Yamamura Ei-Tora
a Technical Department , Kyowa Pharma Chemical Co., Ltd. , Takaoka , Toyama , Japan.
Biosci Biotechnol Biochem. 2018 Aug;82(8):1396-1403. doi: 10.1080/09168451.2018.1463154. Epub 2018 Apr 20.
NADP-dependent aminoalcohol dehydrogenase (AADH) of Rhodococcus erythropolis MAK154 produces double chiral aminoalcohols, which are used as pharmaceuticals. However, the genetic manipulation of Rhodococcus strains to increase their production of such industrially important enzymes is not well studied. Therefore, I aimed to construct Rhodococcus expression vectors, derived from the Rhodococcus-Escherichia coli shuttle vector pRET1102, to express aadh. The plasmid pRET1102 could be transformed into many actinomycete strains, including R. erythropolis. The transformation efficiency for a species closely related to R. erythropolis was higher than that for other actinomycete strains. Promoters of various strengths, hsp, 1200rep, and TRR, were obtained from Gram-positive bacteria. The activity of TRR was stronger than that of hsp and 1200rep. The aadh-expressing plasmid pRET1172 with TRR could be transformed into many actinomycete strains to increase their AADH production. The Rhodococcus expression vector, pRET11100, constructed by removing aadh from the pRET1172 plasmid may be useful for bioconversion.
红平红球菌MAK154的NADP依赖性氨基醇脱氢酶（AADH）可产生双 chiral 氨基醇，这些氨基醇可用作药物。然而，对红球菌菌株进行基因操作以提高其此类具有工业重要性的酶的产量的研究并不充分。因此，我的目标是构建源自红球菌-大肠杆菌穿梭载体pRET1102的红球菌表达载体，以表达aadh。质粒pRET1102可转化到包括红平红球菌在内的许多放线菌菌株中。与红平红球菌密切相关的一个物种的转化效率高于其他放线菌菌株。从革兰氏阳性细菌中获得了各种强度的启动子，即hsp、1200rep和TRR。TRR的活性强于hsp和1200rep。带有TRR的表达aadh的质粒pRET1172可转化到许多放线菌菌株中，以提高它们的AADH产量。通过从pRET1172质粒中去除aadh构建的红球菌表达载体pRET11100可能对生物转化有用。 （注：原文中“double chiral”表述有误，可能是“double chiral”，推测是“双对映体”之类意思，这里按原样翻译）
