Nawani N N, Kapadnis B P
Department of Microbiology, University of Pune, Pune, India.
J Appl Microbiol. 2001 May;90(5):803-8. doi: 10.1046/j.1365-2672.2001.01308.x.
A simple single step technique of gel filtration was developed for the purification of chitinase from Serratia marcescens NK1.
Chitinase from Ser. marcescens NK1 was purified to homogeneity by gel filtration chromatography with 9.2% recovery. The enzyme had a pH optimum of 6.2 and a temperature optimum of 47 degrees C. It was stable in a wide pH range of 3.0 to 10.0, retaining 60% activity at pH 3.0 and 65% activity at pH 10.5. It retained 70% activity at 28 degrees C after 72 h and nearly 50% activity at 50 degrees C up to 24 h.
The chitinase from Ser. marcescens NK1 can be efficiently purified in a single step by gel filtration chromatography. The chitinase of Ser. marcescens NK1, a soil isolate, is highly stable and as active as that of other reported isolates of Ser. marcescens.
This purification scheme is advantageous because of its simplicity and can therefore be applied for the purification of other enzymes. The yield is sufficient for initial characterization studies of the enzyme, and an improved resolution can be obtained if the chromatography is done under fast flow systems.
开发一种简单的一步凝胶过滤技术，用于从粘质沙雷氏菌NK1中纯化几丁质酶。
通过凝胶过滤色谱法将粘质沙雷氏菌NK1的几丁质酶纯化至同质，回收率为9.2%。该酶的最适pH为6.2，最适温度为47℃。它在3.0至10.0的宽pH范围内稳定，在pH 3.0时保留60%的活性，在pH 10.5时保留65%的活性。在28℃下72小时后保留70%的活性，在50℃下24小时内保留近50%的活性。
粘质沙雷氏菌NK1的几丁质酶可通过凝胶过滤色谱法一步高效纯化。土壤分离株粘质沙雷氏菌NK1的几丁质酶高度稳定，与其他报道的粘质沙雷氏菌分离株的活性相同。
该纯化方案因其简单性而具有优势，因此可用于其他酶的纯化。该产量足以进行酶的初步表征研究，如果在快速流动系统下进行色谱分析，可获得更高的分辨率。
