Isolation of Serratia marcescens as a chondroitinase-producing bacterium and purification of a novel chondroitinase AC.
A strain of Serratia marcescens that produced chondroitinase was isolated from soil. It produced a novel chondroitinase AC, which was purified to homogeneity. The enzyme was composed of two identical subunits of 35 kDa as revealed by SDS-PAGE and gel filtration. The isoelectric point for the chondroitinase AC was 7.19. Its optimal activity was at pH 7.5 and 40 degrees C. The purified enzyme was active on chondroitin sulfates A and C and hyaluronic acid, but was not with chondroitin sulfate B (dermatan sulfate), heparin or heparan sulfate. The apparent K(m) and V(max) of the chondroitinase AC for chondroitin sulfate A were 0.4 mg ml(-1) and 85 mmol min(-1) mg(-1), respectively, and for chondroitin sulfate C, 0.5 mg ml(-1) and 103 mmol min(-1) mg(-1), respectively.
Ke
Tao
T
National Laboratory of Grassland Biocontrol Engineering, Sichuan University, Chengdu 610064, China.
Zhangfu
Long
L
Qing
Gao
G
Yong
Tao
T
Hong
Jin
J
Hongyan
Ran
R
Kun
Liu
L
Shigui
Liu
L
D002808:Chondroitin Lyases / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D002809:Chondroitin Sulfates / Q000378:metabolism; D002850:Chromatography, Gel; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D003871:Dermatan Sulfate / Q000378:metabolism; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism; D006820:Hyaluronic Acid / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D021122:Protein Subunits / Q000737:chemistry; D017422:Sequence Analysis, DNA; D012706:Serratia marcescens / Q000145:classification / Q000166:cytology / Q000201:enzymology* / Q000302:isolation & purification*; D012988:Soil Microbiology; D013379:Substrate Specificity; D013696:Temperature
D002808:Chondroitin Lyases; D002809:Chondroitin Sulfates; D004269:DNA, Bacterial; D003871:Dermatan Sulfate; D006493:Heparin; D006497:Heparan Sulfate; D006820:Hyaluronic Acid; D021122:Protein Subunits