Malate dehydrogenase (MDH) is an essential metabolic enzyme

which has high economic value in protein research.To find an efficient method for MDH IBs refolding

the cooperation of osmolytes (sucrose

betaine and trehalose) and artificial chaperone (CTAB/ β-CD) in eMDH IBs refolding was investigated.The results showed that when sucrose and betaine were added with artificial chaperone

the refolding of eMDH was facilitated and the activity was twice higher than the refolded eMDH by dilution.But trehalose did not have any influence.Then

the optimal ratio of CTAB to β-CD was decreased from 1∶8 to 1∶6 by the cooperation.As a result

more artificial chaperone could be used to facilitate the refolding with a higher eMDH concentration.Because of the thermo-protection of osmolytes

the refolding of eMDH at a higher temperature (15℃) was achieved.According to structure analysis

the contents of α-helix and dimer structure in refolded eMDH were increased by the cooperation

and eMDH activity was enhanced.