• Title/Summary/Keyword: In Vitro Protein Solubility

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Food Functionality and In Vitro Bioactivity of Olive Flounder Paralichthys olivaceus Roe Concentrates Prepared by Cook-dried Process (가열-건조처리 넙치(Paralichthys olivaceus) 알 농축물의 식품기능성 및 생리활성)

  • In Seong Yoon;Sang in Kang;Jin-Soo Kim;In Sang Kwon;Hyeung Jun Kim;Min Soo Heu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.1
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    • pp.7-20
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    • 2023
  • Boil-dried concentrate (BDC) and steam-dried concentrate (SDC) were prepared from olive flounder Paralichthys olivaceus roe using the cook-dried process, and their food functionality and in vitro bioactivity were examined. The buffer capacity of BDC and SDC was found to be stronger in the alkaline region than in the acidic region, and the buffer capacity of SDC was superior to that of BDC. The water holding capacities of these concentrates were 7.6 and 7.4 g/g protein, respectively, both of which were significantly lower than that of freeze-dried concentrate (FDC). The solubility of BDC (13.4%) and SDC (12.7%), foaming capacity of BDC (107.7%) and SDC (110.6%), and oil-in-water emulsifying activity index of BDC (7.7 m2/g) and SDC (9.7 m2/g) were all significantly lower than the corresponding values for FDC (P<0.05). The lower food functionality of BDC and SDC compared with FDC can be attributed to the high-temperature denaturation of proteins during the cook-dried process. The 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging activities (IC50) of SDC (2.5 mg protein/mL) was 60.4 ㎍/mL, and the angiotensin I converting enzyme inhibitory activity was 80.9%. Olive flounder roe concentrates have good antioxidant and antihypertensive activities, and can be used as materials or ingredients in the processing of seafood and other foods to enhance protein contents and food functionality.

Neutralization of Human Papillomavirus by Specific Nanobodies Against Major Capsid Protein L1

  • Minaeian, Sara;Rahbarizadeh, Fatemeh;Zarkesh-Esfahani, Sayyed Hamid;Ahmadvand, Davoud;Broom, Oliver Jay
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.721-728
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    • 2012
  • The human papillomavirus (HPV) is the main cause of cervical cancer in developing countries. Rapid diagnosis and initiation of treatment of the HPV infection are critical. Various methods have been employed to reduce the immunogenicity of antibodies targeting HPV serotypes. Nanobodies are the smallest fragments of naturally occurring single-domain antibodies with their antigen-binding site compromised into a single domain. Nanobodies have remarkable properties such as high stability, solubility, and high homology to the human VH3 domain. In this study, a phagemid library was employed to enrich for nanobodies against the L1 protein of the human papilloma virus. Binding reactivity of the selected clones was evaluated using phage enzyme-linked immunosorbent assay (phage-ELISA). Finally, two nanobodies (sm5 and sm8) with the best reactivity against the Gardasil vaccine and the purified HPV-16 L1 protein were expressed and purified using a $Ni^+$-NTA column. The accuracy of expression and purification of the nanobodies was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting assays. In vitro studies demonstrated that neutralization was achieved by the selected nanobodies. The ease of generation and unique features of these molecules make nanobodies promising molecules for the new generation of HPV diagnosis and therapy.

Characteristics of iron powder formulation produced from porcine blood by enzymatic treatment (효소 처리한 돈혈 활용 철분분말제제 특성)

  • Kim, Mi-Yeon;Kim, MinA;Jeong, Yong-Jin
    • Food Science and Preservation
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    • v.23 no.5
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    • pp.753-757
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    • 2016
  • In this study, enzyme (thermoase) hydrolysis was applied to the porcine blood order to increase the iron content and solubility. It was confirmed that content of iron was increase up to 158.11 mg/100 g porcine powders after 0.2% thermoase treatment at $60^{\circ}C$ during 4 hr. The solubility of porcine blood powders was higher than other enzyme (various protease), temperature, reaction time. This optimized conditions were also worked to the in vitro iron bioavailability rate increasement, the bioavailability of hydolyzed porcine powders was 3-fold higher than that of an iron supplement on the market. These results indicate the possibility of porcine blood powder in iron supplements market as natural material. Also utilizing of reduced porcine blood will be possible to improve environmental issues.

Characteristics of Tenebrio molitor larvae and Bombyx mori pupae sequentially fermented with Aspergillus oryzae and Bacillus subtilis (Aspergillus oryzae와 Bacillus subtilis를 순차접종 발효한 갈색거저리유충과 누에번데기 발효물의 품질특성)

  • Kang, Sanghun;Chung, Chang-Ho
    • Korean Journal of Food Science and Technology
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    • v.54 no.1
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    • pp.94-102
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    • 2022
  • Defatted soybean, larvae of brown mealworm (Tenebrio molitor), and powdered pupae of silkworm (Bombyx mori) were fermented in solid and liquid forms using Aspergillus oryzae and Bacillus subtilis. The protein degradation rate (NDR) through solid fermentation was the highest in the fermented soybean control sample (54.69±6.54%), followed by silkworm pupae (34.82±5.99%) and brown mealworm larvae (30.54±3.80%). When these edible insects were fermented in liquid form, solid extraction yield was 37.73-46.88%, and protein yield was 47.47-63.02%. NDR of fermented liquid form products increased to 58.90, 52.62, and 50.13% for soybean, brown mealworm larvae, and silkworm pupae, respectively. SDS-PAGE of the liquid fermented products confirmed that microbial fermentation decomposed higher-molecular-weight proteins into small polypeptides. In vitro digestibility of liquid forms of edible insects increased by 1.26 to 1.53 times after fermentation. The protein solubility, foaming ability, and foam stability of liquid-fermented edible insects all tended to increase through fermentation.

Effects of Heat Treatments on Physicochemical Properties and In Vitro Biological Activities of Quinoa (Chenopodium quinoa Willd.) (퀴노아의 열처리 가공에 따른 이화학적 특성 및 In Vitro 생리활성)

  • Goh, Hye-Kyung;Lee, Young-Tack
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.6
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    • pp.688-694
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    • 2017
  • The effects of heat treatments on the physicochemical properties and in vitro biological activities of quinoa (Chenopodium quinoa Willd.) were investigated. Quinoa grains were subjected to two different heat treatment methods: boiling and steaming plus roasting (steaming/roasting). Compared with raw quinoa, boiled quinoa samples had slightly lower crude protein, crude fat, crude ash, and starch contents. However, steaming/roasting treatment did not cause significant differences in proximate composition. Heat treatments reduced total phenolic and flavonoid contents in quinoa extracts, and higher reduction was detected upon boiling treatment. Heat treatments also reduced lightness and increased yellowness of quinoa samples. Heat treatments increased water absorption index but decreased water solubility index. In vitro starch hydrolysis increased substantially after both heat treatments, and slightly higher values were observed in the boiled quinoa samples. 1,1-Diphenyl-2-picrylhydrazyl free radical scavenging activity and nitrite scavenging activity were reduced by heat treatments, and the boiled quinoa sample showed the lowest activity likely due to loss of activities in cooking water.

Immunostimulating Activity and Characterization of Polysaccharides from Mycelium of Phellinus linteus

  • Lee, Jae Hoon;Soo Muk Cho;Kyung Sik Song;Sang Bae Han;Hwan Mook Kim;Nam Doo Hong;Ick Dong Yoo
    • Journal of Microbiology and Biotechnology
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    • v.6 no.3
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    • pp.213-218
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    • 1996
  • Hot-water extract, Fr. 1, of Phellinus linteus mycelium was fractionated into Fr. 2, 3, 4, and 5 by the difference of solubility in ethanol. The polysaccharide fractions were studied for their immunostimulating activity on in vitro T-independent polyc1onal antibody response to trinitrophenyl-haptened SRBC (sheep red blood cell). The Fr. 4 with the highest immunostimulating activity was subjected to DEAE-cellulose ion exchange chromatography and gave five fractions, 4-I, II, III, IV, and V. The in vitro immunostimulating assay of the five fractions showed that 4-I and 4-III had a similar activity to that of LPS but the other fractions had low activity. By analyses of chemical composition and HPLC, all fractions obtained were found to be heteropolysaccharide-protein complex. The molecular weights ranged from 9, 000 to 15, 000. Sugar analyses showed that glucose, galactose, mannose, arabinose, and xylose were main component. Uronic acid and amino sugar were also detected in the fractions. It should be noted that the molecular weight (15, 000) of 4-III was very small and the structure of 4-III may be different from the known immunostimulating branched $\beta$-(1longrightarrow3)-glucan.

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Effect of Vehicles and Enhancers on the in vitro Skin Penetration of Aspalatone and Its Enzymatic Degradation Across Rat Skins

  • Gwak, Hye-Sun;Chun, In-Koo
    • Archives of Pharmacal Research
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    • v.24 no.6
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    • pp.572-577
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    • 2001
  • The feasibility of skin penetration was studied for aspalatone (AM, acetylsalicylic acid maltol ester), a novel antithrombotic agent. In this studys hairless mouse dorsal skins were used as a model to select composition of vehicle and AM. Based on measurements of solubility and partition coefficient, the concentration of PC that showed the highest flux for AM across the hairless mouse skin was found to be 40%. The cumulative amount permeated at 48 h, however, appear inadequate, even when the PC concentration was employed. To identify a suitable absorption enhancer and its optimal concentration for AM, a number of absorption enhancers and a variety of concentration were screened for the increase in transdermal flux of AM. Amongst these, linoleic acid (LOA) at the concentration of 5% was found to have the largest enhancement factor (i.e., 132). However, a further increase in AM flux was not found in the fatty acid concentration greater than 5%, indicating the enhancement effect is in a bell-shaped currie. In a study of the effect of AM concentration on the permeation, there was no difference in the permeation rate between 0.5 and 1% for AM, below its saturated concentration. At the donor concentration of 2%, over the saturated condition, the flux of AM was markedly increased. A considerable degradation of AM was found during permeation studies, and the extent was correlated with protein concentrations in the epidermal and serosal extracts, and skin homogenates. In rat dorsal skins, the protein concentration decreased in the rank order of skin homogenate > serosal extract > epidermal extract. Estimated first order degradation rate constants were $6.15{\pm}0.14,{\;}0.57{\pm}0.02{\;}and{\;}0.011{\pm}{\;}0.004{\;}h^{-1}$ for skin homogenate, serosal extract and epidermal extract, respectively. Therefore, it appeared that AM was hydrolyzed to some extent into salicylmaltol by esterases in the dermal and subcutaneous tissues of skin. taken together, our data indicated that transdermal delivery of AM is feasible when the combination of PC and LOA is used as a vehicle. However, since AM is not metabolically stable, acceptable degradation inhibitors may be nervessary to fully realize the transdermal delivery of the drug.

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Calcium Solubilization Ability and Anti-Inflammatory Effects of Hydrolyzed Casein

  • Kim, Da Young;Yoo, Jung Sik;Cho, Yoon Ah;Yoon, Ho Sik;Kim, Cheol-Hyun
    • Food Science of Animal Resources
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    • v.41 no.4
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    • pp.687-700
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    • 2021
  • This study performed to evaluate the applicability of functional dairy food materials by comparing the calcium solubilization ability and anti-inflammatory effects of hydrolyzed casein protein. Commercial enzyme (Alcalase®; Neutrase®; Protamex®; Flavourzyme®) was added to the 10% casein solution to prepare the casein hydrolysates. Samples obtained every hour [1:200 (w/v)]. According to results of measuring the degree of hydrolysis (DH), all of four enzymatic hydrolysates increased rapidly from 30 to 40 min, and after 150 min, there were no change. Protamex® and Neutrase® had the highest DH compared to others enzymatic hydrolysates. After that, peptides obtained throughout a preparative liquid chromatography system. In the calcium solubility experiments, neutrase fraction (NF) 4 and NF7 showed similar activities with casein phosphopeptide (CPP). In vitro cell experiments showed that no cytotoxicity except for NF6. Also, the production of nitric oxide (NO) inhibited as the concentration of fraction samples increased. The cytokine (IL-1α, IL-6, and TNF-α) production was lower than lipopolysaccharide (+) group significantly. Therefore, the possibility of anti-inflammatory activity found in the hydrolyzed samples. According to the above experiments, NF3 and Protamex Fraction (PF) 3 selected. Amino acids selected throughout an AccQ-Tag system. As a result, 17 species of amino acids and several species of unknown amino acids identified. Both fractions had the highest content of phenylalanine. This study identified the potential of biologically active and functional peptides derived from casein that affect the food and dairy industry.

Effect of Xylanase on Performance and Apparent Metabolisable Energy in Starter Broilers Fed Diets Containing One Maize Variety Harvested in Different Regions of China

  • O'Neill, H.V. Masey;Liu, N.;Wang, J.P.;Diallo, A.;Hill, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.4
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    • pp.515-523
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    • 2012
  • The objective of this study was to investigate the variability in broiler performance, apparent metabolisable energy (AME) and ileal digestible energy (IDE) between five different maize samples fed with and without xylanase at 16,000 U/kg. Various in vitro characterisations were conducted to determine if any could predict performance or AME. Samples of the maize were harvested in five diverse regions and fed individually in a mash diet as follows (g/kg): test maize 608.3; soya bean meal (SBM) 324.1; poultry fat 25.2; salt 4.6; met 2.6; lys 1.6; thr 0.5; limestone 9.7, dical 18.4; vit/min 5.0; CP 210 and ME (kcal/kg) 3,085. The diets were fed to 720 broilers with 6 replicates, each containing 12 birds per treatment, from 0 to 18 d of age. Maize samples were analysed for starch, protein, crude fibre, fat, protein solubility index (PSI) and vitreousness using near infra red reflectance spectroscopy (NIR). They were also assayed using an in vitro starch digestibility method. The results showed that there was no effect of harvest region on the feed intake (FI), body weight gain (BWG) or feed conversion ratio (FCR) of the broilers over the 18 d period (p = 0.959, 0.926, 0.819 respectively). There was an improvement in all parameters with the addition of xylanase (FI p = 0.011; BWG and FCR p<0.001). There was a significant positive effect of xylanase on IDE, AME, IDE Intake (IDEI) and AME intake (AMEI) (p<0.0001 in all cases). Although there was no significant effect of maize source, there was a strong trend towards variability in IDE (p = 0.066) and AME (p = 0.058). There were no significant correlations (p<0.05) between any proximate or physiochemical values and any performance or AME values. This may suggest that none of those selected were suitable predictors for performance or AME. The broilers performed well according to the breed guidelines, with slightly increased FI, increased BWG and similar FCR prior to the addition of xylanase. When FCR and BWG were analysed with FI as a covariate, xylanase addition remained significant suggesting that the improvement in BWG and FCR was driven by an increase in digestibility and nutrient availability.

Fabrication of PHBV/Keratin Composite Nanofibrous Mats for Biomedical Applications

  • Yuan, Jiang;Xing, Zhi-Cai;Park, Suk-Woo;Geng, Jia;Kang, Inn-Kyu;Yuan, Jiang;Shen, Jian;Meng, Wan;Shim, Kyoung-Jin;Han, In-Suk;Kim, Jung-Chul
    • Macromolecular Research
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    • v.17 no.11
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    • pp.850-855
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    • 2009
  • Keratin is an important protein used in wound healing and tissue recovery. In this study, keratin was modified chemically with iodoacetic acid (IAA) to enhance its solubility in organic solvent. Poly(hydroxybutylate-co-hydroxyvalerate) (PHBV) and modified keratin were dissolved in hexafluoroisopropanol (HFIP) and electrospun to produce nanofibrous mats. The resulting mats were surface-characterized by ATR-FTIR, field-emission scanning electron microscopy (FE-SEM) and electron spectroscopy for chemical analysis (ESCA). The pure m-keratin mat was cross-linked with glutaraldehyde vapor to make it insoluble in water. The biodegradation test in vitro showed that the mats could be biodegraded by PHB depolymerase and trypsin aqueous solution. The results of the cell adhesion experiment showed that the NIH 3T3 cells adhered more to the PHBV/m-keratin nanofibrous mats than the PHBV film. The BrdU assay showed that the keratin and PHBV/m-keratin nanofibrous mats could accelerate the proliferation of fibroblast cells compared to the PHBV nanofibrous mats.