• Title/Summary/Keyword: Artificial enzyme

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A survey of viruses associated with reproductive failure in boar semen in Korean artificial insemination centers (국내 인공수정센터의 웅돈에 대한 번식 관련 바이러스 조사)

  • Kim, Yeong-Hun;Chun, Bong-Su;Kim, Sung-Jae;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.111-116
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    • 2011
  • Artificial insemination (AI) of swine is a very useful reproductive tool and that offers convenience in the Korean swine industry. Since many viruses have been reported to be excreted through boar semen, we investigated the presence of antibodies and antigens against viruses causing reproductive failure in semen of boar in 349 semen samples collected from six Korean AI centers. Viral antigens were detected by polymerase chain reaction (PCR) or reverse transcription-PCR predominantly. The results was as follows. The major reproductive failure causing factor was porcine circovirus type 2 (PCV2), followed by porcine reproductive and respiratory syndrome virus (PRRSV) ($X^2$=166.64, P<0.001). PCV2 and PRRSV, Japanese encephalitis virus (JEV), encephalomyocarditis virus (EMCV) was detected in 73 samples (20.9%), 44 samples (12.6%), 4 samples (1.1%), 3 samples (0.9%), respectively and porcine parvovirus in one sample (0.3%) Classical swine fever virus (CSFV), bovine viral diarrhea virus and Aujeszky's disease virus (ADV) were not detected. Enzyme-linked immunosorbent assay was carried out in 111 serum samples from three AI centers. In most pigs, antibodies response was showed prominently in CSFV (105 sera, 94.6%) ($X^2$=82.580, P<0.001), followed by, in PRRSV (100 sera, 90.1%), PCV2 (92 sera, 90.1%), and PPV (8 sera, 82.9%). ADV antibody was not detected. Thus, the experimental results will be used for the base data, with respect to the state of viral stillbirth in general pig farms, as well as AI centers and breeding farms in Korea.

Bacterial Abundances and Enzymatic Activities under Artificial Vegetation Island in Lake Paldang (팔당호에 설치된 인공식물섬에서의 세균 수와 체외효소 활성도의 변화)

  • Byeon, Myeong-Seop;Yoo, Jae-Jun;Kim, Ok-Sun;Choi, Seung-Ik;Ahn, Tae-Seok
    • Korean Journal of Ecology and Environment
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    • v.35 no.4 s.100
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    • pp.266-272
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    • 2002
  • For analyzing function of a microbial ecosystem which was created under the artificial vegetation island (AVI) installed at Lake Paldang, zooplankton and bacterial numbers and exoenzyme activities (${\beta}$-glucosidase and phosphatase) were measured biweekly from 3 November 2()()1 to 20 April 2002 at AVI site and control site. Under the AVI, the water quality was worse than control site in term of comparing the environmental parameters. But, zooplankton number of AVI site was 25 times higher than that of control site. Respiratory active bacterial numbers were 3-8 times higher at AVI site. In addition, enzymatic activities were higher at AVI site than those of control site. These results suggest that the zooplankton-phytoplankton-bacteria relationships are closely coupled with each other and organic materials are eliminated by respiration of zooplankton and bacterial activities.

Diagnosis of Viral Infection of Pensaeide (보리새우류의 바이러스 감염증의 진단)

  • 허문수;정초록;장경립
    • Journal of Life Science
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    • v.9 no.4
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    • pp.453-459
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    • 1999
  • Baculovirus(WSBV) was isolated from infected Penaeide was collected from shrimp farm at southern sea of Korea from 1993 to 1995. The Infectious virus was purified and used for diagnosis of infected shrimp. Anti-viral serum were used for immunological detection as enzyme linked immunoabsorbent assay(ELISA) and indirect fluorescent antibody technique(IFAT). In IFAT, stomach, lymphoid organ and antenae gland of infected shrimp showed fluorescent reaction. In ELISA, tissues of spontaneously infected shrimp appeared higher O.D. values than in artificial infected shrimp. Primer set was constructed from sequence of 420bp of cloned Baculovirus(WSBV) genome. Specific band for infected shrimp was detected in Polymerase chain reaction(PCR)

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Recent Advances in Nanozyme Research for Disease Diagnostics (질병진단을 위한 나노자임 연구의 최근 동향)

  • Shin, Ho Yun;Yoon, Tae Young;Kim, Moon Il
    • KSBB Journal
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    • v.30 no.1
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    • pp.1-10
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    • 2015
  • Nanomaterial-based artificial enzymes (Nanozymes) have attracted recent attention because of their unique advantageous characteristics such as excellent robustness and stability, low-cost production by facile scale-up, and longterm preservation capability that are critically required as an alternative to natural enzymes. These nanozymes exhibit natural enzyme-like activity, and they have been applied to diverse kinds of detection methods for disease-associated biomolecules such as DNAs, proteins, cells, and small molecules including glucose. To highlight the progress in the field of disease diagnostics using nanozyme, this review discusses many nanozyme-based detection methods categorized by the types of target biomolecules. Finally, we address the current challenges and perspectives for the widespread utilization of nanozyme-based disease diagnostics.

Isolation and Characterization of Marine Bacterial Strain SH-1 Producing Agar-Degrading Enzymes (한천 분해효소를 생산하는 해양 미생물 SH-1의 분리 및 특성 분석)

  • Lee, Jae-Hag;Lee, Soon-Youl
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.324-330
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    • 2014
  • A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.

Studies on tile Resources for the Artificial Diet and Feeding Response of the Silkworm, Bombyx mori (가잠의 인공사료자원과 섭식성에 관한 연구)

  • 김주읍
    • Journal of Sericultural and Entomological Science
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    • v.23 no.2
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    • pp.11-36
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    • 1982
  • Forty-nine plant species as additives to silkworm artificial diet and 5 species as cellulose sources for artificial diet were screened for their economic values as feed-resources for the silkworm. Feeding response to artificial diet was tested on 82 silkworm strains. The effect of rearing conditions on feeding response and enzyme activities in the silkworm was investigated. The results were summarized as follows. 1. Seven species out of 49, Vigna sinensis ENDL, Ipomoea vatatas Lamarck, Cyperus anuricus Var. Laxus, Alnus japonica Stendel, Trifolium repens L, Prunus serrulata Lindley. Var, Glycine max L increased feeding response, compared with the basic formula of artificial diet. 2. The economic values of Vigna sinensis ENDL, Ipomoea vatatas Lamarck, Cyperus anuricus Var. Laxus, Ainus japonica Stendel, Cassia tera L, Erigeron canedensis L as feed-resources for artificiale diet were recognized, through feeding experiment during the entire larval stage. 3. Mulberry cellulose showed the best results in rearing and cocoon characteristics. 4. The extent of feeding response varied according to strains and varieties. Varieties in japanese strains showed higher feeding response than those in chinese and european varieties, with considerable variations among a varieties in strains. 5. The begining of 4th instar seems to be a proper time to convert from mulberry to artificial diet, or artificial diet to mulberry, however the middle of 3rd instar seems acceptable. 6. The optimum temperature for artificial diet rearing is 30$^{\circ}C$ during the period of 1st-3rd instar and 28$^{\circ}C$ for 4th-5th instar. 7. Electrophoretic isozyme patterns of esterase and acid phosphatase on agarose gel, as affected by strain. rearing temperature and feed-resources, were observed as follow. (1) Isozyme patterns of mid-gut esterase varied, depending on instar. One or two more isozyme bands were observed in the larvae than feed on the mulberry fed for the artificial diet. (2) A strain, chinese-15 with a higher feeding response, had 1∼2 more bands than chinese-60 with a lower feeding response. (3) Five bands of mid-gut esterase in 3rd and 4th instar larvae reared at 28$^{\circ}C$. and 4 for 3rd instar and 6∼7 for 4th instar larvae at 35$^{\circ}C$ were observed. (4) No similar esterase bands could be found among mid-gut, blood and silkgland. There are five esterase bands in the midgut, one in blood and three in silkgland. (5) There was rather small digerence in acid phosphatase types of mid-gut and blood according to varieties and rearing temperature. No active band was shown in silkgland. In midgut, there was one acid phosphatase band at 3rd instar, two at 4th instar and three at 5th instar. In blood, one active band at 3rd or 4th instar and three bands at 5th inster wire detected.

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STUDIES ON THE EARLY PREGNANCY DETERMINATION IN COWS BY USING THE ENZYME-IMMUNOASSY AND RADIO-IMMUNOASSAY IN MILK

  • Lee, J.M.;Kim, H.S.;Jeong, S.G.;Jung, J.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.3
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    • pp.299-302
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    • 1996
  • Milk samples(n = 78) were taken 19d, 20d, 21d, 22d after artificial insemination(AI) for early pregnancy diagnosis by using the Enzyme immunoassay(EIA) kit. The progesterone ($P_4$) concentration in the whole milk was measured on the same day of pregnancy diagnosis. Rectal palpation(RP) was accomplished between 60d and 70d after AI to estimate the ovary condition and pregnancy status. Milk progesterone concentrations measured by Radio-immunoassay(RIA) method, in the pregnant cows at 17d, 19d, 21d after insemination were $17.10{\pm}0.91$, $17.60{\pm}0.46$, and $18.43{\pm}0.79nmol/l$, whereas those in the not-pregnant cows were $6.57{\pm}1.03$, $2.63{\pm}0.29$, and $0.67{\pm}0.08nmol/l$, respectively. When the progesterone concentration was less than 7 nmol/l, the color of the EIA kit was lighter and when the progesterone concentration was ${\geq}16nmol/l$, the color of the EIA kit was darker compared to the standard color. The detection rates of error by judging the color differences were 5.1% and 20.7%, respectively. In the early pregnancy diagnosis by the EIA kit and RIA method, the accuracy rates in the pregnancy of cows were 82% and 87%, and those in not-pregnant cows were 86% and 91%, respectively. For ovarian status estimated by the RIA method and certified by RP, the accuracy rates of the ovarian atrophy, follicular cyst and luteal cyst were 80, 91 and 83% and the progesterone concentrations were 2.51, 2.03, and 26.7 nmol/l, respectively.

Effect of Acibenzolar-S-methyl and Rahnella aquatilis (Ra39) on Chitinase and β-1, 3-glucanase Activities and Disease Resistance of Apple Plants

  • Abo-Elyousr, A.M. Kamal;Sallam, M.A.A.;Hassan, M.H.A.;Zeller, W.
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.63-69
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    • 2010
  • The effect of Acibenzolar-S-methyl (ASM) and Rahnella aquatilis Ra39 against apple fire blight disease caused by Erwinia amylovora were tested as a possible alternative to streptomycin. In vitro studies, no inhibition effect against the pathogen was found when ASM was tested. Under greenhouse conditions, application of R. aquatilis Ra39 with the highly susceptible M26 rootstock resulted in a marked disease suppression. Application of ASM and strain Ra39 caused a high decrease of the disease, 82% and 58% respectively; this was correlated with a reduction of the growth of the pathogen within host plants up to 64% and 49.5% respectively. Further studies in the field under artificial infection condition during full bloom revealed that application of ASM and R. aquatilis Ra39 with Gala variety resulted in a control effect up to 21 and 29% respectively. In physiological studies, enhanced activities of PR-proteins (chitinase and $\beta$-1, 3-glucanase) were detected, which are well known as biochemical markers for systemic acquired resistance. Application of ASM to apple shoots caused the highest chitinase activity followed by strain Ra39. The enzyme activity was increased after 2, 4 and 6 days from application. In addition, ASM-treatment caused the higher $\beta$-1, 3-glucanase activity than strain Ra39. Maximum enzyme activity was recorded after 6 days from application and then decreased after 8 and 10 days from application.

Properties of a Fibrinolytic Enzyme Secreted by Bacillus amyloliquefaciens RSB34, Isolated from Doenjang

  • Yao, Zhuang;Liu, Xiaoming;Shim, Jae Min;Lee, Kang Wook;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.9-18
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    • 2017
  • Nine bacilli with fibrinolytic activities were isolated from doenjang, a traditional Korean fermented soy food. Among them, RSB34 showed the strongest activity and was identified as Bacillus amyloliquefaciens by 16S rRNA and recA gene sequencing. During growth on LB up to 96 h, RSB34 showed the highest fibrinolytic activity ($83.23mU/{\mu}l$) at 48 h. Three bands of 23, 27, and 42 kDa in size were observed when the culture supernatant was analyzed by SDS-PAGE and 27 and 42 kDa bands by fibrin zymography. The gene encoding the 27 kDa fibrinolytic enzyme AprE34 was cloned by PCR. BLAST analyses confirmed that the gene was a homolog to genes encoding AprE-type proteases. aprE34 was overexpressed in Escherichia coli BL21(DE3) using pET26b(+). Recombinant AprE34 was purified and examined for its properties. The $K_m$ and $V_{max}$ values of recombinant AprE34 were $0.131{\pm}0.026mM$ and $16.551{\pm}0.316{\mu}M/l/min$, respectively, when measured using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. aprE34 was overexpressed in B. subtilis WB600 using pHY300PLK. B. subtilis transformants harboring pHYRSB34 (pHY300PLK with aprE34) showed higher fibrinolytic activity than B. amyloliquefaciens RSB34.

Screening of Lactic Acid Bacteria as Starter Culture for Making Fermented Sausage (발효 소시지 제조를 위한 기능성 유산균의 선발)

  • Han, Soo-Min;Kim, Young-Joo;Lee, Hong-Chul;Chin, Koo-Bok;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.26 no.4
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    • pp.511-516
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    • 2006
  • The objectives of this study was to compare the probiotic characteristics of lactic acid bacteria (LAB) for their ability to assimilate cholesterol, production of bacteriocin, inhibition of angiotensin I-converting enzyme (ACE), and viability under artificial gastrointestinal fluids. Among tested lactic acid bacteria, L167 strain exhibited the highest ACE inhibitory activity (58.75%). The production of ACE inhibitory peptide derived from fermented milk by L167 strain started at the beginning of stationary phase with maximum activity occurring late of the stationary phase. The highest ACE inhibitory activity was observed at 20 h in 10% skim milk medium. L155 strain exhibited cholesterol assimilation activity compared with probiotic strains such as Lactobacillus acidophilus ATCC 43121. With addition of bacteriocin culture, viable cells of Staphylococcus aureus in fermented sausage were slightly decreased during storage. Among selected strains of LAB, 3 strains weve identified as L. plantarum (L155, L165, L167), and two strains were identified as Pediococcus damnosus (L12) and L. paracasei ssp. paracasei (P113) by use of API carbohydrate fermentation pattern and physiological tests.