Recombinant yeast strain and application thereof
A yeast strain and yeast technology, applied in the field of genetic engineering, can solve problems affecting the production of lycopene
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Embodiment 1
[0027] Example 1: The recombinant yeast strain of the present invention
[0028] 1. Construction of lycopene production strain
[0029] Taking Saccharomyces cerevisiae CEN.PK2-1C as the starting strain, the three genes gal1, gal7 and gal10 were knocked out according to the method of Example 1 in CN105087406A, and then gene fragments 1 and 2 were constructed according to the methods of Example 3 and Example 4 in CN105087406A. When constructing gene fragment 2, crtE derived from Pantoea agglomerans was used, and its nucleotide sequence is shown in SEQ ID NO:1.
[0030] Then, according to the method of Example 7 in CN105087406A, gene fragments 1 and 2 were integrated into Saccharomyces cerevisiae through homologous recombination to obtain a yeast strain producing lycopene, named SyBE_Sc14C07.
[0031] 2. Construction of the recombinant yeast strain of the present invention
[0032] On the basis of SyBE_Sc14C07, the ALD6 promoter was knocked out with reference to the knockout method of ypl...
Embodiment 2
[0034] Example 2: Shake flask fermentation test
[0035] Taking the three strains of SyBE_Sc14C07, SyBE_Sc14C66, and SyBE_Sc14C67 in Example 1 as test objects, the fermentation test was carried out according to the following method:
[0036] The strain was inoculated into 5mL seed culture medium, and cultivated at 30℃, 250rpm for 14-16h, with the initial cell concentration OD 600 =0.2 Transfer to fresh 25mL seed culture medium, cultivate to mid-logarithmic growth at 30℃, 250rpm conditions, with initial cell concentration OD 600 =0.5 were respectively inoculated into 50mL fermentation medium, cultivated at 30°C and 250rpm, and fermented to produce lycopene.
[0037] Wherein, the seed medium is 20g / L or 40g / L glucose, 20g / L peptone, and 10g / L yeast extract; the fermentation medium is 20g / L or 40g / L glucose, 20g / L peptone, 10g / L yeast extract powder and 10g / L D-galactose.
[0038] Lycopene detection method: After 48 hours of fermentation, take two aliquots of the fermentation broth, cen...
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