• Title/Summary/Keyword: pH-stat

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Bitterness and Solubility of Soy Protein, Casein, Gluten, and Gelatin Hydrolysates Treated with Various Enzymes (효소종류에 따른 대두단백, 카제인, 글루텐, 젤라틴 단백질 가수분해물의 쓴맛과 용해도 특성)

  • Kim, Mi-Ryung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.4
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    • pp.587-594
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    • 2010
  • To develop commercially available food protein hydrolysates, the effects of different types of enzymes and substrates on bitterness and solubility of partially hydrolyzed food proteins were investigated. Four types of proteins (casein, isolated soy protein (ISP), wheat gluten, and gelatin) and five types of proteolytic enzymes (a microbial alkaline protease (alcalase), a microbial neutral protease (neutrase), papain, bromelain, trypsin) were used. To profile the pattern of hydrolysis, the degree of hydrolysis (DH) were monitored during 180 min of reaction time by pH-stat method. Casein showed the highest susceptibility to hydrolysis for all five proteases compared to those of ISP, gluten, and gelatin. In addition, the bitter intensity and solubility (nitrogen soluble index, NSI) of each protein hydrolysate were compared at DH 10%. Bitterness and solubility of protein hydrolysates were highly affected by DH and the types of enzymes and substrates. At DH=10%, casein hydrolysate by trypsin, ISP and gluten hydrolysates by either bromelain or neutrase, and gelatin hydrolysates by the five proteases tested in this study were highly soluble and less bitter.

Isolation of Microorganism with HIgh Productivity and Cultivation Optimization for Lactic Acid Production (고생산성 젖산생성균 분리 및 배양 최적화)

  • Cho, Kyu-Hong;Cho, Yun-Kyung;Hong, Seung-Suh;Lee, Hyun-Soo
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.6-11
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    • 1995
  • In order to screen microorganism producing lactic acid with high productivity from nature, we used a medium containing 100 g/l glucose and selected several microorganisms producing more than 80 g/l L-lactic acid. We investigated their physiological characteristics and compared them. The best microorganism was identified as Lactobacillus casei subsp. rhamnosus. The optimum pH for growth and production of lactic acid was 6.0 and this strain showed the highest growth rate at around 30$\circ$C , but the optimum temperature for lactic acid production was 45$\circ$C . The growth was inhibited proportionally from 50 g/l to 300 g/l of glucose and the maximal cell mass increased according to increasing the concentration of corn steep liquor (CSL) protein up to 30 g/l. In batch fermentation for lactic acid production, we produced 128 g/l L-lactic acid with 20 g/l CSL protein and 150 g/l glucose in 35 hours. In pH-stat fed-batch fermentation, we were able to produce 183 g/l L-lactic acid.

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Molecular Characterization and Bitter Taste Formation of Tryptic Hydrolysis of 11S Glycinin

  • Kim, Mi-Ryung;Choi, Sang-Yun;Lee, Cherl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.509-513
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    • 1999
  • The molecular size reduction and the formation of bitterness during a tryptic hydrolysis of soybean 11S glycinin were determined by using quantitative analysis and organoleptic evaluation. The 11S glycinin of 90% purity was prepared by cryoprecipitation and Con A Sepharose 4B affinity chromatography, and hydrolyzed with trypsin in a pH-stat reactor for 4 h. Bitterness was formed within 1 h of hydrolysis, and then slowly increased up to $3.5\times10^{-5}$ M quinine-HCl equivalent. The extent of hydrolysis (DH) was 7% at 1 h and increased up to 12% by the end of the reaction. The -amino nitrogen content increased from an initial 0.7 mM to 7 mM at the end of the period. The SDS-PAGE analysis showed that the acidic subunit of 11S glycinin was mostly hydrolyzed. The GP-HPLC analysis indicated that the bitterness was mainly contributed by the peptide fractions of molecular weights of 360-2,100 Da.

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Purification, Characterization and Immobilization of Lipase from Proteus vulgaris OR34 for Synthesis of Methyl Oleate

  • Misbah, Asmae;Koraichi, Saad Ibnsouda;Jouti, Mohamed Ali Tahri
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.491-505
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    • 2020
  • A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

Comparison of the bovine blood gas parameters produced with three types of portable blood gas analyzers

  • Ro, Younghye;Choi, Woojae;Hong, Leegon;Kim, Eunkyung;Choe, Eunhui;Kim, Danil
    • Journal of Veterinary Science
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    • v.23 no.4
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    • pp.60.1-60.6
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    • 2022
  • Background: A definite diagnosis should be made in the bovine practice field, however, it was difficult to perform laboratory analysis immediately. Currently, three types of portable blood gas analyzers are available in Korea. Objectives: This study aimed to evaluate the correlations among these three analyzers. Methods: Seventy-two plasma samples from Holstein-Friesian cows were used for blood gas analysis, and three instruments (EDAN i15 Vet, VETSCAN i-STAT, and EPOC) were operated simultaneously. Moreover, plasma calcium levels were compared between these portable analyzers and blood chemistry device, which is usually used in a laboratory environment. Pearson analysis was performed to confirm the correlation of each parameter produced with the three instruments and blood chemistry analyzer. Results: As results, high correlation was observed in parameters of pH, pO2, potassium ion, ionized calcium, and glucose (p < 0.001, r > 0.7). In addition, pCO2 showed a moderate correlation among the three analyzers (p < 0.001, r > 0.5), and there was no correlation among all instruments for sodium ions. There was also a high correlation between ionized calcium from the three portable devices and total calcium from the biochemistry analyzer (p < 0.001, r > 0.9). Conclusions: In conclusion, there was a high correlation between results from the three different blood gas analyzers used in the bovine clinical field in Korea. Thus, a consistent diagnosis can be made even with different equipment if the operator is aware of the strengths and weaknesses of each piece of equipment and operates it properly.

High-Level Production of Spider Silk Protein by Fed-Batch Cultivation of Recombinant Escherichia coli and Its Purification

  • Lee, Seok-Jae;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.719-722
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    • 2001
  • Silk proteins from Nephila clavipes are fibrous proteins containing repetitive sequences with both crystalline and amorphous domains. In order to obtain high-level production of silk protein, the synthetic genes had 16 contiguous units of the consensus repeat sequence of the silk protein were expressed in Escherichia coli BL21(DE3) under the strong inducible T7 promoter. For production of recombinant silk protein in large amounts, pH-stat fed-batch cultures were carried out. The recombinant silk protein was produced as soluble forms in E. coli, and the recombinant silk protein content was as high as 11% of the total protein. When cells were induced at $OD_{600}$ of 60, the amount of silk protein produced was 6.49 g/L. After simple purification steps, 9.2 mg of silk protein that was more than 80% pure was obtained from a 50 mL culture, and the recovery yield was 26.3%.

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High Cell Density Culture of Alcaligenes eutrophus and Poly-$\beta$-hydroxybutyrate Production by Optimization of Medium Compositions (배지조성 최적화를 통한 Alcaligenes eutrophus의 고농동 세포배양 및 Poly$\beta$-hydroxybutyrate 생산)

  • 이용우;유영제
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.401-406
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    • 1994
  • The medium compositions of Alcaligenes eutrophus were optimized for increasing PHB productivity. It is very important to optimize the concentrations of inorganic salts and trace eleme- nts as well as carbon and nitrogen sources to maximize cell growth rate and productivity. The fed-batch culture of Alcaligenes eutrophus by dual feeding of ammonia water and glucose under optimized initial medium concentrations was carried out. Glucose was fed manually according to glucose consumption rate and ammonia water by pH-stat. The final cell concentrations and PHB content in 30 hours were 122 g/l and 65% of dry cell weight(yielding 79 g of PHB/l), respectively and 2.64 g/l/hr of PHB production rate was obtained.

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Studies on Salt Injuries of Seedlings Growth-Sand Media Effect of Various Concentration of Added Salts upon 3 Kinds of Seedlings (수묘(樹苗)의 염해(鹽害)에 관(關)한 연구(硏究) -Nacl 농도(濃度)를 달리한 사경(砂耕)에서의 몇 수묘(樹苗)에 미치는 영향(影響))

  • Son, Won Ha
    • Journal of Korean Society of Forest Science
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    • v.8 no.1
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    • pp.3-10
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    • 1968
  • (1) The present work is a study on the tolerance of salt concentration, salt injuries and growth, under varied conditions, of salt concentration of five kind silvicultural species. (2) Pinus thunbergii, which is tolerant of stat, has shown both low fresh leaf's pH and low gravity, while Pinus densiflora, Abies bolophylla and Pinus koraiensis have respectively shown high fresh leaf's pH and high gravity. (3) Pinus thunbergii, which is tolerant otosalt, has shown high water content ind its lead, hbile such speciesras Pinus densiflora an Abies lophylla, which are intolerant of salt have shownlow water content in their leaves. (4) A signifficance has ibeen found, in the nonreatment fresh les's pH and n gravity, among Pinusthunbergiis Pinu rigida, Pinus koraiensis, Pinus densiflora and Abies holophya. (5) The order of thetlolerance of salt among the tested species is as foohws : Pinus thunbergii, P. Korea iensis, Abies holopylla. (6) The signs of salt injuries have been observed at the tops of the trees 2~3 hours after the application of 1.5% salt solution in the case of Abiihs holop hylla which are intolerant of salt, while they have been of served at the same position 4~6 hours after the 1.5% application in the case of Pinus Koraiensis and Pinus thunbergii.

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The role of p21/CIP1/WAF1 (p21) in the negative regulation of the growth hormone/growth hormone receptor and epidermal growth factor/epidermal growth factor receptor pathways, in growth hormone transduction defect

  • Kostopoulou, Eirini;Gil, Andrea Paola Rojas;Spiliotis, Bessie E.
    • Annals of Pediatric Endocrinology and Metabolism
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    • v.23 no.4
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    • pp.204-209
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    • 2018
  • Purpose: Growth hormone transduction defect (GHTD) is characterized by severe short stature, impaired STAT3 (signal transducer and activator of transcription-3) phosphorylation and overexpression of the cytokine inducible SH2 containing protein (CIS) and p21/CIP1/WAF1. To investigate the role of p21/CIP1/WAF1 in the negative regulation of the growth hormone (GH)/GH receptor and Epidermal Growth Factor (EGF)/EGF Receptor pathways in GHTD. Methods: Fibroblast cultures were developed from gingival biopsies of 1 GHTD patient and 1 control. The protein expression and the cellular localization of p21/CIP1/WAF1 was studied by Western immunoblotting and immunofluorescence, respectively: at the basal state and after induction with $200-{\mu}g/L$ human GH (hGH) (GH200), either with or without siRNA CIS (siCIS); at the basal state and after inductions with $200-{\mu}g/L$ hGH (GH200), $1,000-{\mu}g/L$ hGH (GH1000) or 50-ng/mL EGF. Results: After GH200/siCIS, the protein expression and nuclear localization of p21 were reduced in the patient. After successful induction of GH signaling (control, GH200; patient, GH1000), the protein expression and nuclear localization of p21 were reduced. After induction with EGF, p21 translocated to the cytoplasm in the control, whereas in the GHTD patient it remained located in the nucleus. Conclusion: In the GHTD fibroblasts, when CIS is reduced, either after siCIS or after a higher dose of hGH (GH1000), p21's antiproliferative effect (nuclear localization) is also reduced and GH signaling is activated. There also appears to be a positive relationship between the 2 inhibitors of GH signaling, CIS and p21. Finally, in GHTD, p21 seems to participate in the regulation of both the GH and EGF/EGFR pathways, depending upon its cellular location.

Specific Knockdown of Nanog Expression by RNA Interference in P19 Embryonal Carcinoma Stem Cells (P19 배아 암종 줄기세포에서 RNA 간섭에 의한 Nanog 유전자 발현의 특이적 억제)

  • Choi, Seung-Cheol;Lim, Do-Sun
    • Development and Reproduction
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    • v.12 no.2
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    • pp.159-168
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    • 2008
  • Nanog is a newly identified member of the homeobox family of DNA binding transcription factors that functions to maintain the undifferentiated state of stem cells. However, molecular mechanisms underlying the function of Nanog remain largely unknown. To elucidate the regulatory roles of Nanog involved in maintenance of P19 embryonal carcinoma (EC) stem cells, we transfected three small interfering RNA (siRNA) duplexes targeted against different regions of the Nanog gene into P19 cells. The Nanog siRNA-100 duplexes effectively decreased the expression of Nanog up to 30.7% compared to other two Nanog siRNAs, the Nanog siRNA-400 (67.9 %) and -793 (53.0%). When examined by RT-PCR and real-time PCR, the expression of markers for pluripotency such as Fgf4, Oct3/4, Rex1, Sox1 and Yes was downregulated at 48 h after transfection with Nanog siRNA-100. Furthermore, expression of the ectodermal markers, Fgf5 and Isl1 was reduced by Nanog knockdown. By contrast, the expression of other markers for pluripotency such as Cripto, Sox2 and Zfp57 was not affected by Nanog knockdown at this time. On the other hand, the expression of Lif/Stat3 pathway molecules and of the endoderm markers including Dab2, Gata4, Gata6 and the germ cell nuclear factor was not changed by Nanog knockdown. The results of this study demonstrated that the knockdown of Nanog expression by RNA interference in P19 cells was sufficient to modulate the expression of pluripotent markers involved in the self-renewal of EC stem cells. These results provide the valuable information on potential downstream targets of Nanog and add to our understanding of the function of Nanog in P19 EC stem cells.

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