Isolation and characterization of a gene encoding meso-diaminopimelate dehydrogenase fromGlycine max

Wenko, L.K.; Treick, Ronald W.; Wilson, Kenneth G.

doi:10.1007/bf02418236

Cited by 23 publications

(15 citation statements)

References 25 publications

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“…There are other examples in which cloned animal or plant genes have been shown to complement bacterial mutations (31)(32)(33)(34)(35)(36)(37)(38). These examples indicate the feasibility of cloning genes from higher eukaryotes by passing expression libraries through bacterial mutants and selecting for complementation.…”

Section: Discussionmentioning

confidence: 99%

Functional expression of plant acetolactate synthase genes in Escherichia coli

Smith

Schloss²,

Mazur³

1989

Proc. Natl. Acad. Sci. U.S.A.

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Acetolactate synthase (ALS; EC 4.1.3.18) is the first common enzyme in the biosynthetic pathways leading to leucine, isoleucine, and valine. It is the target enzyme for three classes of structurally unrelated herbicides, the sulfonylureas, the imidazolinones, and the triazolopyrimidines. A cloned ALS gene from the small cruciferous plant Arabidopsis thaliana has been fused to bacterial transcription/translation signals and the resulting plasmid has been used to transform Escherichia coli. The cloned plant gene, which includes sequences encoding the chloroplast transit peptide, is functionally expressed in the bacteria. It is able to complement genetically a strain ofE. coli that lacks endogenous ALS activity. An ALS gene cloned from a line of Arabidopsis previously shown to be resistant to sulfonylurea herbicides has been similarly expressed in E. coli. The herbicide-resistance phenotype is expressed in the bacteria, as assayed by both enzyme activity and the ability to grow in the presence of herbicides. This system has been useful for purifying substantial amounts of the plant enzyme, for studying the sequence parameters involved in subcellular protein localization, and for characterizing the interactions that occur between ALS and its various inhibitors.

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Section: Discussionmentioning

confidence: 99%

Functional expression of plant acetolactate synthase genes in Escherichia coli

Smith

Schloss²,

Mazur³

1989

Proc. Natl. Acad. Sci. U.S.A.

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“…The most-known D-AADH is meso-diaminopimelate dehydrogenase (meso-DAPDH; EC 1.4.1.16), which is a key enzyme in the lysine biosynthetic pathway and has been found in bacteria, such as Bacillus sphaericus (17) and Corynebacterium glutamicum (14). In addition, meso-DAPDH has also been isolated from plants, for example, soybeans (Glycine max) (24). meso-DAPDH is NADP ϩ dependent and catalyzes the reversible oxidative deamination on the D-configuration center of meso-2,6-diaminopimelate (meso-DAP) to yield L-2-amino-6-oxopimelate (17).…”

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confidence: 99%

A Novel meso -Diaminopimelate Dehydrogenase from Symbiobacterium thermophilum: Overexpression, Characterization, and Potential for d -Amino Acid Synthesis

Gao

Chen

Liu

et al. 2012

Appl Environ Microbiol

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meso-Diaminopimelate dehydrogenase (meso-DAPDH) is an NADP؉ -dependent enzyme which catalyzes the reversible oxidative deamination on the D-configuration of meso-2,6-diaminopimelate to produce L-2-amino-6-oxopimelate. In this study, the gene encoding a meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum was cloned and expressed in Escherichia coli. In addition to the native substrate meso-2,6-diaminopimelate, the purified enzyme also showed activity toward Dalanine, D-valine, and D-lysine. This enzyme catalyzed the reductive amination of 2-keto acids such as pyruvic acid to generate D-amino acids in up to 99% conversion and 99% enantiomeric excess. Since meso-diaminopimelate dehydrogenases are known to be specific to meso-2,6-diaminopimelate, this is a unique wild-type meso-diaminopimelate dehydrogenase with a more relaxed substrate specificity and potential for D-amino acid synthesis. The enzyme is the most stable meso-diaminopimelate dehydrogenase reported to now. Two amino acid residues (F146 and M152) in the substrate binding sites of S. thermophilum meso-DAPDH different from the sequences of other known meso-DAPDHs were replaced with the conserved amino acids in other meso-DAPDHs, and assay of wild-type and mutant enzyme activities revealed that F146 and M152 are not critical in determining the enzyme's substrate specificity. The high thermostability and relaxed substrate profile of S. thermophilum meso-DAPDH warrant it as an excellent starting enzyme for creating effective D-amino acid dehydrogenases by protein engineering. N ot only do D-amino acids serve as specialized components of many types of machineries in living organisms, such as neural signaling (20), and bacterial cell walls (19), but they also are important components or building blocks in the production of pharmaceuticals and other fine chemicals (4, 5, 12). As such, many methods for the synthesis of D-amino acids and their derivatives have been developed. Just as L-amino acid dehydrogenases are useful for preparation of L-amino acids from the corresponding 2-keto acids (6,7,13,21), the use of D-amino acid dehydrogenase (D-AADH) should offer a straightforward approach, in which the enzyme catalyzes the reductive amination of 2-keto acid to give D-amino acid. However, NAD(P)H-dependent D-amino acid dehydrogenase is much less abundant in nature than its L-amino acid counterpart and largely unexplored. The most-known D-AADH is meso-diaminopimelate dehydrogenase (meso-DAPDH; EC 1.4.1.16), which is a key enzyme in the lysine biosynthetic pathway and has been found in bacteria, such as Bacillus sphaericus (17) and Corynebacterium glutamicum (14). In addition, meso-DAPDH has also been isolated from plants, for example, soybeans (Glycine max) (24). meso-DAPDH is NADP ϩ dependent and catalyzes the reversible oxidative deamination on the D-configuration center of meso-2,6-diaminopimelate (meso-DAP) to yield L-2-amino-6-oxopimelate (17). However, previously reported meso-DAPDHs are generally specific toward meso-DAP and showed only very ...

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“…One unit of enzyme is defined as the amount that catalyses an increase of absorbance of 0.001 OD/min, after attaining maximum velocity. Dihydrodipicolinate reductase (EC 1.3.1.3 C) was measured by the oxidation of N A D P H using a reaction mixture similar to that of Farkas and Gilvarg [25] employing synthetic dihydrodipicolinate. The activity of this enzyme was verified by its susceptibility to inhibition with dipicolinic acid, a known inhibitor of the reductase.…”

Section: Enzyme Assaysmentioning

confidence: 99%

“…This enzyme would be characteristic of the long version of the standard pathway. However, Wenko et al [25] report m-DAP dehydrogenase to be a component of soybean (Glycine max), which would favor the shortened biosynthesis.…”

Section: Introductionmentioning

confidence: 99%

Biosynthesis of lysine in plants: the putative role of meso-diaminopimelate dehydrogenase

Chatterjee

Singh²,

Gilvarg

1994

Plant Mol Biol

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Extracts from Chlamydomonas, corn, soybean and tobacco were tested for enzymes of the lysine biosynthetic pathway. Dihydrodipicolinic acid (DHD) synthase, DHD reductase, diaminopimelate (DAP) epimerase and DAP decarboxylase were present in all. However, in contrast to the report of Wenko et al., meso-DAP dehydrogenase could not be detected in extracts prepared from soybean. Moreover, it was not found in Chlamydomonas, corn and tobacco as well. In order to set an upper limit to the amount of meso-DAP dehydrogenase that might be present, reconstruction experiments were performed with soybean and corn extracts in which the conversion of dihydrodipicolinate to lysine was made dependent on the addition of limited amounts of the meso-DAP dehydrogenase purified from Bacillus sphaericus. The presence of DAP epimerase and the absence of meso-DAP dehydrogenase indicates that the meso-DAP dehydrogenase abbreviated pathway for lysine synthesis is not operative in plants.

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Isolation and characterization of a gene encoding meso-diaminopimelate dehydrogenase fromGlycine max

Cited by 23 publications

References 25 publications

Functional expression of plant acetolactate synthase genes in Escherichia coli

Functional expression of plant acetolactate synthase genes in Escherichia coli

A Novel meso -Diaminopimelate Dehydrogenase from Symbiobacterium thermophilum: Overexpression, Characterization, and Potential for d -Amino Acid Synthesis

Biosynthesis of lysine in plants: the putative role of meso-diaminopimelate dehydrogenase

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