Literature summary for 4.2.1.84 extracted from

Kang, M.S.; Han, S.S.; Kim, M.Y.; Kim, B.Y.; Huh, J.P.; Kim, H.S.; Lee, J.H.
High-level expression in Corynebacterium glutamicum of nitrile hydratase from Rhodococcus rhodochrous for acrylamide production (2014), Appl. Microbiol. Biotechnol., 98, 4379-4387 .

Application

Application	Comment	Organism
synthesis	use of recombinant Corynebacterium cells for the production of acrylamide from acrylonitrile results in a conversion yield of 93% and a final acrylamide concentration of 42.5% within 6 h when the total amount of fed acrylonitrile is 456 g	Rhodococcus rhodochrous

Cloned(Commentary)

Cloned (Comment)	Organism
the nhhBAG gene of Rhodococcus rhodochrous M33 that encodes nitrile hydratase is cloned and expressed in Corynebacterium glutamicum under the control of an ilvC promoter. To overexpress the nitrile hydratase, five types of plasmid variants are constructed by introducing mutations into 80 nucleotides near the translational initiation region of nhhB. Of them, pNBM4 with seven mutations shows the highest NHase activity, exhibiting higher expression levels of NhhB and NhhA than wild-type pNBW33, mainly owing to decreased secondary-structure stability and an introduction of a conserved Shine-Dalgarno sequence in the translational initiation region. In a fed-batch culture of recombinant Corynebacterium cells harboring pNBM4, the cell density reaches 53.4 g dry cell weight/l within 18 h, and the specific and total enzyme activities are estimated to be 0.0373 mM/min/mg dry cell weight and 1.992 mM/min/ml, respectively	Rhodococcus rhodochrous

Cloned (Comment)

Organism

the nhhBAG gene of Rhodococcus rhodochrous M33 that encodes nitrile hydratase is cloned and expressed in Corynebacterium glutamicum under the control of an ilvC promoter. To overexpress the nitrile hydratase, five types of plasmid variants are constructed by introducing mutations into 80 nucleotides near the translational initiation region of nhhB. Of them, pNBM4 with seven mutations shows the highest NHase activity, exhibiting higher expression levels of NhhB and NhhA than wild-type pNBW33, mainly owing to decreased secondary-structure stability and an introduction of a conserved Shine-Dalgarno sequence in the translational initiation region. In a fed-batch culture of recombinant Corynebacterium cells harboring pNBM4, the cell density reaches 53.4 g dry cell weight/l within 18 h, and the specific and total enzyme activities are estimated to be 0.0373 mM/min/mg dry cell weight and 1.992 mM/min/ml, respectively

Rhodococcus rhodochrous

Organism

Organism	UniProt	Comment	Textmining
Rhodococcus rhodochrous	-	-	-
Rhodococcus rhodochrous M33	-	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	the specific enzyme activity of the enzyme from Rhodococcus rhodochrous M33 recombinantly expressed in Corynebacterium glutamicum glutamicum cells is about 0.0136 mM/min/mg dry cell weight (DCW)	Rhodococcus rhodochrous

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
acrylonitrile + H2O	-	Rhodococcus rhodochrous	acrylamide	-	?
acrylonitrile + H2O	-	Rhodococcus rhodochrous M33	acrylamide	-	?