Peroxiredoxins (Prxs) are ubiquitously distributed and play important functions in diverse cellular signaling systems. The proteins are largely classified into three groups, such as typical 2-Cys Prx, atypical 2-Cys Prx, and 1-Cys Prx, that are distinguished by their catalytic mechanisms and number of Cys residues. From the three classes of Prxs, the typical 2-Cys Prx containing the two-conserved Cys residues at its N-terminus and C-terminus catalyzes $H_2O_2$ with the use of thioredoxin (Trx) as an electron donor. During the catalytic cycle, the N-terminal Cys residue undergoes a peroxide-dependent oxidation to sulfenic acid, which can be further oxidized to sulfinic acid at the presence of high concentrations of $H_2O_2$ and a Trx system containing Trx, Trx reductase, and NADPH. The sulfinic acid form of 2-Cys Prx is reduced by the action of sulfiredoxin which requires ATP as an energy source. Under the strong oxidative or heat shock stress conditions, 2-Cys Prx in eukaryotes rapidly switches its protein structure from low-molecular-weight species to high-molecular-weight protein structures. In accordance with its structural changes, the protein concomitantly triggers functional switching from a peroxidase to a molecular chaperone, which can protect its substrate denaturation from external stress. In addition to its N-terminal active site, the C-terminal domain including 'YF-motif' of 2-Cys Prx plays a critical role in the structural changes. Therefore, the C-terminal truncated 2-Cys Prxs are not able to regulate their protein structures and highly resistant to $H_2O_2$-dependent hyperoxidation, suggesting that the reaction is guided by the peroxidatic Cys residue. Based on the results, it may be concluded that the peroxidatic Cys of 2-Cys Prx acts as an '$H_2O_2$-sensor' in the cells. The oxidative stress-dependent regulation of 2-Cys Prx provides a means of defense systems in cells to adapt stress conditions by activating intracellular defense signaling pathways. Particularly, 2-Cys Prxs in plants are localized in chloroplasts with a dynamic protein structure. The protein undergoes conformational changes again oxidative stress. Depending on a redox-potential of the chloroplasts, the plant 2-Cys Prx forms super-molecular weight protein structures, which attach to the thylakoid membranes in a reversible manner.
To improve the productivity of peroxidase (POD) of cell line SP-47 derived from cell suspension cultures of sweet potato (Ipomoea batatas (L) Lam.cv White Star), we optimized culture conditions including the composition and concentration of plant growth regulators and carbon source, and the cell inoculum size. When one g (fr wt) of cells was inoculated into 50 mL TL medium supplemented with l mg/L 2,4-D and 30g/L sucrose in 300 mL Erlenmeyer flask at 25$^{\circ}C$ in the dark (100rpm), the POD activity per g cell dry wt was maximized to be about 6,800 units after 25 days of subculture, which was about 30 times higher than that of intact roots of horseradish plants grown in the greenhouse, but the cell growth was maximum after 15 days of subculture. The protein content per g cell dry wt maintained almost plateau and after 25 days of subculture decreased as culture Proceeded further whereas the POD specific activity (unit/mg protein) was about two times higher after subculture and continuously increased from 12 days to the end of cultures (40 days). The POD isozyme patterns showed almost the same regardless of cell growth stage, but some acidic isozymes were slightly increased after 25 days of subculture. These results indicate that POD activity in suspension cultures of sweet potato is closely associated with cell growth and stresses derived from cell culture renditions and medium depletion. Due to its high POD activity the SPL47cell line seems to be suitable for the mass production of POD.
Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was detected and isolated from diseased 'Fuji' apple (Malus domestica) in Korea. The coat protein (CP) genes of two ApMV strains, denoted as ApMV-Kl and ApMV-K2, were amplified by using the reverse transcription and polymerase chain reaction (RT-PCR) and were analyzed thereafter. The objectives were to define the molecular variability of genomic information of ApMV found in Korea and to develop virus-derived resistant gene source for making virus-resistant trans-genic apple. RT-PCR amplicons for the APMVS were cloned and their nucleotide sequences were determined. The CPs of ApMV-Kl and ApMV-K2 consisted of 222 and 232 amino acid residues, respectively. The identities of the CPs of the two Korean APMVS were 93.1% and 85.6% at the nucleotide and amino acid sequences, respectively. The CP of ApMV-Kl showed 46.1-100% and 43.2-100% identities to eight different ApMV strains at the nucleotide and amino acid levels, respectively. When ApMV-PV32 strain was not included in the analysis, ApMV strains shared over 83.0% and 78.6% homologies at the nucleotide and amino acid levels, respectively. ApMV strains showed heterogeneity in CP size and sequence variability. Most of the amino acid residue differences were located at the N-termini of the strains of ApMV, whereas, the middle regions and C-termini were remarkably conserved. The APMVS were 17.(1-54.5% identical with three other species of the genus Ilarviyus. ApMV strains can be classified into three subgroups (subgroups I, II, and III) based on the phylogenetic analysis of CP gene in both nucleotide and amino acid levels. Interestingly, all the strains of subgroup I were isolated from apple plants, while the strains of subgroups II and III were originated from peach, hop, or pear, The results suggest that ApMV strains co-evolved with their host plants, which may have resulted in the CP heterogeneity.
Ho Yong Shin;Chang Yoon Ji;Ho Soo Kim;Jung-Sung Chung;Sung Hwan Choi;Sang-Soo Kwak;Yun-Hee Kim;Jeung Joo Lee
Journal of Plant Biotechnology
/
v.50
/
pp.1-10
/
2023
Sweet potato (Ipomoea batatas L. Lam) is an economically important root crop and a valuable source of nutrients, processed foods, animal feeds, and pigment materials. However, during post-harvest storage, storage roots of sweet potatoes are susceptible to decay caused by various microorganisms and diseases. Post-harvest curing is the most effective means of healing wounds and preventing spoilage by microorganisms during storage. In this study, we aimed to identify proteins involved in the molecular mechanisms related to curing and study proteomic changes during the post-curing storage period. For this purpose, changes in protein spots were analyzed through 2D-electrophoresis after treatment at 33℃ (curing) and 15℃ (control) for three days, followed by a storage period of eight weeks. As a result, we observed 31 differentially expressed protein spots between curing and control groups, among which 15 were identified. Among the identified proteins, the expression level of 'alpha-amylase (spot 1)' increased only after the curing treatment, whereas the expression levels of 'probable aldo-keto reductase 2-like (spot 3)' and 'hypothetical protein CHGG_01724 (spot 4)' increased in both the curing and control groups. However, the expression level of 'sporamin A (spot 10)' decreased in both the curing and control treatments. In the control treatment, the expression level of 'enolase (spot 14)' increased, but the expression levels of 'chain A of actinidin-E-64 complex+ (spot 19)', 'ascorbate peroxidase (spot 22)', and several 'sporamin proteins (spot 20, 21, 23, 24, 27, 29, 30, and 31)' decreased. These results are expected to help identify proteins related to the curing process in sweet potato storage roots, understand the mechanisms related to disease resistance during post-harvest storage, and derive candidate genes to develop new varieties with improved low-temperature storage capabilities in the future.
With the object of the study id every possible correlation between daily food intake and growth rate of primary school students, aged eleven years old, the author distributed prepared questionnaires through 200 children of 5th grade in both sexes in K private primary school, Seoul. Specific object of this study were included following three points: such as daily food intake records for six days. daily food intake records for six days. school records of sample children at the end of 4th grade. physical growth index calculated by means of weight and height of sample children at the first part of the 5th grade. CR. t. F. test as well as percentage calculations were included for statistical analysis. The result of this study were as follows: 1. For the living circumstances a) Average numbers of brother and sister in sampled family were two to three. b) Approximately 50%of the children were brought up by mother's milk at their infants and about 35% of children were weaned at the age of 7 to 12 months. c) Thirty to forty percent of the respondents revealed irregulity in their meal time due to early school time in the morning. d) Monthly average expenditure for daily food reached 20 to 29% out of total income. e) The most favorit foods reflect as follows: The most favorit main dish is boiled rice mixed with other careals and side dish beaf. Fruits and juice are more popular snack among whole children. 2. Nutrition survey a) The calcium and thiamine were the most limited factors out of daily nutrients consumption. b) The total calory intake were composed of carbohydrate 63%, protein 16% and fat 21% respectively. c) The total food intake were composed of 20 to 24% of animal source of foods and 76 to 80% of plant source of foods (vegetables, grains, fruits) d) The total protein intake were composed of 46% of animal protein and 54% of others. e) The grain stuffs consumed in the largest quantity at the level of 50% out whole food groups. f) Twenty four percent of total food intake came from snack between meal time. 3. The result of statistical analysis were as follows: a) The correlation between the calory intake and plysical growth index revealed statistical significancy at the level of p<0.001. b) The correlation between the protein intake and physical growth index revealed statistical significancy at the level of p<0.001. c) The correlation between the meal time and physical growth index revealed statistical significancy at the level of 0.02
Kim, Eun-Jung;Shin, In-Ung;Kwun, Se-Young;Park, Eun-Hee;Yi, Jae-Hyoung;Kim, Myoung-Dong
Microbiology and Biotechnology Letters
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v.47
no.3
/
pp.434-440
/
2019
Lactic acid bacteria (LAB) isolated from kimchi were studied for their ${\beta}$-glucuronidase activity. Among the 156 strains tested, 52 strains utilized glucuronic acid as a carbon source and their intracellular ${\beta}$-glucuronidase activities were significantly higher than their extracellular activities. Leuconostoc mesenteroides KFRI 73007 isolated from turnip kimchi exhibited the highest intracellular ${\beta}$-glucuronidase activity of $0.77{\pm}0.01U/mg$ protein, which was further increased to $1.14{\pm}0.01U/mg$ protein under optimized reaction conditions (pH 7, $37^{\circ}C$). The activity of ${\beta}$-glucuronidase was notably decreased by the addition of divalent cations, and glucuronic acid was the best carbon source to produce ${\beta}$-glucuronidase in Leu. mesenteroides KFRI 73007.
de Almeida, Renato Goulart;Silva, Osmar Nascimento;de Souza Candido, Elizabete;Moreira, Joao Suender;Jojoa, Dianny Elizabeth Jimenez;Gomes, Diego Garces;de Souza Freire, Mirna;de Miranda Burgel, Pedro Henrique;de Oliveira, Nelson Gomes Junior;Valencia, Jorge William Arboleda;Franco, Octavio Luiz;Dias, Simoni Campos
CELLMED
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v.4
no.1
/
pp.5.1-5.8
/
2014
Healthcare-associated infection represents a frequent cause of mortality that increases hospital costs. Due to increasing microbial resistance to antibiotics, it is necessary to search for alternative therapies. Consequently, novel alternatives for the control of resistant microorganisms have been studied. Among them, plant antimicrobial protein presents enormous potential, with flowers being a new source of antimicrobial molecules. In this work, the antimicrobial activity of protein-rich fractions from flower tissues from 18 different species was evaluated against several human pathogenic bacteria. The results showed that protein-rich fractions of 12 species were able to control bacterial development. Due its broad inhibition spectrum and high antibacterial activity, the protein-rich fraction of Hibiscus rosa-sinensis was subjected to DEAE-Sepharose chromatography, yielding a retained fraction and a non-retained fraction. The retained fraction inhibits 29.5% of Klebsiella pneumoniae growth, and the non-retained fraction showed 31.5% of growth inhibition against the same bacteria. The protein profile of the chromatography fractions was analyzed by using SDS-PAGE, revealing the presence of two major protein bands in the retained fraction, of 20 and 15 kDa. The results indicate that medicinal plants have the biotechnological potential to increase knowledge about antimicrobial protein structure and action mechanisms, assisting in the rational design of antimicrobial compounds for the development of new antibiotic drugs.
Park, Sung-Jin;Chung, Byung-Hee;Choi, Young-Su;Kim, Jong-Dai;Park, Sung-Hye
Journal of Physiology & Pathology in Korean Medicine
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v.21
no.1
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pp.171-180
/
2007
The purpose of this study is to determine the possibility of using Flos Sophora japonica as natural health food source. To accomplish this purpose, the contents of general and antioxidative nutrients of Flos Sophora japonica a were measured. The contents of carbohydrate, crude protein, crude lipid and ash are 67.76%, 19.87%, 4.61% and 7.76%. And the calories of Flos Sophora japonica Linne was 318.32 Kcal. Total dietary fiber was 25.35% of total carbohydrates. The percentages of water soluble dietary fiber to insoluble dietary fiber were 1.80 % and 23.56 %, respectively. The protein were contained total 18 different kinds of amino acids. The contents of non-essential and essential amino acids were 4,898.78mg and 5,953.51mg. The K was the largest mineral followed by Ca, P and Mg, which means Flos Sophora japonica Linne is alkali material. The contents of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids were 29.69%, 34.93% and 35.38%. Therefore, the amount of the total unsaturated fatty acid was higher than that of any other plant. The content of vitamin C in Flos Sophora japonica Linne was higher than that of any other plant, which suggest that it could increase blood elasticity. The content of rutin, which is responsible for capillary vessel permeability, was 22.60%. The contents of water soluble antioxidative materials in 1 mL of water-extracted Flos Sophora japonica Linne were 3.9 ${\mu}$g which is comparable to 1233.0 mmol of vitamin C in antioxidant effect. The general nutrients and other antioxidatant bioactive materials in Flos Sophora japonica Linne were also potential materials for good health food. It is expected that follow up study of Flos Sophora japonica Linne through developing processed food and evaluation of their functional properties would provide useful information as a source of medicinal foods.
Lee, S.S.;Shin, K.J.;Kim, W.Y.;Ha, J.K.;Han, In K.
Asian-Australasian Journal of Animal Sciences
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v.12
no.6
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pp.988-1001
/
1999
The rumen ecosystem is increasingly being recognized as a promising source of superior polysaccharide-degrading enzymes. They contain a wide array of novel enzymes at the levels of specific activities of 1,184, 1,069, 119, 390, 327 and $946{\mu}mol$ Reducing sugar release/min/mg protein for endoglucanase, xylanase, polygalactouronase, amylase, glucanase and arabinase, respectively. These enzymes are mainly located in the surface of rumen microbes. However, glycoside-degrading enzymes (e.g. glucosidase, fucosidase, xylosidase and arabinofuranosidase, etc.) are mainly located in the rumen fluid, when detected enzyme activities according to the ruminal compartments (e.g. enzymes in whole rumen contents, feed-associated enzymes, microbial cell-associated enzymes, and enzymes in the rumen fluid). Ruminal fungi are the primary contributors to high production of novel enzymes; the bacteria and protozoa also have important functions, but less central roles. The enzyme activities of bacteria, protozoa and fungi were detected 32.26, 19.21 and 47.60 mol glucose release/min/mL mediem for cellulose; 42.56, 14.96 and 64.93 mmol xylose release/min/mL medium after 48h incubation, respectively. The polysachharide-degrading enzyme activity of ruminal anaerobic fungi (e.g. Neocallimastix patriciarum and Piromyces communis, etc.) was much higher approximately 3~6 times than that of aerobic fungi (e.g. Tricoderma reesei, T. viridae and Aspergillus oryzae, etc.) used widely in industrial process. Therefore, the rumen ecosystem could be a growing source of novel enzymes having a tremendous potential for industrial applications.
Alzheimer's disease (AD) is a neurodegenerative disorder characterized by neuronal loss and extracellular senile plaques containing $\beta$-amyloid peptide (A$\beta$). The deposition of the A$\beta$ peptide following proteolytic processing of amyloid precursor protein (APP) by $\beta$-secretase (BACE1) and $\gamma$-secretase is a critical feature in the progression of AD. Among the plant extracts tested, the ethanol extract of Petasites japonicus leaves showed novel protective effect on B103 neuroblastoma cells against neurotoxicity induced by A$\beta$, as well as a strong suppressive effect on BACE1 activity. Ethanol extracts of P. japonicus leaves were sequentially extracted with methylene chloride, ethyl acetate and butanol and evaluated for potential to inhibit BACE1, as well as to suppress A$\beta$-induced neurotoxicity. Exposure to A$\beta$ significantly reduced cell viability and increased apoptotic cell death. However, pretreatment with ethyl acetate fraction of P. japonicus leaves prior to A$\beta$ (50 ${\mu}M$) significantly increased cell viability (p<0.01). In parallel, cell apoptosis triggered by A$\beta$ was also dramatically inhibited by ethyl acetate fraction of P. japonicus leaves. Moreover, the ethyl acetate fraction suppressed caspase-3 activity to the basal level at 30 ppm. Taken together, these results demonstrated that P. japonicus leaves appear to be a useful source for the inhibition and/or prevention of AD by suppression of BACE1 activity and attenuation of A$\beta$ induced neurocytotoxicity.
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