• Title/Summary/Keyword: Helix 8

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Purification and Characterization of Bacillus subtilis JS-17 Collagenase. (Bacillus subtilis JS-17이 생산하는 Collagenase의 정제 및 특성)

  • Lim Kyoung-Suk;Son Shung-Hui;Kang Ho Young;Jun Hong-Ki
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.657-663
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    • 2005
  • Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.

Molecular cloning and expression pattern of Metallothionein Gene from the left-handed shell, Physa acuta (왼돌아물달팽이 (Physa acuta) 의 Metallothionein 유전자 클로닝 및 발현양상)

  • Jo, Yong-Hun;Baek, Moon-Ki;Kang, Se-Won;Lee, Jae-Bong;Byun, In-Seon;Choi, Sang-Haeng;Chae, Sung-Hwa;Kang, Jung-Ha;Han, Yeon-Soo;Park, Hong-Seog;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.25 no.3
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    • pp.223-230
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    • 2009
  • Metallothioneins (MTs) play a key role in metallic homeostasis and detoxification in most living organisms. In an attempt to study the biological functions and significance of MT in a snail, we cloned and partially characterized the MT gene from the left-handed snail, Physa acutawhich has been regarded as a potential biomonitering species for fresh water. The complete cDNA sequence of PaMT cDNA was identified from the expressed sequence tag (EST) sequencing project of Physa acuta. The coding region of 180 bp gives 60 amino acid residues including the initiation methionine and termination codon. Clustering and phylogenic analysis of PaMT with other MT amino acid sequences show that it has some identities to Helix pomatia (60%), Arianta arbustorum (58%), Perna viridis (49%), Mytilus edulis (49%), Bathymodiolus azoricus (49%), Bathymodiolus azoricus (48%) and Bathymodiolus sp. FD-2002 (48%). Time dependent induction for PaMT from P. acuta exposed with cadmium (50 ppb) indicated that PaMT was induced at 4-8 hr after exposure. It remains to further develop PaMTas a potential biomarker for water contamination in fresh water.

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Lethal (2) Essential for Life [l(2)efl] Gene in the Two-spotted Cricket, Gryllus bimaculatus (Orthoptera: Gryllidae) (쌍별귀뚜라미(Gryllus bimaculatus)의 l(2)efl cDNA 클로닝과 발현분석)

  • Kwon, Kisang;Lee, Nuri;Kwon, O-Yu
    • Journal of Life Science
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    • v.31 no.7
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    • pp.671-676
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    • 2021
  • A cDNA encoding the protein lethal (2) essential for life [l(2)efl] was cloned from Gryllus bimaculatus and named GBl(2)efl. This protein is composed of 189 amino acids, including an N-glycosylation site and 15 phosphorylation sites. Its predicted molecular mass is 21.19 kDa, with a theoretical isoelectric point of 6.2. The secondary structure of GBl(2)efl was predicted from the identification of random coils (56.08%), alpha helices (22.22%), extended strands (17.99%), and beta turns (3.7%) through sequence analyses. A homology analysis revealed that GBl(2)efl exhibited a high similarity with other species at the amino acid level, ranging from 52% to 69%. While GBl(2)efl mRNA expression was higher in the dorsal longitudinal flight muscle following a three-day starvation and in the Malpighian tubules following a one-day starvation, no changes in expression were detected in other tissues. Furthermore, tunicamycin-induced endoplasmic reticulum (ER) stress resulted in an approximately 1.8-fold higher expression in the fat body compared with the wild type.

Effects of fermentation on protein profile of coffee by-products and its relationship with internal protein structure measured by vibrational spectroscopy

  • Samadi;Xin Feng;Luciana Prates;Siti Wajizah;Zulfahrizal;Agus Arip Munawar;Peiqiang Yu
    • Animal Bioscience
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    • v.36 no.8
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    • pp.1190-1198
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    • 2023
  • Objective: To our knowledge, there are few studies on the correlation between internal structure of fermented products and nutrient delivery from by-products from coffee processing in the ruminant system. The objective of this project was to use advanced mid-infrared vibrational spectroscopic technique (ATR-FT/IR) to reveal interactive correlation between protein internal structure and ruminant-relevant protein and energy metabolic profiles of by-products from coffee processing affected by added-microorganism fermentation duration. Methods: The by-products from coffee processing were fermented using commercial fermentation product, called Saus Burger Pakan, consisting of various microorganisms: cellulolytic, lactic acid, amylolytic, proteolytic, and xylanolytic microbes, for 0, 7, 14, 21, and 28 days. Protein chemical profiles, Cornell Net Carbohydrate and Protein System crude protein and CHO subfractions, and ruminal degradation and intestinal digestion of protein were evaluated. The attenuated total reflectance-Ft/IR (ATR-FTIR) spectroscopy was used to study protein structural features of spectra that were affected by added microorganism fermentation duration. The molecular spectral analyses were carried using OMNIC software. Molecular spectral analysis parameters in fermented and non-fermented by-products from coffee processing included: Amide I area (AIA), Amide II (AIIA) area, Amide I heigh (AIH), Amide II height (AIIH), α-helix height (αH), β-sheet height (βH), AIA to AIIA ratio, AIH to AIIH ratio, and αH to βH ratio. The relationship between protein structure spectral profiles of by-products from coffee processing and protein related metabolic features in ruminant were also investigated. Results: Fermentation decreased rumen degradable protein and increased rumen undegradable protein of by-products from coffee processing (p<0.05), indicating more protein entering from rumen to the small intestine for animal use. The fermentation duration significantly impacted (p<0.05) protein structure spectral features. Fermentation tended to increase (p<0.10) AIA and AIH as well as β-sheet height which all are significantly related to the protein level. Conclusion: Protein structure spectral profiles of by-product form coffee processing could be utilized as potential evaluators to estimate protein related chemical profile and protein metabolic characteristics in ruminant system.

Problem Analysis of the Experiments Illustrating pH Effects on Enzyme Activities in High School Science Textbooks - Focus on Starch-Iodine Reaction - (고등학교 과학 교과서에서 "pH가 효소의 작용에 미치는 영향" 실험의 문제점 분석 -녹말과 요오드 반응을 중심으로-)

  • Ji, Jae-Hwa;Jeong, Dae-Hong
    • Journal of The Korean Association For Science Education
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    • v.29 no.8
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    • pp.923-933
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    • 2009
  • In this study, analysis was carried out on science experiment in high school textbooks, illustrating 'the effect of pH on enzyme activity.' Five of the total 16 science textbooks introduced in this experiment, and the experimental conditions therein were analyzed. Textbook analysis revealed that pH of below 3 was used for 'acidic condition' and that of over 11 was used for 'basic' condition. Using the experimental conditions described in the textbooks, review experiments were performed. Buffering effect with the addition of saliva was found in the pH region around 7 when buffer solution was not used to control pH as was in the textbooks. The enzyme activity experiments were performed controlling pH from pH 2 to 13 with buffer. The color of the sample was blue from pH 2 to 4, and then disappeared from pH 5 to 8, reflecting that starch was digested owing to enzyme activity. In pH 9 light blue color appeared, indicating de-activation of enzyme under this basic condition. However, the blue color of the sample became lighter at pH 10 and disappeared from pH 11, which was different from the expected behavior anticipating dark blue color due to de-activation of enzyme under strong basic condition. These results can wrongly influence students to interpret that enzyme can be activated in this pH condition. So, we analyzed the reason for the color of the sample turning light blue in pH 10 and disappeared from pH 11. The analysis resulted that ${I_3}^-$ and/or ${I_5}^-$ subunits of polyiodides within the starch helix in starch-iodine complex, showing blue, decreases above pH 10 due to disproportionation to HOI, ${IO_3}^-$, and $I^-$ by the reaction with $OH^-$.

Molecular Characterization and Phylogenetic Analysis of Season Influenza Virus Isolated in Busan during the 2006-2008 Seasons (부산지역에서 유행한 계절인플루엔자바이러스의 유전자 특성 및 계통분석('06-'08 절기))

  • Park, Yon-Koung;Kim, Nam-Ho;Choi, Seung-Hwa;Lee, Mi-Oak;Min, Sang-Kee;Kim, Seong-Joon;Cho, Kyung-Soon;Na, Young-Nan
    • Journal of Life Science
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    • v.20 no.3
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    • pp.365-373
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    • 2010
  • To monitor newly emerged influenza virus variants and to investigate the prevalence pattern, our laboratory performed isolation of the viruses from surveillance sentinel hospitals. In the present study, we analysed influenza A/H1N1, A/H3N2, B viruses isolated in Busan during the 2006/07 and 2007/08 seasons by sequence analysis of the hemagglutinin (HA1 subunit) and neuraminidase (NA) genes. The isolates studied here were selected by the stratified random sample method from a total of 277 isolates, in which 15 were A/H1N1, 16 were A/H3N2 and 29 were B. Based on the phylogenetic tree, the HA1 gene showed that A/H1N1 isolates had a 96.7% to 97.7% homology with the A/Brisbane/59/2007, A/H3N2 isolates had a 98.4% to 99.7% homology with the A/Brisbane/10/2007, and B isolates had a 96.5% to 99.7% homology with the B/Florida/4/2006(Yamagata lineage), which are all the vaccine strains for the Northern Hemisphere in 2008~2009 season. In the case of the NA gene, A/H1N1 isolates had 97.8% to 98.5% homologies, A/H3N2 isolates had 98.9% to 99.4% homologies, and B isolates had 98.9% to 100% homologies with each vaccine strain in the 2008~2009 season, respectively. Characterization of the hemagglutinin gene revealed that amino acids at the receptor-binding site and N-linked glycosylation site were highly conserved. These results provide useful information for the control of influenza viruses in Busan and for a better understanding of vaccine strain selection.

Epidermal Structure and Stomatal Types in Some Species of Korean Crassulaceae (한국산 돌나물과(Crassulaceae) 수종의 표피구조와 기공유형)

  • Jeong, Woo-Gyu;Sung, Min-Wung
    • Journal of Plant Biology
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    • v.28 no.2
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    • pp.105-118
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    • 1985
  • This study was carried out to investigate the epidermal structure and types of stomata on the upper and lower surfaces of the leaves in six species and one variety belonging to Korean Crassulaceae. The shape of epidermal cells was polygonal, isodiametric, or elongated. The cell wall was thick, sinuous or deeply sinuous. The subsidiary cell wall was thin or mostly arched. The cytolith-containing cells were found in Orostachys japonicus. The distribution of stomata was more on the upper surface than on the lower surface in the leaves of O. japonicus and Sedum sarmentosum. The stomata in the other species were less on the upper surface than on the lower surface, and stomatal size was different in each species. The great majority of stomatal types was the helico-eumesogenous type. This type was subdivided into two, parahelico-eumesogenous type and diahelico-eumesogenous type by us on the basis of the angle of division of guard mother cell and the long axis of the last-formed subsidiary cells. Sometimes allelo-eumesogenous type as well as aniso-eumesogenous type was found. The tetra-eumesogenous type with four mesogenous subsidiary cells was rarely observed. It was observed that the varieties of aniso-eumesogenous type with one to three subsidiary cells and one helix of subsidiary cells were developed by the secondary division of subsidiary cells of aniso-eumesogenous stoma. The compound shape of helico-eumesogenous stomata was found. Five new kinds of stomatal types were observed as follows; the helico-tetra-eumesogenous type, the coallelohelico-eumesogenous type, the cohelico-allelo-eumesogenous type, the duplotetra-eumesogenous type, and the aniso-euperigenous type. These types were reported here in the vascular plants for the first time.

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Characterization of a cDNA Encoding Transmembrane Protein 258 from a Two-spotted Cricket Gryllus bimaculatus (쌍별귀뚜라미(Gryllus bimaculatus)의 GbTmem258 cDNA 클로닝과 발현분석)

  • Kisang Kwon;Honggeun Kim;Hyewon Park;O-Yu Kwon
    • Journal of Life Science
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    • v.33 no.10
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    • pp.828-834
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    • 2023
  • The cDNA that encodes transmembrane protein 258 (Tmem258) was cloned from Gryllus bimaculatus and named GbTmem258. This protein comprises 80 amino acids, has no N-glycosylation site, and contains five potential phosphorylation sites at two serines, two threonines, and one tyrosine. The predicted molecular mass of GbTmem258 is 9.06 kDa, and its theoretical isoelectric point is 5.5. The tertiary structure of GbTmem258 was predicted using the available secondary structure information, which suggests the presence of alpha helices (52.5%), random coils (22.5%), extended strands (16.25%), and beta turns (8.75%). Homology analysis revealed that GbTmem258 exhibits high similarity at the amino-acid level to Tmem258 found in other species. The effect of starvation and refeeding on GbTmem258 mRNA expression was also examined in this study. It was found that GbTmem258 mRNA expression in the hindgut progressively increased throughout the starvation period, peaking at almost 1.5 times the control level after six days of starvation. However, refeeding for one to two days after the six-day starvation period restored GbTmem258 mRNA expression to the control level. In fat body, GbTmem258 mRNA expression was almost 3-fold higher during starvation compared to the control level. Refeeding for one to two days after the six-day fast resulted in a decline in the expression to about a 2.5-fold increase over the control level. Throughout the starving and refeeding periods, no other tissues showed any discernible alterations in GbTmem258 mRNA expression.

Effects of Evaporative Water-loss from Cultural Pots on Growth of Pot-grown Ornamental Plants (화분(花盆)의 수분증발(水分蒸發)이 분식화훼류(盆植花卉類)의 생육(生育)에 미치는 영향(影響))

  • Suh, Youn-gkyo
    • Korean Journal of Agricultural Science
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    • v.4 no.2
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    • pp.317-343
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    • 1977
  • This study was carried out to obtain the informations about evaporation from pot, soil temperature and soil atmosphere composition in pot, and the effect on the growth of nine ornamental species using seven different containers. The investigated containers were clay pot(CP), clay pot painted in green(CP-P), varnished clay pot(CP-V), polyethylene film inserting in clay pot(CP-PI), clay pot mulched with black polyethylene film(CP-PM), porcelain pot(POP), and plastic pot(PLP). Nine ornamental species were balsam(Impatiens balsamina), chrysanthemum(Chrysanthemum morifolium), cosmos(Cosmos bipinatus), English ivy(Hedera helix), geranium(Pelargonium zonale), kochia(Kochia scoparia var. trichophila), marigold(Tagetes patula), ornamental kale(Brassica oleraceae var. acephala), and salvia (Salvia splendens). The results obtained are summarized as follows: 1. Dry weight of all tested species grown in PLP, POP, CP-P, CP-V and CP-PI was heavier than that of CP. 2. Plant height in nine tested species grown in PLP, POP, CP-P, CP-V, and CP-PI was taller than that of CP. 3. Geranium grown in PLP, POP, CP-P, and CP-V gave more number of leaf than that of CP. 4. The number of flower in balsam grown in PLP, POP, CP-P, CP-V and CP-PI was more than that of CP. The result from marigold was very similar to this tendency. Spike length and floret number in salvia gave the same tendency, but its spike number was not different among containers used. 5. The average diurnal evaporation from PLP and POP was about 43% of that of CP. About two third of total evaporation from CP was through pot wall. 6. The evaporation rate from the slowest to the highest was PLP, POP, CP-P, CP-V, CP-PI, CP-PM and CP. Containers inhibiting evaporation through pot wall hold more soil moisture than CP from one day after water supply. 7. The more evaporative water-loss from containers gave the lower soil temperature. The variation of soil temperature among containers was higher during the day than the night. 8. The $O_2$ concentration of soil atmosphere in CP was higher than that of nonporous containers, and the difference between them was 0.40-1.12%. The range of the $O_2$ concentration 17.95~19.62%. The $CO_2$ concentration of soil atmosphere in CP was lower than that of nonporous containers, and its range was 0.59-1.76%. This deviation in soil atmosphere composition did not influenced on the growth of plants. 9. There was a possitive correlation between the amount of soil water and the plant growth. 10. Plant grown on CP gave more total nitrogen content in top growth than that on PLP. C/N ratio was somewhat low in plant on CP. From the above results, $O_2$ and $CO_2$ concentration in soil atmosphere did not gave enough deviation to the extent which affect the plant growth. The effect of soil moisture on the plant growth using different containers was the far-most significant factor from this investigation. Therefore, it was obious that the utilization of the nonporous container might save the cost for water supply and reduce the production cost of the pot-grown ornamental plant in Korea eventually.

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