title: In situ demonstration and quantitative analysis of the intrinsic properties of glycoside hydrolases.
abstract: Based on digital image analysis techniques and a series of optimizations in native electrophoresis, a new direct method to simultaneously detect the intrinsic properties of each active component in the enzymatic system of glycoside hydrolase was established. The key technique is that the concentration changes of substrate (or product) on the gel can be determined quantitatively by the gray value changes of the corresponding band after electrophoretic separation. In this manner, the catalytic characteristics of each glycoside hydrolase component were demonstrated in situ and were easily determined after immersing the gel in a series of solutions containing substrates or their derivatives. Because of its high throughput, great sensitivity, and convenient operation, this method can be used to demonstrate the natural diversity of glycoside hydrolases and to study spatial and temporal regulation in multienzyme expression systems. Thus, it is an effective approach to study the functional proteomics of glycoside hydrolases.
authors.affiliation: The State Key Laboratory of Microbial Technology, Shandong University, Jinan, PR China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Zhang
authors.forename: Ning
authors.initials: N
authors.lastname: Liu
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.forename: Jinghua
authors.initials: J
authors.lastname: Li
authors.forename: Lushan
authors.initials: L
authors.lastname: Wang
authors.forename: Guanjun
authors.initials: G
authors.lastname: Chen
authors.forename: Peiji
authors.initials: P
authors.lastname: Gao
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism; D041963:Bacteroidetes; D002266:Carboxymethylcellulose Sodium / Q000378:metabolism; D002480:Cellulase / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000032:analysis / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D017510:Protein Folding; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism; D012680:Sensitivity and Specificity; D014242:Trichoderma; D014990:Xylans / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D005656:Fungal Proteins; D011994:Recombinant Proteins; D014990:Xylans; D006026:Glycoside Hydrolases; D002480:Cellulase; D043364:Endo-1,4-beta Xylanases; D002266:Carboxymethylcellulose Sodium