通过无细胞多酶催化生产5-氨基乙酰丙酸。
Production of 5-aminolevulinic acid by cell free multi-enzyme catalysis.
作者信息
Meng Qinglong, Zhang Yanfei, Ju Xiaozhi, Ma Chunling, Ma Hongwu, Chen Jiuzhou, Zheng Ping, Sun Jibin, Zhu Jun, Ma Yanhe, Zhao Xueming, Chen Tao
机构信息
Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, PR China; Key Laboratory of Systems Bioengineering (Ministry of Education), SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, PR China.
Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, PR China.
出版信息
J Biotechnol. 2016 May 20;226:8-13. doi: 10.1016/j.jbiotec.2016.03.024. Epub 2016 Mar 21.
5-Aminolevulinic acid (ALA) is the precursor for the biosynthesis of tetrapyrroles and has broad agricultural and medical applications. Currently ALA is mainly produced by chemical synthesis and microbial fermentation. Cell free multi-enzyme catalysis is a promising method for producing high value chemicals. Here we reported our work on developing a cell free process for ALA production using thermostable enzymes. Cheap substrates (succinate and glycine) were used for ALA synthesis by two enzymes: 5-aminolevulinic acid synthase (ALAS) from Laceyella sacchari (LS-ALAS) and succinyl-CoA synthase (Suc) from Escherichia coli. ATP was regenerated by polyphosphate kinase (Ppk) using polyphosphate as the substrate. Succinate was added into the reaction system in a fed-batch mode to avoid its inhibition effect on Suc. After reaction for 160min, ALA concentration was increased to 5.4mM. This is the first reported work on developing the cell free process for ALA production. Through further process and enzyme optimization the cell free process could be an effective and economic way for ALA production.
5-氨基乙酰丙酸（ALA）是四吡咯生物合成的前体，具有广泛的农业和医学应用。目前，ALA主要通过化学合成和微生物发酵生产。无细胞多酶催化是生产高价值化学品的一种有前景的方法。在此，我们报告了我们利用耐热酶开发无细胞生产ALA工艺的工作。使用廉价底物（琥珀酸和甘氨酸）通过两种酶合成ALA：来自嗜糖莱西氏菌的5-氨基乙酰丙酸合酶（LS-ALAS）和来自大肠杆菌的琥珀酰辅酶A合成酶（Suc）。以多聚磷酸为底物，通过多聚磷酸激酶（Ppk）再生ATP。以补料分批模式向反应体系中添加琥珀酸，以避免其对Suc的抑制作用。反应160分钟后，ALA浓度提高到5.4 mM。这是首次报道的开发无细胞生产ALA工艺的工作。通过进一步的工艺和酶优化，无细胞工艺可能成为生产ALA的一种有效且经济的方式。
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