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KMID : 0545120040140061310
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Journal of Microbiology and Biotechnology
2004 Volume.14 No. 6 p.1310 ~ p.1317
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Process Strategies to Enhance the Production of 5-Aminolevulinic Acid with Recombinant E. coli
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Lee DH
Jun WJ/Yoon JW/Cho HY/Hong BS
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Abstract
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The extracellular production of 5-aminolevulinic acid (ALA) by recombinant E. coli BL21 harboring a fusion gene hemA was investigated in a fermenter. For this purpose the effects of various physiological factors such as isopropylthio- β-D-galactopyranoside (IPTG) concentrations and the time of induction on enzyme activity were studied. Optimum concentrations of glycine and succinic acid were found to be 30 mM and 90 mM respectively. When the cells were permitted to grow for 2 h prior to the addition of 0.1 mM IPTG the activity of ALA synthase was higher than when IPTG was initially added. A 36-fold increase in the activity was observed with only 0.1 mM IPTG added. The pH of the medium also influenced the ALA synthase activity with the maximal activity occurring at pH 6.5. In recombinant E. coli extracts the repeated addition of glycine and D-glucose increased the production of ALA and the inhibited intracellular ALA dehydratase activity with up to 32 mM ALA being produced in the cultivation.
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