恶臭假单胞菌JP-1产腈水合酶的发酵工艺研究

The Study of Fermentation Technology Conditions by Pseudomonas putida JP-1 Producing High Nitrile Hydrotase

70年代以来,国外许多学者开展了用腈水合酶将丙烯腈一步水合生成丙烯酰胺的研究,其反应在常温常压下进行,转化率高(近100%),分离操作简单和产品纯度高。80年代中期日本就有用微生物转化法生产丙烯酰胺的工厂。

The study of fermentation technology coditions by Pseudomonasputida JP-1 producing high nitrile hydratase was in model BF-Ⅱ-10 10L fermenter and MSJ-U3 30 L fermenter. The results suggested thatused 2 or 3 stage fermentation can obtain cells having high nitrile hy-drotase due to used optimum seed culture medium and fermentationculture medium. These cells were enirapped with calcium alginate,thencontinuously feeded acrylonitrile and reacted for 13h, 300g/L of acrula-mide was produced. The half time of the immobilized cells was over1000h. The results also showed that optimum ferm entation technologyconditions are: seed ages is 6--12h, seed volume is 5--10%, culturetemperature is 25--30℃, air volume is 0.25--0.5vvm, agitate speed is 100--200rpm, fermentation cycle is 18--24h. The max nitrile hydrotase ofspecies JP-1 can achieve 200u/ml, and the cell mass can achieve 3 g/L.At the fermeniation process the lowest DO Concentration was over 20% and the average cell specific growth rate is 0.17h^(-1) in logarithmicgrowth phase.