• Journal of Microbiology and Biotechnology
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• v.30 no.9
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• pp.1436-1442
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• 2020

Amylosucrase (ASase, E.C. 2.4.1.4) is capable of efficient glucose transfer from sucrose, acting as the sole donor molecule, to various functional acceptor compounds, such as polyphenols and flavonoids. An ASase variant from Deinococcus geothermalis, in which the 226^th alanine is replaced with asparagine (DgAS-A226N), shows increased polymerization activity due to changes in the flexibility of the loop near the active site. In this study, we further investigated how the mutation modulates the enzymatic activity of DgAS using molecular dynamics and docking simulations to evaluate interactions between the enzyme and phenolic compounds. The computational analysis revealed that the A226N mutation could induce and stabilize structural changes near the substrate-binding site to increase glucose transfer efficiency to phenolic compounds. Kinetic parameters of DgAS-A226N and WT DgAS were determined with sucrose and 4-methylumbelliferone (MU) as donor and acceptor molecules, respectively. The k_cat/K_m value of DgAS-A226N with MU (6.352 mM^-1min^-1) was significantly higher than that of DgAS (5.296 mM^-1min^-1). The enzymatic activity was tested with a small phenolic compound, hydroquinone, and there was a 1.4-fold increase in α-arbutin production. From the results of the study, it was concluded that DgAS-A226N has improved acceptor specificity toward small phenolic compounds by way of stabilizing the active conformation of these compounds.

• Korean Journal of Microbiology
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• v.36 no.3
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• pp.242-248
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• 2000

Since the official allowance of bottled water at Korean domestic market in 1995, Pseudomo~zas aemginosu has been detected from 2.3% and 1.2% of source and products of bottled water sa~nple tested, respectively. according to the nation-wide dala froin May 1995 to December 1996. Therefore, P aeivginosa was the second most important parameter, next to colifoi~ns, anlong the bacieriological parameters regulated for bonled water. The official standard method initially adopted the Japanese officlal method and Standard Methods of the US, w~hich is using aspai-agiii-acetamid mnedia(A-A method). how eve^; the method showed low specificity regardless of the high sensitivity. The $42^{\circ}C$ growth test was the best biochemical featu1-e differentiating the P uelarginosu 6-om P aei-uginosa-like species such as P puririir and P Jhoi.escens amo1zg the other characierisiics such as fluorescence pigment_ pyocyanin, casein hy&olysis, etc. Thel-efore, addition of the $42^{\circ}C$ growth Lest in advance ofthe biochemical identification test, when sainple is positive by A-A method, should strengthen the spec~IiclQ w~tli ~ninin~um addition of testing load.

• Molecules and Cells
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• v.23 no.3
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• pp.280-286
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• 2007

L-asparaginase (EC 3.5.1.1) catalyzes the hydrolysis of the amide group of L-asparagine, releasing aspartate and $NH_4{^+}$. We isolated a low temperature-inducible cDNA sequence encoding L-asparaginase from soybean leaves. The full-length L-asparaginase cDNA, designated GmASP1, contains an open reading frame of 1,258 bp coding for a protein of 326 amino acids. Genomic DNA blotting and fluorescence in situ hybridization showed that the soybean genome has two copies of GmASP1. GmASP1 mRNA was induced by low temperature, ABA and NaCl, but not by heat shock or drought stress. E. coli cells expressing recombinant GmASP1 had 3-fold increased L-asparaginase activity. A possible function of L-asparaginase in the early response to low temperature stress is discussed.

• Korean journal of food and cookery science
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• v.30 no.6
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• pp.785-791
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• 2014

The antioxidative activities of beverages with water or ethanol extracts from Maegmundong (Liriope platyphylla) were investigated with free amino acid and mineral analysis, as well as saponin, total phenolics and flavonoid determination. Antioxidative activity was evaluated by electron donating ability and ABTS radical scavenging activity. There was only a small difference between water extract beverages and ethanol-extract beverages with respect to their electron donating ability and ABTS radical scavenging activity, although the total saponin, total phenolics and flavonoid were found to be greater in the ethanol extract beverages than in the water extract beverages. Three major free amino acids of the Magemundong beverages with water- or ethanol extract were asparagine (58.30, 60.68 mg%), methionine (15.10, 13.95 mg%, respectively) and proline (12.31, 14.00 mg%, respectively). The most abundant mineral in the Maegmundong beverage with water and ethanol extract was potassium (238.68, 244.32 mg%, respectively).

• Journal of Plant Biology
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• v.20 no.3
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• pp.127-134
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• 1977

In order to investigate the changes of free amino acids during the differentiation and development of young spike in naked barley, a typical spring grain, Wanju, and two winter grains, Sedohadaka and Nonsankwa No. 1-6 differing in their spring habits, were analyzed at different growth stages by thin layer chromatography. In all the varieties 22 ninhydrin positive components were detected at the sowing time of March 5 and 20 components in the sowing plots of March 30. In case of the latter plot, β-alanine was identified only in both Wanju and Sedohadaka, whereas pipecolic acid was detected only in Nonsankwa No. 1-6. Particularly, it is interesting that β-alanine was observed only in the case showing the normal heading independent of the varieties and sowing times. Whether these components are directly related to the physiology of spring habits in barley or not is also a question to be answered. Of the major amino acids, alanine and γ-aminobutyric acid were always detected in appreciably large spots, and serine, leucine, aspartic acid, valine and asparagine were somewhat larger. In the plot of march 30, glutamic acid was also detected in very large spot in both Wanju and Sedohadaka at the stage of spikelet differentiation and in Nonsankwa No. 1-6 at the stage of bract differentiation. Histidine, which showed the only qualitative difference among the varieties during seed germination, cannot be observed at all. Proline observed considerably large spot during seed germination was always detected but very small except that it was observed in large spot at the stage of floret differentiation in Nonsankwa No. 1-6 in the plot of March 5.

• Herbal Formula Science
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• v.26 no.4
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• pp.307-318
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• 2018

Objectives : Endoplasmic reticulum (ER) stress designates cellular responses to the accumulation of misfolded and unfolded proteins in ER, which is related to a variety of liver diseases. Present study investigated the inhibitory effects of Litsea japonica flesh water extract (LJE) aganist ER stress. Methods : After HepG2 cells were pretreated with LJE and subsequently exposed to tunicamycin (Tm) or thapsigargin (Tg), expression of C/EBP homologous protein (CHOP), glucose regulated protein 78 kDa (GRP78), asparagine synthetase (ASNS), and endoplasmic reticulum DnaJ homologue 4 (ERDJ4) were determined by immunoblot and real-time PCR analysis. Three canonical signaling pathways in response to ER stress were examined to explore molecular mechanisms involved. Results : Pretreatment of 1 mg/mL LJE inhibited Tm- or Tg-induced CHOP expression, while L. japonica fruit water extract did not. In addition, LJE decreased the levels of GRP78, ASNS, and ERDJ4 mRNA by Tm. Moreover, phosphorylations of eukaryotic translation initiation factor $2{\alpha}$ and inositol-requiring enzyme 1, expression of nuclear form of activating transcription factor $6{\alpha}$, and transactivation of ER stress response element- and unfolded protein response element-harboring luciferase activities were inhibited by LJE pretreatment. Conclusions : Present results suggest that LJE would be a candidate to prevent or treat ER stress-mediated liver injuries.

• Journal of Animal Science and Technology
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• v.61 no.1
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• pp.35-40
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• 2019

This study was conducted to compare the physicochemical traits of dry-cured hams made from two different pig breeds: Berkshire and $Landrace{\times}Yorkshire{\times}Duroc$ (LYD). Pigs were slaughtered at a live weight of approximately 110 kg and cooled at $0^{\circ}C$ for 24 h in a chilling room. Then, the ham portion of the carcasses were cut and processed by dry-curing for physicochemical analyses. The dry-cured hams from Berkshire contain higher crude protein, fat, and ash level than those from LYD, whereas the hams from LYD had higher moisture contents than those from Berkshire(p < 0.05). The pH values of the hams from Berkshire were lower than those from LYD (p < 0.05). The hams from Berkshire had lower $L^*$ and $b^*$ values than those from LYD (p < 0.05). Palmitoleic acid (C16:1), oleic acid (C18:1), elaidic acid (C18:1t), monounsaturated fatty acids, and ratio of n-6 and n-3 fatty acids (n-6/n-3) in the ham from Berkshire were higher than LYD (p < 0.05). Free amino acids such as aspartic acid, threonine, serine, asparagine, glutamic acid, and lysine in hams from Berkshire were higher than those from LYD (p < 0.05). The microbial population had no significant difference between Berkshire and LYD dry-cured ham. The cross sections of dry cured ham showed difference from different breeds using scanning electron microscope and indicates some differences in texture. Considering the meat quality parameters of ham, hams from Berkshire could provide variety of ham for consumer who are seeking various different qualities and stories.

• Journal of Ginseng Research
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• v.43 no.3
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• pp.482-487
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• 2019

Background: The mixed-cultivation of different Panax ginseng cultivars can cause adverse effects on stability of yield and quality. K-1 is a superior cultivar with good root shape and stronger disease resistance. DNA markers mined from functional genes are clearly desirable for K-1, as they may associate with major traits and can be used for marker-assisted selection to maintain the high quality of Korean ginseng. Methods: Five genes encoding pathogenesis-related (PR) proteins of P. ginseng were amplified and compared for polymorphism mining. Primary, secondary, and tertiary structures of PR5 protein were analyzed by ExPASy-ProtParam, PSSpred, and I-TASSER methods, respectively. A coding single nucleotide polymorphism (SNP)-based specific primer was designed for K-1 by introducing a destabilizing mismatch within the 3' end. Allele-specific polymerase chain reaction (PCR) and real-time allele-specific PCR assays were conducted for molecular discrimination of K-1 from other cultivars and landraces. Results: A coding SNP leading to the modification of amino acid residue from aspartic acid to asparagine was exploited in PR5 gene of K-1 cultivar. Bioinformatics analysis showed that the modification of amino acid residue changed the secondary and tertiary structures of the PR5 protein. Primer KSR was designed for specific discrimination of K-1 from other ginseng cultivars and landraces. The developed real-time allele-specific PCR assay enabled easier automation and accurate genotyping of K-1 from a large number of ginseng samples. Conclusion: The SNP marker and the developed real-time allele-specific PCR assay will be useful not only for marker-assisted selection of K-1 cultivar but also for quality control in breeding and seed programs of P. ginseng.

• Food Science of Animal Resources
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• v.43 no.1
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• pp.124-138
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• 2023

This study was carried out to determine the effects of adding pistachio shell (PIS), pomegranate hull (POM), and olive pulp (OP) to the diet on milk amino acid and fatty acid parameters in Awassi sheep. In the study, 40 head of Awassi sheep, which gave birth at least twice, were used as animal material. Sheep were fed a control diet without added byproducts (CON), rations containing PIS, POM, and OP. Milk amino acid profile was determined by liquid chromatography-tandem mass spectrometry, milk fatty acid gas chromatography-flame ionization detection device. There was a dramatic reduction in alanine, citrulline, glutamine, glutamic acid, glycine, leucine, ornithine and alphaaminoadipic acid in the research groups. In the PIS group, argininosuccinic acid, gammaminobutyric acid, beta-alanine and sarcosine; In the POM group, asparagine, gammaminobutyric acid, beta-alanine, and taurine; In the OP group, a significant positive increase was found in terms of alanine, histidine, gammaminobutyric acid, and taurine amino acids. The applications in the study did not have a statistically significant effect on the ratio of short, medium and long chain fatty acids in milk (p>0.05). In the presented study, it was determined that PIS, POM, and OP, which were added to the sheep rations at a rate of 5%, caused significant changes in the milk amino acid profiles. In this change in milk amino acid profiles, the benefit-harm relationship should be considered.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.14.1-14.11
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• 2022

Background: Carnivore protoparvovirus 1, also known as canine parvovirus type 2 (CPV-2), is the main pathogen in hemorrhagic gastroenteritis in dogs, with a high mortality rate. Three subtypes (a, b, c) have been described based on VP2 residue 426, where 2a, 2b, and 2c have asparagine, aspartic acid, and glutamic acid, respectively. Objectives: This study examined the presence of CPV-2 variants in the fecal samples of dogs diagnosed with canine parvovirus in Bogotá. Methods: Fecal samples were collected from 54 puppies and young dogs (< 1 year) that tested positive for the CPV through rapid antigen test detection between 2014-2018. Molecular screening was developed for VP1 because primers 555 for VP2 do not amplify, it was necessary to design a primer set for VP2 amplification of 982 nt. All samples that were amplified were sequenced by Sanger. Phylogenetics and structural analysis was carried out, focusing on residue 426. Results: As a result 47 out of 54 samples tested positive for VP1 screening, and 34/47 samples tested positive for VP2 980 primers as subtype 2a (n = 30) or 2b (n = 4); subtype 2c was not detected. All VP2 sequences had the amino acid, T, at 440, and most Colombian sequences showed an S514A substitution, which in the structural modeling is located in an antigenic region, together with the 426 residue. Conclusions: The 2c variant was not detected, and these findings suggest that Colombian strains of CPV-2 might be under an antigenic drift.