Method for producing xylo-oligosaccharide and xylose by using immobilized enzyme
A technology of xylo-oligosaccharides and immobilized enzymes, applied in directions such as immobilization on/in organic carriers, fermentation, etc., can solve the problems of unreported research on application and few researches on application, so as to improve the utilization rate, Good economy and the effect of improving utilization efficiency
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Embodiment 1
[0027] Embodiment 1, take corn cob steam explosion liquid as raw material preparation xylo-oligosaccharide
[0028] 1. Preparation of corn cob steam explosion liquid
[0029] Weigh 100g corncobs (20-40 mesh) (containing 32.4% xylan), add 800mL distilled water according to the ratio of 1:8, soak at room temperature for 10h, add to the high-pressure tank, cover and seal, heat until the reaction temperature reaches Start timing at 170°C, keep the temperature at around 170°C for 30 minutes, and immediately open the ball valve to release the material into the collection tank. Suction filtration is performed when the temperature of the collected material drops to 60-70°C, and the obtained filtrate is corncob steam explosion liquid. After the corncob steam explosion liquid was cooled to room temperature, the total sugar content was determined by the orcinol-hydrochloric acid method, and the direct reducing sugar content was determined by the Somogyi method. The results showed that ...
Embodiment 2
[0058] Embodiment 2, using corn cob steam explosion liquid as raw material to prepare xylose
[0059] 1. Preparation of corn cob steam explosion liquid
[0060] For the specific preparation method, refer to Step 2 of Example 1.
[0061] 2. Preparation of immobilized xylanase-xylosidase
[0062] (1) Preparation of enzyme solution to be immobilized
[0063] Paecilomyes thermophila (Paecilomyes thermophila) J18 seeds were streaked and inoculated from the slant of preserved PDA on a newly prepared PDA plate, and then the plate was placed in a 40-60°C incubator for static culture for 2-8 days.
[0064] Liquid fermentation medium: corn cob powder carbon source 40g / L, tryptone 10g / L, CaCl 2 0.3g / L, MgSO 4 ·7H 2 O 0.3g / L, FeSO 4 0.3g / L; Sterilize at 121°C for 20min, and the pH of the medium is 7.
[0065] Liquid fermentation: Scrape 1 cm from the above Paecilomyces thermophila J18 plate 2 The mycelia of Paecilomyces thermophilia were inoculated in a conical flask containing 60...
Embodiment 3
[0085] Embodiment 3, prepare xylose with birch xylan solution as raw material
[0086] 1. Preparation of immobilized xylanase-xylosidase
[0087] Concrete preparation method is with embodiment 2. The following experiments were all carried out using immobilized xylanase-xylosidase with xylanase activity of 53.9 U / g and xylosidase activity of 34.5 U / g.
[0088] 2. Preparation of xylose by hydrolyzing birch xylan solution with immobilized xylanase-xylosidase
[0089] Inner diameter is the stainless steel jacketed chromatographic column of 12mm, add the xylanase enzyme activity of 40g above-mentioned step 1 preparations and be 53.9U / g, the enzyme activity of xylosidase is the immobilized xylanase of 34.5U / g - Xylosidase to a bed height of 30 cm. Connect the peristaltic pump and the partial collector, equilibrate with 50mM citric acid buffer solution with pH 6.0 for 30min, and set the temperature of the reaction column bed at 60°C. Pump 400mL of 2% (mass percentage) birch xylan...
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