Koo Ok Kyung, Jeong Do-Won, Lee Jung Min, Kim Min Jung, Lee Jong-Hoon, Chang Hae Choon, Kim Jeong Hwan, Lee Hyong Joo
School of Agricultural Biotechnology, and Center for Agricultural Biomaterials, Seoul National University, Seoul 151-742, Republic of Korea.
Biotechnol Lett. 2005 Apr;27(7):505-10. doi: 10.1007/s10529-005-2541-z.
A bifunctional alcohol/acetaldehyde dehydrogenase (AdhE) gene (adhE) was cloned from Leuconostoc mesenteroides C7 (LMC7), which is the dominant lactic acid bacterium produced during heterofermentation of kimchi. The nucleotide sequence of the DNA fragment containing putative adhE, which is 2685 bp long and encodes an 886 amino acid polypeptide, exhibits 99% homology with Leu. mesenteroides sp. cremoris. The deduced AdhE comprises two conserved domains: alcohol dehydrogenase (Adh) and acetaldehyde dehydrogenase (Aldh). Moreover, two NAD-binding sites were observed, based on the presence of the GXGXXG motif. A pADHE containing the adhE gene expressed AdhE at the translational level in Escherichia coli BL21, which was at a higher level than in E. coli DH5alpha and E. coli JM109. The AdhE of LMC7 showed Adh and Aldh activities that, when expressed in E. coli. BL21, were 7.5 and 5.7 U mg(-1) , respectively.
从泡菜异型发酵过程中占主导地位的乳酸菌肠系膜明串珠菌C7（LMC7）中克隆了一个双功能乙醇/乙醛脱氢酶（AdhE）基因（adhE）。包含假定adhE的DNA片段长度为2685 bp，编码一个886个氨基酸的多肽，其核苷酸序列与肠系膜明串珠菌乳脂亚种具有99%的同源性。推导的AdhE包含两个保守结构域：乙醇脱氢酶（Adh）和乙醛脱氢酶（Aldh）。此外，基于GXGXXG基序的存在，观察到两个NAD结合位点。含有adhE基因的pADHE在大肠杆菌BL21中翻译水平表达AdhE，其表达水平高于大肠杆菌DH5α和大肠杆菌JM109。LMC7的AdhE在大肠杆菌BL21中表达时显示出Adh和Aldh活性，分别为7.5和5.7 U mg(-1)。
