Roy Nityananda, Ray Lalitagauri, Chattopadhyay Parimal
Department of Food Technology & Biochemical Engineering, Jadavpur University, Kolkata 700 032, India.
Indian J Exp Biol. 2004 Feb;42(2):202-7.
Extracellular Corynebacterium lipase was produced using a 2.5 L Chemap fermentor using 1300 ml fermentation medium at temperature 33 degrees C, agitator speed 50 rpm, aeration rate 1 VVM having KLa 16.21 hr(-1). Crude lipase was purified by salting out method followed by dialysis and immobilized using calcium alginate gel matrix followed by glutaraldehyde cross linking Purification process increased specific activity of enzyme from 2.76 to 114.7 IU/mg. Activity of immobilized enzyme was 107.31 IU/mg. Optimum temperature for purified and immobilized enzyme activity were 65 degrees and 50 degrees C respectively. Optimum pH was 8.0 in both the cases, Km and Vmax value for purified lipase were 111.1 micromol/min and 14.7% respectively. Ca2+ (5 mM) was found to be stimulator for enzyme activity. Immobilized lipase retained 68.18% of the original activity when stored for 40 days.
使用2.5升Chemap发酵罐，在33℃的温度、50转/分钟的搅拌速度、1体积空气每分钟（VVM）且传质系数（KLa）为16.21小时⁻¹的条件下，以1300毫升发酵培养基生产细胞外棒状杆菌脂肪酶。粗脂肪酶通过盐析法纯化，随后进行透析，并用海藻酸钙凝胶基质固定，接着进行戊二醛交联。纯化过程使酶的比活性从2.76提高到114.7国际单位/毫克。固定化酶的活性为107.31国际单位/毫克。纯化酶和固定化酶活性的最适温度分别为65℃和50℃。两种情况下的最适pH均为8.0。纯化脂肪酶的米氏常数（Km）和最大反应速度（Vmax）值分别为111.1微摩尔/分钟和14.7%。发现Ca²⁺（5毫摩尔）是酶活性的刺激剂。固定化脂肪酶在储存40天时保留了68.18%的原始活性。
