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Literature summary extracted from

  • Li, Y.; Liu, S.; You, C.
    Permeabilized Escherichia coli whole cells containing co-expressed two thermophilic enzymes facilitate the synthesis of scyllo-inositol from myo-inositol (2020), Biotechnol. J., 15, e1900191 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.18 gene GK1898, recombinant expression in Escherichia coli strain BL21(DE3) Geobacillus kaustophilus

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.18 additional information thermophilic myo-inositol 2-dehydrogenase (IDH) and scyllo-inositol 2-dehydrogenase (SIDH, EC 1.1.1.370) from Geobacillus kaustophilus are co-expressed in Escherichia coli strain BL21(DE3). The Escherichia coli cells containing the two enzymes are permeabilized by heat treatment (heat treatment at 70°C for 20 min for cell permeabilization) as whole-cell catalysts to convert myo-inositol (MI) to scyllo-inositol (SI). After condition optimizations about permeabilized temperature, reaction temperature, and initial MI concentration, about 82 g/l of SI is produced from 250 g/l of MI within 24 h without any cofactor supplementation. The whole-cell catalytic pathway for SI synthesis is initiated by oxidation of MI to scyllo-inosose catalyzed by cofactor NAD+-dependent IDH. Scyllo-inosose is subsequently reduced to SI by SIDH in the presence of NADH. Recycling of NAD+/NADH is achieved in the whole pathway. The specific activity of SIDH is lower than that of IDH, so SIDH is the rate-limiting step in the two-step cascade reaction. The optimal pH of SIDH is 7.0 and IDH does not become the rate-limited enzyme at pH 7.0. The optimal reaction temperature for SI production is set at 60°C, instabilty and loss of activity at 75°C and above. Method development, overview Geobacillus kaustophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.18 myo-inositol + NAD+ Geobacillus kaustophilus
-
 2,4,6/3,5-pentahydroxycyclohexanone + NADH + H+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.18 Geobacillus kaustophilus Q5KYQ3
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.18 myo-inositol + NAD+
-
 Geobacillus kaustophilus 2,4,6/3,5-pentahydroxycyclohexanone + NADH + H+
-
 r

Synonyms

EC Number Synonyms Comment Organism
1.1.1.18 GK1898
-
 Geobacillus kaustophilus
1.1.1.18 IDH
-
 Geobacillus kaustophilus
1.1.1.18 myo-inositol 2-dehydrogenase
-
 Geobacillus kaustophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.18 55 75 assay at Geobacillus kaustophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.18 7
-
 assay at Geobacillus kaustophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.18 NAD+
-
 Geobacillus kaustophilus
1.1.1.18 NADH
-
 Geobacillus kaustophilus