Functional peptide-reinforced healthcare food
A health food and functional peptide technology, applied in food preparation, food science, application, etc., can solve problems such as gene application reports that have not been seen
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Embodiment 1
[0033] Example 1: Glutathione high-yielding strain Saccharomyces cerevisiae BY-GSW ( Saccharomyces cerevisiae BY-GSW) build
[0034] 1. Cloning of bifunctional glutathione synthase gene GSW and construction of yeast strain BY-GSW
[0035] 1.1 Construction of homologous recombination fragments: GSW sequence was synthesized and cloned according to the Genbank database sequence GQ848551 (see sequence table SEQ ID NO.1), DNA was obtained from Invitrogen’s Yeast Deletion Clones series products as a template, and the following were obtained by PCR technology: Fragment: promoter ADH1, terminator sequence term, marker gene KanMX4 and homologous recombination sequence Trp1, connect the loxP sequence derived from P1 phage to both ends of the KanMX4 fragment, apply preset endonuclease sites and ligase, press figure 1 Design and construct the recombinant fragment, insert the fragment into the vector pBlueScriptIISK to obtain the recombinant plasmid pBlueScript-KanMX-GSW, transform Sacc...
Embodiment 2
[0047] Embodiment two: the preparation example one of glutathione preparation
[0048] Inoculate the transformant Saccharomyces cerevisiae BY-GSW in YPD medium, shake the flask overnight at 30°C to enter the logarithmic phase, insert it as a seed with 2% inoculum, put it in 100ml YPD medium (500ml large Erlenmeyer flask), 30 ℃ shaker (230rpm) culture, at 12, 24, 36, 48, 60, and 72 hours, take 30ml of bacterial liquid, centrifuge at 6000rpm for 5 minutes to collect the bacteria, wash twice with sterile water, and dry in an oven at 60℃ until constant Heavy. Resuspend the cells at the ratio of 0.05g stem cells to 1ml sterile water, put them at -20°C for 12 hours, then boil them in a water bath for 5 minutes, then centrifuge them at 12000rpm for 1 minute after cooling, and take the supernatant to detect the glutathione content by DTNB method. Cultivate the starting strain under the same conditions and detect its glutathione. The result showed that after cultivating for 60 hours,...
Embodiment 3
[0049] Embodiment three: Preparation example two of glutathione preparation
[0050] The overnight strain of the transformant Saccharomyces cerevisiae BY-GSW was inoculated into 30 liters of medium composed of sterilized peptone, yeast powder and glucose in a 50 liter tank, cultured with stirring at 30 °C for 55 hours, and centrifuged to obtain The cells were suspended in 42% ethanol (dry cell weight: ethanol = 1: 8), extracted at 25°C for 1.5 hours, centrifuged to get the supernatant, and the solvent was removed under reduced pressure to obtain 35g of glutathione preparation.
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