The reversed micelles were formed with cationic cetyltrimethylammonium bromide (CTAB) as surfactant and n-hexanol as cosolvent in the CTAB (50mmol.L-1)/hexanol (15% by volume)/hexane system. Cibacron Blue 3GA (CB) as ...The reversed micelles were formed with cationic cetyltrimethylammonium bromide (CTAB) as surfactant and n-hexanol as cosolvent in the CTAB (50mmol.L-1)/hexanol (15% by volume)/hexane system. Cibacron Blue 3GA (CB) as an affinity ligand in the aqueous phase was directly introduced to the reversed micelles with electrostatic interaction between anionic CB and cationic surfactant. High molecular weight (Mr) protein, yeast alcohol dehydrogenase (YADH, Mr = 141000) from baker's yeast, has been purified using the affinity reversed micelles by the phase transfer method. Various parameters, such as CB concentration, pH and ionic strength, on YADH forward and backward transfer were studied. YADH can be transferred into and out from the reversed micelles under mild conditions (only by regulation of solution pH and salt concentration) with the successful recovery of most YADH activity. Both forward and backward extractions occurred when the aqueous phase pH＞pI with electrostatic attraction between YADH and CTAB. The recovery of YADH activity and purification factor have been improved with addition of a small amount of affinity CB. The recovery of YADH activity obtained was ～99% and the purification factor was about 4.0-fold after one cycle of full forward and backward extraction. The low ionic strength in the initial aqueous phase might be responsible for the YADH transfer into the reversed micellar phase.展开更多
Ethyl caproate and isoamyl alcohol are important for the quality of Yakju, one of the Korean traditional alcoholic beverages. From Korean traditional fermentation agent, Nuruk, we have isolated 8 yeasts which produced...Ethyl caproate and isoamyl alcohol are important for the quality of Yakju, one of the Korean traditional alcoholic beverages. From Korean traditional fermentation agent, Nuruk, we have isolated 8 yeasts which produced rich aromatic compounds using YEPD agar plates (1% yeast extract, 2% peptone, 2% dextrose, 2% agar) containing 50 uL cerulenin at 30℃. The isolated aromatic yeasts are identified as Pichia anomala (4 strains), Pichia fabianii (2 strains), Pichia farinose (1 strains), Geotrichum candidum (1 strains). We conducted alcohol fermentation with each of the aromatic yeasts and the compounds (ethyl caproate and isoamyl alcohol) producing range were 59.5 - 193.2 ppm and 10.8 - 91.6 ppm respectively. As a control, Fermivin®, famous aromatic wine yeast, was 89.4 ppm and 16.2 ppm respectively. We also find that the isolated Pichia anomala could produce higher level ethanol (4.2% - 5.0%, v/v) than other species (0.4% - 2.2%, v/v). Using the aromatic yeasts in fermentation industries, we expect to improve the quality of traditional alcoholic beverages.展开更多
基金the National Natural Science Foundation of China (No. 29836130).
文摘The reversed micelles were formed with cationic cetyltrimethylammonium bromide (CTAB) as surfactant and n-hexanol as cosolvent in the CTAB (50mmol.L-1)/hexanol (15% by volume)/hexane system. Cibacron Blue 3GA (CB) as an affinity ligand in the aqueous phase was directly introduced to the reversed micelles with electrostatic interaction between anionic CB and cationic surfactant. High molecular weight (Mr) protein, yeast alcohol dehydrogenase (YADH, Mr = 141000) from baker's yeast, has been purified using the affinity reversed micelles by the phase transfer method. Various parameters, such as CB concentration, pH and ionic strength, on YADH forward and backward transfer were studied. YADH can be transferred into and out from the reversed micelles under mild conditions (only by regulation of solution pH and salt concentration) with the successful recovery of most YADH activity. Both forward and backward extractions occurred when the aqueous phase pH＞pI with electrostatic attraction between YADH and CTAB. The recovery of YADH activity and purification factor have been improved with addition of a small amount of affinity CB. The recovery of YADH activity obtained was ～99% and the purification factor was about 4.0-fold after one cycle of full forward and backward extraction. The low ionic strength in the initial aqueous phase might be responsible for the YADH transfer into the reversed micellar phase.
文摘Ethyl caproate and isoamyl alcohol are important for the quality of Yakju, one of the Korean traditional alcoholic beverages. From Korean traditional fermentation agent, Nuruk, we have isolated 8 yeasts which produced rich aromatic compounds using YEPD agar plates (1% yeast extract, 2% peptone, 2% dextrose, 2% agar) containing 50 uL cerulenin at 30℃. The isolated aromatic yeasts are identified as Pichia anomala (4 strains), Pichia fabianii (2 strains), Pichia farinose (1 strains), Geotrichum candidum (1 strains). We conducted alcohol fermentation with each of the aromatic yeasts and the compounds (ethyl caproate and isoamyl alcohol) producing range were 59.5 - 193.2 ppm and 10.8 - 91.6 ppm respectively. As a control, Fermivin®, famous aromatic wine yeast, was 89.4 ppm and 16.2 ppm respectively. We also find that the isolated Pichia anomala could produce higher level ethanol (4.2% - 5.0%, v/v) than other species (0.4% - 2.2%, v/v). Using the aromatic yeasts in fermentation industries, we expect to improve the quality of traditional alcoholic beverages.
