Chaubey Asha, Parshad Rajinder, Koul Surrinder, Taneja Subhash C, Qazi Ghulam N
Regional Research Laboratory CSIR, Canal Road, Jammu Tawi 180001, India.
Appl Microbiol Biotechnol. 2006 Dec;73(3):598-606. doi: 10.1007/s00253-006-0520-5. Epub 2006 Aug 5.
Arthrobacter sp. lipase (ABL, MTCC no. 5125) is being recognized as an efficient enzyme for the resolution of drugs and their intermediates. The immobilization of ABL on various matrices for its enantioselectivity, stability, and reusability has been studied. Immobilization by covalent bonding on sepharose and silica afforded a maximum of 380 and 40 IU/g activity, respectively, whereas sol-gel entrapment provided a maximum of 150 IU/g activity in dry powder. The immobilized enzyme displayed excellent stability in the pH range of 4-10 and even at higher temperature, i.e., 50-60 degrees C, compared to free enzyme, which is unstable under extreme conditions. The resolution of racemic auxiliaries like 1-phenyl ethanol and an intermediate of antidepressant drug fluoxetine, i.e., ethyl 3-hydroxy-3-phenylpropanoate alkyl acylates, provided exclusively R-(+) products ( approximately 99% ee, E=646 and 473), compared to cell free extract/whole cells which gave a product with approximately 96% ee (E=106 and 150). The repeated use (ten times) of covalently immobilized and entrapped ABL resulted in no loss in activity, thus demonstrating its prospects for commercial applications.
节杆菌脂肪酶（ABL，MTCC编号5125）被认为是一种用于拆分药物及其中间体的高效酶。人们已经研究了将ABL固定在各种基质上以提高其对映选择性、稳定性和可重复使用性。通过共价键合固定在琼脂糖和硅胶上时，活性分别最高可达380和40 IU/g，而溶胶-凝胶包埋法在干粉中活性最高可达150 IU/g。与在极端条件下不稳定的游离酶相比，固定化酶在pH值4-10的范围内以及甚至在较高温度（即50-60摄氏度）下都表现出优异的稳定性。与无细胞提取物/完整细胞生成的对映体过量值约为96%（E=106和150）的产物相比，外消旋助剂如1-苯乙醇和抗抑郁药物氟西汀的中间体即3-羟基-3-苯基丙酸乙酯烷基酰化物的拆分仅生成R-(+)产物（对映体过量值约为99%，E=646和473）。共价固定和包埋的ABL重复使用（十次）后活性没有损失，因此证明了其商业应用前景。
