Purification and characterization of cellulase from a Vibrio isolate
Date Issued
2014
Date
2014
Author(s)
Cheng, Shih-Shuan
Abstract
In this study, an isolate from sea water of Rueifang, Xinbei, had been tested for its cellulase activity. After the 16S rRNA was sequenced to establish phylogenetic tree of this isolate, which belonged to the genus Vibrio. Cellulase of this isolate was purified and characterized. Optimal cellulase yielding conditions for the culture were determined. For the best cellulase production, the culture was incubated at 30 °C with 50 mL LB broth (pH 6) in 250 mL flask and shaken for 72 hours (75 rpm). CMC (1%) was added as an cellulase inducer in LB broth. The cellulase was purified by ion exchange chromatography after ammonium sulfate precipitation, dialysis, and desalting. The molecular mass of the cellulases determined by SDS-PAGE was approximately 60 kDa, 36 kDa and 20 kDa. The cellulase showed optimum activity at pH 6 and 50 °C, and was stable at pH 4-9 and 4-40°C. When the cellulase was treated with various surfactants, 70 % activity was inhibited by 1% (g/mL) SDS and 1% Tween 80 , and had a lost of 55 percent under the effect of Tween 20. The cellulase was inactivated by 5 mM phenylmethanesulfonylfluride (PMSF) and 5 mM ethylenediaminetetraacetic acid (EDTA), but was not influenced by 2-Mercaptoethanol. Finally, in the industrial application tests, the cellulase showed better ability to decompose microcrystalline cellulose than to Ulva and paper, but showed weaker ability to break down sugar cane bagasse than to any others substrates.
Subjects
纖維素
纖維素分解酶
弧菌屬
酵素抑制劑
界面活性劑
陰離子交換層析
Type
thesis
File(s)![Thumbnail Image]()
Loading...
Name
ntu-103-R99241208-1.pdf
Size
23.54 KB
Format
Adobe PDF
Checksum
(MD5):5a9f8dd08592e92666d95acb0b0cc7f6