Habe Hiroshi, Sato Shun, Fukuoka Tokuma, Kitamoto Dai, Yakushi Toshiharu, Matsushita Kazunobu, Sakaki Keiji
Research Institute for Innovation in Sustainable Chemistry, National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki.
J Oleo Sci. 2011;60(9):489-94. doi: 10.5650/jos.60.489.
Acetobacter tropicalis NBRC16470 can produce highly enantiomerically pure D-glyceric acid (D-GA; >99 % enantiomeric excess) from glycerol. To investigate whether membrane-bound alcohol dehydrogenase (mADH) is involved in GA production in A. tropicalis, we amplified part of the gene encoding mADH subunit I (adhA) using polymerase chain reaction and constructed an adhA-disrupted mutant of A. tropicalis (ΔadhA). Because ΔadhA did not produce GA, we confirmed that mADH is essential for the conversion of glycerol to GA. We also cloned and sequenced the entire region corresponding to adhA and adhB, which encodes mADH subunit II. The sequences showed high identities (84-86 %) with the equivalent mADH subunits from other Acetobacter spp.
热带醋杆菌NBRC16470能够从甘油生产出对映体纯度极高的D-甘油酸(D-GA;对映体过量>99%)。为了研究膜结合醇脱氢酶(mADH)是否参与热带醋杆菌中GA的产生,我们使用聚合酶链反应扩增了编码mADH亚基I(adhA)的部分基因,并构建了热带醋杆菌的adhA缺失突变体(ΔadhA)。由于ΔadhA不产生GA,我们证实mADH对于甘油转化为GA至关重要。我们还克隆并测序了与adhA和adhB对应的整个区域,adhB编码mADH亚基II。这些序列与其他醋杆菌属的等效mADH亚基具有高度同源性(84-86%)。