Inclusion Body를 형성한 B-Glucosidase의 Chaperonin에 의한 활성 향상

B-Glucosidaes from Cellvibrio gilvus(CG) was successfully overproduced in soluble form in E. coli with the coexpression of GroEL/ES/. Without the GroEL/ES protein, the B-glucosidase overexpressed in E. coli constituted a huge amount(80%) of total cellular protein, but was localized in the insoluble...

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Bibliographic Details
Published inKSBB Journal Vol. 14; no. 4; pp. 429 - 433
Main Authors 김종덕, 하순덕, 공재열
Format Journal Article
LanguageKorean
Published 30.08.1999
Subjects
chaperonin system
Cellvibrio gilvus
GroEL/ES
B-glucosidase; pET28(a) vector
Agrobacterium tumefaciens
coexpression
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Summary:B-Glucosidaes from Cellvibrio gilvus(CG) was successfully overproduced in soluble form in E. coli with the coexpression of GroEL/ES/. Without the GroEL/ES protein, the B-glucosidase overexpressed in E. coli constituted a huge amount(80%) of total cellular protein, but was localized in the insoluble fraction, and little activity was detected in the soluble fraction. Coexpression of the E. coli GroEL/ES had a drastic impact on the proper folding of the B-glucosidase; 20% of the overexpressed enzyme was recovered in the soluble fraction in active form. Similar effects of GroEL/ES were also observed on the overexpressed -glucosidase from Agrobacterium tumefaciens(AT). And pET28(a)-RGRAR, partially deleted mutant lacking 5-amino acid residues at carboxy teminus also could be folded into an active form when expressed with the molecular chaperonin GroEL/ES, and its activity was higher than that of the without GroEL/ES system, In addition, the synergistic effect of GroEL/ES and the low induction temperature were important factors for solubilization of the inclusion body from overproduced -glucosidases.
ISSN:1225-7117