Preparation method of polyvinylpyrrolidone modified wheat esterase
A technology of polyvinylpyrrolidone and wheat esterase, which is applied in the direction of enzyme stabilization and the like, can solve the problems of harsh storage conditions and difficult to obtain materials, and achieves the effects of easy source, increased activity and simple modification process.
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Embodiment 1
[0023] 1) Add 0.07mol / L NaCl solution to wheat flour, the volume ratio of material to liquid is 1:2 (m:v), shake and extract at 29°C for 120min, then filter through double-layer gauze, and the filtrate obtained is at 6000r / min , centrifuge at 4°C for 10min, collect the supernatant; then salt out with 20%-70% ammonium sulfate, centrifuge at 10,000r / min at 4°C for 15min to collect the precipitate, use a dialysis bag with a molecular weight cut-off of 8,000-14,000Da, put the dialysis bag dialyzed into distilled water to remove ammonium sulfate. It is the prepared wheat esterase.
[0024] 2) Sodium periodate and polyvinylpyrrolidone (2:1, m:m) were dissolved in 40 mL of distilled water, and after stirring for 20 hours at 25°C in the dark, the resulting product was dissolved in distilled water with a dialysis bag with a molecular weight cut-off of 8,000 to 14,000 Da. Dialyzed for 20 hours to obtain an oxidized solution of polyvinylpyrrolidone.
[0025] 3) Mix the prepared wheat e...
Embodiment 2
[0028] 1) Add 0.15 mol / L NaCl solution to wheat flour, the volume ratio of material to liquid is 1:6 (m:v), shake and extract at 40°C for 40 minutes, and then filter through double-layer gauze. / min, centrifuge at 4°C for 5min, collect the supernatant; then salt out with 20%-70% ammonium sulfate, centrifuge at 10,000r / min at 4°C for 19min to collect the precipitate, and use a dialysis bag with a molecular weight cut-off of 8,000-14,000Da to dialyze the precipitate The bag was put into distilled water and dialyzed to remove ammonium sulfate. It is the prepared wheat esterase.
[0029] 2) Sodium periodate and polyvinylpyrrolidone (4:1, m:m) were dissolved in 45 mL of distilled water, and after stirring for 35 hours in the dark at 25°C, the resulting product was dissolved in distilled water with a dialysis bag with a molecular weight cut-off of 8,000-14,000 Da. Dialyzed in medium for 19 hours to obtain an oxidized solution of polyvinylpyrrolidone.
[0030] 3) Mix the prepared w...
Embodiment 3
[0033] 1) Add 0.07mol / L NaCl solution to wheat flour, the volume ratio of material to liquid is 1:3 (m:v), shake and extract at 30°C for 60min, then filter through double-layer gauze, and the filtrate obtained is at 30000r / min , centrifuged at 4°C for 7 minutes, and collected the supernatant; then salted out with 20%-70% ammonium sulfate, centrifuged at 10,000 r / min at 4°C for 16 minutes to collect the precipitate, and used a dialysis bag with a molecular weight cut-off of 8,000 to 14,000 Da, and put the dialysis bag into dialyzed into distilled water to remove ammonium sulfate. It is the prepared wheat esterase.
[0034] 2) Sodium periodate and polyvinylpyrrolidone (2.5:1, m:m) were dissolved in 90 mL of distilled water, and after stirring for 30 hours in the dark at 25°C, the resulting product was dissolved in distilled water with a dialysis bag with a molecular weight cut-off of 8,000-14,000 Da. Dialyzed in medium for 19 hours to obtain an oxidized solution of polyvinylpyr...
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