• BMB Reports
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• v.32 no.2
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• pp.210-213
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• 1999

In Serratia marcescens, acetolactate produced by the catabolic acetolactate synthase (ALS) is converted into acetoin, its physiological role of which is to maintain intracellular pH homeostasis. In this study, the expression mode of catabolic ALS by aeration and branched-chain amino acids was examined by the ELISA method. The amount of catabolic ALS decreased approximately 93% under aerobic conditions. We also showed that the expression of catabolic ALS decreased approximately 34 % and 65 % in the presence of 2.5 mM and 10 mM leucine, respectively. The repression of catabolic ALS by leucine has not been reported previously. In contrast to leucine, catabolic ALS levels increased approximately 13% and 38% by treatment with 2.5 mM and 10 mM isoleucine, respectively, while valine alone did not have any significant effect on the synthesis of catabolic ALS. The amount of catabolic ALS was also reduced to approximately 32% and 45% in the presence of 10 mM Leu+Ile and Leu+Ile+Val, respectively. The regulatory mode of the Serratia catabolic ALS suggests that catabolic ALS may also have a role in supplying acetolactate as an intermediate of valine and leucine biosynthesis in addition to the maintenance of internal pH.

• BMB Reports
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• v.31 no.1
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• pp.37-43
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• 1998

The catabolic ${\alpha}$-acetolactate synthase was purified to homogeneity from Serratia marcescens ATCC 25419 using ammonium sulfate fractionation, DEAE-Sepharose, Phenyl-Sepharose, and Hydroxylapatite column chromatography. The native molecular weight of the enzyme was approximately 150 kDa and composed of two identical subunits with molecular weights of 64 kDa each. The N-terminal amino acid sequence of the enzyme was determined to be Ala-Gln-Glu-Lys-Thr-Gly-Asn-Asp-Trp-Gln-His-Gly-Ala-Asp-Leu-Val-Val-Lys-Asn-Leu. It was not inhibited by the branched chain amino acids and sulfometuron methyl herbicide. The optimum pH of the enzyme was around pH 5.5 and the pI value was 6.1. The catabolic ${\alpha}$-acetolactate synthase showed weak immunological relationships with recombinant tobacco ALS, barley ALS, and the valine-sensitive ALS isozyme from Serratia marcescens.