Production of fructo-oligosaccharides from chicory or Jerusalem artichoke by endo-inulinase
A technology of fructo-oligosaccharide and polyfructo-oligosaccharide, applied in the field of production of fructo-oligosaccharide, which can solve the problems of restricting the application of sugar-controlling people
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Embodiment 1
[0056] The artificially synthesized nucleotide sequence shown in SEQ ID NO: 2 contains EcoR I and not I restriction site. in, EcoR I is located at the 5' end, not I is located at the 3' end. In addition, 8 bp of protective bases are connected at the 3' end, and the sequence is CCCCAAAA. The fragment after the whole gene synthesis was constructed in the pGH plasmid, the size was 1507bp, and the vector was Amp resistance. Then transform DH5α competent cells; the obtained transformants are detected by PCR to see if a 1.5kb band can be amplified; then the plasmid DNA is extracted and digested to verify whether the plasmid contains a 1.5kb fragment insert; finally, the cloned plasmid DNA is selected for sequence The results showed that the DNA sequence of the endonuclease gene in the selected clone was completely correct. So far, the plasmid pGH / Inu2 containing the nucleotide sequence shown in SEQ ID NO:2 was obtained.
[0057] After extracting the plasmids synthesized by...
Embodiment 2
[0060] Fresh Jerusalem artichoke is washed, peeled, sliced, dried, and crushed to become Jerusalem artichoke powder.
[0061] Weigh Jerusalem artichoke powder, add 10 times the mass of water, heat and stir at 95°C for half an hour, centrifuge to take the supernatant, add the supernatant of endo-inulinase prepared in Example 1 according to the final concentration of 1 u / ml, pH 7.0, react at 60°C for 3 hours (stirring is required during the period, 60r / min), after cooling, the above enzymatic hydrolysis products are centrifuged at high speed to remove sediment, diluted 10 times (total sugar concentration is about 5ug / ul), and detected by HPLC The content of various sugars in the standard product and the diluted product, the results are shown in Table 1:
[0062] Table 1: Content of various sugars in the product
[0063] type of sugar Content (before dilution) Mass fraction of total sugar (%) fructose 5.88mg / ml 11.76 glucose + sucrose 13.99mg / ml 27....
Embodiment 3
[0066] The tubers of Jerusalem artichoke were squeezed with a drum juice extractor, and the total amount of slag obtained was 1792g, and the total amount of juice was 3411g; the proportion of juice was 65.6%.
[0067] Weigh the Jerusalem artichoke juice, add water of equal quality, heat at 90°C and stir for half an hour, centrifuge to take the supernatant, add the supernatant of endo-inulinase prepared in Example 1 according to the final concentration of 1 u / ml, pH 7.5 React at 60°C for 3 hours (stirring is required during the period, 60r / min). After cooling, the above enzymatic hydrolysis products are centrifuged at high speed to remove the precipitate, diluted 10 times (the total sugar concentration is about 5ug / ul), and the standards are detected by HPLC. The content of various sugars in the product and the diluted product, the results are shown in Table 2:
[0068] Table 2: Content of various sugars in the product
[0069] type of sugar content Mass fraction...
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