quenches the incubation of the test vials containing the enzyme or enzyme mixture.

18. The process of claim 17, wherein the reagent comprises dinitrosalicylic acid.

19. The process of claim 15, wherein the optical absorbance of the liquid fraction of the at least one test vial is analyzed at a unique wavelength.

20. The process of claim 16, wherein the optical absorbance of the liquid fraction of the at least one test vial is analyzed at a unique wavelength.

21. The process of claim 17, wherein the optical absorbance of the liquid fraction of the at least one test vial is analyzed at a unique wavelength.

22. The process of claim 18, wherein the optical absorbance of the liquid fraction of the at least one test vial is analyzed at a unique wavelength.