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Hydrolysis Characteristics of Goat Milk $\beta-Casein$ by Enzyme and Angiotensin Converting Enzyme Inhibition Effects of Hydrolysate (산야유 $\beta-Casein$의 효소 가수분해 특성과 가수분해물의 Angiotensin Converting Enzyme 저해 효과)

  • Park Yong-Kuk;Kwon Il-Kyoung;Kim Gur-Yoo
    • Food Science of Animal Resources
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    • v.25 no.2
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    • pp.238-243
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    • 2005
  • This study was carried out to understand hyrolytic characteristics of $\beta-casein$ by enzyme in goat milk and to measure the inhibition effect of the ACE of the hydrolysate. In order to conduct the experiment, $\beta-casein$ of goat milk was separated using Mono S HR 5/5, a cation exchange column. The separated $\beta-casein$ was treated with trypsin of animal hydrolysis enzymes, in an effort to verify the characteristics of hydrolysis. The inhibition activity of ACE was measured and the results are as follows. By analyzing the hydrolysate separated from the trypsin-processed $\beta-casein$ of goat milk, the inhibition effect of the ACE was measured trypsin-hydrolyzed $\beta-casein$ demonstrated a $25.36\pm0.79\%$ of inhibition effect and the $IC_{50}$ of the hydrolysate from the trypsin-processed $\beta-casein$ reached $308.7\pm2.77({\mu}g/mL)$.

Biosynthesis of recombinant human prominiinsulin in E. coli and plant systems (대장균과 식물시스템에서 재조합 인간 prominiinsulin 생합성 분석)

  • Choi, Yu Jin;Park, Su Hyun;Kim, Ji Su;Wi, Soo Jin;Park, Ky Young
    • Journal of Plant Biotechnology
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    • v.40 no.3
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    • pp.169-177
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    • 2013
  • Recently, the number of people with diabetes is rapidly increasing, coupled with the fact that the insulin market is remarkably increasing. Therefore, molecular farming for plant-derived pharmaceutical protein production is reported as becoming more attractive than ever. In this study, we carried out experiments step by step for development of recombinant insulin constructs, which were transformed into E. coli system, in vitro transcription and translation system, and tobacco cells. At first, recombinant proinsulin protein was successfully produced in in vitro transcription and translation system with wheat germ extract. After which, recombinant construct of prominiinsulin encoded a fusion protein of 7.8 kDa with trypsin cleavage sites at N terminus and C terminus of minimized C-peptide was tried to in vitro expression using E.coli culture. After purification with His-tag column, the resulting recombinant prominiinsulin protein was processed with trypsin, and then checked insulin biosynthesis by SDS-PAGE and western blot analysis with anti-insulin monoclonal antibody. The immunoreactive product of trypsin-treated miniinsulin was identical to the predicted insulin hexamer. The construct of 35S promoter-driven preprominiinsulin recombinant gene with signal peptide region for ER-targeting and red fluorescence protein gene [N terminus ${\rightarrow}$ tobacco E2 signal peptide ${\rightarrow}$ B-peptide (1-29 AA) ${\rightarrow}$ AAK ${\rightarrow}$ A-peptide (1-21 AA) ${\rightarrow}$ RR ${\rightarrow}$ His6 ${\rightarrow}$ KDEL ${\rightarrow}$ C terminus] was transformed into BY-2 tobacco cells. A polypeptide corresponding to the 38-kDa molecular mass predicted for fusion protein was detected in total protein profiles from transgenic BY-2 cells by western analysis. Therefore, this recombinant preprominiinsulin construct can be used for generation of transgenic tobacco plants producing therapeutic recombinant insulin.

Isolation and comparison of somatic and excretory-secretory antigens for serological diagnosis in Trichuris suis (혈청학적 진단을 위한 돼지 편충의 체항원, 배설/분비항원의 분리 및 비교)

  • Jee, Cha-ho;Lee, Chul-soon;Park, Seung-jun
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.159-168
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    • 1999
  • Swine whipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swine gastrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the reasonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follows; 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values($mean{\pm}SD$) of adult somatic antigen against positive(negative) sera were $0.30{\pm}0.12(0.09{\pm}0.006)$ and third-stage larva of somatic antigen were $0.28{\pm}0.038(0.10{\pm}0.005)$. And OD values of excretory-secretory antigens of adult and third-stage larva were $0.24{\pm}0.031(0.11{\pm}0.005)$ and $0.08{\pm}0.013(0.10{\pm}0.003)$, respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p<0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were $0.30{\pm}0.120(0.09{\pm}0.006)$ and $0.25{\pm}0.141(0.09{\pm}0.003)$. These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.

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Socio-economic Features of One Slice of Chicagoland Using A Geo-Spatial Information System (시카고 부분지역의 사회경제적 특성에 대한 지형공간정보체계의 이용)

  • Oh, Jong-Woo
    • Journal of Korean Society for Geospatial Information Science
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    • v.1 no.2 s.2
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    • pp.223-235
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    • 1993
  • This study associates with socio-economic status in a slice section area of Chicago metropolitan to get spatial patterns of urban windows. GSIS(Geo-spatial Information System) has been monitored with several statistic methods, and geo-spatial map presentations. From the grouping analysis, the result displays that most suburban town have high income values, such as Elmhurst, Melrose Park, North Lake(Income ranges between $25,000${\sim}$30,000 : 1980 Sensus data). The factors produced form both analyses of SAS and BMDP are socio-ethnic, economic, hispanic, black, life expectancy, and multiple car ownership. In the study area the socio-ethnic factor is striking, and is composed of nine out of the fourteen varialbles. Geo-spatial 3-D mapping represents a socio-economic configuration of the study area. The high income value areas are Elmhurst and North Lack, and a spot between Belmont Ave. and the Lake Shore. Economic configuration is a vital importance of socio-economic activities in the urban areas. In the study area a minimum average income level is about $4,364 and maximun is $30,311.

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Sequence analysis and expression of groE gene encoding heat shock proteins of Brucella abortus isolates (Brucella abortus 국내 분리주의 Heat Shock Protein 암호 groE 유전자의 염기서열 분석과 발현)

  • Kim, Tae-Yong;Kim, Ji-Young;Chang, Kyung-Soo;Kim, Myung-Cheol;Park, Chang-Sik;Han, Hong-Ryul;Jun, Moo-Hyung
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.45-53
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    • 2005
  • GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains. In comparison of the sequences of nucleotides and amino acids among the strains, GroES showed 100% identity in both sequences. GroEL was evaluated 99.0~99.9% in nucleotides and 98.0~100% homology in amino acids. The groE gene including groES and groEL was inserted into pET29a vector and constructed pET29a-GroE recombinant plasmids. The inserted groE was confirmed by digestion with Nco1 and EcoR1 endonucleases and nucleotide sequencing. E. coli BL (DE3) was transformed with pET29a-GroE, named as E. coli BL (DE3)/pET29a-GroE. In SDS-PAGE, it was evident that the recombinant plasmid effectively expressed the polypeptides for GroES (10 kDa) and GroEL (60 kDa) in 0.5, 1 and 2 hours after IPTG induction. The immuno-reactivity of the expressed proteins were proved in mouse inoculation and Western blot analysis.

Functional and Immunological Properties of Ryanodine Receptor in the Eel Skeletal Muscle (뱀장어 근육내 Ryanodine Receptor의 기능 및 면역학적 성질)

  • Seok, Jeong-Ho;Lee, Yeon-Soo;Nam, Jang-Hyeon;Choi, Suk-Jeong;Hong, Jang-Hee;Lee, Jae-Heun
    • The Korean Journal of Pharmacology
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    • v.31 no.2
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    • pp.207-217
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    • 1995
  • To investigate the functional and immunological properties of the Ca-release channel in the sarcoplasmic reticulum(SR) of the eel skeletal muscle, $[^3H]ryanodine$ binding, SDS gel electrophoresis, $^{45}Ca\;release$ studies, and immunoblot assay were carried out in the SR of the eel skeletal muscle. Maximal binding sites(Bmax) and $K_D$ values of $[^3H]ryanodine$ for Ca-release channel of the SR of the eel skeletal muscle were $19.44{\pm}1.40\;pmole/mg$ protein and $15.55{\pm}1.69\;nM$, respectively. $[^3H]Ryanodine$ binding to RyR was increased by calcium and AMP. The SR of the eel skeletal muscle has two high molecular weight bands on the SDS PAGE. The mobility of upper band was more slower than the single band of the rabbit skeletal muscle, and that of the lower band was similar with the single band of canine cardiac muscle. Vesicular $^{45}Ca-release$ was activated by calcium. Ca-induced $^{45}Ca-release$ was significantly inhibited by $MgCl_2(2\;mM)$, ruthenium red$(10\;{/mu}M)$ or tetracaine(1 mM), but not by high concentration of calcium itself. AMP-induced $^{45}Ca-release$ was slightly occurred only in the absence of calcium, it was not inhibited by $MgCl_2$ or ruthenium red. Caffeine also increased $^{45}Ca-release$ from the SR vesicles, but it was not affected by $MgCl_2$ or ruthenium red. Polyclonal Ab against rat skeletal muscle RyR is reacted with that of rabbit, but not reacted with that of the eel skeletal muscle. These results suggested that ryanodine receptor of the SR of the eel skeletal muscle is showing some similar properties with that of mammalian skeletal muscle, but might be an another isotype channel having two bands which is less sensitive to AMP, not cross-reacted with antisera against rat RyR, and not inhibited by high concentration of calcium.

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Physicochemical Characteristics of Silk Fibroin Degummed by Protease in Bacillus licheniformis II. Behavior in Aqueous Solution of Silk fibroin (Bacillus licheniformis 단백질 분해 효소에 의한 정련 견사의 특성 III. 견 피브로인 수용액의 거동)

  • 김영대;남중희
    • Journal of Sericultural and Entomological Science
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    • v.35 no.1
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    • pp.60-68
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    • 1993
  • It has been known that the silk degumming treated by hot alkali solution is easy to handle but is liable to yield poor-quality silk due to the degree of degumming loss, incomplete-degumming or over-degumming. Therefore, many studies have been carried out on the silk degumming by enzyme in order to improve the quality of silk. However, no attention has been paid to the physicochemical analysis of enzymatic degummed silk. In this paper, two different degumming methods, soap and enzymatic, are compared in aqueous solution state of silk fibroin. The results can be summarized as follows: There was no significant difference between two solutions on the bases of polarizing microscopy, TEM observation and SDS-PAGE. Spherulite of silk fibroin was not observed in polarizing microscopy, however the leaf-shape fibril structure was developed upon solidification. The size of spherulites of silk fibroin in TEM observation were 30~120nm with a wide range of size distribution. The intrinsic viscosity of enzymatic degummed fibroin solution was lower than that of soap degummed solution. This can be explained that the silk fibroin was more degraded by enzymatic degumming method compared with the soap degumming method. SDS-polyacrylamide gel electrophoresis showed that the fibroin molecule was composed of large component of molecule weight above 50 kd and small component of molecule weight about 20 kd. There was no difference in crystallinity between two degumming methods on the bases of results of DSC thermograms and IR spectra.

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Identification of Brassinosteroid-Related Protein, BAK1 from Nutrition Deficient Tomato Cultivated by Soilless Cultivation System (수경재배 영양결핍토마토에서 브레시노스테로이드관련 신호전달 단백질 BAK1의 동정)

  • Shin, Pyung-Gyun;Chang, An-Cheol;Hong, Sung-Chang;Lee, Ki-Sang
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1729-1733
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    • 2007
  • Brassinolide insensitive associated receptor kinase 1(BAK1) is a critical component that play an important roles in signaling of brassinosteroid biosynthesis. Brassinosteroid-deficient and -insensitive mutants showed the characteristic of dwarf symptom. The nutrient deficient tomato showing stunt phenomenon was selected from soiless cultivation system using modified Sonneveld hydroponic solution. Twenty eight protein spots showing different expression levels compared to the control were isolated from extracts of stunted tomato leaves by 2D PAGE analyses. Significantly down-regulated 6 protein spots out of 28 protein spots were analyzed and sequenced by MALDI-TOF mass spectrometry. The protein spot having pI=4.5 and MW=24 kDa was identified as a signal protein, BAK1, which is directly related to brassinosteroid biosynthesis. In addition, five other protein spots were identified as BCK1, cystein proteinase, sulfutase, peroxidase and zinc finger factor respectively, and they were also signal proteins related to brassinosteroid biosynthesis. Furthermore, amplification of 500bp of BAK1 mRNA by RT-PCR using a primer set of peptide matched regions was inhibited conpared to that of the wild type. The results sugested that the BAK1 might be regulated at the transcription level in response to nutrition applications.

Changes in Allergenicity of Porcine Serum Albumin by Microwave, Sonication, and High Hydrostatic Pressure (Microwave, 초음파 및 초고압 처리에 의한 돼지 혈청 알부민의 항원성 변화)

  • Kim, Koth-Bong-Woo-Ri;Kim, Seo-Jin;Lee, So-Young;Song, Eu-Jin;Ahn, Dong-Hyun
    • Food Science of Animal Resources
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    • v.28 no.4
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    • pp.499-504
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    • 2008
  • Even though pork have frequently induecd allergic reactions in Korea, few papers have been published on pork allergy. This study was carried out to investigate the changes in allergenicity of porcine serum albumin (PSA) by microwave, sonication, and high hydrostatic pressure (HHP). The binding ability of p-IgG to PSA treated with microwave (1,5, or 10 min) directly decreased with increasing treatment time. Particularly, the binding ability of PSA treated 10 min was about 30%. Immunoblotting assay with p-IgG showed that band of PSA treated microwave directly disappeared at 5 and 10 min. However, the binding ability of PSA was not changed by the microwave treatment without heat. Also the reduction of allergenicity by sonication or HHP treatment was not found. In conclusion. allergenicity of PSA treated with microwave directly decreased with increasing time, therefore these results may be used for development of hypoallergenic pork.

The Association of Institutional Information on Websites with Present and Future Financial Performance (웹사이트에 게시된 기업의 소개글 분석을 통한 기업의 현재 및 미래 가치 예측 분석 방법)

  • Na, Hyung Jong;Choi, Sukjae;Kwon, Ohbyung
    • The Journal of Society for e-Business Studies
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    • v.23 no.4
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    • pp.63-85
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    • 2018
  • The "About Us" page on the website of a corporation provides information regarding the organization's vision, philosophy, and values. We examine the association between institutional information provided on corporate websites (i.e., the "About Us" section) with present and future financial performance. Utilizing a text mining technique, we analyze the institutional information of S&P500 firms in the year 2016. We conduct a factor analysis including words that are intentionally repeated in the introductory text of corporate websites. The results of the analysis reveal that keywords from this institutional information can be grouped into six factors. We then carry out an ordinary least squares regression analysis to determine the associations between these six factors and present financial performance. The results show that keywords in Factor 2 (those related to Purchasing experience) are positively associated with ROE, a variable representing present financial performance, while keywords in Factor 1 (those related to Note to customers) show a negative relationship with ROE. On the other hand, keywords in Factor 1 have a positive relationship with Tobin's Q, a variable representing future financial performance. These results indicate that there is some relationship between the words used in the institutional information in this section of corporate websites and firms' financial performance. Hence, the institutional information on a website may be a useful indicator of current firm performance and future firm value.