title: Purification and characterization of an endo-D: -arabinase produced by cellulomonas.
abstract: An extracellular endo-D: -arabinase enzyme produced by the bacterial strain of Cellulomonas was purified 77.1-fold with 0.20% recovery for protein by DEAE Sepharose anion exchange, Sephacryl S-300 gel filtration and blue Sepharose affinity chromatography, and designated as CEDAase. The apparent molecular mass of CEDAase was 45 kDa determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. CEDAase is an endoenzyme for arabinogalactan with the main and specific product of hexa-arabinofuranoside. It reacts optimally with its substrate, arabinogalactan, at approximately pH 8.0 and at 40 °C. CEDAase shows stability in the pH range of 6.0-9.0 and at the temperature below 50 °C. The K(m) measured for the CEDAase was 55.6 μM, with an apparent V(max) of 0.083 μmol/min. To our knowledge, for the first time, the current work obtains an extracellular Cellulomonas endo-D: -arabinase enzyme that might be potentially served as a tool enzyme for hydrolyzing specific cell wall such as Mycobacterium cell. It is purified as an important potential initial material basis for mass spectrometric sequencing and chemical gene synthesis. It may make it possible to clone and express this valuable endo-D: -arabinase and make it available to the mycobacteria scientific community.
authors: Department of Biotechnology, Dalian Medical University, 9 West Lvshun Southern Road, 116044, Dalian, China.
authors: Ming-Zhong
authors: MZ
authors: Sun
authors: Xiao-Ying
authors: XY
authors: Zhang
authors: Yi
authors: Y
authors: Xin
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D040143:Cellulomonas / Q000737:chemistry / Q000201:enzymology*; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000302:isolation & purification*; D007700:Kinetics; D008970:Molecular Weight; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; C074451:Endo-arabinase; D006026:Glycoside Hydrolases