• BMB Reports
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• 제32권2호
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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.

• Food Science and Biotechnology
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• 제16권3호
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• pp.343-348
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• 2007

Starch contents play important roles in determining the fruit quality. Stawberry accumulates starch in the early stages and then mobilized into soluble sugars during fruit ripening. To date the molecular studies on the ADP-glucose pyrophosphorylase (AGPase), a key enzyme of starch biosynthesis, were not reported. cDNAs encoding small (FagpS) and large (FagpL1 and FaspL2) AGPase subunits were isolated from strawberry (Fragaria ${\times}$ ananassa Duch. cv. Niyobou). Both FagpS and FagpL1 cDNAs have open reading frames deriving 55-58 kDa polypeptides, where FagpL2 contains a partial fragment. Sequence analyses showed that FagpS has a glutamate-threonine-cysteine-leucine (ETCL) instead of a glutamine-threonine-cysteine-leucine (QTCL) motif found in all the dicot plants except for Citrus. In fruits, FagpS and FagpL1 were expressed in all stages with a little change in the amounts of transcripts. In the case of FagpL2, we were not able to detect any signal from all stages of fruit development and all tissues except for very a weak signal from the leaf. The results indicate that FagpL1 and FagpL2 show ubiquitous and leaf-specific expression patterns, respectively. The studies suggest that the starch contents in strawberry might be controlled by the expression of AGPase gene at both the transcriptional and post-transcriptional levels during fruit development.

• Journal of Nutrition and Health
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• 제27권9호
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• pp.910-923
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• 1994

The effects of individual fatty acids, differing in their degree of unsaturation(18:0, 18:1, 18:2 and 18:3) on the biosynthesis and secretion and lipids were investigated in Hep-G2 cells. Synthesis of apolipoprotein was measured by the incorporation of 3H-leucine into apolipoprotein(d<1.21g/ml) and synthesis of lipids was measured by the incorporation of 3H-glycerol and 14C-acetate into various lipid classes. Inclusion of 1.0mM of each fatty acids into the culture medium significantly increased the synthesis of total apolipoprotein and Apo B(p<0.05). However, addition of fatty acid did not affect the synthesis of cellular and medium protein. Among different fatty acids tested, oleic acid had the greatest effect on Apo B synthesis. While stearic, linoleic and linolenic acid, all had similar effects. The secretion of triglyceride into the medium markedly increased in all fatty acid groups being 5-6 times over the albumin control. The triglyceride secretion was the highest int he oleic acid group. The secretion of phospholipid and cholesterol also increased with triglyceride output. A positive relationship existed between the output of lipoprotein-triglyceride and Apo B. Since the synthesis of Apo B was significantly increased when various fatty acids were included into the culture medium, part of the apparently stimulated synthesis of the apolipoprotein may be in response to the increased formation and secretion of lipoprotein lipids.

• BMB Reports
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• 제30권4호
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• pp.274-279
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• 1997

Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of branchedchain amino acids, valine, leucine, and isoleucine. ALS is the target site for several structually diverse classes of herbicides including sulfonylureas, imidazolinones. and triazolopyrimidines. We have purified ALS from etiolated barley shoots to homogeneity. The five major purification steps are ammonium sulfate fractionation, DEAE anion exchange, hydroxylapatite, Bio-Gel A gel filtration, and low pressure Mono-Q chrornatoqraphy. Approximately 170-fold purification was achieved and the yield was 0.45% of initial activity in the crude extract. Both SDS-PAGE and Western blot analysis showed a single polypeptide of ALS with an apparent molecular mass of 64 kDa. The result of nondenaturing gel electrophoresis with activity staining indicated that the molecular mass of its native form is approximately 225 to 250 kDa. The values of $K_m$ for pyruvate. pl. and optimum pH of ALS were determined to be 2.0 mM, 5.2. and 7.0. respectively Feedback inhibition studies showed that ALS is more susceptible to leucine than valine. And $IC_{50}$ value of Cadre, a class of irnidazolinones, is about $1.5\mu{M}$ for ALS.

• BMB Reports
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• 제28권6호
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• pp.471-476
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• 1995

Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of valine, leucine, and isoleucine. ALS is the target enzyme for several classes of structually diverse herbicides. We have synthesized 4,6-dimethoxypyrimidine derivatives as ALS inhibitors, and their inhibitory activities on barley ALS were determined. $IC_{50}$ values for the derivatives are 0.2~200 ${\mu}m$. K11570, the most potent ALS inhibitor with $IC_{50}$ of 0.2 ${\mu}m$, showed mixed-type inhibition with respect to substrate pyruvate, and the progress curves for ALS inhibition by K11570 indicated that the amount of inhibition increased with time. Inhibition-competition experiments were carried out and indicated that three different classes of inhibitors, K11570, a sulfonylurea Ally, and leucine, bind to ALS in a mutually exclusive manner. Chemical modification of tryptophanyl and tyrosyl residues of ALS decreased the sensitivity of ALS to K11570, while cysteine modification did not affect the sensitivity. These results suggest that tryptophanyl and tyrosynyl residues are probably located at or near the inhibitor binding site.

• 한국미생물·생명공학회지
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• 제20권2호
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• pp.115-121
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• 1992

최소 배지에 여러가지 아미노산들을 첨가하여 배양한 Serratia marcescens ATCC 25419 세포 추출물에서 acetolactate syhthase(ALS)의 비활성도를 0.5mM에서 40 증가시킨 반면 8mM에서 60, 20mM에서 90 감소시켰다. Valine은 효소의 비활성도를 2-4 mM에서 20-40 정도 증가시켰고 20mM의 높은 농도에서 10 정도 감소시켰다.

• Journal of Plant Biotechnology
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• 제48권4호
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• pp.228-235
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• 2021

The amino acids found in plants play important roles in protein biosynthesis, signaling processes, and stress responses, and as components in other biosynthesis pathways. Amino acid degradation helps maintain plant cells' energy states under certain carbon starvation conditions. Branched-chain amino acid transferases (BCATs) play an essential role in the metabolism of branched-chain amino acids (BCAAs) such as isoleucine, leucine and valine. In this paper, we performed genome-wide RNA-seq analysis using CsBCAT7-overexpressing Arabidopsis plants. We observed significant changes in genes related to flowering time and genes that are germination-responsive in transgenic plants. RNA-seq and RT-qPCR analyses revealed that the expression levels of some BCAA catabolic genes were upregulated in these same transgenic plants, and that this correlated with a delay in their senescence phenotype when the plants were placed in extended darkness conditions. These results suggest a connection between BCAT and the genes implicated in BCAA catabolism.

• 한국축산식품학회지
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• 제43권6호
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• pp.1044-1054
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• 2023

Growing evidence indicates a crucial role of the gut microbiota in physiological functions. Gut-brain axis imbalance has also been associated with neuropsychiatric and neurodegenerative disorders. Studies have suggested that probiotics regulate the stress response and alleviate mood-related symptoms. In this study, we investigated the effects of the probiotic Lacticaseibacillus rhamnosus IDCC3201 (L3201) on the behavioral response and fecal metabolite content in an unpredictable chronic mild stress (UCMS) mouse model. Our study shows that chronic stress in mice for three weeks resulted in significant changes in behavior, including lower locomotor activity, higher levels of anxiety, and depressive-like symptoms, compared to the control group. Metabolomic analysis demonstrated that disrupted fecal metabolites associated with aminoacyl-tRNA biosynthesis and valine, leucine, and isoleucine biosynthesis by UCMS were restored with the administration of L3201. Oral administration of the L3201 ameliorated the observed changes and improved the behavioral alterations along with fecal metabolites, suggesting that probiotics play a neuroprotective role.

• 한국미생물·생명공학회지
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• 제51권4호
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• pp.551-554
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• 2023

We report the draft genome sequence of Sporobolomyces phaffii RJAF-17, a basidiomycetous yeast strain producing lipoamino acid surfactants, N-palmitoyl leucine and N-parmitoleyl glutamine. The annotation and classification of protein-coding genes provided the basic information for the genome of strain RJAF-17, including prediction of abundant genes as well as detection of genes involved in the biosynthesis of lipoamino acids. With the molecular importance of lipoamino acids as promising alternatives to chemical surfactants, the genomic information of strain RJAF-17 can help us understand the role of biomolecules in yeasts and explore possibilities of large-scale synthesis for industrial applications.

• 한국식품영양학회지
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• 제9권4호
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• pp.372-378
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• 1996

최소 배지에 여러 아미노산을 첨가하여 배양시킨 Serratia marcescens ATCC 25419 세포 추출물에서 threonine dehydratase의 비활성을 조사한 결과 이소루신은 1~4mM에서 효소의 비활성을 100~140% 정도 증가시킨 반면 15~30mM에서 25~80% 정도 감소시켰다. 루신은 효소의 비활성을 1~120mM에서 60% 이상 감소시켰다. 한편, 발린은 효소의 비활성을 1~4mM에서 20% 정도, 8~10mM에서 10% 정도 감소시켰으나 20mM에서 효소의 비활성을 원래대로 회복시켰다. 각 2mM 농도의 이소루신과 발린 그리고 이소루신과 루신은 효소의 비활성을 각각 75, 50%씩 증가시킨 반면 세 아미노산을 각각 2과 10mM씩 동시 첨가했을 경우에는 각각 25, 58%씩 감소시켰다. 글리옥실산은 효소의 비활성을 75%, 이마다졸, 피루브산, $\alpha$-아미노부티르산 그리고 AMP는 모두 50%씩, 호모세린, $\alpha$-케토글루타르산, $\alpha$-케토부티르산 및 IPTG를 첨가했을 경우는 모두 25%씩 감소시켰다. cAMP는 효소의 비활성을 1~2mM에서 10~40% 정도 감소시킨 반면 10mM에서 80% 증가시켰다. 이러한 결과들로부터 S. marcescens threonine dehydratase는 E. coli K-12 효소와는 다르게 낮은 농도의 이소루신에 의해 효소의 비활성이 증가되고, 높은 농도의 이소루신 (10~30mM)에 의해서는 감소되는 등 이소루신 생합성 과정에서 조절 역할을 하는 것으로 생각된다.