Identification and characterization of a novel 2,3‐butanediol dehydrogenase/acetoin reductase from Corynebacterium crenatum SYPA5‐5
Identification and characterization of a novel 2,3‐butanediol dehydrogenase/acetoin reductase from Corynebacterium crenatum SYPA5‐5
复制标题
DOI：
10.1111/lam.12495
复制
发表时间：
2015-12
影响因子：
2.4
通讯作者：
X. Zhao;X. Zhang;Z. Rao;T. Bao;X. Li;M. Xu;T. Yang;S. Yang
中科院分区：
文献类型：
--
作者：
X. Zhao;X. Zhang;Z. Rao;T. Bao;X. Li;M. Xu;T. Yang;S. Yang
乙偶姻和2，3-丁二醇广泛用于化学和制药工业。这种酶，2，3-丁二醇脱氢酶/乙偶姻还原酶（2，3-BDH/AR），通过催化乙偶姻和2，3-丁二醇之间的可逆反应，在乙偶姻和2，3-丁二醇的微生物生产中起着重要作用。迄今为止，尚未从钝齿棒杆菌中表征2，3-BDH。从Coryne克隆2，3-BDH。crenatum SYPA 5 - 5，并在大肠杆菌BL 21中表达。序列分析表明2，3-BDH来自Coryne。crenatum SYPA 5 - 5属于短链脱氢酶/还原酶超家族。在35°C时比活性最高，但当温度高于35°C时，比活性变得很不稳定。其还原反应的最适pH为4.0，氧化反应的最适pH为10.0。Ca 2+、Mg 2+、Zn 2+和Mn 2+离子在一定程度上提高了2，3-BDH的活性。特别是，Ca 2+诱导约1.5倍的增加。双乙酰和乙偶姻的kcat/Km值高于2，3-丁二醇，表明2，3-BDH可以在较低pH条件下容易地将双乙酰或乙偶姻还原为2，3-丁二醇。来自Coryne的2，3-BDH的特征。crenatum SYPA 5 - 5将为进一步研究用工程化Coryne生产乙偶姻和2，3-丁二醇提供指导。crenatum SYPA 5 - 5.
Acetoin and 2,3‐butanediol are widely used in the chemical and pharmaceutical industries. The enzyme, 2,3‐butanediol dehydrogenase/acetoin reductase (2,3‐BDH/AR), plays a significant role in the microbial production of acetoin and 2,3‐butanediol by catalysing a reversible reaction between acetoin and 2,3‐butanediol. To date, a 2,3‐BDH has not been characterized from Corynebacterium crenatum. 2,3‐BDH was cloned from Coryne. crenatum SYPA5‐5 and expressed in Escherichia coli BL21. Sequence analysis suggested that the 2,3‐BDH from Coryne. crenatum SYPA5‐5 belongs to the short‐chain dehydrogenase/reductase superfamily. Its maximum specific activity was obtained at 35°C, however, it became very unstable when the temperature was above 35°C. Its optimal pH was 4·0 for reduction reaction and 10·0 for oxidation reaction. The 2,3‐BDH activity was increased to some extent by Ca2+, Mg2+, Zn2+ and Mn2+ ions. In particular, Ca2+ induced about 1·5‐fold increase. The value of kcat/Km for diacetyl and acetoin are higher than for 2,3‐butanediol indicating that 2,3‐BDH can easily reduce diacetyl or acetoin to 2,3‐butanediol under lower pH conditions. The characteristics of 2,3‐BDH from Coryne. crenatum SYPA5‐5 will give guide to further studies for the production of acetoin and 2,3‐butanediol with engineered Coryne. crenatum SYPA5‐5.