• Title/Summary/Keyword: Iron-containing protein

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Optimization of the Expression of the Ferritin Protein Gene in Pleurotus eryngii and Its Biological Activity (큰느타리버섯에서 석충 페리틴 단백질 유전자의 발현 최적화 및 생물학적 활성)

  • Woo, Yean Jeong;Oh, Si Yoon;Choi, Jang Won
    • The Korean Journal of Mycology
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    • v.47 no.4
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    • pp.359-371
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    • 2019
  • To optimize the expression and secretion of ferritin protein associated with ion storage in the mushroom, Pleurotus eryngii, a recombinant secretion vector, harboring the ferritin gene, was constructed using a pPEVPR1b vector under the control of the CaMV 35S promoter and signal sequence of pathogen related protein (PR1b). The ferritin gene was isolated from the T-Fer vector following digestion with EcoRI and HindIII. The gene was then introduced into the pPEVPR1b secretion vector, and it was then named pPEVPR1b-Fer. The recombinant vector was transferred into P. eryngii via Agrobacterium tumefaciens-mediated transformation. The transformants were selected on MCM medium supplemented with kanamycin and its expression was confirmed by SDS-PAGE and western blotting. Expression of ferritin protein was optimized by modifying the culture conditions such as incubation time and temperature in batch and 20 L airlift type fermenter. The optimal conditions for ferritin production were achieved at 25℃ and after incubating for 8 days on MCM medium. The amount of ferritin protein was 2.4 mg/g mycelia, as measured by a quantitative protein assay. However, the signal sequence of PR1b (32 amino acids) seems to be correctly processed by peptidase and ferritin protein may be targeted in the apoplast region of mycelia, and it might not be secreted in the culture medium. The iron binding activity was confirmed by Perls' staining in a 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in P. eryngii mycelia. Mycelium powder containing ferritin was tested as a feed additive in broilers. The addition of ferritin powder stimulated the growth of young broilers and improved their feed efficiency and production index.

Effect of Protease on the Morphological Properties and Dyeability of Human Hair (프로테아제 처리가 모발의 염색성 및 형태적 특성에 미치는 영향)

  • Kim, Hong-Hee;Kwon, Tae-Jong
    • Textile Coloration and Finishing
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    • v.20 no.2
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    • pp.59-65
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    • 2008
  • The tannin acid and the enzymes have been used in order to improve the ruggedness in laundry and the absorption of dyes and pigments in the textile industry for several years. The enzyme processing on the protein fiber minimizes the damage of the entire fiber and improves the dyeability by effectively modifying only the hydrophobic surface. This study tried out the structural observation by applying the Castanea crenata sieb. et. zucc. containing abundant tannin to the hair dyeing as the natural dyeing pigment along with Protease of Rhizopus sp. The dyeability was improved as compared to the dyeing using only the synthetic tanning and iron mordant. When the depth of pigment was higher in accordance with the surface observation, the enzyme dissolution had impact on dyeing and so the keratin layer on the hair surface. Accordingly, it was found that the appropriate depth was between 0.01 and 0.03%. It was estimated that 0.1% protease would treated within 30min. Consequently, it would cause the good reaction with the functional group of tannin pigment.

The Riboflavin Status in Insulin-Dependent Diabetes Mellitus During Growing Period (성장기 인슐린 의존성 당뇨병 환자의 리보플라빈 영양상태)

  • 이정희
    • Journal of Nutrition and Health
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    • v.27 no.7
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    • pp.729-739
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    • 1994
  • Riboflavin status of 17 insulin-dependent diabetic mellitus(IDDM) patients in growing period was evaluated as a function of energy intake and expenditure, biochemical nutritional status and diabetic control indicators. Compared with recommended dietary allowances for Koreans(RDA, 1989), only 35.3% of subjects was at good levels of all nutrients intakes and 52.9% of subjects was below normal level of height and weight. Nutrients consumed below RDA levels were energy(=88.5% of subjects), niacin(64.7%), iron(52.9%) and protein(23.5%) respectively. The riboflavin status was within normal range by urinary riboflavin excretion but 17.6% of subjects was evaluated as showed riboflavin deficiency by erythrocyte glutathione reductase activity coefficient(EGRAC). Correlation between riboflavin intake, urinary riboflavin excretion, EGRA level and diabetic duration were not statistically significant. Correlation analyses indicated that EGRA level was inversely correlated with thiamin, niacin and cabohydrate intake. No significant correlations were found between the EGRA and glycosylated hemoglobin A1(HbA1) (r=-0.464, p=0.129). From this study, it is suggested that IDDM subjects need to maintain balanced diet containing nutrients above RDA for individual activity during growing period. It needs more study whether the current recommended riboflavin allowance is adequate for diabetic patients.

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Relationship between Red Pepper Intake, Capsaicin Threshold, Nutrient Intake, and Anthropometric Measurements in Young Korean Women (젊은 한국여성에서 붉은고추의 섭취량, 캡사이신 역치, 영양소 섭취량 및 신체계측치간의 관련성)

  • Kim Seok-Young;Park Gyeong-Min
    • Journal of Nutrition and Health
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    • v.38 no.1
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    • pp.76-81
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    • 2005
  • The effects of the amount of red pepper intake per a day on the capsaicin threshold, nutrient intake, and anthropo-metric measurements were investigated in 100 female students of university. Three 24-hour recalls were performed to estimate usual nutrient and red pepper intake of free-living participants. The solutions containing capsaicin at concentration of 0.05, 0.1, 0.2, 0.3, 0.4, 0.5ppm used for the evaluation of hot-taste detection threshold. Subjects were asked to recognize the burning sensation after tasting l0ml of each test solution in ascending order of capsaicin concentrations. Mean intake of red pepper was 4.6 g/d and the capsaicin threshold was 0.27 ppm. The detection threshold for capsaicin was correlated with the amount of pepper intake per a day. Red pepper intake was correlated with energy, protein, lipid, carbohydrate, Iron, vitamin A, vitamin B$_2$ niacin, and vitamin E intakes. However, it was not correlated with intake of calcium, vitamin B$_1$ or vitamin C. The red pepper intake was negatively correlated with fat mass and waist girth and the capsaicin threshold was also negatively correlated with pulse. In conclusion, red pepper intake was associated with nutrient intake and capsaicin threshold whereas it was negatively correlated with fat mass and waist girth.

Evaluation of systemic and mucosal immune responses in mice administered with recombinant Salmonella Typhimurium expressing IutA protein

  • Oh, In-Gyeong;Choi, Minsu;Lee, John Hwa
    • Korean Journal of Veterinary Research
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    • v.53 no.3
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    • pp.163-167
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    • 2013
  • Avian pathogenic Escherichia coli (APEC) are known to cause extraintestinal disease in poultry, leading to substantial losses in the industry. IutA, iron-regulated aerobactin receptor is firmly associated with APEC. To assess the potential of IutA to induce protective immune responses, attenuated Salmonella Typhimurium strain expressing IutA was constructed and administered orally to BALB/c mice. The IutA-specific immune responses were measured with sera, vaginal and fecal samples by an enzyme-linked immunosorbent assay. We found that the Salmonella-IutA vaccine induced significantly higher immune responses as compared to the control inoculated with the attenuated S. Typhimurium containing the plasmid only. The IutA-specific immune responses were increased by second immunization at third week after initial immunization, whereas triple immunization induced lower immune responses than those induced by the double immunization. The Salmonella-IutA vaccine induced a nature of immunity biased to the Th1-type, as judged by the ratio of IutA-specific IgG isotypes (IgG2a/IgG1). Overall, these results suggest that the Salmonella-IutA vaccine appear to be suitable candidate for a vaccine against APEC.

Molecular characterization and docking dynamics simulation prediction of cytosolic OASTL switch cysteine and mimosine expression in Leucaena leucocephala

  • Harun-Ur-Rashid, Md.;Masakazu, Fukuta;Amzad Hossain, Md.;Oku, Hirosuke;Iwasaki, Hironori;Oogai, Shigeki;Anai, Toyoaki
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.36-36
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    • 2017
  • Out of twenty common protein amino acids, there are many kinds of non protein amino acids (NPAAs) that exist as secondary metabolites and exert ecological functions in plants. Mimosine (Mim), one of those NPAAs derived from L. leucocephala acts as an iron chelator and reversely block mammalian cell cycle at G1/S phases. Cysteine (Cys) is decisive for protein and glutathione that acts as an indispensable sulfur grantor for methionine and many other sulfur-containing secondary products. Cys biosynthesis includes consecutive two steps using two enzymes-serine acetyl transferase (SAT) and O-acetylserine (thiol)lyase (OASTL) and appeared in plant cytosol, chloroplast, and mitochondria. In the first step, the acetylation of the ${\beta}$-hydroxyl of L-serine by acetyl-CoA in the existence of SAT and finally, OASTL triggers ${\alpha}$, ${\beta}$-elimination of acetate from OAS and bind $H_2S$ to catalyze the synthesis of Cys. Mimosine synthase, one of the isozymes of the OASTLs, is able to synthesize Mim with 3-hydroxy-4-pyridone (3H4P) instead of $H_2S$ for Cys in the last step. Thus, the aim of this study was to clone and characterize the cytosolic (Cy) OASTL gene from L. leucocephala, express the recombinant OASTL in Escherichia coli, purify it, do enzyme kinetic analysis, perform docking dynamics simulation analysis between the receptor and the ligands and compare its performance between Cys and Mim synthesis. Cy-OASTL was obtained through both directional degenerate primers corresponding to conserved amino acid region among plant Cys synthase family and the purified protein was 34.3KDa. After cleaving the GST-tag, Cy-OASTL was observed to form mimosine with 3H4P and OAS. The optimum Cys and Mim reaction pH and temperature were 7.5 and $40^{\circ}C$, and 8.0 and $35^{\circ}C$ respectively. Michaelis constant (Km) values of OAS from Cys were higher than the OAS from Mim. Inter fragment interaction energy (IFIE) of substrate OAS-Cy-OASTL complex model showed that Lys, Thr81, Thr77 and Gln150 demonstrated higher attraction force for Cys but 3H4P-mimosine synthase-OAS intermediate complex showed that Gly230, Tyr227, Ala231, Gly228 and Gly232 might provide higher attraction energy for the Mim. It may be concluded that Cy-OASTL demonstrates a dual role in biosynthesis both Cys and Mim and extending the knowledge on the biochemical regulatory mechanism of mimosine and cysteine.

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Transgenic Siberian Ginseng Cultured Cells That Produce High Levels of Human Lactoferrin (인체 락토페린 생산 형질전환 가시오갈피 배양세포)

  • Jo Seung-Hyun;Kwon Suk-Yoon;Kim Jae-Whune;Lee Ki-Teak;Kwak Sang-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.209-215
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    • 2005
  • Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

Nutritional Properties, as Food Resources for Menu Development, of Cubed Snailfish, Shaggy Sea Raven, and Two Kinds of Wild Vegetables that are Staple Products in Samcheok (삼척지역 특산물인 곰치, 풍덕구이, 곰취, 곤드레의 향토식품 소재화를 위한 영양적 특성 연구)

  • Surh, Jeong-Hee;Kim, Jong-Ok;Kim, Mi-Hyun;Lee, Jae-Choel;Yang, Hee-Wan;Lee, Byung-Yong;Kim, Mi-Young;Yun, Sun-Ju;Jeong, Hye-Ryeon
    • Korean journal of food and cookery science
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    • v.25 no.6
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    • pp.690-702
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    • 2009
  • Liparis tessellatus(cubed snailfish), Hemitripterus villosus(shaggy sea raven), Ligularia fischeri, and Cirsiu setidens, which are staple products in Samcheok, were studied as food resources for the development of a representative local menu. Thus, their nutritional compositions were analyzed, and the resulting values were used to assess the nutritional characteristics of the newly-developed menu. In Liparis tessellatus and Hemitripterus villosus, the organic components including protein and fat tended to be lower than the corresponding values referred to as the 'Seventh Revision Food Composition Table' (2006), which was presumably linked to the spawning period of the fish. The contents of Zn, Cu, and Mg, which are nutritionally important even though the consumption of these minerals by Koreans is not sufficient, were determined to 0.5, 0.2, and 34.3 mg/100 g for Ligularia fischeri, and 26.2, 1.1, and 147.7 mg/100 g for dried Cirsiu setidens. Cirsiu setidens cultivated in Samcheok contained noticeably higher amounts of calcium, zinc, and iron, and its calcium level was more than an order of magnitude higher than that provided by the Food Composition Table. This might be associated with the fact of that Samcheok is abundant in lime which consists of calcium-containing inorganic materials. n-3 fatty acids including docosahexaenoic acid(DHA) accounted for 21% and 13% of the total fatty acids, for Liparis tessellatus and Hemitripterus villosus, respectively, and the major amino acids were glutamic acid, aspartic acid, glycine, and alanine for both fish. One of the newly developed menus, Cirsiu setidens hae-jang-gook, was nutritionally superior to the corresponding menu that used dried radish leaves, a typical ingredient in hae-jang-gook, in terms of the protein, iron, and zinc contents. Rice pottage cooked with Liparis tessellatus instead of abalone was high in protein, calcium, iron and zinc, where one serving provided 29.1, 15.4, 31.0, and 27.8% of their Reference Intake (RI)s, respectively.

Relationship between atopic dermatitis and the Korean Healthy Eating Index score of adults: based on the 7th (2016-2018) Korea National Health and Nutrition Examination Survey (성인의 아토피 피부염과 식생활평가지수와의 관계: 국민건강영양조사 제7기 (2016-2018년) 자료 이용)

  • Kim, Hye Won;Kim, Ji-Myung
    • Journal of Nutrition and Health
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    • v.55 no.5
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    • pp.558-571
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    • 2022
  • Purpose: The purpose of this study was to analyze the dietary intake of Korean adults with atopic dermatitis (AD) to determine whether the risk of developing AD was related to their diet. Methods: Among the participants in the 7th National Health and Nutrition Survey (2016 to 2018), 10,571 adults aged 19-64 were divided into AD (AD group, n = 366) and control groups (non-AD group, n = 10,205) to compare and analyze their nutrient intake status and Korean Healthy Eating Index (KHEI) score. Depending on the prevalence of AD, the nutrient intake, nutrient intake per 1,000 kcal, acceptable macronutrient distribution range, and KHEI score were tested for significance by correcting for age, sex, body mass index, education, marriage, occupation, residence area, physical activity, and energy intake variables that were derived for confounding factors. Results: The acceptable macronutrient distribution range for protein was lower in the AD group than in the non-AD group. Comparing the nutrient intake per 1,000 kcal, the intakes of protein, phosphorus, iron, and potassium of the AD group were significantly lower than those of the non-AD group. In the KHEI, the scores of total vegetables, vegetables excluding kimchi and pickles, meat, fish, eggs, and legumes of the AD group were significantly lower than those of the non-AD group. In addition, as these food intake scores increased, the risk of AD significantly decreased. Conclusion: In conclusion, adult AD patients had low intakes of vegetables, meat, fish, eggs, and legumes, and it was confirmed that high intakes of these foods may be associated with low AD risk. Therefore, we suggest that an adequate intake of vegetables and foods containing protein would be necessary for the management and treatment of AD in adults.

Effects of Salicornia herbacea L. Supplementation on Lipid Peroxidation and Mineral Levels in Streptozotocin-Induced Diabetic Rats (함초 보충시 당뇨유발 흰쥐의 지질과산화와 무기질 함량에 미치는 영향)

  • Kim, Myung-Wha
    • Journal of Nutrition and Health
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    • v.40 no.5
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    • pp.403-412
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    • 2007
  • This study was designed to examine the effects of Salicornia herbacea L. (glasswort: GW) on the lipid peroxidation and mineral levels in diabetic rats. Diabetes mellitus was induced in male Sprague-Dawley rats weighing 200-220 g by an injection of streptozotocin (STZ) dissolved in a citrate buffer into the tail vein at a dose of 45 mg/kg of body weight. Sprague-Dawley rats were fed an AIN-93 recommended diet and the experimental groups were fed a modified diet containing 10% and 20% of glasswort powder for 4 weeks. The experimental groups were divided into 6 groups which consisted of normal (N)-control group, N-GW 10% and N-GW 20% treated groups, STZ-control, STZ-GW 10% and STZ-GW 20% treated groups. The rats' liver and muscle glycogen, liver and kidney protein, cholesterol and triglyceride (TG) in liver, malondialdehyde (MDA) in liver and kidney values were measured, along with the hepatic of chromium (Cr), iron (Fe), and zinc (Zn) content. The liver glycogen levels was significantly affected in N-GW 20% group among all the experimental groups. The liver MDA levels of the STZ-GW 10% and STZ-GW 20% groups were significantly lower than for the STZ-control group. There were significant differences between the N-control group and the STZ-control group in the hepatic of Zn levels. The hepatic of Cr levels in the N-GW 20% and STZ-GW 10% and STZ-GW 20% groups were significanly higher than for the each control groups. These results exhibited dose related effect of glasswort and it may have favorable influence on lipid peroxidation in the liver.