Production of lipase in a fermentor using a mutant strain of Corynebacterium species: its partial purification and immobilization.
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.
Roy
Nityananda
N
Department of Food Technology & Biochemical Engineering, Jadavpur University, Kolkata 700 032, India.
Ray
Lalitagauri
L
Chattopadhyay
Parimal
P
D000464:Alginates / Q000737:chemistry / Q000378:metabolism; D003352:Corynebacterium / Q000201:enzymology* / Q000235:genetics; D003432:Cross-Linking Reagents; D003956:Dialysis; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005285:Fermentation; D020723:Glucuronic Acid / Q000737:chemistry / Q000378:metabolism; D005976:Glutaral / Q000378:metabolism; D006603:Hexuronic Acids / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007218:Industrial Microbiology; D007700:Kinetics; D008049:Lipase / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature
D000464:Alginates; D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D008049:Lipase; D005976:Glutaral