• Title/Summary/Keyword: LEU

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Association of the X-linked Androgen Receptor Leu57Gln Polymorphism with Monomelic Amyotrophy

  • Park, Young-Mi;Lim, Young-Min;Kim, Dae-Seong;Lee, Jong-Keuk;Kim, Kwang-Kuk
    • Genomics & Informatics
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    • v.9 no.2
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    • pp.64-68
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    • 2011
  • Monomelic amyotrophy (MA), also known as Hirayama disease, occurs mainly in young men and manifests as weakness and wasting of the muscles of the distal upper limbs. Here, we sought to identify a genetic basis for MA. Given the predominance of MA in males, we focused on candidate neurological disease genes located on the X chromosome, selecting two X-linked candidate genes, androgen receptor (AR ) and ubiquitin-like modifier activating enzyme 1 (UBA1). Screening for genetic variants using patients' genomic DNA revealed three known genetic variants in the coding region of the AR gene: one nonsynonymous single-nucleotide polymorphism (SNP; rs78686797) encoding Leu57Gln, and two variants of polymorphic trinucleotide repeat segments that encode polyglutamine (CAG repeat; rs5902610) and polyglycine (GGC repeat; rs3138869) tracts. Notably, the Leu57Gln polymorphism was found in two patients with MA from 24 MA patients, whereas no variants were found in 142 healthy male controls. However, the numbers of CAG and GGC repeats in the AR gene were within the normal range. These data suggest that the Leu57Gln polymorphism encoded by the X-linked AR gene may contribute to the development of MA.

The Purification and Characterization of a Bacillus stearothermophilus Methionine Aminopeptidase (MetAP)

  • Chung, Jae-Min;Chung, Il-Yup;Lee, Young-Seek
    • BMB Reports
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    • v.35 no.2
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    • pp.228-235
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    • 2002
  • Methionine aminopeptidase (MetAP) catalyzes the removal of an amino-terminal methionine from a newly synthesized polypeptide. The enzyme was purified to homogeneity from Bacillus stearothermophilus (KCTC 1752) by a procedure that involves heat precipitation and four sequential chromatographs (including DEAE-Sepharose ion exchange, hydroxylapatite, Ultrogel AcA 54 gel filtration, and Reactive red 120 dye affinity chromatography). The apparent molecular masses of the enzyme were 81,300 Da and 41,000 Da, as determined by gel filtration chromatography and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. This indicates that the enzyme is comprised of two identical subunits. The MetAP specifically hydrolyzed the N-terminal residue of Met-Ala-Ser that was used as a substrate, and exhibited a strong preference for Met-Ala-Ser over Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The enzyme has an optimal pH at 8.0, an optimal temperature at $80^{\circ}C$, and pI at 4.1. The enzyme was heat-stable, as its activity remained unaltered when incubated at $80^{\circ}C$ for 45 min. The Km and Vmax values of the enzyme were 3.0mM and 1.7 mmol/min/mg, respectively. The B. stearothernmophilus MetAP was completely inactivated by EDTA and required $Co^{2+}$ ion(s) for activation, suggesting the metal dependence of this enzyme.

The Effects of Lactic Acid Bacteria on the Growth and Aflatoxin $B_1$ Production of Aspergillus parasiticus (유산균이 Aspergillus parasiticus의 성장과 Aflatoxin $B_1$ 생성에 미치는 영향)

  • Lee Kyung-Min;Kim Jong-Gyu
    • Journal of Environmental Health Sciences
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    • v.31 no.2 s.83
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    • pp.127-133
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    • 2005
  • This study was performed to investigate the competitive effect of Leuconostoc mesenteroides and Lactobacillus plantarum on the growth and aflatoxin $B_1$ production of Aspergillus parasiticus. Leu. mesenteroides and Lac. plantarum were grown with A. parasiticus in a modified APT broth at $28^{\circ}C$ for 9 days. It was observed that the growth of A. parasiticus got lower when the two bacteria were inoculated simultaneously than when the mold grew alone. The pH of mixed culture of Leu. mesenteroides and Lac. plantarum showed significantly lower value than the pH of the pure culture of A. parasiticus (p<0.05). The acidity of the mixed culture group significantly increased compared with the control group (A. parasiticus alone group) (p<0.05). The aflatoxin $B_1$ production was significantly decreased in the mixed culture group than in the A. parasiticus alone group (p<0.05). Leu. mesenteroides showed the more efficient effect than Lac. plantarum. These results indicate that the two lactic acid bacteria have inhibitory effect on the growth and aflatoxin $B_1$ production of Aspergillus parasiticus.

Cultivation of Psychrotrophic Lactic Acid Bacteria Isolated from Kimchi in Korean Cabbage-Juice (김치에서 분리한 저온성 젖산균의 배추즙에서의 배양)

  • 소명환;오현진
    • The Korean Journal of Food And Nutrition
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    • v.7 no.4
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    • pp.392-398
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    • 1994
  • The purpose of this study was to predict the actions and roles of 10 strains of representative psychrotrophic lactic acid bacteria, in kimchi fermentation, which were Isolated from kimchi and identified as Leu. mesenteroides subsp. mesenteroides, Leu. mesenteroides subsp. dextranicum, Leu. lactic, Leu. paramesenteroides, Lac. bavaricus and Lac. homahiochii. For this, 0.01% of tactic starters were inoculated in germ free Korean cabbage-juice containing 2.5% NaCl, and then cultivated for 14 days at 1$0^{\circ}C$. All strains grew actively, and reached their stationary phase in 4 days. In death phase, the slopes of curves were much different each other by strains. The acidity increased rapidly between 2 and 4 days, and the pH decreased rapidly between 2 and 3 days. The total acidity was 0.5B~0.75%, the volatile acidity 0.04~0.18% and the pH 3.55~3.85, in final cultures. The cultures of Leuconostocs were better than those of lactobacilli on flavor test. It was thought that the ripening periods of kimchi would be much reduced, and that the over ripening would be also somewhat avoided, when these strains were used as starter bacteria for kimchi and the kimchi was fermented at low temperature.

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Physicochemical Stability of Leucine Enkephalin and $[D-Ala^2]$-Leucine Enkephalinamide in Buffered Aqueous Solution (완충 수용액중 로이신엔케팔린 및 [D-알라$^2]$-로이신엔케팔린아미드의 물리화학적 안정성)

  • Park, In-Sook;Chun, In-Koo
    • YAKHAK HOEJI
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    • v.38 no.5
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    • pp.488-495
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    • 1994
  • To evaluate the feasibility of transmucosal delivery of leucine enkephalin (Leu-Enk) and its synthetic analog, $[D-Ala^2]$-leucine enkephalinamide (YAGFL), their physicochemical stabilities in aqueous buffered solutions were first investigated using a stability indicating high performance liquid chromatography. The degradation of Leu-Enk and YAGFL followed the pseudo-first-order kinetics. From the pH-rate profiles, it was found that the maximal stability of the two pentapeptides was at the pH of about 5.0. The shelf lives $(t_{90%})$ for the degradation of Leu-Enk and YAGFL at pH 5.0 and $40^{\circ}C$ were found to be 48.13 and 50.9 days, respectively. From the temperature dependence of the degradation, activation energies for Leu-Enk and YAGFL were calculated to be 13.61 and 13.47 kcal/mole, respectively. A higher ionic strength and a higher initial peptide concentration in buffered solution slowed the degradation of the two pentapeptides. The addition of 2-hydroxypropyl-${\beta}$-cyclodextrin into the peptide solution did not affect the stability significantly.

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Importance of Leu-5 and Pro-6 in the Inhibitory Activity of the Serratia marcescens Metalloprotease Inhibitor (SmaPI)

  • Bae, Kwang-Hee;Kim, Dong-Min;Kim, Sun-Taek;Kim, Tae-Hoon;Shin, Yong-Chul;Byun, Si-Myung
    • BMB Reports
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    • v.34 no.2
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    • pp.109-113
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    • 2001
  • The Serratia marcescens metalloprotease inhibitor (SmaPI) is a proteinase inhibitor toward Serratia marcescens metalloprotease (SMP). The three-dimensional structure of SmaPI was calculated by computer modeling using the structure complex between SMP and the Erwinia chrysanthemi inhibitor as a template. Based on this model structure, the substitution of the amino acid residues, Ala4, Leu-5, Pro-6, and Thr-7, were located at the hinge region of the N-terminal segment by site-directed mutagenesis. Although the A4R and T7A mutant SmaPIs showed a nearly full inhibitory activity, the inhibitory activity of SmaPI decreased significantly when the Leu-5 was converted to Ala, Gly, Ile, or Val. Surprisingly, the L5I and L5V mutant SmaPIs showed less inhibitory activities than the L5A mutant. From these results, we suggested that the orientations and positions of respective aliphatic groups in the side-chain of position 5 mainly affected the inhibitory activity of SmaPI. The overall side-chain hydrophobicity was only slightly affected. The side-chain of the Leu-5 residue contributed approximately 0.79 kcal/mol out of 8.44 kcal/mol to the binding of SmaPI with SMP The inhibitory activities of P6A and F6G were also severely decreased. The Pro-6 may have a critical role in maintaining the strict conformation of the N-terminal portion that may be important in the inhibitory activity of SmaPI. In conclusion, Leu-5 and Pro-6 have crucial roles in the inhibitory function of SmaPI toward SMP.

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Characteristics of Transformants in Pleurotus florida (사철느타리버섯 형질전환주(形質轉換珠)의 특성(特性))

  • Byun, Myung-Ok;Kim, Kyung-Soo;Cha, Dong-Yeol
    • The Korean Journal of Mycology
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    • v.20 no.3
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    • pp.222-228
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    • 1992
  • Pleurotus florida was transformed by complementation of auxotrophic mutant using chimeric plasmid containing Flammulina velutipes leu 2 gene and pBR 322 replicon. Mycelial morphology of transformants was grown and compared on mushroom complete and minimal medium. Transformants were mated with monokaryon and their genetic recombination was investigated for the morphology of fruitbody and spore analysis. $Leu^+$ transformant showed same mating type of $A_1B_1$ as to the untransformed mutant. The transformant and the untransformed mutant were mated with monokaryon of which mating type is $A_2B_1$, respectively. Although fruitbody of the untransformed mutant was not produced, $leu^+$ transformant produced fruitbody. Spore analysis showed that leucine requiring spores from fruitbody of $leu^+$ transformant were diminished when compared with those of untransformed mutant.

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Effect of Lactic Acid Bacteria and Temperature on Kimchi Fermentation (II) (젖산균과 온도가 김치 발효에 미치는 영향(II))

  • Cho, Young;Rhee, Hei-Soo
    • Korean journal of food and cookery science
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    • v.7 no.2
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    • pp.89-95
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    • 1991
  • The effects of lactic acid bacteria on the chemical and microbial changes of fermented kimchi at various temperatures were studied. Kimchi was homogenized and was sterilized by ultra violet (UV), then Lactobacillus plantarum, Leuconostoc mesenteroides, Pediococous acidilactici, Lactobacillus brevis and the mixture of these bacteria inoculated on sterilized kimchi, respectively. The measurement of alcohol by gas chromatography, and changes of sugar content and total viable count were investigated and palatability test was carried while inoculated kimchi was fermented at $30^{\circ}C$, $21^{\circ}C$ and $7^{\circ}C$. Ethyl alcohol was detected by GC in sample I (original Kimchi homogenate), III(inoculated Leu. mesenteroides), V(inoculated with Lac. brevis), then especially, more content were detected at $14^{\circ}C$. Sugar content reduced in accordance with fermentation proceeding. Total viable count increased at early fermentation stage, but thereafter decreased slowly. In the result of palatability test, sample I was the highest at all temperatures, sample III and IV (inoculated with mixed lactic acid bacteria) was the following in that kimchi odor and taste and the temperature.

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Interaction between Lactobacillus acidophilus and Kluyveromyces fragilis on the Metabolism of Amino Acids in Soymilk (Lactobacillus acidophilus와 Kluyveromyces fragilis의 혼합배양에 의한 두유의 젖산발효중 아미노산 대사의 상호작용)

  • 류인덕;박정길;유주현
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.287-292
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    • 1988
  • The interaction between Lactobacillus acidophilus and Kluyveromyces fragilis on the utilization of amino acids in soymilk was investigated. K. fragilis assimilated relatively well various amino acids such as Met., ILe., Phe., Leu., Thr., Lys., Val., Arg., Tyr., Ser., Asp., Ala. and Glu. that existed only in trace amounts in soymilk. K. fragilis did not utilized Gly., while hi accumulated His. L. acidophilus hydrolyzed soyprotein to liberate various amino acids. Among various amino acids, it utilized Met., ILe., Thr., Tyr., Ser., Val. and His. as growth factors and accumulated Leu., Phe., Lys., Arg., Glu., Asp. and Ala. among the essential amino acids required by K. fragilis and Gly. These results implied that K. fragilis grew on amino acids that existed only in trace amounts in soymilk, but it's growth was stimulated by amino acids such as Leu., Phe., Lys., Arg., Glu., Asp. and Ala. ac-cumulated by L. acidophilus.

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Selection of Probiotic Bacteria from Yulmoo Kimchi Using a Stimulated Human Intestinal Model System (인체장모델시스템에 의한 열무김치로부터 프로바이오틱스 균주 선발)

  • Kang, Mi-Ran;Kim, Da-Ram;Kim, Tae-Woon;Park, Sung-Hee;Kim, Hyun-Ju;Jang, Ja-Young;Han, Eung-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.3
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    • pp.396-401
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    • 2012
  • To select potent probiotics from lactic acid bacteria in Yulmoo Kimchi, an in vitro and stimulated human intestinal model system (SHIMS) test were performed. One Leuconostoc mesenteroides strain from five strains of Yulmoo Kimchi and one Lactobacillus plantarum from 12 strains of KCTC and KCCM were selected according to survival in acidic and bile salts conditions. Between the two species, Leu. mesentroides displayed higher survival activity in a SHIMS test. The strain was identified as Leu. mesentroides by 16S rRNA sequencing and was designated as Leu. mesentroides K01.