• Title, Summary, Keyword: $^3H$-leucine

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Studies on the Production of Lysine by Fermentation Process (2) -Lysine Production by Auxotrophs- (발효에 의한 라이신(L-Lysine) 생산에 관한 연구 (2) -영양요구성 변이주에 의한 Lysine 생산-)

  • Min, Tae-Ick;Kim, Hang-Mook;Kwon, Tai-Wan
    • Korean Journal of Food Science and Technology
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    • v.4 no.2
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    • pp.123-133
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    • 1972
  • Over 90 of lysine producing auxotrophs were obtained from Corynebacterium sp. S-27-12, Brevibacterium flavum ATCC 15168 and Micrococcus glutamicus ATCC 13032 by UV light, $Co^{60}$ irradiation and N-methyl-N'-nitro-N-nitrosoguanidine treatment. One of the mutant, Brev. flavum U46-N59, was identified as a leucine auxotroph and accumulated lysine during flask (500 ml) cultivation (180 strokes/min.) up to 21.6 mg per ml of broth at pH 7.5 and $28^{\circ}C$ after 4 days. The medium consisted of glucose, 100; urea, 10; corn steep liquor, 40; $KH_2PO_4,\;2;\;K_2HPO_4,\;0.5;\; MgSO_4.\;7H_2O,\;0.4;\;antifoam\;S-57,\;1g;\;Fe_2(SO_4)_3.XH-2O,\;10;\; MnCl_2,\;4H_2O,\;10mg;\;biotin,\;30;\;thiamine-HCl,\;100{\mu}g$in 1l of distilled water, and 40 U/ml of penicillin was added after 36 hrs fermentation.

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Computational Analysis of Apolipophorin-III in Hyphantria cunea

  • Chandrasekar R.;Dhanalakshmi R.;Krishnan M.;Kim H. J.;Jeong H. C.;Seo S. J.
    • International Journal of Industrial Entomology
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    • v.10 no.1
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    • pp.25-33
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    • 2005
  • Recently a cDNA clone of apoLp-III from Hyphantria cunea was isolated and subjected to computational analysis to compare with other available sequences. Multiple sequence alignments were carried out using the amino acid sequences of apoLp-III from six insects. It was found that the H. cunea apoLp-III has relatively high sequence identities to Spodoptera litura ($69.5\%$), Manduca sexta ($66.8\%$), Galleria mellonella ($65.1\%$), Bombyx mori N4 ($54.3\%$) but less identity to Locusta migratoria ($18.3\%$). The amino acid composition was compared with other insects using EXPASY tools; it shows that alanine (Ala), glutamine (Gln), leucine (Leu) and lysine (Lys) are the major amino acid components of apoLp-III in H. cunea as well as other lepidopterans. Homology modeling performed using PSI-BLAST (PDB template M. sexta) reveals that the apoLp-III molecules consist of five, long amphipathic alpha helical bundles with short loops connecting the helices and shows homology with other insects. Phylogenetic analysis shows that the orthopteran apoLp-III represented by locust was most distantly related to the lepidopteran insects.

Analysis of fusogenic activity of autographa californica nuclear polyhedrosis virus (Ac NPV) gp64 envelope glycoprotein

  • Kim, Hee-Jin;Yang, Jai-Myung
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.7-14
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    • 1996
  • Teh baculovirus gp64 glycoprotein is a major component of the envelope of budded virus (BV) and has been shown that it plays an essential role in the infection process, especially virus-cell membrane fusion. We have cloned Autographa californica Nuclear Polyhedrosis Virus (AcNPV) gp64 protein were examined for membrane fusion activity by using a synchtium formation assay under various conditions. The optimal conditions required for inducing membrane fusion are 1) form pH 4.0 to 4.8 2) 15 min exposure of cells to acidic pH 3) at least 1 .mu.g of gp64 cloned plasmid DNA per 3 * 10$^{6}$ cells 4) and an exposure of cells to acidic pH at 72 h post-transfection. In order to investigate the role of hydrophobicity of the gp64 glycoprotein for the membrane fusion, the two leucine residues (amino acid position at 229 and 230) within hydrophobic region I were substituted to alanine by PCR-derived site-directed mutagenisis and the membrane fusion activity of the mutant was anlaysed. The gp64 glycoprotein carrying double alamine substitution mutation showed no significant difference in fusion activity. This result suggested that minor changes in hydrophobicity at the amino acid position 229 and 230 does not affect the acid-induced membrane fusion activity of the gp64 glycoprotein.

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Amino acids profiles of six dinoflagellate species belonging to diverse families: possible use as animal feeds in aquaculture

  • Lim, An Suk;Jeong, Hae Jin;Kim, So Jin;Ok, Jin Hee
    • ALGAE
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    • v.33 no.3
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    • pp.279-290
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    • 2018
  • Microalgae have been utilized in diverse industries including aquaculture. Among the microalgae, dinoflagellates are known to have various bioactive compounds, and thus the interest in their application to industry has increased. In order to test their potential as food materials for aquaculture animals, the crude protein contents and compositions of amino acids of six dinoflagellates Heterocapsa rotundata (family Heterocapsaceae), Ansanella granifera (Suessiaceae), Alexandrium andersonii (Ostreopsidaceae), Takayama tasmanica (Brachidiniaceae), Takayama helix, and Gymnodinium smaydae (Gymnodiniaceae) belonging to diverse families were analyzed. The percentage of the amount of the crude protein relative to dry weight of T. tasmanica was the highest (65%) and that of A. andersonii was the lowest (26%). However, the highest percentage of total detected amino acids in crude protein was found in A. andersonii (98.2%). In all six dinoflagellates, glutamic acid was the most dominant amino acid in crude protein. However, the second main amino acid was aspartic acid for H. rotundata, A. granifera, T. helix, and G. smaydae, but were arginine and leucine for A. andersonii and T. tasmanica, respectively. Furthermore, T. tasmanica and T. helix did not have taurine and gamma-aminobutyric acid, whereas the other dinoflagellates possessed them. The percentages of essential amino acid contents of the dinoflagellates met the requirement levels for juvenile shrimps. In addition, the dinoflagellates were not toxic to the brine shrimp Artemia salina. Compared with the other microalgae reported so far, H. rotundata and A. andersonii can be used for arginine-rich diets, T. tasmanica for valine and leucine-rich diets, A. granifera for histidine-rich diets, T. helix for threonine-rich diets, and G. smaydae for lysine-rich diets. Therefore, based on their biochemical composition and toxicity to Artemia, the dinoflagellates could be used as essential amino acid sources for cultivating animals in the aquaculture industry.

Preparation and characterization of protein isolate from Yellowfin tuna Thunnus albacares roe by isoelectric solubilization/precipitation process

  • Lee, Hyun Ji;Lee, Gyoon-Woo;Yoon, In Seong;Park, Sung Hwan;Park, Sun Young;Kim, Jin-Soo;Heu, Min Soo
    • Fisheries and aquatic sciences
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    • v.19 no.3
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    • pp.14.1-14.10
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    • 2016
  • Isoelectric solubilization/precipitation (ISP) processing allows selective, pH-induced water solubility of proteins with concurrent separation of lipids and removal of materials not intended for human consumption such as bone, scales, skin, etc. Recovered proteins retain functional properties and nutritional value. Four roe protein isolates (RPIs) from yellowfin tuna roe were prepared under different solubilization and precipitation condition (pH 11/4.5, pH 11/5.5, pH 12/4.5 and pH 12/5.5). RPIs contained 2.3-5.0 % moisture, 79.1-87.8 % protein, 5.6-7. 4 % lipid and 3.0-3.8 % ash. Protein content of RPI-1 and RPI-2 precipitated at pH 4.5 and 5.5 after alkaline solubilization at pH 11, was higher than those of RPI-3 and RPI-4 after alkaline solubilization at pH 12 (P < 0.05). Lipid content (5.6-7.4 %) of RPIs was lower than that of freeze-dried concentrate (10.6 %). And leucine and lysine of RPIs were the most abundant amino acids (8.8-9.4 and 8.5-8.9 g/100 g protein, respectively). S, Na, P, K as minerals were the major elements in RPIs. SDS-PAGE of RPIs showed bands at 100, 45, 25 and 15 K. Moisture and protein contents of process water as a 2'nd byproduct were 98.9-99.0 and 1.3-1.8 %, respectively. Therefore, yellowfin tuna roe isolate could be a promising source of valuable nutrients for human food and animal feeds.

Quality Characteristics of Salt-Fermented Anchovy Sauce and Sandlance Sauce (멸치액젓 및 까나리액젓의 품질 특성)

  • OH Kwang-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.3
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    • pp.252-255
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    • 1999
  • The quality characteristics of the Korean traditional salt-fermented fish sauces, the traditional anchovy sauce (TAS) and the sandlance sauce(TKS) were evaluated comparing to the commercial anchovy sauce (CAS) and sandlance sauce (CKS). The acidity was higher in TKS than in TAS, whereas the contents of VBN, total-N and amino-N were higher levels in TAS. In color values, L and b values in TAS were generally higher than those in TKS, whereas a and ${\Delta}E$ values were higher in TKS. The contents of total free amino acids in TAS and TKS were $12.40\;g\%$ (w/v) and $9.549 g\%$ (w/v), respectively. The contents or six amino acids, alanine, glutamic acid, leucine, isoleucine, valine and Iysine were higher in TAS, whereas the contents of arginine, glutamic acid, leucine, alanine and valine were higher in TKS. Nucleotides such as IMP and hypoxanthine were principal components in both TAS and TKS. The nitrogen related compounds, TMAO, TMA and total creatinine were determined to be $108.8\;mg\%$ (w/v), $60.5\;mg\%$ (w/v), $62.4\;mg\%$ (w/v) in TAS, and those in TKS were $60.1\;mg\%$ (w/v), $24.1\;mg\%$ (w/v), $67.6\;mg\%$ (w/v), respectively.

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Change of Free Amino Acid and Nucleotide Compound of Puffer Fish Fillet under Storage Condition (저장조건에 따른 복어육의 유리아미노산 및 핵산 화합물의 변화)

  • Mun, Seung-Kweon;Sung, Ki-Hyup;Yoo, Seung-Seok
    • Korean journal of food and cookery science
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    • v.28 no.3
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    • pp.249-255
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    • 2012
  • The objectives of this study were to evaluate the physicochemical characteristics of puffer fish under storage conditions. Free amino acids were identified in the order of taurine > alanine > lysine > leucine > glutamic acid > valine. Glutamic acid, lysine, histidine, arginine, proline, and aspartic acid increased over time and with increased temperature, and valine and tyrosine were affected by temperature. ATP decreased dramatically during 36 h of storage at $4^{\circ}C$, 24 h of storage at $12^{\circ}C$, and 16 h of storage at $20^{\circ}C$. IMP reached its highest level when puffer fish was stored for 36 h at 4 and $12^{\circ}C$ and 24 h at $20^{\circ}C$, and hypoxanthine levels grew steeply at 60 h at $4^{\circ}C$, 24 h at $12^{\circ}C$ and 20 h at $20^{\circ}C$. In terms of K value, the puffer fish was available for sliced raw fish within 60 h at $4^{\circ}C$, 24 h at $12^{\circ}C$ and 12 h at $20^{\circ}C$, and the fish can be taken in after cooking within 72 h at $4^{\circ}C$ and $12^{\circ}C$ and 36 h at $20^{\circ}C$. The physicochemical quality characteristics showed that puffer fish is available for sliced raw fish within 36 h at $4^{\circ}C$, 16 h at $12^{\circ}C$ and 12 h at $20^{\circ}C$, and that the fish can be taken after cooking within 72 h at $4^{\circ}C$ and $12^{\circ}C$ and 36 h at $20^{\circ}C$.

Effects of Amino Acids on Larval Settlement and Metamorphosis in Haliotis discus hannai

  • Kang, Kyoung-Ho;Kim, Byoung-Hak;Kim, Jae-Min;Kim, Young-Hun
    • The Korean Journal of Malacology
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    • v.19 no.2
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    • pp.95-106
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    • 2003
  • The compositions of amino acid in 6 monocultured benthic diatoms used in aquaculture of Haliotis discus hannai were analyzed, and effects of 15 artificial synthetic amino acids on the settlement and metamorphosis of H. discus hannai larvae. Results showed that the content of L-aspartic acid was highest in all diatoms, and that of L-glutamic acid was second high. In experiment using settlement slat without benthic diatom attached, the highest settlement rate (33.3 ${\pm}$ 8.8%) was obtained with L-glutamic acid, and a higher value (16.7 ${\pm}$ 3.3%) was found with L-aspartic acid at 24 h after experimental commencement, compared to that of control (8.6 ${\pm}$ 5.1%). 80 h later the metamorphosis rates of L-glutamic acid (86.7 ${\pm}$ 10.7%) and L-aspartic acid (80.0 ${\pm}$ 3.3%) groups were higher than control group(0) and other amino acids significantly. The response rate of L-glutamic acid was the highest (62.0%), and those of L-aspartic acid (30.0%) and L-threonine (25.3%) groups were also significantly higher than control group. In the experiment using settlement slat with benthic diatom attached, the best effect of various amino acids on induction of larval settlement was obtained with L-glutamic (82.0 ${\pm}$ 6.9%) and L-aspartic acid (78.7 ${\pm}$ 5.1%) at 24 h after experimental commencement. The settlement rates of L-histidine, L-leucine, L-lysine, L-methionine, L-phenylalanine, and L-tyrosine groups were significantly lower than control group. The same differences in the metamorphosis rate at 56 h after experimental commencement and in the response rate were found. It should be noted that after 80 h the metamorphosis rates of L-histidine (74.0 ${\pm}$ 12.0%) and L-lysine (87.0 ${\pm}$ 8.8%) declined rapidly compared to those of 56 h (8.0 ${\pm}$ 12.0%; 7.7 ${\pm}$ 12.0%).

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Changes of Proteolytic Activity and Amino Acid Composition of the Tissue Extract from Sea Cucumber Entrails during Fermentation with Salt (해삼내장(內臟)젓갈 숙성중(熟成中) 단백질분해효소(蛋白質分解酵素)의 활성(活性)과 아미노산(酸) 조성(組成)의 변화(變化))

  • Lee, Gi Chan;Cho, Deuk Moon;Byun, Dae Seok;Joo, Hyen Kyu;Pyeun, Jae Hyeung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.12 no.4
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    • pp.342-349
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    • 1983
  • This study was undertaken to ascertain food and nutritional evaluating data on the processing of fermented sea cucumber (Stichopus japonicus) entrails. In the experiment, the crude proteolytic enzyme from the entrails tissue of raw and fermented sea cucumber during the days of ripening was extracted. The optimal activity condition for the crude enzyme and the compositional changes of amino acid of the protein and free amino acid in the raw and fermented sample were also investigated. 1. Less than three kinds of proteolytic enzymes that each enzyme has optimal activity condition at pH 3.1 $50^{\circ}C$(A-enzyme), pH 5.7 $50^{\circ}C$(B-enzyme) and pH 7.7 $45^{\circ}C$(C-enzyme), respectively were believed to be exist in the entrails tissue of sea cucumber. 2. A-enzyme and C-enzyme were strongly inhibited with the increase of the salt concentration, and B-enzyme was activated at the 1% salt concentration and was inhibited above the 5% salt concentration. 3. The result of the effect of several salt ions on the proteolytic activity showed that A-enzyme was slightly inhibited in the presence of all salt ions added, B-enzyme was activated in the presence of the all salt ions except $Cu^{2+}$ and C-enzyme was activated in the presence of $Ca^{2+}$ and $Mn^{2+}$, and inhibited by $Cu^{2+}$, $Co^{2+}$ and $Mg^{2+}$. 4. When the effects of the ripening days on the proteolytic activity of the crude enzymes were analysed, the activity of the A-enzyme was slightly weakened with the lapse of the fermentation days, whereas the B-enzyme was not influenced by the fermentation days. 5. In the analysis of amino acid composition of the protein of the samples, the 8 days fermented sea cucumber entrails showed the diminution of all kinds of amino acid. Apparently diminished amino acids were arginine, alanine, glutamic acid, glycine, serine, valine, threonine and lysine etc., and methionine, histidine and isoleucine were slightly decreased. 6. In the analysis of free amino acid composition of the 8 days fermented sample, glutamic acid, aspartic acid, leucine and lysine were rich, while histidine, methionine, proline and tyrosine were poor. The most of free amino acids were increased during the fermentation procedure and especially in lysine, histidine, threonine, glutamic acid, methionine, valine and leucine.

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Onion peel extract and its constituent, quercetin inhibits human Slo3 in a pH and calcium dependent manner

  • Wijerathne, Tharaka Darshana;Kim, Ji Hyun;Kim, Min Ji;Kim, Chul Young;Chae, Mee Ree;Lee, Sung Won;Lee, Kyu Pil
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.5
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    • pp.381-392
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    • 2019
  • Sperm function and male fertility are closely related to pH dependent $K^+$ current (KSper) in human sperm, which is most likely composed of Slo3 and its auxiliary subunit leucine-rich repeat-containing protein 52 (LRRC52). Onion peel extract (OPE) and its major active ingredient quercetin are widely used as fertility enhancers; however, the effect of OPE and quercetin on Slo3 has not been elucidated. The purpose of this study is to investigate the effect of quercetin on human Slo3 channels. Human Slo3 and LRRC52 were co-transfected into HEK293 cells and pharmacological properties were studied with the whole cell patch clamp technique. We successfully expressed and measured pH sensitive and calcium insensitive Slo3 currents in HEK293 cells. We found that OPE and its key ingredient quercetin inhibit Slo3 currents. Inhibition by quercetin is dose dependent and this degree of inhibition decreases with elevating internal alkalization and internal free calcium concentrations. Functional moieties in the quercetin polyphenolic ring govern the degree of inhibition of Slo3 by quercetin, and the composition of such functional moieties are sensitive to the pH of the medium. These results suggest that quercetin inhibits Slo3 in a pH and calcium dependent manner. Therefore, we surmise that quercetin induced depolarization in spermatozoa may enhance the voltage gated proton channel (Hv1), and activate non-selective cation channels of sperm (CatSper) dependent calcium influx to trigger sperm capacitation and acrosome reaction.