Red hydrophobic pigment as well as preparation method and application thereof
A hydrophobic and pigmented technology, applied in the field of bioengineering, which can solve the problems of complex structure, difficult degradation and limited content of synthetic dyes
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Embodiment 1
[0028] Embodiment 1 prepares compound (I)
[0029] The concrete steps that present embodiment prepares red natural dye (compound I) are as follows:
[0030] 1) using the front primer shown in SEQ ID NO.3 and the back primer shown in SEQ ID NO.4 to amplify the DeBI gene shown in SEQ ID NO.1 by polymerase chain reaction, the amplified The increased DeBI gene was connected to the pYES2 plasmid vector through restriction endonucleases BamHI and EcoRI to obtain the recombinant plasmid pYES2-DeBI; then the recombinant plasmid pYES2-DeBI was transformed into S. cerevisiae engineering strain INVSc1 to obtain the transformed engineering bacterium, the applicant named the transformed engineering bacterium as engineering bacterium S;
[0031] 2) Pick a single colony from the solid culture medium with engineering bacteria S in 2ml of SC medium, and culture it in a shaker at 28°C and 200rpm for 24 hours to obtain the seed culture solution of engineering bacteria S; draw 1ml of engineering...
Embodiment 2
[0038] The test of embodiment 2 compound (I) solubility
[0039] At 25°C, weigh 100 mL of the solutions: ethanol, methanol, acetone, ethyl acetate, chloroform, n-hexane, chloroform, glycerin, oil and DMSO, and put them into beakers. Take by weighing 20g of the red pigment solid powder (i.e. compound I) that embodiment 1 obtains respectively, and add respectively in the beaker that fills solution ethanol, methanol, acetone, ethyl acetate, chloroform, normal hexane, chloroform, glycerin, grease and DMSO . Gradually add the solute of the red pigment solid powder, stir constantly, and fully dissolve. Ensure that the solution is supersaturated, filter out the undissolved solid red pigment, and dry at 55°C for 24 hours to ensure that the solvent is completely evaporated. Weigh the total mass of undissolved solids respectively, and use the formula (20g-mass of undissolved pigment=mass of dissolved pigment) to obtain the mass of solids dissolved in 100mL of solvent, which is the sol...
Embodiment 3
[0044] Analysis of Compound I Production in Example 3 Saccharomyces cerevisiae Engineering Bacteria INVSc1
[0045] 1. The Saccharomyces cerevisiae engineering strain INVSc1 containing the blank plasmid pYES2 was cultured for 72 hours by the same preparation method as in Example 1, and the culture was taken for later use.
[0046] 2. Get the culture of engineering bacterium S in the preparation method and the above-mentioned culture, carry out liquid phase detection, 420nm wavelength, mobile phase is acetonitrile-water (0.1% formic acid), flow velocity is 1ml / min, and elution method is 10- 90% acetonitrile gradient elution for 60min.
[0047] Test results such as Figure 4 Shown, the culture of the Saccharomyces cerevisiae engineering bacterium INVSc1 comprising blank plasmid pYES2 is as ( Figure 4 A) shows that the synthesis of compound I is not detected; the culture of engineering bacteria S is as ( Figure 4 As shown in B), it shows that compound I is detected; it prove...
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