Sugar solution production method
A manufacturing method and technology of sugar solution, applied in biofuels, fermentation, etc., can solve the problems of low enzyme reusability, increased manufacturing cost of sugar solution, high price of cellulase, etc., to achieve efficient recovery, control of manufacturing costs, and reduction of The effect of usage
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reference example 1
[0066] (Reference Example 1) Measurement of Protein Concentration
[0067] For the measurement of the protein concentration, a commercially available protein concentration measurement reagent (Quick Start Bradford Protein Assay, manufactured by Bio-Rad) was used. Add 5 μL of diluted cellulase solution to 250 μL of the protein concentration measurement reagent returned to room temperature, and after standing at room temperature for 5 minutes, measure its concentration at 595 nm with a microplate reader (POWERSCAN HT, manufactured by Dainippon Sumitomo Pharmaceutical Co., Ltd.). Absorbance. The bovine serum albumin aqueous solution was used as a standard solution, and the protein concentration of the cellulase solution was calculated with reference to the standard curve.
reference example 2
[0068] (Reference example 2) Measurement of sugar concentration
[0069] The concentrations of glucose and xylose contained in the sugar solution were quantified by comparison with standard products under the HPLC conditions shown below.
[0070] Column: Luna NH 2 (manufactured by Phenomenex)
[0071] Mobile phase: Milli-Q:acetonitrile=25:75 (flow rate 0.6mL / min)
[0072] Detection method: RI (differential refractive index)
[0073] Temperature: 30°C.
reference example 3
[0074] (Reference Example 3) Method for Measuring Cellulase Activity Derived from Filamentous Fungi
[0075] Cellulase activity, that is, the activity of an enzyme group involved in the decomposition of cellulose, was measured and evaluated by the method shown below (1) 4-nitrophenyl-β-D-lactopyranoside (pNP-Lac) and (2) the decomposing activity of 4-nitrophenyl-β-D-glucopyranoside (pNP-Glc); as the activity of enzyme group related to the decomposition of xylan which is the main component of hemicellulose, by (3) The decomposition activity of 4-nitrophenyl-β-D-xylopyranoside (pNP-Xyl) was measured and evaluated by the method shown below. In addition, the substrates of (1) to (3) above are collectively referred to as pNP-sugar.
[0076] Add 0.1 mL of enzyme solution to 0.9 mL of 55 mM acetate buffer (pH 5.0) containing each substrate at a concentration of 1.1 mM, and react at 30°C (final concentration of substrate 1 mM, final concentration of buffer 50mM). In the case of sub...
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