• Title/Summary/Keyword: cultivation activities

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Raw Starch-digesting Amylase is Comprised of two Distinct Domains of Catalytic and Substrate-Adsorbable Domain: Role of the C- Terminal Region in Raw-Starch-Binding

  • Kim, Cheorl-Ho
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.40-45
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    • 2001
  • Raw starch-digesting amylase (BF-2A, M.W. 93, 000 Da) from Bacillus circulans F-2 was converted to two components during digestion with subtilisin. Two components were separated and designated as BF-2A' (63, 000 Da) and BF-2B (30, 000 Da), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in $K_{m}$, Vmax for, and in their specific activity for soluble starch hydrolysis. However, its adsorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was greatly different from that of the BF-2A. A smaller peptide (BF-2B) showed adsorb ability onto raw starch. By these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption. A similar phenomenon is observed during limited proteinase K, thermolysin, and endopeptidase Glu-C proteolysis of the enzyme. Fragments resulting from proteolysis were characterized by immunoblotting with anti-RSDA. The proteolytic patterns resulting from proteinase K and subtilisin were the same, producing 63- and 30-kDa fragments. Similar patterns were obtained with endopeptidase Glu-C or thermolysin. All proteolytic digests contained a common, major 63-kDa fragment. Inactivation of RSDA activity results from splitting off the C-terminal domain. Hence, it seems probable that the protease sensitive locus is in a hinge region susceptible to cleavage. Extracellular enzymes immunoreactive toward anti-RSDA were detected through whole bacterial cultivation. Proteins of sizes 93-, 75-, 63-, 55-, 38-, and 31-kDa were immunologically identical to RSDA. Of these, the 75-kDa and 63-kDa proteins correspond to the major products of proteolysis with Glu-C and thermolysin. These results postulated that enzyme heterogeneity of the raw starch-hydrolysis system might arise from the endogeneous proteolytic activity of the bacterium. Truncated forms of rsda, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional amylase that did not bind to raw starch. This indicates that the conserved region of RSDA constitutes a raw starch-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.d.

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Isolation and Characterization of Bacillus Species Possessing Antifungal Activity against Ginseng Root Rot Pathogens (인삼 뿌리썩음병에 길항력이 있는 Bacillus 균의 분리 동정 및 특성 조사)

  • Kim, Byung-Yong;Ahn, Jae-Hyung;Weon, Hang-Yeon;Song, Jaekyeong;Kim, Sung-Il;Kim, Wan-Gyu
    • The Korean Journal of Pesticide Science
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    • v.16 no.4
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    • pp.357-363
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    • 2012
  • Ginseng (Panax ginseng C. A. Meyer) is an economically important crop in Korea. While the consumption of the crop is gradually increasing, the yield is decreasing due to the injury of continuous cultivation or infection of soil-borne fungal pathogens such as Cylindrocarpon destructans, Fusarium solani, Rhizoctonia solani and Sclerotinia nivalis. In order to find promising biocontrol agents, we have isolated 439 soil bacteria from ginseng cultivated soil and tested their antifungal activities against ginseng rot pathogens. Among them, 3 strains were finally selected and tested for the elucidation of their genetic and biochemical properties. They were identified as Bacillus amyloliquefaciens using phylogenetic analysis based on 16S rRNA gene sequences. Moreover, all selected strains showed positive reaction for PCR detection targeting biosynthetic gene sequences of iturin A and surfactin. The results provided promising evidences that the bacterial strains isolated from ginseng cultivated soil can be novel biocontrol agents for ginseng cultivaion.

Effect of Mixed Cultivation of Green Manure Crops on Mineral Nutrients, Microbial Activities, Weed Occurrences and Yields of Red-pepper (토양 무기양분, 미생물활성, 잡초발생 및 고추수량에 대한 녹비작물 혼파효과)

  • Sung, Jwa-Kyung;Lee, Sang-Min;Lee, Yong-Hwan;Lee, Youn;Lee, Ju-Young;Jang, Byong-Choon
    • Proceedings of the Korean Society of Organic Agriculture Conference
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    • 2009.12a
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    • pp.301-301
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    • 2009
  • 겨울철 농경지 이용율 제고 및 자연경관 조성을 위하여 녹비작물을 재배하는 면적이 지속적으로 증가하고 있는 추세이다. 그 동안 녹비작물에 대한 연구는 주로 파종적기, 작물에 대한 양분공급효과 및 타감 효과 등에 집중되어왔다. 본 연구에서는 토양 중 양분공급 및 미생물활성의 경시적 변화, 잡초억제 및 작물생육에 대한 녹비작물의 투입효과를 알아보기 위하여 수행하였다. 국립농업과학원 구내포장에서 2007년 9월 하순에 헤어리베치(8kg/10a)와 호밀(10kg/10a) 종자를 파종한 후, 이듬해 4월 하순에 수확하여 토양에 환원하였다. 질소공급량은 화학비료(15.2kgN/10a) 대비 헤어리베치(32.2kg/10a, 212%), 호밀(9.2kg/10a, 60%) 및 헤어리베치+호밀(15.5kg/10a, 102%)로 헤어리베치의 질소공급효과가 매우 높았다. 토양 무기태질소 함량은 녹비 처리 후 30일경에 최대에 도달하였으나, 호밀 처리구는 조사기간 동안 큰 변화가 없었다. 토양 유기물 함량의 변화는 시험 전(10DBI)과 비교하여 큰 차이를 보이지 않았으나, 녹비환원 후 10일경에 일시적으로 감소하는 경향을 보였는데 이는 녹비환원 후 토양경운에 의한 영향으로 판단되며, 무기태질소와는 달리 호밀처리구의 유기물함량이 지속적으로 증가하는 경향을 보였다. 토양 탄수화물(수용성 당)은 시험 전에 가장 높았으며, 시간의 경과와 함께 서서히 감소되는 양상을 보였으며 토양 미생물탄소 및 미생물질소의 양은 녹비처리구가 화학비료 처리구에 비해 높았으며 녹비환원 후 50일 까지는 증가하다가 그 이후에 감소하는 경향을 보였는데 이는 토양의 무기태질소 및 탄수화물과 밀접한 관계가 있는 것으로 판단되었다. 녹비처리 후 85일경에 잡초발생량을 조사한 결과, 화학비료 처리구에 비하여 헤어리베치 처리구는 40%, 호밀 처리구는 68%, 헤어리베치+호밀 처리구는 40%의 잡초억제효과가 있었다. 고추과의 수량은 화학비료 처리구(702kg/10a)>헤어리베치(694kg>10a)>헤어리베치+호밀(361kg/10a)>호밀(179kg/10a)의 순으로 나타났다.

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Antithrombotic Activity of Hermetia illucens (Black Soldier Fly) (동애등에의 항혈전 활성)

  • Pyo, Su-Jin;Won, Jun;Kang, Deok-Gyeong;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.30 no.4
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    • pp.386-393
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    • 2020
  • Insect industry has been focused as production of food, animal feed, pollinator and for environmental remediation. Hermetia illicens, called as black soldier fly (BSF) is famous as nutritive feed. In this study, to evaluate the antithrombotic activities of BSF, the larvae (instar 2~6), pupae, residue after adult emergence [RAAE] and adult of Hermetia illicens [black soldier fly, BSF] were collected and their ethanol extracts were prepared. Growth of BSF larvae was very rapid and the weight of larvae was increased to 25-folds during 10 days cultivation. The ethanol extraction ratios showed from 1.0% (pupae) to 18.5%(adults). The highest total polyphenol, total flavonoid, and total sugar contents were observed in RAEE (17.2 mg/g), pupae (3.4 mg/g), and instar 6 (37.6 mg/g), suggesting that metabolic changes occur during the life cycle of the BSF. Anti-coagulation assay showed that extracts of RAEE, instar 6 and pupa of BSF significantly inhibited thrombin, prothrombin, and blood coagulation factors. Furthermore, the extracts of RAEE, instar 3 and adult of BSF showed a strong platelet aggregation inhibitory activity. Our results suggest that pupae and RAEE of BSF have potential as antithrombotic agents. This is the first study to provide evidence of the antithrombotic activity of the BSF and bioactivity alterations during its life cycle.

Collection of Soil Actinomycetes from Cheju Island and Screening for their Antibacterial Activities (제주도 토양방선균의 수집과 항균물질 생산균의 선별)

  • Chung, Wan-Seok;Kim, Chang-Jin;Ko, Young-Hwan
    • Applied Biological Chemistry
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    • v.42 no.2
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    • pp.99-104
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    • 1999
  • Soil actinomycetes of 703 strains were isolated from 25 sampling points in Cheju Island using 4 different media. Arginine glycerol salts agar containing soil extract was found to be the best medium for the isolation of soil actinomycetes. Soil samples from pasture land showed higher population and diversity of the actinomycetes than those from citrus field, forest, island, hill or valley. When the antibacterial activity of the 526 isolates was tested against three bacterial strains, Escherichia coli, Staphylococcus aureus and Pseudomonas solanacearum the frequency of the isolates with antibacterial activity varied much depending upon the media used for isolation and cultivation. BL106Ba, one of the 10 isolates that showed antibacterial activity against all the above 3 test strains, was chosen based upon the pH and heat stability of its antibacterial metabolites, and was identified as Streptomyces sp. based upon its cultural, morphological and physiological characteristics. The partially purified white crystalline substance obtained from the culture supematant of BL1063a through cation exchange chromatography(AG MP-50) and three times consecutive gel filtration(Sephadex G-10) showed high antimicrobial activity against gram positive and negative bacteria, but low activity against yeasts. The partially purified substance was found to contain at least four different compounds with antibacterial activity by both thin layer chromatography and high performance liquid chromatography.

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Influence of Soil Flooding on Control of Pink Root Disease in Onion Crop (양파 연작지의 분홍색뿌리썩음병 방제를 위한 담수처리 효과)

  • Moon, Jin-Seong;Lee, Jong-Tae;Ha, In-Jong;Whang, Seon-Gyeong;Song, Wan-Doo;Cheon, Mi-Geon;Lee, Chan-Jung
    • Research in Plant Disease
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    • v.13 no.2
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    • pp.104-109
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    • 2007
  • Pink root disease of onion, a known worldwide constraint upon onion production, significantly reduces crop levels in the main Korea cultivation area. In order to examine the effect of flooding on incidence of pink root disease caused by Pyrenochaeta terrestris and on onion growth and on populations of soil fungi, field experiments were conducted during two seasons, 2003/04 and 2004/05 at Onion Research Institute. Populations of soil fungi from fields were assayed on selective media. Flooding treatment was effective in reducing populations of P. terrestris, Pythium spp., and Rhizoctonia sp. in soil; fungal populations in soils flooded for 90 days were reduced to 1/2 to 1/3 of those in non-flooded soils. In nursery bed, protective activities of soils flooded for over 60 days were 93.5 to 99.2% and their pink root incidences were less than 5%, which were 1/11 to 1/18 of that in control. Increased yield of onion bulbs was associated with control of soil borne pathogenic fungi by flooding treatments. As flooding period was prolonged, bulb grade showed the tendency to increase. Soil flooding for over 60 days resulted in effective disease control, facilitated accurate planning of plant population in the field, extension of the growing-season and consequently higher yields of better quality.

Varietal characteristics of cross-bred Cordyceps militaris 'Dowonhongcho' (교배육종에 의한 밀리타리스 동충하초 '도원홍초'의 품종특성)

  • Lee, Byung-joo;Lee, Mi-Ae;Kim, Yong-Gyun;Lee, Kwang-won;Choi, Young-sang;Lee, Byung-eui
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.151-156
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    • 2015
  • Cordyceps militaris is being studied and cultivated as a medicinal mushroom having many valuable biological and pharmaceutical activities. In the breeding of new C. militaris mushroom, single ascospores were isolated and examined their mycelial growth, mycelial density, and production of stroma and perithecia. Among them selected isolates were crossed and hybrids were produced showing high quality fruiting bodies on artificial media. Mycelial growth rate of new strain 'Dowonhongcho' was higher than that of better on 'Yedang 3' on SDAY at $10-25^{\circ}C$. The stromata of new strain were club-shaped and bright orange-red. Its height was 6.1 cm and the cordycepin content was 0.34% on average. The new strain showed 9% higher yield than 'Yedang 3' with producing firmer fruit bodies. The optimum temperature for mycelial growth was $22{\sim}25^{\circ}C$ and the optimum temperature for stroma development was $18{\sim}22^{\circ}C$. Fruiting bodies were began to produce 45 days later after inoculation. This new cultivar may serve as a valuable one for artificial cultivation and industrial-scale production of C. militaris.

Evaluation of the anti-Helicobacter pylori and cytotoxic properties of the antimicrobial substances from Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 (Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 균주가 생산한 항균물질의 anti-Helicobacter pylori 활성 및 위장상피세포에 대한 세포독성 평가)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.156-168
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    • 2015
  • The objective of this study is to investigate the anti-Helicobacter pylori and anti-cancer activities of the live cells (LC), cell-free culture supernatants (CFCS), and bacteriocin solution (BS) obtained from Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 strains. After incubation for 30 h in MRS broth, the concentration of lactic acid produced by L. paracasei BK57 ($155.9{\pm}10.2mM$) was higher than in MRS broth using L. acidophilus BK13 ($126.8{\pm}7.9mM$). Maximum bacteriocin activity (128 AU/ml) of BK13 strain was observed after 30 h of cultivation at $37^{\circ}C$, however its magnitude was significantly lower than that of BK57 strain (256 AU/ml). The LC of L. acidophilus BK13 and L. paracasei BK57 were able to inhibit the growth of H. pylori ATCC 43504 at different incubation times, depending on the initial inoculum of the LAB. These CFCS and BS obtained from BK13 and BK57 strains dramatically inhibited the growth, adhesive ability, and enzymatic activity of H. pylori. Meanwhile, the anti-cancer effect of the lactic acid from L. acidophilus BK13 and L. paracasei BK57 strains on AGS cells had significant differences with the control group. Therefore, these antagonistic substances-producing strains are potentially useful as new potential antimicrobial agents for the management and prevention of H. pylori infections.

Physicochemical properties, bioactive composition, and antioxidant activity of different coffee beans dependent on the cultivation region (원산지에 따른 커피의 이화학적 특성, 생리활성 성분 및 산화방지 활성)

  • Lee, Kyung Soo;Kim, Ja Min;Yoon, Kyung Young
    • Korean Journal of Food Science and Technology
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    • v.49 no.5
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    • pp.474-479
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    • 2017
  • Five types of coffee bean, which are usually imported and consumed in Korea, were roasted at $250^{\circ}C$ for 15 min and extracted by using a filter coffee machine. The physicochemical properties, functional components, and radical scavenging activity of coffee bean extracts were investigated. The pH of extract was the highest among the extracts and the soluble solid contents of extracts were $0.9-1.0^{\circ}Bx$. The acidity of the extracts was in the range from 0.46-0.55%, which was not significantly different from the control. Indonesian coffee bean extract showed the highest brown color intensity and contained the highest amounts of caffeine and chlorogenic acid. The highest total polyphenol content was found in Kenyan coffee bean extract. Coffee bean extracts from Indonesia and Kenya showed significantly higher radical scavenging activities than the other extracts. This study showed that coffee bean extracts from Indonesia and Kenya contained a large number of bioactive compounds and high antioxidant activity.

Antioxidant effect of ethanol extract from Poria cocos depending on cultivation methods (재배방법을 달리한 복령 에탄올추출물의 항산화 효과)

  • Kim, Yoo-Jin;Park, Hae-Jin;Lee, Jin Sang;Do, Eunju;Sohn, Hyeong Rack;Jeon, Seon Man;Yeum, Jeong Hyun;Kim, Mi Ryeo
    • The Korea Journal of Herbology
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    • v.31 no.5
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    • pp.107-114
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    • 2016
  • Objectives : Poria cocos Wolf has been widely used in Korean medicine as a medicinal fungus. In this study, we investigated that comparative anti-oxidant effects of ethanol extract from wild Poria cocos (WP) and plastic bag-cultivated Poria cocos (PBP).Methods : Total polyphenol and flavonoid contents in WP and PBP extract were monitored. And DPPH, ABTS, hydroxyl (·OH) free radical scavenging capacity, reducing power and xanthine oxidase inhibition activities of them were determined at 5, 1, 0.5 mg/ml concentrations.Results : Higher total polyphenol contents were found in the PBP extract (52.07±0.6 mg/TAEg) than in the WP extract (28.44±0.26 mg/TAEg). The flavonoid contents in WP and PBP extract were 17.29±0.30 mg/ RUEg, 21.36±0.40 mg/ RUEg, respectively. Also, DPPH, ABTS and ·OH free radical scavenging capacity of PBP extract in treated concentrations (5, 1, 0.5 mg/ml) significantly increased compared to those of WP-treated group. In particular, DPPH, ABTS free radical scavenging capacity and reducing power of PBP extract at 5 mg/ml concentration were similar to positive control (BHA or vit. C). Xanthine oxidase (XO) inhibition rate in both extract increased dose dependently. But it was significantly increased in PBP-treated group, only at 5 mg/ml, compared to those of WP-treated group. Then, their inhibition rate by PBP was similar to positive control (BHA).Conclusions : These results suggest that PBP extract is superior to WP extract in anti-oxidant capacity thus PBP can be applied in variable antioxidative products as a substitute for WP.