STRATEGY FOR THE ASSAY OF ENZYME AND ENZYME MIXTURES FOR THE HYDROLYSIS LIGNO-CELLULOSIC BIOMASS
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Abstract
The invention is to a novel method for assaying most or all of the activities of an enzyme or enzyme solution that could be used to convert ligno-cellulosic material into monomeric sugars. The method characterizes enzymes and enzyme mixtures for the hydrolysis of ligno-cellulosic biomass by measuring the protein concentration of an enzyme or enzyme mixture, adding the enzyme or enzyme mixture to a plurality of test vials creating a liquid fraction in the plurality of test vials wherein each test vial in the plurality of test vials contains a different substrate, incubating each test vial for a period of time sufficient to establish the conversion of the substrate in each test vial of the plurality of test vials at the same pH and temperature, while, if necessary, providing at least enough agitation to keep the substrate in suspension, quenching the reaction, and analyzing the optical absorbance of the liquid fraction of at least one test vial of the plurality of test vials.
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Citations
30 Claims
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1-15. -15. (canceled)
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16. A method for characterizing enzyme and enzyme mixtures for the hydrolysis of ligno-cellulosic biomass comprising the steps of
a. Adding the enzyme or the enzyme mixture to a plurality of test vials creating a liquid fraction in the plurality of test vials, wherein there is at least one test vial containing a filter paper substrate, at least one test vial containing a CMC substrate, at least one test vial containing a salicin substrate, at least one test vial containing a Xylan substrate, at least one test vial containing an Avicell substrate, and at least one test vial containing at least one PNP-substrate selected from the group consisting of α - -L-arabinofuranoside, β
-D-galactopyranoside, β
-D-glucuronide, α
-D-glucopyranoside, β
-D-mannopyranoside, α
-D-galactopyranoside,b. Incubating simultaneously each test vial for a period of time sufficient to establish the hydrolysis of the substrate in each test vial of the plurality of test vials at the same pH and temperature, while, if necessary, providing at least enough agitation to keep the substrate in suspension, c. Quenching the reaction, d. Analyzing the optical absorbance of the liquid fraction of at least one test vial of the plurality. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
- -L-arabinofuranoside, β
Specification