Fast-fermenting bread yeast strain and construction method thereof
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A technology of baker's yeast and baker's yeast is applied in the field of bioengineering to achieve the effects of high dough rising ability, improved economic benefit and shortened fermentation time
Active Publication Date: 2014-10-01
TIANJIN UNIV OF SCI & TECH
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[0006] At present, many studies have improved the rate of maltose metabolism through the modification of genes directly related to the maltose metabolism pathway, thereby improving the fermentation power of the dough. The research on H / ACA small nucleolar RNA is basically focused on theoretical research, and there are only a few reports on the application of H / ACA small nucleolar RNA, especially in the research of breeding rapid fermentation strains.
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Embodiment 1
[0042] Embodiment 1: Construction of quick-fermenting baker's yeast strain
[0043] (1) Construction of expression plasmid Yep-KPS
[0044] The construction process of the expression plasmid Yep-KPS is as follows: figure 1 shown;
[0045] Using the total DNA of yeast strain CICC31616 as a template, the complete sequence of the SNR84 gene was amplified by PCR. The reaction system is shown in Table 1:
[0046] Upstream primer: 5'-GGA AGATCT ATTGCACAACTTAAGTTTGTCGAGG-3' (SEQ ID NO: 3);
[0047] Downstream primer: 5'-GGA AGATCT TAATGTGTCTCTTTGAGTCATGTTCCTT-3' (SEQ ID NO: 4); the underlined part is the enzyme cutting site;
[0048]Table 1 PCR amplification of SNR84 gene
[0049]
[0050] The PCR product was connected to the pUC-PGK1 vector containing a strong promoter to obtain pUC-PGKS; using pUC-PGKS as a template, PCR amplification obtained PGK inserted into the SNR84 gene P -SNR84-PGK T (PS) fragment, the reaction system is shown in Table 2;
[0072] (1) No-sugar-added dough fermentation experiments of recombinant strains and starting strains
[0073] Pick a ring of yeast cells in YEPD medium, 30 ℃, static culture for 24h; transfer to molasses medium with 10% (v / v) inoculation amount (add 0.5g / L Ammonium sulfate, 5g / L yeast powder), 30°C, 180r / min, cultivated for 24h to the stationary phase (OD 600 1.5 or so); statically cultivated for 2 hours, centrifuged at 4000r / min for 5 minutes, washed twice with sterile water, and collected the thalli for subsequent use.
[0074] Weigh 8.0 g of the above-mentioned yeast collected by centrifugation, mix it with 4.0 g of NaCl, 150 mL of water, and 280 g of standard flour, and put it into a preheated fermenter for fermentation. Record 1h CO 2 Gas production and the time to reach the maximum gas production.
[0075] (2) No-sugar-added dough fermentation experiment results of recombinant stra...
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Abstract
The invention discloses a fast-fermenting bread yeast strain and a construction method thereof, and belongs to the technical field of a bioengineering technology. The construction method comprises the following steps: knocking out phosphoglucomutase genes PGM2 completely in the parent bread yeast strain; meanwhile, choosing a strong promoter PGK1 to over-express the complete sequence of H / ACA small nucleolus RNA (SNR84) to acquire the fast-fermenting yeast strain. The CO2 gas production of the fast-fermenting bread yeast strain in a non-sugaring paste for 1 hour reaches to 818mL; compared with the parent strain, the CO2 gas production is improved by 21.2% (the CO2 gas production of the parent strain fermenting in the non-sugaring paste for 1 hour reaches to 675mL); meanwhile, the fermenting time is shortened by 18.0%. The bread yeast strain has a good fermentability during fermenting in the non-sugaring paste, so that the requirement for the 'fast' yeast in a cooked wheaten food processing market is met. Therefore, the bread yeast strain has an extensive application prospect.
Description
Technical field: [0001] The invention belongs to the technical field of bioengineering, and relates to the breeding of industrial microorganisms, in particular to a fast-fermenting baker's yeast and a construction method thereof. Background technique: [0002] Nowadays, there are many kinds of pasta, and the production methods are also various, but no matter what production method is adopted, the fermentation process of dough is an important step in the whole process, which directly affects the quality of pasta products. Developed countries such as Europe and the United States generally take salty bread as the staple food in consideration of health factors, and bread is generally sugar-free or low-sugar (sugar content is less than 6%). my country has a large population, of which about half of the population is dominated by pasta, and most of them are unsweetened dough fermented pasta such as steamed buns. According to statistics, about 70% of the wheat flour in the north is ...
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