Benov Ludmil, Al-Ibraheem Jameela
The Department of Biochemistry, Faculty of Medicine, Kuwait University, P. O. Box 24923, Safat 13110, Kuwait.
J Biochem Mol Biol. 2002 Jul 31;35(4):428-31. doi: 10.5483/bmbrep.2002.35.4.428.
The often-encountered problem of disrupting bacteria for the purpose of extracting soluble protein has generated various methods. Many require specialized equipment. Very often, especially during preliminary studies, investigators need a simple, fast, and inexpensive method for cell disruption that preserves biological activity. This paper compares some simple and inexpensive methods for cell disruption, such as bead-vortexing, freezing-thawing, French pressing, and sonication. It also provides some tips to increase protein yield and preserve biological activity. If performed under optimal conditions, bead-vortexing gives protein yields that are comparable to French pressing and sonication. It also preserves the activities of labile enzymes and releases periplasmic enzymes. Vortexing with glass beads appears to be the simplest method for cell disruption.
为提取可溶性蛋白质而破坏细菌这一经常遇到的问题催生了各种方法。许多方法需要专门的设备。尤其是在初步研究期间，研究人员常常需要一种简单、快速且廉价的细胞破碎方法，同时还要保留生物活性。本文比较了一些简单且廉价的细胞破碎方法，如珠磨法、冻融法、高压匀浆法和超声破碎法。还提供了一些提高蛋白质产量并保留生物活性的技巧。如果在最佳条件下进行，珠磨法获得的蛋白质产量与高压匀浆法和超声破碎法相当。它还能保留不稳定酶的活性并释放周质酶。用玻璃珠进行涡旋似乎是最简单的细胞破碎方法。
