• Title/Summary/Keyword: solid and protein

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Comparative Monthly Analysis of Goat Milk Components by Individual Farms (우리나라 산양유의 농장별 및 계절별 성분 비교분석)

  • Ahn, Jong-Ho;Park, Woong-Yeoul
    • Korean Journal of Organic Agriculture
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    • v.16 no.3
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    • pp.321-330
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    • 2008
  • In order to investigate any differences of the characteristics of goat milk according to seasons and individual farms, we analysed and compared the components of fat, protein lactose, total solid, solid-not fat (SNF) and cells of goat milk collected from 8 individual farms between December 2006 and June 2007. Milk fat content has shown higher values in December to March than in other seasons, and SNF appeared especially higher in February. However, lactose content was not different according to seasons. Regional differences of milk components were not big at all either. Milk components from goats were in general similar to cow milk except a little lower fat content, but appeared higher than mare milk. Production of goat milk is in difficulty in aspects of balancing demand and supply due to its seasonal reproductive system. For the future of goat milk industry, it is necessary to develop various products of longer storage life.

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Solid-phase Refolding of Immobilized Enterokinase for Fusion Protein Cleavage (융합단백질 절단반응을 위한 고정화된 enterokinase의 고체상 재접힘)

  • 서창우;나세진;박신혜;박승국;이은규
    • KSBB Journal
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    • v.18 no.4
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    • pp.306-311
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    • 2003
  • Solid-phase refolding of immobilized proteins can be an effective way to reuse an immobilized enzyme column. Oriented immobilization methods are known to provide higher activity of the immobilized enzymes. In this study, using recombinant EK (enterokinase) as a model enzyme and a fusion protein, that consisted of recombinant human growth hormone and six His tag that was linked by the peptide of EK-specific recognition sequence, as a model substrate, we evaluated two oriented immobilization methods, i. e., reductive alkylation of N-terminus ${\alpha}$-amine and affinity interaction between poly-histidine tag and Ni-NTA (nickel-nitrilotriacetic acid). The immobilization yield, activity and cleavage of the immobilized enzymes, and the yield of solid-phase refolding were compared. The Ni affinity immobilization and the covalent immobilization yields were about 100% and 65%, respectively. But the specific activities were the same, about 50% of that of the soluble enzyme. The cleavage rate by the covalently immobilized EK was higher than the soluble enzyme and the side reaction of cryptic cleavage was significantly decreased. Covalently immobilized EK showed almost 100% refolding yield but the affinity immobilized EK showed only 70% yield, which suggested the covalent conjugation provided more rigid ‘reference structure’ for the solid-phase refolding. The monomeric hGH could be easily obtained by capturing the cleaved poly Histidine tag by the Ni affinity column.

Production of protein-bound polysaccharides by solid-state fermentation of Coriolus versicolor (구름버섯의 고체발효에 의한 단백다당류 생산)

  • Park, Kyung-Sook;Park, Shin;Jung, In-Chang;Ha, Hyo-Cheol;Kim, Seon-Hee;Lee, Jae-Sung
    • The Korean Journal of Mycology
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    • v.22 no.2
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    • pp.184-189
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    • 1994
  • The possibility of solid-substrate fermentation of Coriolus versicolor for the production of protein-bound polysaccharides(PBP) was studied. Zeolite and orchid-pot soil were used as solid materials for the culture because of the desirable physical properties. Glucose, sucrose and starch showed to be good carbon sources for the production of PBP by the solid-substrate fermantation of C. versicolor. Among the nitrogen sources, bactosoyton and peptone were very effective for the PBP production. The optimum pH for solid-substrate culture for the production of PBP was at the range of 5-6. The yields of PBP reached to 5-6 mg per 100 g solid-substrate.

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A Biotin-avidin Labeled Enzyme Immunoassay for the Quantitation of Serum TSH Using Protein-layered Solid Phase

  • Choi, Myung-Ja;Song, Eun-Young;Chung, Tai-Wha
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.231-235
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    • 1998
  • A sensitive enzyme immunoassay for serum TSH has been developed utilizing the tight binding between biotin and avidin, and three layered protein polystyrene beads as solid phase. To increase binding capacity of TSH and sensitivity of the assay, the polystyrene beads were coated sequentially with mouse immunoglobulin as first layer, rabbit antimouse immunoglobulin as second layer and monoclonal anti-TSH as third layer. A serum sample was incubated simultaneously with a monoclonal anti-TSH immobilized polystyrene beads and a second monoclonal anti-TSH covalently attached to biotin. After washing, the antibody bound serum TSH-anti-TSH-biotin complex is reacted with horseradish peroxidase (HRP)-labeled avidin. Following second wash, the bound HRP activity was measured calorimetrically. Reproducible results were obtained within 4 hours for serum TSH in the range between $0{\mu}\textrm{IU}$ml and ${50}{\mu}\textrm{IU}$ml with detection limit of $0.1{\mu}\textrm{IU}$ per test.

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Fabrication and Characterisation of a Novel Pellicular Adsorbent Customised for the Effectvie Fluidised Bed Adsorption of Protein Products

  • Sun, Yam;Pacek, Andrzej W.;Nienow, Alvin W.;Lyddiatt, Andrew
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.419-425
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    • 2001
  • A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zironia-silica(ZS) spheres by watr-in-oil emulsification . The agarose evenly laminated the ZS bead to a depth of 30㎛, and the resultin gpellicular assembly was characterised by densities up to 2.39g/mL and a mean particle dimeter of 136 ㎛. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark ad-sorbents necessary to achieve common values of bed expansion. Furthermore, the perlicular parti-cles were characterised by improved qualities of chromatographic behaviour, particularly with re-spect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorp-tion were attributed to the physical design and pellicular deployment of the reactive surface in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks(whole fermentation broths, cell disruptates and biological extracts).

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Effects of Processing Conditions on the Protein Quality of Fried Anchovy Kamaboko Engraulis japonica

  • Ramos, Leny R. Ordonez;Choi, Nam-Do;Ryu, Hong-Soo
    • Fisheries and Aquatic Sciences
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    • v.15 no.4
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    • pp.265-273
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    • 2012
  • The effects of processing and frozen storage conditions on the quality of anchovy Engraulis japonica fried surimi gels were investigated. Protein content decreased after surimi gel processing from 19.6% (raw meat) to 12.1% (kamaboko) due to the added ingredients and change in water content. Lipid content decreased from 2.8% (raw meat) to 1.3% in minced and 0.5% in surimi, but fried kamaboko showed a 6.9 % lipid level. Thiobarbituric acid values and thiobarbituric acid reactive substances levels were highest in kamaboko samples, 89.5 and 1.9 mg/g solid, and increased gradually with storage time to 101.8 and 4.6 mg/g solid, respectively. In vitro protein digestibility increased from 79.2% in raw anchovy to 88.5% in kamaboko samples. Levels of trypsin inhibitor decreased gradually with processing and during storage time from 2.43 in raw anchovy to 0.31 mg/g solid in the kamaboko sample after 60 days of frozen storage. No noticeable changes in total essential amino acid was observed during processing conditions. Computed protein efficiency ratio for kamaboko was highest (2.59) compared with whole anchovy (1.96), minced (1.94) and surimi (2.50). Fresh fried anchovy kamaboko showed similar values of hardness, springiness, gumminess and chewiness to commercial surimi gel, but a higher values were seen for fracturability and adhesiveness, and lower values for cohesiveness and resilience. The frozen and thawed anchovy kamaboko showed higher values for all of these rheological parameters compared with fresh and commercial kamaboko. Anchovy kamaboko showed the lowest lightness (62.9) and redness (0.16) and similar yellowness (11.9) compared with commercial kamaboko. Frozen storage and vacuum packaging were effective maintaining the shelf life of anchovy kamaboko within 30 days, but were not effective after 45 days due to fat oxidation.

Structure Determination of Syndecan-4 Transmembrane Domain using PISA Wheel Pattern and Molecular Dynamics simulation

  • Choi, Sung-Sub;Jeong, Ji-Ho;Kim, Ji-Sun;Kim, Yongae
    • Journal of the Korean Magnetic Resonance Society
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    • v.18 no.2
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    • pp.58-62
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    • 2014
  • Human transmembrane proteins (hTMPs) are closely related to transport, channel formation, signaling, cell to cell interaction, so they are the crucial target of modern medicinal drugs. In order to study the structure and function of these hTMPs, it is important to prepare reasonable amounts of proteins. However, their preparation is seriously difficult and time-consuming due to insufficient yields and low solubility of hTMPs. We tried to produce large amounts of Syndecan-4 transmembrane domain (Syd4-TM) that is related to the healing wounds and tumor for a long time. In this study, we performed the structure determination of Syd4-TM combining the Polarity Index at Slanted Angle (PISA) wheel pattern analysis based on $^{15}N-^1H$ 2D SAMPI-4 solid-state NMR of expressed Syd4-TM and Molecular Dynamics (MD) simulation using Discovery Studio 3.1.

Physico-chemical Properties and In Vivo Anti-cancer Effects of Potato Kimchi Prepared by adding Hot Water Extracts of Potato (생감자의 열탕 추출물을 첨가한 감자 김치의 이화학적 특성 및 In Vivo에서의 항암 효과)

  • Chang, Sang-Keun;Kim, Hee-Joo
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.3
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    • pp.302-310
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    • 2008
  • In this study, potato kimchi was prepared by applying heat to raw potatoes, and then the physico-chemical properties and anti-cancer effects of the kimchi were analyzed. The texture results indicated the potato kimchi had very good hardness and springiness attributes. During th late storage period, total vitamin C content of the kimchi slowly increased. In addition, the potato kimchi had non-volatile organic acid changes that promoted early aging; however, after the complete aging period, it was comparatively similar to other types of kimchi. Using the methanol extracts of various kimchi samples, the potato kimchi(solid 100%) showed the highest anti-carcinogenic effects in terms of anti-tumor activity in tumor bearing Balb/c mice with sarcoma-180 cells. In addition, the effects of the methanol extracts on hepatic glutathione S-transferase content were $289.76\;{\mu}mol/mg$ protein/min, $250.97\;{\mu}mol/mg$ protein/min, $251.20\;{\mu}mol/mg$ protein/min, $219.53\;{\mu}mol/mg$ protein/min, $183.79\;{\mu}mol/mg$ protein/min, for control kimchi, mul kimchi, and two potato kimchis [(solid 100%) and(solid 60%+kimchi juice 40%)], respectively. The in vivo anti-cancer effects of the potato kimchi were investigated using AGS human gastric adenocarcionoma cells and HT-29 human colon adenocarcionoma cells. Overall, an MTT assay revealed that the methanol extract of the potato kimchi showed the highest anti-carcinogenic effects.

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Protein Quality Evaluation of Cooked Monkfish (Lophiomus setigerus) Meats

  • Jeung Young-Ae;Ryu Hong-Soo;Shin Eun-Soo;Mun Sook-Im
    • Fisheries and Aquatic Sciences
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    • v.6 no.4
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    • pp.165-171
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    • 2003
  • To investigate the effect of cooking methods on protein quality of domestic fresh monkfish meat (FMM) and imported frozen monkfish meat (IMM), in vitro protein qualities were determined by amino acid anlysis, trypsin indigestible substrate (TIS) formation, and protein digestibility using the four-enzyme method. Crude protein contents of the boiled FMM and IMM were $90\%$ of the dry base, which were higher than fresh FMM $(82\%)$ and IMM $(84\%)$. Profiles of total amino acid in FMM and IMM were not changed by cooking methods. Total free amino acid contents decreased to $ 29.0-33.6\%$ for boiled $(l00^{\circ}C,\;10 min)\;and\;24\%$ for steamed $(100^{\circ}C,\;10\;min)$ samples. In vitro protein digestibilities of boiled and steamed FMM incnased $86.6-86.8\%$, compared to raw IMM $(82.9\%)$, boiled and steamed IMM $85.1-85.5\%$ and raw IMM $(83.6\%)$. TIS of FMM (23.6 mg/g solid) and IMM (15.9 mg/g solid) showed no significant (p<0.05) difference in cooking methods. The C-PERs (computed protein efficiency ratio) of boiled FMM (2.63) and IMM (2.50) were significantly higher (<0.05) than raw (1.97) and steamed FMM(1.97) and IMM(1.94). These results demonstrate that boiling of FMM and IMM improves protein digestibility and C-PER when compared to steamed FMM and IMM. Therefore, boiling could be an excellent means to maintain high-protein quality of monkfish meat. Also, the cooking method may be applicable to the preparation of monkfish stew without any loss of free amino acids.

Solid-Phase Refolding of Poly-Lysine fusion Protein of hEGF and Angiogenin (Poly-lysine이 연결된 hEGF와 angiogenin의 융합단백질의 고체상 재접힘)

  • Park, Sang-Joong;Ryu, Kang;Suh, Chang-Woo;Chai, Young-Gyu;Kwon, Oh-Byung;Park, Seung-Kook;Lee, Eun-Kyu
    • KSBB Journal
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    • v.17 no.2
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    • pp.153-157
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    • 2002
  • A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pl values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six Iysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similat result was also obtained when an Ni-NTA metal affinity resin was used.