Purification and characterization of hydrosoluble components from the sap of Chinese lacquer tree Rhus vernicifera.
Continuous gradient elution chromatography (CGEC) was employed to purify and separate enzymes and polysaccharides from the sap of Rhus vernicifera Chinese lacquer tree. There are three different molecules with laccase enzyme activity. Two are enzymes of each other (L1, and L2), whereas the third (RL) is an entirely separate entity. Two polysaccharides (GP1 and GP2) were also found. The Rhus laccase (RL), and isoenzymes L1 and L2, have peak molecular masses of 109,100, 120,000, 103,000 respectively; each has four copper atoms per molecule, and the pI values were 8.2, 8.6, and 9.1, respectively. The structure of the laccases was studied by Fourier-transform infrared (FT-IR) and Matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) mass spectrometry. The typical amide I (1646cm(-1)) and amide II (1545cm(-1)) bands were observed. The results from MALDI-TOF were similar to those from CGEC, but the molecular mass from the MALDI-TOF was significantly different from that obtained from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).
Yun-Yang
Wan
W
College of Resource and Environmental Science, Wuhan University, Wuhan 430072, Hubei, China.
Yu-Min
Du
D
Fang-Xing
Yang
Y
Ying
Xu
X
Rong-Zhi
Chen
C
Kennedy
John F
JF
D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003300:Copper / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D046911:Macromolecular Substances / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D010936:Plant Extracts / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D020033:Protein Isoforms; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017550:Spectroscopy, Fourier Transform Infrared; D014197:Trees / Q000378:metabolism*
D046911:Macromolecular Substances; D010936:Plant Extracts; D010940:Plant Proteins; D011134:Polysaccharides; D020033:Protein Isoforms; D003300:Copper