Thermal stability and conformational changes of transglutaminase from a newly isolated Streptomyces hygroscopicus.
Thermal stability and conformational changes of transglutaminase (TGase) from a newly isolated Streptomyces hygroscopicus were investigated in this study. The inactivation kinetics of the microbial transglutaminase (MTGase) was fitted using one-step inactivation model. It was much more stable under 40 degrees C. The half-lives for the MTGase at 50 degrees C and 60 degrees C were only 20 min and 8 min, respectively. Spectroscopic studies of the enzyme suggested conformational transition from ordered secondary structural elements (alpha/beta-protein) to unordered structure during thermal denaturation. Some polyols could improve the thermal stability of the enzyme. Among the polyols examined, the prolonged half-lives of 40 min at 50 degrees C and 20 min at 60 degrees C were gained by adding 10% glycerol. The results of differential scanning calorimetric (DSC) analysis showed a distinct transition peak with a significant greater Tm and DeltaH for the MTGase mixed with polyols in comparison with the control, which indicated that the polyols could maintain the natural structure of the enzyme to some extent. The SDS-PAGE electrophoresis of cross-linked casein confirmed that the stabilizers could protect the MTGase from thermal denaturation.
Cui
Li
L
Key Laboratory of Science and Technology of Eco-Textile, Ministry of Education, Southern Yangtze University, Wuxi 214122, China.
Du
Guocheng
G
Zhang
Dongxu
D
Chen
Jian
J
D002152:Calorimetry, Differential Scanning; D002364:Caseins / Q000032:analysis; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability / Q000187:drug effects; D006358:Hot Temperature; D007700:Kinetics; D011108:Polymers / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013302:Streptomyces / Q000187:drug effects / Q000201:enzymology* / Q000302:isolation & purification*; D013816:Thermodynamics; D011503:Transglutaminases / Q000737:chemistry* / Q000378:metabolism*; D044366:Transition Temperature
D002364:Caseins; D011108:Polymers; C024617:polyol; D011503:Transglutaminases