During the process of producing α-ketoglutaric acid by whole cell transformation, the biotransformation solution contains impurities, such as residual substrates, macromolecular proteins, organic heteroacids and pigments, which is difficult to remove by a single separation method. In order to purify α-ketoglutaric acid from whole cell transformation solution, the calcium salt method and ion exchange method were together used for separation and extraction process of α-ketoglutaric acid. The pretreated whole cell transformation stock solution was acidified to pH 2.0, and calcium α-ketoglutarate was formed by precipitation of α-ketoglutaric acid with Ca(OH)2 as the precipitant. The ion-exchange master batch was obtained by treatment of calcium α-ketoglutarate using concentrated H2SO4. The purity of α-ketoglutaric acid increased from 78.79% to 93.66%, analyzed by HPLC. The D301 anion resin was selected as ion exchange filler by ion exchange static test. The optimal parameters for dynamic loading showed as follows: the height to diameter ratio of 2.5∶1, loading concentration of 30 g/L and loading flow rate of 2.4 BV/h. The optimal elution conditions were: 2 BVs eluted by 0.02% mass fraction of HCl at a flow rate of 4 BV/h, 4 BVs eluted by 5% mass fraction of NaOH at a flow rate of 2 BV/h. The total yield of α-ketoglutaric acid after separation and purification process was 80.32% and the chromatographic purity reached 97.32%.
YANG Xuechen
,
FU Shaoping
,
WANG Lixia
,
XU Ning
,
XU Chao
,
LIU Guanghui
,
MA Zhenping
,
LIU Jun
. Separation, purification and process optimization of α-ketoglutaric acid[J]. Food and Fermentation Industries, 2022
, 48(23)
: 65
-70
.
DOI: 10.13995/j.cnki.11-1802/ts.030468
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