Mori Ayana, Hirata Yuuki, Kishida Mayumi, Mori Yutaro, Kondo Akihiko, Noda Shuhei, Tanaka Tsutomu
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo 657-8501, Japan.
Center for Sustainable Resource Science, RIKEN, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan; Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo 657-8501, Japan.
Enzyme Microb Technol. 2023 Dec;171:110321. doi: 10.1016/j.enzmictec.2023.110321. Epub 2023 Sep 6.
Nitroaromatic compounds are widely used in industry, but their production is associated with issues such as the hazardousness of the process and low regioselectivity. Here, we successfully demonstrated the production of p-nitrobenzoate (PNBA) from glucose by constructing p-aminobenzoate N-oxygenase AurF-expressing E. coli. We generated this strain, which we named PN-1 by disrupting four genes involved in PNBA degradation: nfsA, nfsB, nemA, and azoR. We then expressed AurF from Streptomyces thioluteus in this strain, which resulted in the production of 945 mg/L PNBA in the presence of 1 g/L p-aminobenzoate. Direct production of PNBA from glucose was achieved by co-expressing the pabA, pabB, and pabC, as well as aurF, resulting in the production of 393 mg/L PNBA from 20 g/L glucose. To improve the PNBA titer, we disrupted genes involved in competing pathways: pheA, tyrA, trpE, pykA, and pykF. The resultant strain PN-4Ap produced 975 mg/L PNBA after 72 h of cultivation. These results highlight the potential of using microorganisms to produce other nitroaromatic compounds.
硝基芳香族化合物在工业中广泛使用，但其生产与诸如过程危险性和区域选择性低等问题相关。在此，我们通过构建表达对氨基苯甲酸N-加氧酶AurF的大肠杆菌，成功证明了从葡萄糖生产对硝基苯甲酸酯（PNBA）。我们通过破坏参与PNBA降解的四个基因：nfsA、nfsB、nemA和azoR，构建了该菌株，我们将其命名为PN-1。然后我们在该菌株中表达来自硫黄链霉菌的AurF，在存在1 g/L对氨基苯甲酸的情况下，这导致产生了945 mg/L的PNBA。通过共表达pabA、pabB和pabC以及aurF，实现了从葡萄糖直接生产PNBA，从20 g/L葡萄糖中产生了393 mg/L的PNBA。为了提高PNBA的滴度，我们破坏了参与竞争途径的基因：pheA、tyrA、trpE、pykA和pykF。所得菌株PN-4Ap在培养72小时后产生了975 mg/L的PNBA。这些结果突出了利用微生物生产其他硝基芳香族化合物的潜力。
