Geraniol-producing hanseniaspora uvarum and fermentation method thereof
A technology of Hansenula yeast and geraniol is applied in the field of fermentation engineering to achieve the effects of increasing geraniol content, high capacity, flavor and nutritional value
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Embodiment 1
[0030] Embodiment 1: the screening and identification of the Hanseniaspora uvarum of high yield geraniol
[0031] Take 10g of highland barley Daqu and dissolve it in 90mL of sterile saline, shake it on a shaker for 30 minutes, and then carry out gradient dilution. The selected dilution is 10 -2 、10 -3 、10 -4 、10 -5 Take 50 μl of each bacterial solution to coat the WL solid plate, and after culturing at 30°C for 48 hours, the colony morphology of the Hanseniaspora uvarum plate is as follows: figure 1 shown.
[0032] Pick a single colony conforming to the phenotypic characteristics of Hanseniaspora uvarum into a 25mL test tube filled with 5mL barley juice medium, and culture it at natural pH, 30°C, 200rpm for 36h to become a seed culture solution. The cultivated seed culture liquid was inoculated in a 250 mL Erlenmeyer flask containing 50 mL of highland barley juice medium with a 5% inoculation amount, and fermented for 48 h at natural pH at 30° C. Three strains with higher...
Embodiment 2
[0036]Example 2: Comparison of Geraniol produced by Hanseniaspora uvarum A14 and other yeasts
[0037] The bacterial strain that present embodiment comprises has Saccharomyces cerevisiae (Sc), Hyphopichiaburtonii (Hb), Kazachstania unispora (Ku), Wickerhamomyces anomalus (Wa), Torulaspora delbrueckii (Td), Pichia kudriavzevi (Pku), Pichia scaptomyzae (Psc), Pichia membranifaciens (Pme) and Pichia manshurica (Pma), and the strain of the invention Hanseniaspora uvarum A14 (Hu A14). These 10 strains all come from the brewing environment of highland barley wine.
[0038] Pick a single colony of the above 10 strains into a 25mL test tube filled with 5mL barley juice medium, cultivate at natural pH, 30°C, 200rpm for 36h to become a seed culture solution. Inoculate the cultivated seed culture solution with 5% inoculum in a 250mL Erlenmeyer flask containing 50g of sterilized highland barley, at natural pH, 30°C, 200rpm, ferment for 48h, and detect each strain by HS-SPME and GC-MS tec...
Embodiment 3
[0042] Example 3: Detection of Physiological and Biochemical Properties of Hanseniaspora uvarum A14
[0043] Temperature tolerance test: Pick the Hanseniaspora uvarum A14 strain obtained in Example 1, inoculate it in 5ml YPD liquid liquid medium, and culture it at 30°C for 36h to OD 800 In 1.2-1.4. Dilute medium OD with sterile saline 800 To 1, inoculate 0.5mL bacterial liquid into 50ml YPD liquid medium. They were cultured at different temperature gradients of 20°C, 25°C, 30°C, 37°C, 40°C, 42°C and 46°C for 36 hours. The results show that the Hanseniaspora uvarum A14 bacterial strain of the present invention can grow in the temperature range of 20-46 DEG C, and the optimum temperature is 25-35 DEG C (OD 800 Greater than 1).
[0044] Acidity tolerance test: Pick the Hanseniaspora uvarum A14 strain obtained in Example 1, inoculate it in 5ml YPD liquid medium, and culture it at 30°C for 36h to OD 800 In 1.2-1.4. Dilute medium OD with sterile saline 800 To 1, inoculate 0.5...
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