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Cloning of two 5-aminolevulinic acid synthase isozymes HemA and HemO from Rhodopseudomonas palustris with favorable characteristics for 5-aminolevulinic acid production

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Abstract

5-Aminolevulinic acid (ALA) synthase (ALAS) HemA from non-sulfur photosynthetic bacteria has been used for the ALA bioproduction, whereas the isoenzyme HemT/HemO is less studied and not used for ALA production. Two ALAS-encoding genes, hemA and hemO from Rhodopseudomonas palustris were cloned, purified and characterized. The ALASs had very high specific activity, 3.6 and 2.7 U/mg, respectively, and strong affinity for one of its substrates, succinyl-CoA, K m with values of 11 and 4.4 μM, respectively. HemO retained up to 60 % maximum activity within a broad range of concentrations of hemin, while HemA kept only 20 % at 10 μM hemin. Escherichia coli overexpressing HemA or HemO produced 5.7 and 6.3 g ALA/l, respectively, in a 5 l bioreactor.

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Acknowledgments

This work was supported by the Knowledge Innovation Program of the Chinese Academy of Sciences (Grant No. KSCX2-E-W-Q-13), Tianjin Research Program of Application Foundation and Advanced Technology (No. 10JCZDJC16200), National Natural Science Foundation of China (No. 31070037), and Key Technologies Research and Development Program of Tianjin (No. 11ZCZDSY08500).

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Authors and Affiliations

  1. College of Biotechnology, Tianjin University of Science and Biotechnology, Tianjin, 300457, People’s Republic of China

    Lilu Zhang & Ning Chen

  2. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People’s Republic of China

    Lilu Zhang, Jiuzhou Chen, Jibin Sun & Ping Zheng

  3. National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Xiqi Road 32#, Tianjin Airport Economic Area, Tianjin, 300308, People’s Republic of China

    Lilu Zhang, Jiuzhou Chen, Jibin Sun, Ping Zheng & Yanhe Ma

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  1. Lilu Zhang
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Correspondence to Ping Zheng.

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Lilu Zhang, Jiuzhou Chen dedicated equally to this manuscript.

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Zhang, L., Chen, J., Chen, N. et al. Cloning of two 5-aminolevulinic acid synthase isozymes HemA and HemO from Rhodopseudomonas palustris with favorable characteristics for 5-aminolevulinic acid production. Biotechnol Lett 35, 763–768 (2013). https://doi.org/10.1007/s10529-013-1143-4

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  • DOI: https://doi.org/10.1007/s10529-013-1143-4

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