Mutation Analysis of the Feedback Inhibition Site of Aspartokinase III of Escherichia coli K-12 and its Use in L-Threonine Production

Ogawa-Miyata, Yuri; Kojima, Hiroyuki; Sano, Konosuke

doi:10.1271/bbb.65.1149

Cited by 49 publications

(29 citation statements)

References 13 publications

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“…Escherichia coli strains were grown at 37 C in L broth with appropriate antibiotics, 15) and L-lysine production was evaluated in medium 1, as described previously. 16) Medium 1 contained 50 g/l glucose, 30 g/l CaCO 3 , and 100 mg/l streptomycin. C. glutamicum was grown at 31.5 C in CM2G medium, 17) and L-lysine production was evaluated in modified CM2G medium, which contained 100 g/l glucose, 50 g/l CaCO 3 , and, optionally, 5 mg/l kanamycin.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Unique MutantlysEGene, Originating fromCorynebacterium glutamicum, Encoding a Product That InducesL-Lysine Production inMethylophilus methylotrophus

Gunji

Ito

Hara

et al. 2006

Bioscience, Biotechnology, and Biochemistry

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Section: Methodsmentioning

confidence: 99%

Characterization of a Unique MutantlysEGene, Originating fromCorynebacterium glutamicum, Encoding a Product That InducesL-Lysine Production inMethylophilus methylotrophus

Gunji

Ito

Hara

et al. 2006

Bioscience, Biotechnology, and Biochemistry

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“…The ACT domain has been found in enzymes that are regulated by amino acids or purines and is involved in regulation by these effectors (Chipman and Shaanan, 2001). In Figure 5, only aspartate kinases for which single mutations affecting enzymatic activity or regulation have been described (Arévalo-Rodríguez et al, 1999;Bareich and Wright, 2003;Farfán, 1997;Follettie et al, 1993;Hernando-Rico et al, 2001;Kalinowski et al, 1991;Kikuchi et al, 1999;Marco-Marin et al, 2003;Martín-Rendón et al, 1993;Ogawa-Miyata et al, 2001;Ohnishi et al, 2002;Omori et al, 1993;Paris et al, 2003) are shown. Most but not all of those mutations affecting regulation are located in the ACT domains.…”

Section: Sequencing Of the Hom3-ts31d Hom3-r7 And Hom3-87wt Allelesmentioning

confidence: 99%

“…A few of them are in the amino acid kinase domain, viz. E282D and E279A in S. cerevisiae (this work; Bareich and Wright, 2003) (12) I522A (12) Q524 (12) AtAK-HD S301Y (8) G345D (8) AlAK E164K (6) K8R (5) E119D (5) R198K (5) D202E (5) E250K (7) G323D (7,6) M318I (6) L325F (6) T352I (7,6) V347M (6) T344M (7) S345L (7,6) EcAKIII S301Y (9) T311Y ( Bareich and Wright, 2003;(2) Arévalo-Rodríguez et al, 1999;(3) Martín-Rendón et al, 1993;(4) Farfán, 1997;(5) Marco-Marin et al, 2003;(6) Ogawa-Miyata et al, 2001;(7) Kikuchi et al, 1999;(8) Hernando-Rico et al, 2001;(9) Kalinowski et al, 1991;(10) Ohnishi et al, 2002;(11) Follettie et al, 1993;(12) Paris et al, 2003;(13) Omori et al, 1993Omori et al, et al, 1999Marco-Marin et al, 2003;O...…”

Section: Sequencing Of the Hom3-ts31d Hom3-r7 And Hom3-87wt Allelesmentioning

confidence: 99%

A new mutation in the yeast aspartate kinase induces threonine accumulation in a temperature‐regulated way

Velasco

Arévalo-Rodrı́guez

Marina

et al. 2005

Yeast

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In Saccharomyces cerevisiae, aspartate kinase (the HOM3 product) regulates the metabolic flux through the threonine biosynthetic pathway through feedback inhibition by the end product. In order to obtain a strain able to produce threonine in a controlled way, we have isolated a mutant allele (HOM3-ts31d ) that gives rise to a deregulated aspartate kinase. This allele has been isolated as an extragenic suppressor of ilv1, which confers an Ilv + phenotype at 37 • C but not at 22 • C. We have stated that at high temperature the mutant aspartate kinase is slightly more deregulated and shows a higher specific activity, inducing threonine accumulation. The HOM3-ts31d allele carries a mutation that leads to a Ser399 → Phe substitution in the postulated regulatory region of the enzyme. We have detected other changes in the nucleotide sequence but they are also present in the parental strain, reflecting the genetic differences between different wild-type strains. A sequence comparison among all the reported mutant aspartate kinases suggests that not all residues involved in regulation of the activity are clustered in the so-called regulatory domain, as is the case of that mutated in AK-R7, another deregulated aspartate kinase obtained with the same strategy of ilv1 suppression.

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“…Asterisks show the known mutation sites of AKs desensitized to L-lysine. 46,48,49) The sequence motifs, 36) and D-P-R, 35) are indicated above the sequences by bold lines. The black triangle indicates the mutation found in G49.…”

Section: Characterization Of Enzymes Of the L-lysine Biosynthetic Patmentioning

confidence: 99%

“…By mutation analysis of AKIII of E. coli, OgawaMiyata et al 49) showed that two areas (corresponding to amino acid residues 318-325 and 345-352) in the Cterminal domain of AKIII are highly involved in feedback regulation by L-lysine. Also, in Serratia marcescens aspartokinase I-homoserine dehydrogenase I (AKI-HDI), it has been reported that mutations in the region corresponding to amino acid residues 300-449 of E. coli AKIII resulted in the AKI-HDI insensitive to L-threonine as allosteric effector.…”

Section: )mentioning

confidence: 99%

Characterization of theL-Lysine Biosynthetic Pathway in the Obligate MethylotrophMethylophilus methylotrophus

Gunji

Tsujimoto

Shimaoka

et al. 2004

Bioscience, Biotechnology, and Biochemistry

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The L-lysine biosynthetic pathway of the gramnegative obligate methylotroph Methylophilus methylotrophus AS1 was examined through characterization of the enzymes aspartokinase (AK), aspartsemialdehyde dehydrogenase, dihydrodipicolinate synthase (DDPS), dihydrodipicolinate reductase, and diaminopimelate decarboxylase. The AK was inhibited by L-threonine and by a combination of L-threonine and L-lysine, but not by L-lysine alone, and the activity of DDPS was moderately reduced by L-lysine. In an L-lysine producing mutant (G49), isolated as an S-(2-aminoethyl)-Lcysteine (lysine analog) resistant strain, both AK and DDPS were partially resistant to feedback inhibition. The ask and dapA genes encoding AK and DDPS respectively were isolated from the parental strain, AS1, and its G49 derivative. Comparison of the sequences revealed a point mutation in each of these genes in G49. The mutation in the ask gene altered aspartic acid in a key region involved in the allosteric regulation common to AKs, while a novel mutation in the dapA gene altered tyrosine-106, which was assumed to be involved in the binding of L-lysine to DDPS.

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Mutation Analysis of the Feedback Inhibition Site of Aspartokinase III of Escherichia coli K-12 and its Use in L-Threonine Production

Cited by 49 publications

References 13 publications

Characterization of a Unique MutantlysEGene, Originating fromCorynebacterium glutamicum, Encoding a Product That InducesL-Lysine Production inMethylophilus methylotrophus

Characterization of a Unique MutantlysEGene, Originating fromCorynebacterium glutamicum, Encoding a Product That InducesL-Lysine Production inMethylophilus methylotrophus

A new mutation in the yeast aspartate kinase induces threonine accumulation in a temperature‐regulated way

Characterization of theL-Lysine Biosynthetic Pathway in the Obligate MethylotrophMethylophilus methylotrophus

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