Process for producing food containing gamma-aminobutyric acid and yeast having high ability to produce gamma-aminobutyric acid
An aminobutyric acid and yeast technology, which is applied in the directions of microorganism-based methods, methods using fungi, biochemical equipment and methods, etc., can solve the problem that the effect of γ-aminobutyric acid cannot be obtained and the γ-aminobutyric acid cannot be produced in large quantities. , not suitable for large-scale production in factories, etc.
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[0065] Hereinafter, the present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto.
reference example 1
[0067] Pichia anomaly MR-1 is a marine yeast isolated from the seawater of Hachinohe, Japan by the following procedure.
[0068] [separated from sea water]
[0069] First, about 50 liters of seawater at a depth of 5 meters is collected from a ship on the Hachinohe Sea, about 15 kilometers from the coast of Aomori, with a sterile water collector. Transport seawater at refrigerated temperatures. On the next day, seawater was filtered through a sterile filter (pore size: 0.45 μm). The microbial cells retained on the filter membrane were suspended in 15ml of sterile water, and used as the bacterial cell samples collected from seawater in subsequent experiments.
[0070] Take 200 μl each time from the above-mentioned collected bacterial sample (15ml), and spread it on the YPD agar medium (glucose 2%, peptone 2%, yeast extract powder 1%, chlorinated Sodium 3%, agar 2% and chloramphenicol 0.01%), cultured at 25°C for 5 days. Bacteria were collected from yeast-like colonies grown ...
reference example 2
[0106] [Reconstitution of preserved strains]
[0107] Thaw MR-1 strains stored in a frozen or lyophilized state at room temperature. After collecting the bacteria with a platinum ring, disperse the bacteria in about 1ml of sterile water, then draw a line and inoculate them on YPD agar medium, and culture them at 25°C for 5 days. MR-1 cells are obtained in the form of milky white round colonies on the culture medium. The bacterium can be directly used for pre-cultivation as follows. In addition, as long as the bacterial strain on the agar medium is stored under refrigeration at 5° C. to 10° C. for no more than 2 months, it can be used for preculture and the like.
[0108] [pre-culture]
[0109] After the bacterial cells were collected from the colonies of the agar medium, the bacterial cells were inoculated into sterile YPD liquid medium (200 ml) in a Erlenmeyer flask, and cultured with shaking at 25° C. for 2 to 3 days. During the cultivation period, 1 ml of the culture so...
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