대사공학 기법을 이용한 초고온 고세균에서의 2,3-BDO 생산

Abstract

2,3-butanediol (BDO) is a precursor for various plastics and pesticides. Many reports has been come out about biological 2,3-BDO production by mesophilic microorganisms. In hyperthermophiles, it has been reported that acetoin, a precursor of 2,3-BDO, is produced from Pyrococcus furiosus. However, there are still no reports on 2,3-butnadiol production in hyperthermophiles. In this study, we first attempted to produce 2,3-BDO from hyperthermophilic archaeon, Thermococcus onnurineus NA1. These organism has no enzyme that turn out acetolactate, acetoin or 2,3-butanediol. So, we express two heterologous genes (acetolactate synthase (alsS) from Pyrococcus sp. NA2 and alcohol dehydrogenase (adh) from Thermococcus guaymacensis) to produce target metabolite in T. onnurineus NA1. The mutant strain produced about 4.0 g/L meso-form 2,3-BDO. In addition, to improve 2,3-BDO production, we have constructed acetyl-CoA synthetase III (TON_1001) deletion mutant and the mutant strain showed a 1.24-fold increase in 2,3-BDO productivity. Also, when carbon monoxide (CO) gas was added as a reductant, the 2,3-BDO productivity increased 1.25-fold.f 2,3-BDO, is produced from Pyrococcus furiosus. However, there are still no reports on 2,3-butnadiol production in hyperthermophiles. In this study, we first attempted to produce 2,3-BDO from hyperthermophilic archaeon, Thermococcus onnurineus NA1. These organism has no enzyme that turn out acetolactate, acetoin or 2,3-butanediol. So, we express two heterologous genes (acetolactate synthase (alsS) from Pyrococcus sp. NA2 and alcohol dehydrogenase (adh) from Thermococcus guaymacensis) to produce target metabolite in T. onnurineus NA1. The mutant strain produced about 4.0 g/L meso-form 2,3-BDO. In addition, to improve 2,3-BDO production, we have constructed acetyl-CoA synthetase III (TON_1001) deletion mutant and the mutant strain showed a 1.24-fold increase in 2,3-BDO productivity. Also, when carbon monoxide (CO) gas was added as a reductant, the 2,3-BDO productivity increased 1.25-fold.