Composite sweetening agent
A compound sweetener and concentration technology, applied in the field of food additives, can solve the problems of low calorie, poor solubility and high cost, and achieve the effects of reducing production cost, good water solubility and simple method
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Embodiment 1
[0026] 1. Preparation of recombinant microorganisms:
[0027] Rice (Oryza sativa) leaf total RNA was extracted and rice cDNA was obtained by reverse transcription. According to the EUGT11 gene sequence (Accession No. AK121682) in the GeneBank database, PCR amplification primers were designed, and the upstream and downstream primers were respectively introduced into the BamH I and Hind III sites, and the EUGT11 coding gene was obtained by PCR amplification.
[0028] Stevia total RNA was extracted and rice cDNA was obtained by reverse transcription. According to the EUGT11 gene sequence (Accession No. AK121682) in the GeneBank database, PCR amplification primers were designed, and the upstream and downstream primers were respectively introduced into the Nde I and Xho III sites, and the UGT76G1 coding gene was amplified by PCR.
[0029] The EUGT11 fragment and the expression vector pETDuet were double digested with BamH I and Hind III respectively, and the target fragment was re...
Embodiment 2~4
[0042] Using the method of Example 1, only part of the parameters during conversion were changed to obtain the product, wherein the changed parameters are shown in Table 2.
[0043] Table 2
[0044] Reaction parameters
Example 1
Example 2
Example 3
Example 4
OD600
100
80
120
110
Concentration of Rebaudioside A (g / L)
5
1
3
10
Concentration of trisodium citrate (mmol / L)
60
50
70
80
Zinc chloride concentration (mmol / L)
1
0.5
1.5
2
Sucrose concentration (%, w / v)
40
30
40
50
pH value
8.0
8.5
7.5
8.0
Reaction temperature (°C)
37
35
38
40
Reaction time (h)
24
20
28
30
[0045] Measure the amount of RD and RM in the products prepared in Examples 2-4, and record the weight ratio of RD and RM. The results are shown in Table 3.
[0046] table 3
[0047] Example number
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