Separation of alpha-amylase by reversed micellar extraction. Effect of solvent type and cosolvent concentration on the transfer process.
The recovery of alpha-amylase from the crude enzyme preparation by the reversed micellar liquid-liquid extraction was investigated. The reversed micellar solution was formed by dissolving a cationic surfactant Aliquat 336 in six different alkanes (cyclohexane, n-hexane, isooctane, n-octane, n-decane, and n-dodecane) respectively with addition of a cosolvent n-octanol. It was found that a minimal quantity of n-octanol was needed for Aliquat 336 to dissolve in apolar solvent and form reversed micelles. Furthermore, this minimal amount of n-octanol needed was found to be different when Aliquat 336 was dissolved in different alkanes. It tended to increase with the number of carbon atoms in alkane and also depended on the solvent structure. During the forward extraction process, it was revealed that a high value of solubilization of protein in Aliquat 336 reversed micelles could be achieved when four out of the six alkanes (cyclohexane, n-hexane, isooctane, n-octane) were used as the solvent for Aliquat 336. After a full forward and backward extraction cycle, however, a high recovery of both the protein mass and alpha-amylase activity in the stripping solution could be obtained only when two out of the six alkanes (n-hexane and isooctane) were used as the solvent for Aliquat 336. When n-hexane and isooctane were used as the solvent for Aliquat 336, up to 80% of the total alpha-amylase activity in the crude enzyme preparation could be recovered at the end of extraction cycle, meanwhile alpha-amylase could be concentrated about 1.4-fold. In the cases of other four alkanes (cyclohexane, n-octane, n-decane, and n-dodecane) as solvent, most of the alpha-amylase activity in the crude enzyme preparation would be denatured after an extraction cycle.
Chang
Q L
QL
Institute of Chemical Metallurgy, Chinese Academy of Sciences, Beijing, China.
Chen
J Y
JY
D020003:1-Octanol; D006863:Hydrogen-Ion Concentration; D008823:Micelles; D000442:Octanols; D011506:Proteins / Q000302:isolation & purification; D000644:Quaternary Ammonium Compounds; D012997:Solvents; D000516:alpha-Amylases / Q000302:isolation & purification*
D008823:Micelles; D000442:Octanols; D011506:Proteins; D000644:Quaternary Ammonium Compounds; D012997:Solvents; C037759:trioctylmethylammonium; D000516:alpha-Amylases; D020003:1-Octanol