Abstract Background 2R,3R-butanediol dehydrogenase (R-BDH) and other BDHs contribute to metabolism of 3R/3S-Acetoin (3R/3S-AC) and 2,3-butanediol (2,3-BD), which are important bulk chemicals used in different industries. R-BDH is responsible for oxidizing the hydroxyl group at their (R) configuration. Bacillus species is a promising producer of 3R/3S-AC and 2,3-BD. In this study, R-bdh gene encoding R-BDH from Bacillus sp. DL01 was isolated, expressed and identified. Results R-BDH exerted reducing activities towards Diacetyl (DA) and 3R/3S-AC using NADH, and oxidizing activities towards 2R,3R-BD and Meso-BD using NAD+, while no activity was detected with 2S,3S-BD. The R-BDH showed its activity at a wide range of temperature (25°C to 65°C) and pH (5.0–8.0). The R-BDH activity was increased significantly by Cd2+ when DA, 3R/3S-AC, and Meso-BD were used as substrates, while Fe2+ enhanced the activity remarkably at 2R,3R-BD oxidation. Kinetic parameters of the R-BDH from Bacillus sp. DL01 showed the lowest Km, the highest Vmax, and the highest Kcat towards the racemic 3R/3S-AC substrate, also displayed low Km towards 2R,3R-BD and Meso-BD when compared with other reported R-BDHs. Conclusions The R-BDH from Bacillus sp. DL01 was characterized as a novel R-BDH with high enantioselectivity for R-configuration. It considered NAD+ and Zn2+ dependant enzyme, with a significant affinity towards 3R/3S-AC, 2R,3R-BD, and Meso-BD substrates. Thus, R-BDH is providing an approach to regulate the production of 3R/3S-AC or 2,3-BD from Bacillus sp. DL01. How to cite: Elmahmoudy M, Elfeky N, Zhongji P, et al. Identification and characterization of a novel 2R,3R-Butanediol Dehydrogenase from Bacillus sp. DL01. Electron J Biotechnol 2021;49. https://doi.org/10.1016/j.ejbt.2020.11.002

中文翻译：

---

来自芽孢杆菌的新型 2R,3R-丁二醇脱氢酶的鉴定和表征。DL01

摘要 背景 2R,3R-丁二醇脱氢酶 (R-BDH) 和其他 BDH 有助于 3R/3S-乙偶姻 (3R/3S-AC) 和 2,3-丁二醇 (2,3-BD) 的代谢，它们是重要的原料用于不同行业的化学品。R-BDH 负责氧化其 (R) 构型的羟基。芽孢杆菌属是一种很有前途的 3R/3S-AC 和 2,3-BD 生产者。在这项研究中，R-bdh 基因编码来自芽孢杆菌的 R-BDH。DL01 被分离、表达和鉴定。结果 R-BDH 使用 NADH 对双乙酰 (DA) 和 3R/3S-AC 产生还原活性，使用 NAD+ 对 2R,3R-BD 和 Meso-BD 产生氧化活性，而使用 2S,3S-BD 未检测到活性。R-BDH 在很宽的温度范围（25°C 至 65°C）和 pH 值（5.0-8.0）下显示出其活性。当 DA、3R/3S-AC、和Meso-BD作为底物，而Fe2+显着增强了2R,3R-BD氧化的活性。来自芽孢杆菌的 R-BDH 的动力学参数。与其他报道的 R-BDH 相比，DL01 对外消旋 3R/3S-AC 底物显示出最低 Km、最高 Vmax 和最高 Kcat，对 2R、3R-BD 和 Meso-BD 也显示出低 Km。结论来自芽孢杆菌的 R-BDH。DL01 被表征为一种新型 R-BDH，对 R 构型具有高对映选择性。它考虑了 NAD+ 和 Zn2+ 依赖性酶，对 3R/3S-AC、2R、3R-BD 和 Meso-BD 底物具有显着的亲和力。因此，R-BDH 提供了一种调节芽孢杆菌产生 3R/3S-AC 或 2,3-BD 的方法。DL01。如何引用：Elmahmoudy M、Elfeky N、Zhongji P 等。新型 2R 的鉴定和表征，来自芽孢杆菌的 3R-丁二醇脱氢酶。DL01。电子 J 生物技术 2021；49。https://doi.org/10.1016/j.ejbt.2020.11.002