Pichia pastoris 3-j15 and its application
A technology of 3-J15 and yeast, which is applied in the field of microbiology, can solve the problems of few varieties of yeast with aroma-enhancing potential and insufficient aroma of soy sauce, etc., and achieve the effects of strong application potential, improved flavor and quality, and good growth
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Embodiment 1
[0035] Isolation, cultivation and molecular biology identification of Pichia mongolica strain 3-J15.
[0036] Under aseptic conditions, 5 g of fermented sauce samples were taken and added to 45 mL of sterilized saline with glass beads in advance, and shaken on a shaker at 37 °C for 15 min. Take 1 mL of the sample diluent for 10-fold serial dilution, and select an appropriate dilution to spread on the PDA medium containing 10% sodium chloride. After culturing at 28 °C for 72 h, characteristic colonies were selected, and after repeated streaking and purification, more than 20 strains of yeast were isolated from the fermented sauce. Considering the food fermentation background of different strains and the aroma-enhancing effect of yeast on the product, the main Focusing on the middle and late stages of fermentation, a strain of yeast 3-J15 isolated from the fermented sauce grains in the third year was finally screened out. The isolated yeast 3-J15 was inoculated into the corresp...
Embodiment 2
[0045] Exploration on the salt tolerance of strain 3-J15:
[0046] The strains were activated using YPD liquid medium, and then the growth of the strains in YPD medium containing different gradient concentrations of NaCl was determined. After oscillating on a shaker at 120 rpm at 30 °C for 24 h, the absorbance value of the culture solution was measured at a wavelength of 660 nm by a spectrophotometer, and the tolerance curve of the strain to NaCl was drawn, as shown in Figure 7 As shown, the results indicated that Pichia quaterii 3-J15 was able to tolerate at least 15% salt concentration.
Embodiment 3
[0048] Pichia pastoris was applied to simulate the fermentation of sauce fermented grain liquid medium:
[0049] 1. Simulated sauce fermented grains liquid medium: 10 g soybean flour, 2.5 g wheat flour, 50 g fresh sauce fermented grains, buffer system Na 2 HPO 4 / KH 2 PO 4 Configured as a 1 L system, boiled and stirred, filtered through four layers of gauze, pH = 5-5.2, and sterilized. According to the experimental design, it was divided into salt stress type with additional addition of 150 g / L sodium chloride and non-salt stress type without addition of sodium chloride.
[0050] 2. Put the Jiyemeng yeast preserved in the glycerol tube into the YPD liquid medium for activation, centrifuge at 4000 rpm, take sterile physiological saline to wash the bacteria repeatedly, and put 1 mL 5×10 6 Add the CFU / mL bacterial cell suspension into 30 mL of the simulated sauce fermented grain liquid medium, shake the flask at 120 rpm at the optimum growth temperature until the stable grow...
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