Preparation method of inulin source recombinant allophycocyanin
A technology of allophycocyanin and inulin, which is applied in the field of microbial preparation of recombinant allophycocyanin from inulin sources, can solve the problems of difficulty in establishing standardized and unified quality standards, high preparation costs, and unstable effects, and achieve low cost, Energy-saving, easy-to-extract effects
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Embodiment 1
[0023] Example 1: Construction of strains
[0024] Escherichia coli P2 is derived from Escherichia coli strain P1 capable of producing active recombinant allophycocyanin, and constructed according to the prior art records to obtain Escherichia coli P2.
[0025] Among them, the construction method of Escherichia coli strain P1 capable of producing active recombinant phycobiliprotein is described in the literature of the application number 9612023.x, the name of the invention, the microorganism capable of producing active recombinant phycobiliprotein and its preparation method. P1 is now deposited at the General Microbiology Center (CGMCC) of the China Microorganism Collection Management Committee, with the deposit number CGMCC No. 0261, and the deposit date is April 17, 1996.
[0026] Take E. coli strain P1 as the starting strain, knock out the ampicillin resistance gene in it by molecular biological means, insert the kanamycin resistance gene in the original position, and get the kan...
Embodiment 2
[0027] Example 2 Preparation of shake flask of allophycocyanin from inulin
[0028] 1) Preparation of seed liquid: Fill the seed liquid culture medium into the Erlenmeyer flask according to the proportion of 20% (v / v) of the liquid volume, pick an appropriate amount of the above-mentioned Escherichia coli P1 slant bacteria in the inoculation loop, and then inoculate it at 37°C, Incubate for 8 hours under the condition of a shaker rotating speed of 200 rpm.
[0029] Among them, the seed medium (take the preparation of 1L medium as an example): 10g peptone, 5g yeast powder, 10g NaCl, 1L tap water, adjust the pH to 7.0 with 5mol / L NaOH solution, 115°C, sterilize for 20min, then add 1ml of 60mg / ml kanamycin solution.
[0030] 2) Preparation of allophycocyanin shake flask: Fill the fermentation medium into the Erlenmeyer flask according to the proportion of 20% of the filling volume, insert the seed solution, control the inoculum volume at 5% (v / v), and the fermentation temperature at 37...
Embodiment 3
[0035] Example 3 Fermentation tank preparation of allophycocyanin derived from inulin
[0036] 1) Preparation of the first-level seed solution: fill the seed solution medium into the Erlenmeyer flask according to the proportion of 20% (v / v) of the filling amount, pick an appropriate amount of the above-mentioned Escherichia coli P1 slant bacteria in the 37 Cultivate for 8 hours at a temperature of 200 rpm on a shaker. To
[0037] Said, the first-level seed medium (take the preparation of 1L medium as an example): 10g peptone, 5g yeast powder, 10g NaCl, 1L tap water, adjust the pH to 7.0 with 5mol / L NaOH solution, 115°C, sterilize for 20min , And then add 1ml of 60mg / ml kanamycin solution.
[0038] 2) Preparation of secondary seed liquid (take the preparation of 1L medium as an example): Fill the secondary seed liquid medium into the Erlenmeyer flask according to the proportion of 20% of the filling volume, connect the primary seed liquid, and control the inoculum 5% (v / v), and the...
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