• Journal of Microbiology and Biotechnology
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• 제22권8호
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• pp.1044-1053
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• 2012

The gene encoding an esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est5S) was 1,026 bp in length, encoding a protein of 366 amino acid residues with a calculated molecular mass of 40,168 Da. The molecular mass of the enzyme was estimated to be 40,000 Da. The Est5S protein contains the Gly-X-Ser-X-Gly motif found in most bacterial and eukaryotic serine hydrolases. However, the Asp or Glu necessary for the catalytic triad [Ser-Asp-(Glu)-His] was not present, indicating Est5S represents a novel member of the GHSQG family of esterolytic enzymes. BlastP in the NCBI database analysis of Est5S revealed homology to hypothetical proteins and it had no homology to previous known lipases and esterases. Est5S was optimally active at pH 7.0 and $40^{\circ}C$. Among the p-nitrophenyl acylesters tested, high enzymatic activities were observed on the short-chain p-nitrophenyl acylesters, such as p-nitrophenyl acetate, etc. The conserved serine residue ($Ser_{190}$) was shown to be important for Est5S activity. The primers that amplified the est5S gene did not show any relative band with 49 species of culturable rumen bacteria. This implies that a new group esterase gene, est5S, may have come from a noncultured cow rumen bacterium.

• 농업생명과학연구
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• 제46권3호
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• pp.109-118
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• 2012

The gene encoding an esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est2R) was 2,120 bp in length, encoding a protein of 516 amino acid residues with a calculated molecular weight of 57,286 Da. The molecular weight of the enzyme was estimated to be 57,000 Da by SDS-PAGE. Est2R shared 35.6% amino acid identity with esterase (CAH19079) of uncultured prokaryote. The Est2R was most active at $20-40^{\circ}C$, and showed optimum at $30^{\circ}C$ and pH 8.0. The most activity of Est2R for the different chain length of p-nitrophenyl ester group as substrate was p-nitrophenyl acetate. Moreover, the enzyme was found to be most active without organic solvent, followed by 98% active with ethanol, and the enzyme activity was highly affected by the acetonitrile. The enzyme was significantly inhibited by $Zn^{2+}$ but stimulated by $Ca^{2+}$. So, novel esterase gene est2R is likely to obtain from cow rumen metagenome and supposed to use for industrial purpose.

• The Journal of Korean Physical Therapy
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• 제26권6호
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• pp.436-441
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• 2014

Purpose: The objective of this study was to offer clinical primary data that it's aims to examine effects of transcranial direct current stimulation (tDCS) on cognitive function and biochemical change of rat with alzheimer's disease(AD) induced by injecting scopolamine. Methods: Subjects were instructed cognitive dysfunction model, rat of Sprague-Dawley system was injected with scopolamine and each experimental group was classified into three; group I (n=16) is non-treatment groups; group II (n=16) is applied with the tacrine; group III (n=16) is applied with the tDCS. The ziggurat task test was conducted to observe behavioral changes and cognitive function ability and 7, 14, 21, 28 days after the model. Acetylcholine Esterase (Ach E) activity was examined for biochemical assessment of which the results are followed. Results: Participants showed as to behavioral change, tacrine application group was the most significantly responded, following tDCS application group. As to biochemical change, same as above, tacrine application group was the most significantly responded, following tDCS application group. Conclusion: From these results, confirm that tDCS application to rat with alzheimer's disease leads to positive effects on behavioral, cognitive function changes, and biochemical changes, lasting for certain period of time. This study, in particular, tDCS, which can change excitability of brain cells non-invasively, could provide basic data that is useful as a new treatment way for the patients with cognitive dysfunction.

• Journal of Plant Biology
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• 제29권4호
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• pp.233-242
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• 1986

In order to pursue some physiological studies on organogenesis in ginseng tissue culture, ginseng root explants were cultured on a modified MS medium containing NAA and kinetin. The activities of peroxidase and some enzymes were investigated and their isoenzyme patterns were also observed. The activity of peroxidase decreased by 20% in one week's culture and increased thereafter by 80% in culturing for 7 weeks compared with the control group. Glucose-6-phosphate dehydrogenase activity increased by 400% after culturing for 5 weeks and increased during the days preceeding root formation. The activities of glutamate dehydrogenase and acid phosphatase also increased during the culture. After 3 weeks' culture, new peroxidase isozyme (pH 7.6) appeared and 7 weeks' culture, another new peroxidase isozyme (pH unidentified) appeared. These patterns were also identified by using FPLC. After 7 weeks' culture, a new esterase isozyme of pH 8.5 appeared and isozyme patterns of acid phosphatase were quite changed compared with the isozyme patterns of tissue cultured for 5 weeks. In so far as these new isoenzymes appear distinctively after 7 weeks' culture, root differentiation is supposed to be induced after 7 weeks' culture.

• 한국잠사곤충학회지
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• 제39권2호
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• pp.119-133
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• 1997

Isozyme was used to characterize general protein patterns of genetic relationships among 303 silkworm stocks preserved in National Sericultural and Entomology Research Institute, RDA. Six isozymes (esterase, acid phosphatase, alkaline phosphatase, amylase, glucose-6-phosphate dehydrogenase and sucrase) from hemolymph, midgut, and digestive juice were employed to construct dendograms(UPGMA method) using a polycrylamide gel electrophoresis. A cluster analysis revealed four major group, which were divided into several subgroups within each group, contained assemglages of Japanese and Chinese races. Especially, genetic differentiation in the first and second group was greatest rather than within Japanese and Chinese races repectively and was concordant with the hypothesis of phyletic sorting of initial variability in China many years ago. Hypothesized recent introgression between groups was also plausible, but the eviednce suggested bidirectional gene flow between the Chinese and the Japnaese lineages. Interpreting the results in light of evidence from the current study, the genetic diversity and relationship showed in Korean silkworm race, Hansammyun reflected early and independent evolution from the Chinese ancestor, limited addition of new variability and phyletic sorting within Korean peninsula more than 4,000 years.

• 생명과학회지
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• 제20권9호
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• pp.1306-1313
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• 2010

한우의 반추위에서 게놈 DNA를 분리하여 메타게놈 은행을 구축한 다음 carboxylesterase를 암호화하는 유전자를 클로닝 및 유전자를 선별하였다. 선별된 유전자의 DNA 염기서열 및 아미노산 서열을 분석하고 유전산물의 생화학적인 특성을 조사하였다. est1R 유전자는 2,465 bp로 366개의 아미노산 잔기를 가진 단백질을 암호화하였으며 이 단백질의 이론적인 분자량은 61,166 Da이었다. Est1R단백질은 PNB carboxylesterase (P37967), acetylcholinesterase (1EEAA) 및 chain A (1H23A)와 각각 5.9%, 6.1%, 6.1% 상동성을 가지고 있었다. 이러한 검색 결과 기존의 알려진 lipase 및esterase와의 상동성이 낮은 것으로 보아 새로운 그룹의 효소로 추정된다. Est1R효소의 최적 pH는 7.0 근방이었으며 최적 온도는 $40^{\circ}C$ 부근이었다. 한편 10% 유기용매를 함유한 기질의 효소활성측정에서 대조구에 비해 methanol은 95%의 상대적인 활성을 가진 반면에 hexane 용액에서는 그 활성이 반으로 감소하였다. 따라서 유기용매 농도의 작용성에 따라 이 효소의 산업적 이용성도 가능하리라 추정된다.

• 한국동물학회지
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• 제16권2호
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• pp.79-96
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• 1973

붕어(Carassius carassius)에 미치는 동의 영향을 밝히기 위하여 다음과 같은 연구를 하였다. 1) 셀루로오스 아세테이트 전기영동법에 의한 젖산수소이탈효소 아이소자임 상, 2) 분광비색법에 의한 LDH활성과 LDH효소계에 대한 동의 영향, 3) 한천 박층 전기영동법에 의한 에스테라아제 아이소자임 상, 4) 전분 겔 전기영동법에 의한 혈색소 상, 및 5) 조직 학적 연구. 1. LDH 아이소자임 영동대는 정상 붕어의 아가미에 2개 (LDH-3 및 LDH-5), 간에 3개(LDH-2, LDH-4, 및 LDH-5), 그리고 근육에서 2개 (LDH-3 및 LDH-4) 나타났다. LDH-1은 이상의 세가지 조직에서 나타나지 않았다. 동을 처리한 붕어의 간에서는 LDH-3이 나타났고 근육에서는 LDH-5가 나타났다. 그러나 아가미에서는 새로운 영동대가 나타나지 않았다. 2. LDH활성은 정상 붕어의 아가미, 간, 및 근육 중 아가미에서 가장 낯았고 근육에서 가장 높았으며, 아가미를 제외한 간과 근육에서는 1일부터 10일간의 동처리에 따라 점차적으로 감소되었다. 3. 동을 처리한 붕어의 간과 근육의 LDH활성 감소는 주로 체내 M-LDH에 대한 동의 억제에 기인된다. 4. 정상 붕어의 아가미, 간, 근육, 혈액, 뇌, 및 신장의 에스테라아제 아이소자임 영동대 수는 각각 3,6,2,2,2 및 2개 였고 이들은 동을 처리한 경우에서도 같았다. 동처리군의 아가미, 간, 혈액 그리고 신장의 에스테라아제 영동대의 상대이동도는 대조군의 그것들과 상이하였다. 5. 정상 붕어의 혈색소 영동대는 양극에 1개 있었다. 동을 처리한 붕어의 혈색소 영동대의 이동도는 정상 붕어의 그 것보다 약간 빠른 것으로 보였다. 6. 동(20ppm)에 5일간 처리한 붕어의 조직학적 연구는 다음과 같았다. 1) 아가미에서는 처음에 표피층이 분리되었다가 분해된 후 완전히 파괴되었다. 2) 간에서는 많은 지방립이 나타났다. 3) 정상 붕어와 동에 처리된 붕어의 근육 사이에서는 조직학적 차이가 없었다. 4) 신장족직은 기부요세관에 손상을 받았다. 요세관세포의 높이가 작아진 후 요세관의 내강이 확대되었다. 많은 기부요세관이 분홍빛의 과립성 형태와 여러 단계의 변성을 나타내었다.

• 한국미생물·생명공학회지
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• 제32권3호
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• pp.224-229
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• 2004

토양으로부터 강력한 CAH activity를 갖는 균주를 분리하여, 생화학적, 배양학적, 전자현미경 동정을 한 결과, Bacillus sphaericus로 확인되었으며, 이를 B. sphaericus 366M-9로 명명하였다. 또한 이 균으로부터 최초로 cephalosporin-C deacetylase(CAH)를 분리 정제하였다. 정제수율은 약 7.5% 였으며, B. sphaericus 366M-9에서 분리한 CAH-9의 최적활성 온도는 $50^{\circ}C$였으며, 효소안정 온도구간은 30～$50^{\circ}C$이다. 또한 최적 활성 pH는 7.0이었으며, 효소안정 pH구간은 pH 6.0～8.0으로 90% 이상의 잔존 활성도를 나타내었다. 효소반응속도에 미치는 기질의 영향에서는 $K_{m}$ 값은 0.87 mM 이며,$ V_{max}$는1.22 unit/ml이었다.다.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2009년도 추계학술대회 및 정기총회
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• pp.3-13
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• 2009

90 wild Agaricus strains from China, including 44 Agaricus bisporus strains identified preliminarily by isozyme electrophoresis, were studied by the techniques of SRAP and ISSR. 18 special SRAP bands and 12 special ISSR bands were analyzed, the strains were clustered and a demdrogram was obtained. The results showed that the strains were divided into 2 groups, wild A. bisporus group and the other Agaricus group. It is similar to the result of isozyme electrophoresis. 41 wild A. bisporus strains from Sichuan and Tibet were divided into 4 groups based on their growing places, suggesting the regionally difference of the strains to be quite obvious. Some white wild A. bisporus strains from Xinjiang and Tibet had special patterns, resulting in lower coefficient values with other wild A. bisporus strains. The biological characteristics of three wild A. bisporus strains were analyzed, and the results showed: 1. The wild strains grew slowly on PDA medium with weak appressed mycelia, and grew normally in kernel or fermented cottonseed shell substrate. 2. They grew faster than control strain As2796 under lower temperature of $16^{\circ}C$, and higher temperature of $32^{\circ}C$, with optimum growing temperature of $20-24^{\circ}C$, which was $4^{\circ}C$ lower than that of control strain. 3. In the cultivation with manure compost via twice fermentation, the mycelia grew normally in compost and quite slowly in casing soil, and the fruitbodies occurred less and late with easily opening and low production. 4. The fruitbody was off-white with flat and scaled cap, long stipe and dark gill. The bisporus basidia occupied 70-80% and trisporus basidia 20-30% of the total basidia. 5. Heterokaryotic monospore isolates could fruit in cultivation, and the homokaryotic isolates could cross with those derived from overseas wild A.bisporus strains. 6. The electrophoresis phenotype of isozymes such as esterase etc. belonged to high production type (H type). 7. The RAPD patterns made much difference from those of high production, good quality or hybrid strains, which indicated that the wild strains produce a new kind of RAPD type.