• Title/Summary/Keyword: protein recovery

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Determination of L-Carnitine in Infant Powdered Milk Samples after Derivatization

  • Park, Jung Min;Koh, Jong Ho;Kim, Jin Man
    • Food Science of Animal Resources
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    • v.41 no.4
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    • pp.731-738
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    • 2021
  • Herein, a novel analytical method using a high-performance liquid chromatography-fluorescence detector (HPLC/FLD) is developed for rapidly measuring an L-carnitine ester derivative in infant powdered milk. In this study, solid-phase extraction cartridges filled with derivatized methanol and distilled water were used to effectively separate L-carnitine. Protein precipitation pretreatment was carried out to remove the protein and recover the analyte extract with a high recovery (97.16%-106.56%), following which carnitine in the formula was derivatized to its ester form. Precolumn derivation with 1-aminoanthracene (1AA) was carried out in a phosphate buffer using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) as the catalyst. Method validation was performed following the AOAC guidelines. The calibration curves were linear in the L-carnitine concentration range of 0.1-2.5 mg/L. The lower limit of quantitation and limit of detection of L-carnitine were 0.076 and 0.024 mg/L, respectively. The intra- and interday precision and recovery results were within the allowable limits. The results showed that our method helped reduce the sample preparation time. It also afforded higher resolution and better reproducibility than those obtained by traditional methods. Our method is suitable for detecting the quantity of L-carnitine in infant powdered milk containing a large amount of protein or starch.

Changes in Biochemical Composition of the Digestive Gland of the Female Purple Shell, Rapana venosa, in Relation to the Ovarian Developmental Phases

  • Chung, Ee-Yung;Kim, Sung-Yeon;Park, Kwan-Ha
    • The Korean Journal of Malacology
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    • v.17 no.1
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    • pp.27-33
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    • 2001
  • The Ovarian developmental phases of the reproductive cycle of Rapana venosa can be classified into five successive stages by histological study: early active stage (September to February), late active stage (December to April), ripe stage (March to July), partially spawned stage (May to August), and recovery stage (June to September). To understand the characteristics of nutrient storage and utilization in the digestive gland cells with ovarian developmental phases, we examined the digestive gland - which is the major nutrient supply organ associated with ovarian development of the female purple shell - by biochemical methods. Total protein contents in the digestive gland tissues increased in March (late active stage) and reached the maximum in May (ripe and partially spawned stages), and then their levels sharply decreased in July (partially spawned and recovery stages). Total lipid contents in the digestive gland tissues reached the maximum in January (early active stage). Thereafter, their levels rapidly decreased from May (ripe and partially spawned stages) and reached a minimum in July (partially spawned and recovery stages). The total DNA contents did not significantly change regardless of the different developmental stages of the ovary. However, it was also found from biochemical analysis that changes in total RNA content follow the same seasonal cycling to protein. These results indicate that the digestive gland is an important energy storage and supply organ in purple shells, and that the nutrient contents of the digestive gland change in response to gonadal energy needs.

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Effects of Nerve Regeneration by Bogijetong-tang Treatment on Peripheral Nerves Damaged by Taxol and Crush Injury (보기제통탕이 말초신경병증 모델에서 신경 손상 회복에 미치는 영향)

  • Park, Sang-Woo;Kim, Chul-Jung;Cho, Chung-Sik
    • The Journal of Internal Korean Medicine
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    • v.34 no.4
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    • pp.384-404
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    • 2013
  • Objectives : Effects of Bogijetong-tang (BJT) on peripheral nerve regeneration have been reported in a previous study on BJT but additional study on a damaged peripheral neuropathy where its damage level is physically and chemically more severe was needed. Plus, this study was conducted because there haven't been any studies for BJT on central nerve regeneration. Methods : In order to check the effect on central nerve regeneration, the study on cerebellum cells was started and the sciatic nerve was used to observe the effects on a peripheral nerve which was severely damaged both physically and chemically. Nerve recovery effects were observed by analyzing target proteins such as phospho-extracellular signal-regulated kinase, ${\beta}1$ integrin, neurofilament 200, growth-associated protein-43, cyclin-dependent kinase 1, phospho-vimentin, phospho-Smad, and caspase 3. Results : The significant changes of target protein in cerebellum neurons have been observed. The changes of index protein on the axon regeneration and the nerve recovery in the sciatic nerve have been observed and the effects on cell protection were observed, as well. Conclusions : This study confirmed that BJT made a significant influence on nerve protection and recovery of a damaged peripheral neuropathy and it also made a possibility of its regeneration in a damaged central nerve injury.

Development of Immunoassay Systems for the Assay of Soy Protein in Meat Products; The Assay of Soy Protein in Meat Blends and Commercial Product by Enzyme-linked Immunosorbent Assay(ELISA) (육제품에 첨가된 대두단백 정량을 위한 면역분석법 개발에 관한 연구 ; ELISA에 의한 고기유화물 및 시판육제품에 첨가된 대두단백 정량)

  • Kim, Cheon-Jei;Kim, Jong-Bae;Kim, Byung-Cheol;Lee, Seoung-Bae;Jung, Sung-Won;Choe, Doo-Young;Ko, Won-Sick
    • Korean Journal of Food Science and Technology
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    • v.24 no.5
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    • pp.397-403
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    • 1992
  • This study was carried out for the development of assay method to quantify the soy protein content in meat homogenate, emulsion-type sausage and commercial meat products by Enzyme-linked Immunosorbent Assay(ELISA). The standard antigen was extracted before and after heat treatment. It was observed that the degree of reaction was not varied significantly according to the heating temperature. The recovery rate in meat homogenate and emulsion-type sausage was not varied significantly according to the heating temperature. The reaction was not interfered with fat and spices of the samples. Samples with 10% soy protein showed lower correlation than those with 2% and 5% soy protein. The recovery rate in commercial meat products showed difference individually. The correlation of some products with raw vegetable and wheat starch was relatively low.

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Effects of High-Protein Diet and/or Resveratrol Supplementation on the Immune Response of Irradiated Rats

  • Kim, Kyoung Ok;Park, Hyunjin;Kim, Hyun-Sook
    • Preventive Nutrition and Food Science
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    • v.19 no.3
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    • pp.156-163
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    • 2014
  • We investigated the effects of a high-protein diet and resveratrol supplementation on immune cells changes induced by abdominal irradiation in rats. Female Wistar rats were divided into 5 groups: 1) control diet, 2) control diet with irradiation 3) 30% high-protein diet with irradiation, 4) normal diet with resveratrol supplementation and irradiation, and 5) 30% high-protein diet with resveratrol supplementation and irradiation. We measured blood protein and albumin concentrations, lipid profiles, white blood cell (WBC) counts, proinflammatory cytokine production, and splenocyte proliferation in rats that had been treated with a 17.5 Gy dose of radiation 30 days prior. A high-protein diet affected plasma total cholesterol and very low density lipoprotein-cholesterol levels, which were increased by the radiation treatment. In addition, the lymphocyte percentage and immunoglobulin M (IgM) concentration were increased, and the neutrophil percentage was decreased in rats fed a high-protein diet. Resveratrol supplementation decreased the triglyceride (TG) level, but increased the IgM concentration and splenocyte proliferation. Proinflammatory cytokine production was lower in rats fed a high-protein diet supplemented with resveratrol than in rats fed a control diet. The results of the present study indicate that high-protein diets, with or without resveratrol supplementation, might assist with recovery from radiation-induced inflammation by modulating immune cell percentages and cytokine production.

Effect of regular exercise during recovery period following steroid treatment on the atrophied hindlimb muscles induced by steroid in rats (스테로이드 치료 후 회복기의 규칙적인 운동이 쥐의 스테로이드 유발성 위축 뒷다리근에 미치는 영향)

  • Choe, Myoung-Ae;Byun, Young-Soon;Hwang, Ae-Ran;Kim, Hee-Seung;Hong, Hae-Sook;Choi, S-Mi;Seo, Wha-Sook;Lee, Kyung-Sook;Park, Mi-Jung;Shin, Gi-Soo;An, Gyeong-Ju;Lee, Yoon-Kyoung;Im, Ji-Hae
    • Journal of Korean Biological Nursing Science
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    • v.4 no.1
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    • pp.85-99
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    • 2002
  • This study was conducted to determine whether low intensity regular exercise following steroid treatment could attenuate steroid-induced muscle atrophy. Thirty-eight Sprague-Dawley rats weighing $165{\sim}175g$ were divided into six groups ; control group(C), dexamethasone administration group(D), sedentary normal saline administration group(C+Se), exercise after normal saline administration group(C+Ex), sedentary group after dexamethasone administration(D+Se), exercise group after dexamethasone administration(D+Ex). Either dexamethasone(5mg/kg) or normal saline was injected for 7days accordingly. Exercise was started at 10m/min on the $10^{\circ}$ grade treadmill and gradually increased up to 15m/min by the 7th day for 60minutes/day($20min{\times}3$). The data were analyzed by Kruskal-Wallis test and Mann-Whitney U test using the SPSS WIN 9.0 program. Body weight, muscle weight and myofibrillar protein content of both plantaris and gastrocnemius, Type I, II muscle fiber cross-sectional area of plantaris, and Type II muscle fiber cross-sectional area of gastrocnemius in D group were significantly lower than those of C group(p<0.05) respectively. Hindlimb muscle weight, myofibrillar protein content of both plantaris and gastrocnemius. Type I muscle fiber cross-sectional area of soleus and Type I, II muscle fiber cross-sectional area of plantaris in D+Ex group tended to increase compared to those of D+Se group. Myofibrillar protein content of both plantaris and gastrocnemius, Type I muscle fiber cross-sectional area of plantaris in D+Ex group tended to increase compared to those of C+Se group. Based on these results, it is suggested that regular low-intensity exercise during recovery period after steroid treatment might facilitate the recovery from steroid-induced muscle atrophy.

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Heterologous Expression and Optimized One-Step Separation of Levansucrase via Elastin-like Polypeptides Tagging System

  • Kang, Hye-Jin;Kim, Jin-Hee;Chang, Woo-Jin;Kim, Eung-Soo;Koo, Yoon-Mo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1751-1757
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    • 2007
  • Elastin-like polypeptides (ELPs) undergo a reversible inverse phase transition upon a change in temperature. This thermally triggered phase transition allows for a simple and rapid means of purifying a fusion protein. Recovery of ELPs-tagged fusion protein was easily achieved by aggregation, triggered either by raising temperature or by adding salt. In this study, levansucrase has been used as a model enzyme in the development of a simple one-step purification method using ELPs. The levansucrase gene cloned from Pseudomonas aurantiaca S-4380 was tagged with various sizes of ELPs to functionally express and optimize the purification of levansucrase. One of two ELPs, ELP[V-20] or ELP[V-40], was fused at the C-terminus of the levansucrase gene. A levansucrase-ELP fusion protein was expressed in Escherichia coli $DH5{\alpha}$ at $37^{\circ}C$ for 18 h. The molecular masses of levansucrase-ELP[V-20] and levansucrase-ELP[V-40] were determined as 56 kDa and 65 kDa, respectively. The phase transition of levansucrase-ELP[V-20] occurred at $20^{\circ}C$ in 50 mM Tris-Cl (pH 8) buffer with 3 M NaCl added, whereas the phase transition temperature ($T_t$) of levansucrase-ELP[V-40] was $17^{\circ}C$ with 2 M NaCl. Levansucrase was successfully purified using the phase transition characteristics of ELPs, with a recovery yield of higher than 80%, as verified by SDS-PAGE. The specific activity was measured spectrophotometrically to be 173 U/mg and 171 U/mg for levansucrase-ELP[V-20] and levansucrase-ELP[V-40], respectively, implying that the ELP-tagging system provides an efficient one-step separation method for protein purification.

The Degumming and Sericin Recovery of the Silk fabric Using the Electrolytic Water (전해수를 이용한 견섬유 정련 및 세리신 회수 (I))

  • 배기서;하헌주;박광수
    • Textile Coloration and Finishing
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    • v.14 no.4
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    • pp.249-258
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    • 2002
  • Natural silk is formed by two proteins : the crystalline fibroin (inside the silk thread) and amorphous sericin (as a tube outside the thread). The degumming process is used to eliminate the external sericin prior to dyeing ; generally it makes use of soaps at about pH 10. Sericin is the protein constituent that "gums"together the fibroin filaments of cocoon silk. It constitutes about 25% of the weight of the cocoon, is soluble in hot water and "gels" on cooling. The removal of sericin from raw silk, known as degumming, is a simple but important process usually employing hot dilute soap or alkaline solution and occasionally dilute acids or enzymic methods. During degumming, alkali is taken up by the sericin and the free acid from the soap is formed ; this may be deposited on the fiber, reducing the rate of degumming and protecting it from hydrolysis. Alkali is often added to maintain or restore the pH of the baths, but it is rarely used alone, since it leaves the silk rather harsh in handle. If complete sericin removal is required as for printing, sodium carbonate may be added. If the pH of the bath exceeds 11, the fibroin is attacked. Recently, According to the development of electrolysis, we can be obtained the electrolytic reduction water(above pH 11.5) and electrolytic oxidation water (below pH 3). The aim of this work was to study a degumming process using electrolytic water and a possibility of sericin recovery. The new degumming process used electrolytic water operates at $95^\circ{C}$ for 2hr. without any reagents. The wastewater of this process are formed by a solution of sericin in water. This conditions suggest the study of a possible recovery of this protein (sericin) which has an amino acid composition suitable for many used in cosmetics, textile finishing agents, animal feeding, etc. The degumming process using electrolytic water is available to reduce treatment costs and pollute and at the same time to recover sericin.

Expression of orf7(oxi III) as dTDP-Glucose 4,6-Dehydratase Gene Cloned from Streptomyces antibioticus Tu99 and Biochemical Characteristics of Expressed Protein

  • Yoo, Jin-Cheol;Han, Ji-Man;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.206-212
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    • 1999
  • The gene orf7(oxi III) was expressed using an E. coli system in anticipation that it would encode dTDP-glucose 4,6-dehydratase which is involved in the biosynthesis of the olivose moiety of chlorothricin produced from Streptomyces antibioticus Tu99. The solubility of the expressed protein increased up to 20% under optimal induction conditions. The expressed protein was purified from the E. coli BL 21(DE3) cell lysate by a 28.5-fold purification in two chromatography steps with a 38% recovery to near homogeneity. The molecular weight and N-terminal amino acid sequence of the purified protein correlated with the predicted mass and sequence deduced from the orf7 gene. The purified protein was a homodimer with a subunit relative molecular weight of 38,000 Dalton. The expressed protein was found to exhibit dTDP-glucose 4,6-dehydratase activity and be highly specific for dTDP-glucose as a substrate. The values of K'm and V'max for dTDP-glucose were 28 $\mu$M and 295 nmol $min^{-1} (mg protein)^{-1}$, respectively. dTTP and dTDP were strong inhibitors of this enzyme.$NAD^+$, the coenzyme for dTDP-glucose 4,6-dehydratase, was tightly bound to the expressed protein.

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Effects of Number of Washes and Salt Treatment on the Quality Characteristics of Protein Recovered from Alaska Pollock and Pork Leg

  • Jin, Sang Keun;Lee, Seung Yun;Yim, Dong Gyun;Hur, Sun Jin
    • Food Science of Animal Resources
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    • v.39 no.3
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    • pp.503-509
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    • 2019
  • The aim of this study was to compare the effects of number of washes and salt addition on the meat quality in protein recovered from Alaska Pollock compared with pork leg. Various properties of protein recovered from Alaska Pollock (C, washed twice, no salt) and pork leg (T1, washed twice, no salt; T2, two washes, salt added; T3, washed four times, no salt; and T4, washed four times, salt added) were assessed in this study. Pork leg samples exhibited better color (more whiteness, less yellowness) than Alaska Pollock samples. In pork leg samples, four washes (T3, T4) during processing yielded whiter, less yellow protein than two washes (T1, T2). Overall, the textural property measures were higher in pork leg samples (T2, T3, and T4) than in other samples. Breaking force, jelly strength, and folding resistance were significantly higher in salt-treated pork leg samples (T2, T4) than in the other samples. Our findings demonstrate that protein recovered from pork leg has better color parameters, and physical strength compared with Alaska Pollock-derived protein. A higher number of wash steps and treatment with salt during processing were furthermore found to yield better color, and physical strength in the protein samples.