Purification and Characterization of the Anabolic Acetolactate Synthase III from Serratia marcescens ATCC 25419

The anabolic acetolactate synthase III was purified to homogeneity from Serratia marcescens using DEAE-Sepharose, Phenyl-Sepharose, and hydroxylapatite column chromatography The native molecular weight of the enzyme was approximately 165 kDa. The enzyme is composed of two large and two small subunits with molecular weights of 64 and 15 kDa, respectively. The N-terminal sequence of the large and small subunit of the enzyme was Ser-Ala-Thr-Pro-Gln-Pro-Ser-Thr-Arg-Phe-Thr-Cys-Ala-Gln-Leu-Ile-Ala-His-Leu and Met-Leu-Gln-Pro-Gln-Asp-Lys-Pro-Gln-Val-Ile-Leu-Glu-Leu-Ala-Val-Arg-Asn-His-Pro-Gly-Val-Met-Ser-His-Val, respectively. The optimum pH and pI value were 7.5 and 5.5, respectively The $IC_{50}$ values were $20\;{\mu}M$ and $14\;{\mu}M$ for valine and herbicide SU7, respectively. The substrate specificity ratio, R value, was determined to be approximately 40, which suggests that this enzyme prefers the formation of $\alpha$-aceto-$\alpha$-hydroxybutyrate leading to the synthesis of isoleucine.