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Enzyme-linked Immunosorbent Assay for the Detection of Hen's Egg Proteins in Processed Foods

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Kim, Soo-Ho;Kwak, Bo-Yeon
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.36-41
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    • 2010
  • The Hen's egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30 ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries ($mean{\pm}SD$) were $129{\pm}13.7%$, $73.9{\pm}12.5%$, and $65.5{\pm}13.6%$, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen's egg proteins in processed foods.

Protein quality, hematological properties and nutritional status of albino rats fed complementary foods with fermented popcorn, African locust bean, and bambara groundnut flour blends

  • Ijarotimi, Oluwole Steve;Keshinro, Oluremi Olufunke
    • Nutrition Research and Practice
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    • v.6 no.5
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    • pp.381-388
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    • 2012
  • The objective of this study was to determine protein quality and hematological properties of infant diets formulated from local food materials. The food materials were obtained locally, fermented, and milled into flour. The flours were mixed as 70% popcorn and 30% African locust bean (FPA), 70% popcorn and 30% bambara groundnut (FPB), and 70% popcorn, 20% bambara groundnut, and 10% African locust bean (FPAB). Proximate analysis, protein quality, hematological properties, and anthropometric measurements of the animals fed with the formulations were investigated. The protein contents of the formulated diets were significantly higher than that of Cerelac (a commercial preparation) ($15.75{\pm}0.01g$/100 g) and ogi (traditional complementary food) ($6.52{\pm}0.31g$/100 g). The energy value of FPAB ($464.94{\pm}1.22\;kcal$) was higher than those of FPA ($441.41{\pm}3.05\;kcal$) and FPB ($441.48{\pm}3.05\;kcal$). The biological value (BV) of FPAB (60.20%) was the highest followed by FPB (44.24%) and FPA (41.15%); however, BV of the diets was higher than that of ogi (10.03%) but lower than that of Cerelac (70.43%). Net protein utilization (NPU) of the formulations was 41.16-60.20%, whereas true protein digestibility was 41.05-60.05%. Metabolizable energy (232.98 kcal) and digestible energy (83.69 kcal) of FPAB were the highest, whereas that of FPA had the lowest values. The protein digestibility values corrected for amino acid score of the diets (0.22-0.44) were lower than that of Cerelac (0.52), but higher than that of ogi (0.21). The growth patterns and hematological properties (packed cell volume, red blood cells, hemoglobin, mean corpuscular hemoglobin concentration, mean corpuscular hemoglobin, and mean corpuscular volume) of the formulated diets were higher than those of ogi, but lower than those of Cerelac. In conclusion, we established that the FPAB food sample was rated best in terms of protein quality over the other formulated diets. Therefore, a FPAB blend may be used as a substitute for ogi.

Proteome Analysis of Pigs Fed with Tissue Culture Medium Waste after Harvest of Korean Wild Ginseng (산삼배양액을 급여한 돼지에서 근육의 프로테옴 분석)

  • Seol, Jae-Won;Chae, Joon-Seok;Kang, Hyung-Sub;Kang, Chun-Seong;Ihn, Dong-Chul;Park, Sang-Youel
    • Journal of Veterinary Clinics
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    • v.28 no.1
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    • pp.75-80
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    • 2011
  • Proteomics is a useful approach to know protein expression, post-translational modification and protein function. We investigated the protein expression pattern and identity in pigs fed with the tissue culture medium waste after harvest of Korean wild ginseng (TCM-KWG) (Panax ginseng). Two groups (n = 30/group) of pigs were administered with 0 (control) and 16 ml/L (treatment) TCM-KWG through drinking water. After 4 weeks, we examined the protein expression pattern of longissimus dorsi muscle by Two-dimensional electrophoresis analysis. TCM-KWG treatment significantly increased two spot's density, and markedly reduced one spot's density in the muscles. We identified 3 proteins (heat shock protein 90-alpha, myosin binding protein and cofilin 2) by the ESI-MS/MS (Q-TOF2, Micromass). These results demonstrate that TCM-KWG treatment may play a protection role against physiological stress in pigs, like as increased heat shock protein 90-alpha.

Functional analysis of the rice BRI1 receptor kinase (벼 Brassinosteroid Insensitive 1 Receptor Kinase의 기능에 관한 연구)

  • Yeon, Jinouk;Kim, Hoy-Taek;Nou, Ill-Sup;Oh, Man-Ho
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.30-36
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    • 2016
  • Brassinosteroids (BRs) are essential plant steroid hormones required for cell elongation, plant growth, development and abiotic and biotic stress tolerance. BRs are recognized by BRI1 receptor kinase that is localized in the plasma membrane, and the BRI1 protein will eventually autophosphorylate in the intracellular domain and transphosphorylate BAK1, which is a co-receptor in Arabidopsis thaliana. However, little is known of the role OsBRI1 receptor kinase plays in Oryza sativa, monocotyledonous plants, compared to that in Arabidopsis thaliana, dicotyledonous plants. As such, we have studied OsBRI1 receptor kinase in vitro and in vivo with recombinant protein and transgenic plants, whose phenotypes were also investigated. A OsBRI1 cytoplasmic domain (CD) recombinant protein was induced in BL21 (DE3) E.coli cells with IPTG, and purified to obtain OsBRI1 recombinant protein. Based on Western blot analysis with phospho-specific pTyr and pThr antibodies, OsBRI1 recombinant protein and OsBRI1-Flag protein were phosphorylated on Threonine residue(s), however, not on Tyrosine residue(s), both in vitro and in vivo. This is particularly intriguing as AtBRI1 protein was phosphorylated on both Ser/Thr and Tyr residues. Also, the OsBRI1 full-length gene was expressed in, and rescued, bri1-5 mutants, such as is seen in normal wild-type plants where AtBRI1-Flag rescues bri1-5 mutant plants. Root growth in seedlings decreased in Ws2, AtBRI1, and 3 independent OsBRI1 transgenic seedlings and had an almost complete lack of response to brassinolide in the bri1-5 mutant. In conclusion, OsBRI1, an orthologous gene of AtBRI1, can mediate normal BR signaling for plant growth and development in Arabidopsis thaliana.

Biological Activity of Dandelion (Taraxacum officinale) extracts Fermented with Raw Sugar (당 첨가 민들레(Taraxacum officinale) 발효물의 생리활성)

  • Kim, Kyung-Min;Kim, Young-Nam;Choi, Byoung-Kon;Oh, Deog-Hwan
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.607-612
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    • 2012
  • Dandelions were fermented for 120 days at $15-20^{\circ}C$ in batches containing thirty and fifty percent raw sugar (FD30 and FD50). The total phenolic concentration of FD30 and FD50 were about $78.9{\pm}2.17$ and $59.35{\pm}2.56$ mg/g, respectively, both being higher than the 2$54.{\pm}1.49$ mg/g determined for DWE (dandelion water extract). The DPPH radical scavenging activity of FD30 and FD50 showed $IC_{50}$ values of $118.7{\pm}2.03$ and $123.40{\pm}2.15{\mu}g/mL$, respectively, and FD30 displayed the highest antioxidant activity. 2 mg/mL of FD30 and FD50 showed $8.8{\pm}1.72$ and $11.8{\pm}2.87{\mu}M$ production of NO, respectively, compared with $4.9{\pm}1.20{\mu}M$ of the dandelion extract. The protease, ${\alpha}$-amylase, and lipase activity of FD50 was the highest. The fibrinolytic activity of FD30 and FD50 were $0.56{\pm}0.28$ and $1.39{\pm}0.20$ unit/mg protein, respectively, which was substantially higher than the 0.28 unit/mg protein of DWE. In conclusion, the dandelions fermented by sugar showed improved antioxidant, anti-inflammatory and enzymatic activities.

Chicken Insulin-Like Growth Factor-I Stimulates Protein Synthesis of Chicken Embryo Myoblasts Cultured in Serum-Free Medium

  • Kita, K.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.1
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    • pp.17-20
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    • 2001
  • The effect of chicken IGF-I on protein synthesis of chicken embryo myoblasts cultured in serum-free medium was examined. When myoblasts were expanded to approximate 20-30% of well, the medium was changed to the serum-free medium including 0, 2, 20, 200 or 2000 ng/ml of recombinant chicken IGF-I. The culture medium including 10% fetal calf serum (FCS) was used as positive control. After 1 day of incubation, protein synthesis was measured by the incorporation of [$^3H$]-L-leucine. Thereafter cells were continued to incubate for further 18 hours, and the radioactivity in the protein was measured as an index of protein synthesis. The values for protein synthesis cultured in the serum-free medium without chicken IGF-I or with 2000 ng/ml of chicken IGF-I were the lowest. Protein synthesis was elevated with increasing chicken IGF-I concentration from 0 to 20 ng/ml. The values for protein synthesis in the 20 ng/ml and 200 ng/ml IGF-I groups were about half of that of the FCS group. The present study revealed that the potency of chicken IGF-I at the levels of 20 to 200 ng/ml to stimulate myoblast protein synthesis was about half of that of 10% FCS.

Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein

  • Yang, Zhaoshou;Lee, Jihoo;Ahn, Hye-Jin;Chong, Chom-Kyu;Dias, Ronaldo F.;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.54 no.2
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    • pp.239-241
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    • 2016
  • Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection.

Preparation of Diphtheria Toxin A Chain from Escherichia coli

  • Lee, Jong-Soo;Yoon, Kyoung-Bum;Park, Jong-Won;Choi, Suk-Jung
    • BMB Reports
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    • v.30 no.2
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    • pp.144-149
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    • 1997
  • An expression vector was constructed containing the gene encoding diphtheria toxin A (DTA) which was placed after a T7 promoter. Cytoplasmic expression of the DTA gene resulted in the formation of an insoluble inclusion body. The inclusion body was collected after the complete lysis of the cell, and subsequent washing with 0.1% Triton X-100 released 16~30% of DTA protein from the inclusion body along with other contaminating proteins. The released DTA protein was purified by dialysis. The remaining pellet was dissolved in 8 M urea containing 5% ${\beta}-mercaptoethanol$, and the denatured DTA was renatured by the dilution-dialysis method. The total yield was 35%, and about 5 mg DTA was obtained from 1 L culture. The DTA protein has a free sulfhydryl group exposed to the protein surface, and was shown to have a tendency to dimerize through disulfide formation in the absence of ${\beta}-mercaptoethanol$. The utility of the sulfhydryl group was tested for the construction of recombinant toxins.

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Enzymatic preparation and antioxidant activities of protein hydrolysates from hemp (Cannabis sativa L.) seeds

  • Hyeon-Ji Yoon;Gyu-Hyeon Park;Yu-Rim Lee;Jeong-Min Lee;Hyun-Lim Ahn;Syng-Ook Lee
    • Food Science and Preservation
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    • v.30 no.3
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    • pp.434-445
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    • 2023
  • Hemp (Cannabis sativa L.) seeds have recently been attracting attention as a new high-value-added food material owing to their excellent nutritional properties, and research on the development of functional food materials using hemp seeds is actively progressing. This study aimed to evaluate the antioxidant properties of hemp seed protein hydrolysates. Protein hydrolysates were prepared from defatted hemp seed powder (HS) by enzymatic hydrolysis using five different proteases (alcalase, bromelain, flavourzyme, neutrase, and papain). 2,4,6-trinitrobenzene sulfonic acid (TNBS) assay and SDS-PAGE analysis revealed that HS showed a high degree of hydrolysis after treatment with each enzyme except papain. The total polyphenol content of the protein hydrolysates (<3 kDa) and the RC50 values obtained from two different antioxidant tests showed that alcalase hydrolysate (HSA) had a relatively high level of antioxidant capacity. In addition, treatment with HSA (25-100 ㎍/mL) significantly inhibited linoleic acid peroxidation. These results suggest that hemp seed protein hydrolysates are potential sources of natural antioxidants. Future studies will focus on the identification of active peptides from HSA.

Analysis of Bone Mineral Density, Biochemical Index and Nutrient Intakes of 30-70 Years Old Women -Based on 2011 KNHANES- (30~70대 여성의 골밀도, 생화학적 특성 및 영양소 섭취 분석 -2011년 국민건강영양조사를 중심으로-)

  • Koo, Jae Ok;Kim, Myung Sook
    • Korean Journal of Community Nutrition
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    • v.19 no.4
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    • pp.328-341
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    • 2014
  • Objectives: This study was carried out to investigate bone mineral density (BMD), blood pressure, age and biochemical index and nutrient intakes, and to analyze the relationship between BMD, blood pressure, biochemical index, nutrient intakes of Korean women. Methods: The study subjects were 499 women with a age range of 30-79 years living in Seoul and Kyunggi area who participated in 2011 KNHANES. The study subjects were divided into 5 age groups : 30 years (145), 40 years (110), 50 years (102), 60 years (85) and 70 years (57), and 3 BMD groups: normal 258 (50.4%), osteopenia 163 (32.9%) and osteoporosis 78 (16.7%). Results: The average waist circumference, BMI and body fat increased with age, but fat free mass decreased with age. Average BMD and T-score was decreased from 0.84, $0.74g/cm^2$ in 30 years to 0.05, $-1.05g/cm^2$ in 70 years. The rates of osteopenia and osteoporosis increased significantly 22% in 30~40 years, 47% in 50 years and 56% in 60~70 years. Systolic blood pressure and cholesterol were significantly increased with age. The rates of hypertension was significantly increased 2.1% in 30 years, 30.4% in 50 years 89.5% in 70 years. Average nutrient intakes such as protein, calcium, iron, vitamin A and C were significantly decreased with age. There were significant negative correlations between age and calcium, systolic blood pressure, triglyceride. Conclusions: In this study, we found the risk of osteoporosis and hypertension were significantly increase with over 50 years age. Calcium and protein intake decreased with age. Therefore, in order to prevent osteoporosis and hypertension, adult women need to be educated regarding the importance of protein, calcium and other nutrients in their diet.