UNASSIGNED: Three combinations of molecular chaperones from @entity526 , i.e., DnaK-DnaJ-GrpE- @entity26540 - @entity26541 , @entity26540 - @entity26541 and DnaK-DnaJ-GrpE, were overproduced in @entity13724 , and their in vitro stabilizing effects on a @entity7575 hydratase (NHase) were assessed. The optimal gene-combination, @entity526 @entity26540 - @entity26541 (ecgroEL-ES), were introduced into Rhodococcus ruber TH3. A novel engineered strain, R. ruber TH3G was constructed with the native NHase gene on its chromosome and the heterologous ecgroEL-ES genes in a shuttle plasmid. In R. ruber TH3G, NHase activity was enhanced 37.3% compared with the control, TH3. The in vivo stabilizing effect of ecGroEL-ES on the NHase was assessed using both @entity3323 immersion and heat shock experiments. The inactivation behavior of the in vivo NHase after immersion in a solution of dynamically increased concentrations of @entity3323 was particularly evident. When the @entity3323 concentration was increased to 500 g/L (50%), the remaining NHase activity in TH3G was 38%, but in TH3, activity was reduced to 10%. Reactivation of the in vivo NHases after varying degrees of inactivation was further assessed. The activity of the reactivated NHase was more than twofold greater in TH3G than in TH3. The hydration synthesis of @entity3323 catalyzed by the in vivo NHase was performed with continuous acrylonitrile feeding. The final concentration of @entity3323 was 640 g/L when catalyzed by TH3G compared with 490 g/L @entity3323 by TH3. This study is the first to show that the chaperones ecGroEL-ES work well in Rhodococcus and simultaneously possess protein-folding assistance functions and the ability to stabilize and reactivate the native NHases.

Overproduction of the @entity526 chaperones @entity26540 - XXXX in Rhodococcus ruber improves the activity and stability of cell catalysts harboring a @entity7575 hydratase.

@entity26540 :: ('13903475', 'Gene') :: ['GroEL', 'groEL'] @entity7575 :: ('CHEBI:18379', 'Chemical') :: ['nitrile'] @entity26541 :: ('13876916', 'Gene') :: ['groES', 'GroES'] @entity3323 :: ('MESH:D020106', 'Chemical') :: ['acrylamide'] @entity13724 :: ('511693', 'Species') :: ['E. coli BL21'] @entity526 :: ('562', 'Species') :: ['Escherichia coli', 'E. coli']
@entity26541 :: (13876916,Gene) :: ['GroES']
