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

Inactive Publication Date: 2017-06-27
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The expression level of the gene ALD6 directly affects the accumulation of acetate i

Method used

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  • Recombinant yeast strain and application thereof
  • Recombinant yeast strain and application thereof

Examples

Experimental program
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Effect test

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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Abstract

The invention relates to the technical field of genetic engineering and discloses a recombinant yeast strain and application thereof. The recombinant yeast strain is a yeast strain for fermenting production of lycopene, and the ALD6 promoter expression intensity of the yeast strain is lowered. According to the recombinant yeast strain and the application thereof, the ALD6 promoter of the yeast strain is deeply studied, it is discovered that the lycopene yield of the yeast strain can be improved remarkably by constructing the recombinant yeast strain for fermentation production of the lycopene in the mode that the ALD6 promoter expression intensity is lowered through full knockout, lower expression promoter replacement or the like, the recombinant yeast strain can be taken as an optimizing means for a yeast engineering bacterium synthesis route, and recombinant yeast strains with more excellentproductioncapacity can be constructed.

Description

Technical field [0001] The invention relates to the technical field of genetic engineering, and more specifically to a recombinant yeast strain and its application. Background technique [0002] In the world, with the improvement of economic level and health requirements, the safety, nutrition and functionality of food have received more and more attention. Therefore, functional nutritional chemicals have become a development trend, representing the development of contemporary food The new trend has broad market prospects. Lycopene is a kind of fat-soluble natural food coloring. It belongs to carotenoid in chemical structure and possesses strong antioxidant capacity. It is a hot spot in the research of functional food ingredients in the world in recent years. [0003] The preparation of lycopene mainly relies on plant extraction, chemical synthesis and microbial synthesis. The first two methods have their own shortcomings. Microbial synthesis is considered the most promising metho...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12P5/02C12R1/865
CPCC12N9/0008C12P5/026C12Y102/0101
Inventor 周晓陈艳王颖肖文海姚明东李博刘宏元英进
Owner TIANJIN UNIV
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