OPTIMIZATION OF PROCESS PARAMETERS FOR THE PRODUCTION OF GLUTAMINASE ENZYME FROM ERWINIAAT FLASK LEVEL.

This purpose of study was to determine parameter optimization for the<br>production of Glutaminase enzyme from Erwinia at flask level. This study was carried<br>out at Department of Biotechnology, Mata Gujri College, Fatehgarh Sahib, Punjabi<br>University Patiala For maximum production of glutaminase from free cells of Erwinia,<br>various physical and chemical parameters were optimized. The microorganism Erwinia<br>which is a source of glutaminase was procured from Institute of Microbial Technology,<br>Chandigarh and was grown on specified medium. The media was sterilized by<br>autoclaving for 15 minutes at 15 psi/cm2. The microorganism was maintained on agar<br>slants and kept in refrigerator at 4.0 + 0.5 0C for further use and was sub-cultured after<br>every 20 days. The viability check of microorganism was done by gram staining and<br>tryphan blue. The work utilized Beef extract, Peptone, Yeast extract, Sodium chloride<br>and Water as composition of growth medium for the production of glutaminase enzyme<br>from Erwinia. The parameters such as Temp., Ph, Time and Inducer concentration were<br>optimized.The enzyme activity was determined by Nessler's reagent method and protein<br>concentration was determined by Bradford method. It was concluded that Maximum<br>production of glutaminase enzyme from Erwinia was observed at temp.-25?C, pH-6.5 &amp;<br>Time after-48 hours. At these optimized parameters enzyme activity and Protein<br>concentration was found maximum本研究旨在确定摇瓶水平下欧文氏菌（Erwinia）生产谷氨酰胺酶（Glutaminase）的参数优化方案。 本研究于旁遮普大学帕提亚拉分校法特加尔萨希布校区的马塔古吉里学院生物技术系开展。 为从欧文氏菌游离细胞中最大化谷氨酰胺酶产量，研究人员对多项物理及化学参数进行了优化。 作为谷氨酰胺酶来源的欧文氏菌菌株购自昌迪加尔微生物技术研究所，将其接种于指定培养基中培养。 培养基采用15 psi/cm²压力高压蒸汽灭菌15分钟。 菌种于琼脂斜面培养基上保藏，置于4.0±0.5℃冰箱中备用，每20天传代一次。 采用革兰氏染色与台盼蓝染色对菌种进行活力验证。 本研究使用牛肉浸膏、蛋白胨、酵母提取物、氯化钠与纯水作为欧文氏菌生产谷氨酰胺酶的生长培养基组分。 研究对温度、pH值、培养时间及诱导剂浓度等参数进行了优化。 采用奈斯勒试剂法测定酶活性，布拉德福德（Bradford）法测定蛋白质浓度。 研究结果表明，在温度25℃、pH值6.5及培养48小时的优化参数下，欧文氏菌谷氨酰胺酶的产量达到最高，此时酶活性与蛋白质浓度均为最优。