• BMB Reports
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• v.30 no.5
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• pp.332-336
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• 1997

The stimulatory effects of leucine on the activities of two soluble forms of brain glutamate dehydrogenase isoproteins (GDH I and GDH II) have been studied at various conditions. There were significant differences between GDH I and GDH II in their sensitivities to the action of leucine. When the effects of varied leucine concentrations on GDH activities were studied in the direction of reductive amination of 2-oxoglutarate with NADPH as a coenzyme, a marked activation was observed for both isoproteins at leucine concentrations up to 10 mM, whereas both isoproteins showed activation to a lesser extent with NADH as a coenzyme. The stimulatory effects of leucine on GDH activities in the direction of the oxidative deamination of glutamate were also observed, but to a much lesser extent. Leucine relieved the inhibition of GDH I by GTP and this resulted in an increase in the apparent activation by leucine in the presence of GTP. 2-Oxoglutarate was found to give rise to high substrate inhibition and leucine significantly reduced the substrate inhibition in the presence of $200\;{\mu}M$ NADH. Thus, the effects of leucine might be composed of a direct effect on the enzyme together with a relief of high substrate inhibition.

• Journal of Microbiology
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• v.44 no.1
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• pp.50-53
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• 2006

Cys-29 and Cys-251 of Streptomyces albus valine dehydrogenase(ValDH) were highly conserved in the corresponding region of $NAD(P)^+$-dependent amino acid dehydroganase sequences. To ascertain the functional role of these cysteine residues in S. albus ValDH, site-directed mutagenesis was performed to change each of the two residues to serine. Kinetic analyses of the enzymes mutated at Cys-29 and Cys-251 revealed that these residues are involved in catalysis. We also constructed mutant ValDH by substituting valine for leucine at 305 by site-directed mutagenesis. This residue was chosen, because it has been proposed to be important for substrate discrimination by phenylalanine dehydrogenase (PheDH) and leucine dehydrogenase (LeuDH). Kinetic analysis of the V305L mutant enzyme revealed that it is involved in the substrate binding site. However it displayed less activity than the wild type enzyme toward all aliphatic and aromatic amino acids tested.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.30 no.4
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• pp.405-411
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• 1985

The buffer soluble proteins were extracted from six cultivars of barley grains and analyzed by various electrophoresis; 7.5% polyacrylamide slab gel, 2-30% polyacrylamide porosity gradient tube gel, isoelectric focusing (pH4-9) and starch gel electrophoresis. The proteins, esterase, acid phosphatase, malate dehydrogenase, glutamate dehydrogenase and leucine aminopeptidase were investigated to find out the best method to differentiate barley cultivars. The result were that protein and esterase bands in 2-30% polyacrylamide porosity gradient tube gel electrophoresis and protein bands in 7.5% polyacrylamide slab gel electrophoresis showed typical varietal differences. Therefore, those methods were suitable for differentiation of barley cultivars. It was difficult to differentiate the cultivars by the other methodes and patterns of the other enzymes.

• Molecules and Cells
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• v.45 no.7
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• pp.495-501
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• 2022

Leucine dehydrogenase (LDH, EC 1.4.1.9) catalyzes the reversible deamination of branched-chain L-amino acids to their corresponding keto acids using NAD⁺ as a cofactor. LDH generally adopts an octameric structure with D4 symmetry, generating a molecular mass of approximately 400 kDa. Here, the crystal structure of the LDH from Pseudomonas aeruginosa (Pa-LDH) was determined at 2.5 Å resolution. Interestingly, the crystal structure shows that the enzyme exists as a dimer with C2 symmetry in a crystal lattice. The dimeric structure was also observed in solution using multiangle light scattering coupled with size-exclusion chromatography. The enzyme assay revealed that the specific activity was maximal at 60℃ and pH 8.5. The kinetic parameters for three different amino acid and the cofactor (NAD⁺) were determined. The crystal structure represents that the subunit has more compact structure than homologs' structure. In addition, the crystal structure along with sequence alignments indicates a set of non-conserved arginine residues which are important in stability. Subsequent mutation analysis for those residues revealed that the enzyme activity reduced to one third of the wild type. These results provide structural and biochemical insights for its future studies on its application for industrial purposes.

• The Korean Journal of Mycology
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• v.17 no.1
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• pp.35-38
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• 1989

Gene libraries of DNA from Flammulina velutipes were constructed using Escherichia coli plasmid pBR 322. Leu 2 gene coding ${\beta}-isopropylmalate$ dehydrogenase from F. velutipes was cloned by complementation of leucine requiring mutant of E. coli. The size of inserted DNA fragment of this clone is about 1 Kbp. The fragment has Bam H1 and Ava 1 restriction sites.

• The Korean Journal of Mycology
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• v.17 no.1
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• pp.27-30
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• 1989

F. velutipes Leu 2 gene (${\beta}-isopropylmalate$ dehydrogenase gene) was used for transformation of P. florida leucine requiring auxotrophic mutant P101. Transformation frequency was very low but the transformed colony can grow on minimal medium very slowly. Transformation was identified by Southern hybridization and reverse transformation into E. coli using chromosome DNA isolated from transformed P. florida.

• Nutrition Research and Practice
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• v.11 no.2
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• pp.121-129
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• 2017

BACKGROUND/OBJECTIVES: The study was conducted to evaluate the effects of dietary leucine supplementation on mitochondrial biogenesis and energy metabolism in the liver of normal birth weight (NBW) and intrauterine growth-retarded (IUGR) weanling piglets. MATERIALS/METHODS: A total of sixteen pairs of NBW and IUGR piglets from sixteen sows were selected according to their birth weight. At postnatal day 14, all piglets were weaned and fed either a control diet or a leucine-supplemented diet for 21 d. Thereafter, a $2{\times}2$ factorial experimental design was used. Each treatment consisted of eight replications with one piglet per replication. RESULTS: Compared with NBW piglets, IUGR piglets had a decreased (P < 0.05) hepatic adenosine triphosphate (ATP) content. Also, IUGR piglets exhibited reductions (P < 0.05) in the activities of hepatic mitochondrial pyruvate dehydrogenase (PDH), citrate synthase (CS), ${\alpha}$-ketoglutarate dehydrogenase (${\alpha}$-KGDH), malate dehydrogenase (MDH), and complexes I and V, along with decreases (P < 0.05) in the concentration of mitochondrial DNA (mtDNA) and the protein expression of hepatic peroxisome proliferator-activated receptor-${\gamma}$ coactivator $1{\alpha}$ (PGC-$1{\alpha}$). Dietary leucine supplementation increased (P < 0.05) the content of ATP, and the activities of CS, ${\alpha}$-KGDH, MDH, and complex V in the liver of piglets. Furthermore, compared to those fed a control diet, piglets given a leucine-supplemented diet exhibited increases (P < 0.05) in the mtDNA content and in the mRNA expressions of sirtuin 1, PGC-$1{\alpha}$, nuclear respiratory factor 1, mitochondrial transcription factor A, and ATP synthase, $H^+$ transporting, mitochondrial F1 complex, ${\beta}$ polypeptide in liver. CONCLUSIONS: Dietary leucine supplementation may exert beneficial effects on mitochondrial biogenesis and energy metabolism in NBW and IUGR weanling piglets.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.1
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• pp.53-58
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• 2012

The aim of this study was to investigate the antimicrobial, antiperspirant and deodorant effect of herb mixtures including Schisandra chinensis(SC). This herb mixtures including Schisandra chinensis was made of Astragalus membranaceus(AM), Triticum aestivum(TA), Atractylodes japonica(AJ) and peppermint essential oil extracted from Mentha piperita(MP). We examined antimicrobial activity, antiperspirant activity and L-leucine dehydrogenase activity of the herbal extracts. We found that the mixture of SC and MP have the best antimicrobial effects on B. subtilis subsp. spizizenii and mixture of SC, AJ, AM and TA have best antiperspirant effect. Moreover, the combination of SC and AM showed the most inhibitory effect on L-leucine dehydrogenase activity. These results suggested that the herb mixtures Including Schisandra chinensis have good antimicrobial and antiperspirant effects.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.331-340
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• 1995

Interstrain polymorphisms of isoenzyme profiles and mitochondrial (Mt) DNA fingerprints were observed among seven strains of Acnnthnmoeba isolated from different sources and morphologically assigned to A. polvphngn. Mt DNA ringerprints by eight restriction endonucleases (Bgl II, Sca I, Cla I, EcoR I, Xbo I, Kpn I, Sal I, and Sst I) revealed considerable interstrain polymorphisms . Isoenzyme profiles revealed considerable interstrain polymorphisms for acid phosphatase, lactate dehydrogenase, and glucose-6- phosphate dehydrogenase while those for glucose phosphate isomerase , leucine aminopeptidase , and malate dehydrogenase showed similarity Despite of the interstrain polymorphisms, the isoengyme profiles and Mt DNA fingerprints of the strain Ap were found to be identical with those of the strain .tones . Mt DNA fingerprinting was found to be highly applicable for the strain identification, characterization, and differentiation. Key words: Acanthnmoebn polyphcga, interstrain polymorphism, isoenzyme profiles , Mt DNA fingerprints, strain differentiation, strain identification.

• Journal of Nutrition and Health
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• v.42 no.8
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• pp.714-722
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• 2009

The principal objective of this study was to determine the effects of leucine on body weight reduction in high fat diet-induced overweight rats. To induce overweight, six-month-old male Sprague-Dawley rats (n = 80) were divided into 8 groups; one group of 10 rats was fed on a normal fat diet and the remaining 70 rats were fed on a high-fat diet (40% of energy as fat) for 14 weeks. Then, 10 rats fed on the normal fat diet and another 10 rats fed on the high fat diet were sacrificed to identify overweight induction. The remaining 60 rats were divided randomly into 6 groups according to body weight and fed on one of the diets with different dietary fat levels (9.6% or 40% of energy as fat) and leucine levels (0, 0.6 or 1.2 g/kg BW) for the following 5 weeks of experiments. The body weight loss in the Leu-administered groups (0.6 g, 1.2 g/kg BW) was significantly higher than those of Leu non-administered groups. The perirenal fat pad weights in the Leu-administered groups were significantly lower than those of the Leu non-administered groups. Of the hepatic enzymes, glucose-6-phosphate dehydrogenase (G6PDH) activities were reduced significantly in the Leu-administered groups than in the Leu non-administered groups. With the oral glucose tolerance test (OGTT), the incremental areas under the curve of the glucose response (IAUC) of the Leu-administered groups were significantly lower than those of the Leu non-administered groups. The fasting glucose concentration and HOMA-IR of the Leu-administered groups were significantly lower than those of the Leu non-administered groups. In conclusion, the results of this study suggest that one of the possible mechanisms of leucine in the observed body weight reduction might involve the inhibition of lipogenic enzyme activities such as glucose-6-phosphate dehydrogenase, rather than the activation of lipolysis enzymes. Additionally, leucine adminstration resulted in improved glucose metabolism.