Preparation of oligomeric xylose and alpha-glucuronylic enzyme used therefor
A technology of glucuronidase and xylooligosaccharides, which is applied in the field of preparation of xylooligosaccharides, can solve problems such as inability to combine and decompose, affect the quality of xylooligosaccharides, and affect the efficiency of enzymatic hydrolysis
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Embodiment 1
[0030] Embodiment 1: refer to Figure 6And other accompanying drawings, the concrete operation of the present invention is as follows:
[0031] (1) Extract corn cob xylan with alkali.
[0032] Take the air-dried corn cob fiber with a particle diameter of about 3mm, first soak it in clean water and then centrifuge to remove the water, add sodium hydroxide solution according to the solid-liquid ratio of 7-10:1, and the final concentration is 1%-3%, at room temperature or high temperature Then, the filtrate was collected and neutralized to slightly acidic with acid, and then precipitated with three times the volume of ethanol to obtain xylan.
[0033] (2) Preparation of enzymes
[0034] Two pairs of primers were designed according to the sequences of xynB and aguA in Genbank, and the genomic DNA of T. maritima was used as a template for PCR amplification. Tm-xynB-N-terminal primer is 5'-CCGTTCCATGGACTACAGGATGTGC-3', Tm-xynB-C-terminal primer is 5'-CCGCTCGAGCGGATATATCTTTCTTCCCT...
Embodiment 2
[0043] Embodiment 2: substantially the same as Example 1, the difference is that xylan is treated with 25mmol / L pH6.3Na 2 HPO 4 -NaH 2 PO 4 The buffer solution is formulated at a concentration of 20%, and the enzyme dosage is: 200 U / g of thermostable xylanase, 15 U / g of thermostable α-glucuronidase, hydrolysis temperature of 80° C., hydrolysis for 12 hours.
Embodiment 3
[0044] Embodiment 3: substantially identical with embodiment 1, difference is that xylan is used 40mmol / L pH6.0Na 2 HPO 4 -NaH 2 PO 4 The buffer solution is prepared at a concentration of 15%, and the enzyme dosage is: 300 U / g of thermostable xylanase, 30 U / g of thermostable α-glucuronidase, hydrolysis temperature of 75° C., hydrolysis for 5 hours.
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