Expression, purification and characterisation of chondroitinase AC II with glyceraldehyde-3-phosphate dehydrogenase tag and chaperone (GroEs-GroEL) from Arthrobacter sp. CS01.

In this study, chondroitinase (ChSase) AC II from Arthrobacter sp. CS01 was cloned, expressed in Escherichia coli BL21 (DE3), purified and characterised. To assist in protein folding and improve on high protein aggregation rates, two strategies involving chaperones and fusion tags were chosen to increase enzyme activity and improve enzymatic properties. ChSase AC II enzyme activity increased from 3.12 to 9.15 U/ml with chaperone GroEs-GroEL, and the specific activity increased from 19.8 to 25.74 U/mg with the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) tag. ChSase AC II and GAPDH-ChSase AC II displayed maximum activities at 37 °C and 40 °C, at pH 6.5 and 7.0, respectively. GAPDH-ChSase AC II activity remained above 69.8% after incubation at 40 °C for 120 min, and ChSase AC II activity remained approximately 32.1% under the same conditions, indicating that ChSase AC II thermostability was enhanced by the GAPDH tag. These properties suggested that the enzymes are promising prospects in medical and industrial applications.