미생물을 이용하여 pyridine계 보효소인 NAD를 생산하기 위하여, 12종의 효모로부터 NAD 함량이 높은 Saccharomyces sake KBA No. 6을 선별하였으며, NAD 생산에 미치는 여러 가지 효과를 검토하였다. NAD 생산에 있어서 탄소원으로는 4% glucose, 질소원으로는 2% bactopeptone이 가장 좋았으며 최적 PM와 온도는 각각 5.0과 $30^{circ}$C$이었다. 한편 NAD 전구물질로서 nicotinamide와 adenine을 동시에 사용한 경우 NAD 생산이 가장 좋았으며 이때의 농도는 각각 4 mg/ml과 3 mg/ml이었다. 또한 NAD 생산을 증가시키기 위하여 배양시 2가 금속이온들을 사용하였으며 이들 중 $Zn^{2+}$가 매우 효과적이었다. 계면활성제의 영향은 음이온 계면활성제인 sodium dodecyl sulfate(SDS)를 사용하였을 때 효과적이었다. 상기의 최적 조건하에서, 144시간 배양 후에 최대 NAD 생산량은 배지 100 ml당 35 mg이었으며 전체량의 89%인 31mg의 NAD가 배지 중에 유리되었다.
In order to produce nicotinamide adenine dinucleotide (NAD) which is a pyridine nucleotide coenzyme, Saccharomyces sake KBA No. 6 having high NAD content was selected from 12 strains of yeast and various factors affecting the production of NAD were investigated. For NAD production, 4% of glucose was effective as a carbon source and 2% of bactopeptone was the best nitrogen source. The optimum pH and temperature was 5.0 and $30^{circ}$, respectively. Also, when 4 mg/ml of nicotinamide and 3 mg/ml adenine were used as precursors simultaneously, NAD production was the best. To increase NAD production, 2 valence metal ions were used during cultivation and $Zn^{2+}$ was very efficient. Among the surface active agents, anionic sodium dodesyl sulfate (SDS) was effective. Under the optimum conditions, the maximum amount of produced NhD was 35 mg/100 ml medium after cultivation of 144 hrs and 89% of total NAD amount, 31 mg of NAD, was leaked into culture broth.
