• Title/Summary/Keyword: Bacterial Production

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Identification of Antagonistic Bacteria, Pseudomonas aurantiaca YC4963 to Colletotri­chum orbiculare Causing Anthracnose of Cucumber and Production of the Antibiotic Phenazine-l-carboxylic acid (Colletotrichum orbiculare에 대한 길항세균 Pseudomonas aurantiaca YC4963의 분리 동정 및 항균물질 Phenazine-1-carboxylic acid의 생산)

  • Chae Hee-Jung;Kim Rumi;Moon Surk-Sik;Ahn Jong-Woong;Chung Young-Ryun
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.342-347
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    • 2004
  • A bacterial strain YC4963 with antifungal activity against Colletotrichum orbiculare, a causal organism of cucumber anthracnose was isolated from the rhizosphere soil of Siegesbeckia pubescens Makino in Korea. Based on physiological and biochemical characteristics and 16S ribosomal DNA sequence analysis, the bac­terial strain was identified as Pseudomonas aurantiaca. The bacteria also inhibited mycelial growth of several plant fungal pathogens such as Botrytis cinerea, Fusarium oxysporum and Rhizoctonia solani on PDA and 0.1 TSA media. The antifungal activity was found from the culture filtrate of this isolate and the active compound was quantitatively bound to XAD adsorption resin. The antibiotic compound was purified and identified as phenazine-l-carboxylic acid on the basis of combined spectral and chemical analyses data. This is the first report on the production of phenazine-l-carboxylic acid by Pseudomonas aurantiaca.

A novel cold-active lipase from Psychrobacter sp. ArcL13: gene identification, expression in E. coli, refolding, and characterization (새로운 Psychrobacter sp. ArcL13 유래 저온활성 지질분해효소 : 유전자 분리동정, 대장균에서의 발현, refolding 및 특성 연구)

  • Koo, Bon-Hun;Moon, Byung-Hern;Shin, Jong-Suh;Yim, Joung-Han
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.192-201
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    • 2016
  • Recently, Psychrobacter sp. ArcL13 strain showing the extracellular lipase activity was isolated from the Chuckchi Sea of the Arctic Ocean. However, due to the low expression levels of the enzyme in the natural strain, the production of recombinant lipase is crucial for various applications. Identification of the gene for the enzyme is prerequisite for the production of the recombinant protein. Therefore, in the present study, a novel lipase gene (ArcL13-Lip) was isolated from Psychrobacter sp. ArcL13 strain by gene prospecting using PCR, and its complete nucleotide sequence was determined. Sequence analysis showed that ArcL13-Lip has high amino acid sequence similarity to lipases from bacteria of some Psychrobacter genus (84-90%) despite low nucleotide sequence similarity. The lipase gene was cloned into the bacterial expression plasmid and expressed in E. coli. SDS-PAGE analysis of the cells showed that ArcL13-Lip was expressed as inclusion bodies with a molecular mass of about 35 kDa. Refolding was achieved by diluting the unfolded protein into refolding buffers containing various additives, and the highest refolding efficiency was seen in the glucose-containing buffer. Refolded ArcL13-Lip showed high hydrolytic activity toward p-nitrophenyl caprylate and p-nitrophenyl decanoate among different p-nitrophenyl esters. Recombinant ArcL13-Lip displayed maximal activity at $40^{\circ}C$ and pH 8.0 with p-nitrophenyl caprylate as a substrate. Activity assays performed at various temperatures showed that ArcL13-Lip is a cold-active lipase with about 40% and 73% of enzymatic activity at $10^{\circ}C$ and $20^{\circ}C$, respectively, compared to its maximal activity at $40^{\circ}C$.

Characterization of Physiological Properties in Vibrio fluvialis by the Deletion of Oligopeptide Permease (oppA) Gene (Vibrio fluvialis oligopeptide permease (oppA) 유전자 deletion에 의한 생리적 특성)

  • Ahn Sun Hee;Lee Eun Mi;Kim Dong Gyun;Hong Gyoung Eun;Park Eun Mi;Kong In Soo
    • Journal of Life Science
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    • v.16 no.1
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    • pp.131-135
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    • 2006
  • Oligopeptide is known to be an essential nitrogen nutrient for bacterial growth. Oligopeptide can be transported into cytoplasm by a specific transport system, Opp system. Opp system is composed of five proteins, which are transcribed by an operon. These are responsible for oligopeptide binding protein (OppA), permease (OppB and OppC) and energy generation system (OppD and OppF), respectively. Previously, we isolated the opp operon from Vibrio fluvialis and constructed the oppA mutant by allelic exchange method. In this study, we investigated the growth pattern and biofilm production under the different growth condition. When the cells were cultivated using brain heart infusion(BHI) medium, the wild type was faster than the mutant in growth during the exponential phase. However, it showed that the growth pattern of two strains in M9 medium is very similar. The growth of wild type showed better than that of the mutant grown at pH 8. At pH 7, there was no an obvious difference in growth. After 5 mM $H_2O_2$ was treated to the cells $(OD_{600}=1.2)$, the cell survival was examined. The oppA mutation did not affect in survivability. In the presence of $10{\mu}g/ml$ polymyxin B, the biofilm production of the oppA mutant was higher than that of the wild type.

Investigation of bacteria in the agricultural by-products imported for the use as media materials in mushroom cultivation (버섯재배 배지재료용 수입 농업부산물에서의 세균 조사 연구)

  • Kim, Jun Young;Kim, Susan;Kim, Seong Hwan
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.410-419
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    • 2018
  • It is urgently required to construct safety data on agricultural by-products imported for use as medium materials for domestic mushroom production. However, research on microorganisms is insufficient. This study was conducted to investigate the presence of bacteria that have the possibility of harmful effects on human, plants and mushroom in wheat straw, peatmoss, cottonseed hull, cottonseed meal, and beet pulp imported from Australia, Canada, China, Egypt, Germany. Bacteria were found in the range of $1.35{\times}10^2$ to $8.34{\times}10^6CFU/g$. As a result of 16S rDNA sequence analysis, total of 19 genera and 45 species of bacteria were identified. Bacillus genus was dominant, followed by Paenibacillus genus. At the species level, diverse species was in the order of Firmicute, Proteobacteria and Actinobacteria. Regarding the agricultural by-products, straw and peat moss had more diverse bacteria than other agricultural by-products. Among the indentified bacteria, 6 species of 5 genera (Enterobacter asburiae, Enterobacter ludwigii, Stenotrophomonas maltophilia, Pseudomonas monteilii, Bacillus anthracis, and Cellulosimicrobium funkei) were present as potent harmful bacteria to human. Surprisingly, both the human and plant pathogenic Klebsiella pneumoniae subsp. pneumonia was present. Bacillus altitudinis was present as a plant pathogen. Lysinibacillus sphaericus, an insect pathogen, and Ochrobactrum pseudogrignonense, a mushroom pathogen, were also present. The results of this study confirmed that several kinds of pathogenic bacteria were present in the agricultural by-products for the mushroom cultivation medium imported into Korea. Our work suggests that hygiene inspection and management is urgently needed for imported agricultural by-products to be safely used for mushroom production.

'Neulchan', a Middle-seed, Disease-resistant, and High-yield Soybean Cultivar for Soy-paste and Tofu (중립 내병 다수성 장류·두부용 콩 '늘찬')

  • Kim, Hyun Tae;Ko, Jong Min;Baek, In Youl;Han, Won Young;Yun, Hong Tai;Lee, Byoung Won;Shin, Sang Ouk;Seo, Jeong Hyun;Kim, Hong Sik;Kwak, Do Yeon
    • Korean Journal of Breeding Science
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    • v.51 no.4
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    • pp.475-481
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    • 2019
  • The soybean cultivar 'Neulchan' was developed for production of soy-paste and tofu. SS91501-9-1-1 and SS96205 (F2) were crossed in 1998, and F3 to F7 were selected by the pedigree method. A preliminary yield trial (PYT) and an advanced yield trial (AYT) were conducted from 2006 to 2008, and a regional yield trial (RYT) in nine regions was conducted from 2009 to 2011. In the RYT, 'Neulchan' was stable in variable environments and generated high yield. 'Neulchan' was determinate with white flower, light brown pod color, yellow spherical seed, and yellow hilum. Its flowering date and maturity date were Jul. 30 and Oct. 9, respectively. The plant height was shorter than that of 'Daewonkong' (a standard cultivar). 'Neulchan' had the same node number (14), higher first-pod height (12 cm), and lighter seed weight (21.7 g/100-seed weight) than those of 'Daewonkong' (14, 11, and 24.2 g/100-seed weight, respectively). 'Neulchan' had high resistance to bacterial pustule, and its resistance to soybean mosaic virus was similar to that of 'Daewonkong'. The yield and color of 'Neulchan' tofu were similar to those of 'Daewonkong' tofu, but the hardness was lower than that of 'Daewonkong' tofu. The soybean malt scent, fermented soybean yield, and γ-polyglutamic acid (γ-PGA) of 'Neulchan' were 3, 215%, and 24.6 mg/g, respectively. Its yield in adaptable regions was 307 kg/10a, higher than that of 'Daewonkong'. 'Neulchan' was expected to be cultivated and used widely for soy-paste and tofu production. (Registration No. 4904).

Selection and appropriate culture conditions of antagonistic bacterium Bacillus altitudinis HC7 against button mushroom cobweb disease caused by Cladobotryum mycophilum (양송이버섯 솜털곰팡이병균(Cladobotryum mycophilum)에 대한 길항미생물 Bacillus altitudinis HC7의 선발 및 적정 배양조건)

  • Chan-Jung Lee;Hye-Sung Park;Seong-Yeon Jo;Gi-Hong An;Ja-Yun Kim;Kang-Hyo Lee
    • Journal of Mushroom
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    • v.22 no.2
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    • pp.60-66
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    • 2024
  • This study was conducted to selection and investigate appropriate conditions for mass production of antagonistic microbes to control cobweb disease caused by Cladobotryum mycophilum. A grampositive bacterium was isolated from spent substrate of Agaricus bisporus and showed significant antagonistic activity against Cladobotryum mycophilum. The bacterium was identified as Bacillus altitudinis. based on the cultural, biochemical and physiological characteristics, and 16S rRNA sequence. The isolate is saprophytic, but not parasitic nor pathogenic to cultivated mushroom whereas it showed strong inhibitory effects against C. mycophilum cells in vitro. The control efficacy of B. altitudinis HC7 against cobweb disease of C. mycophilum was up to 78.2% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol system. To define the appropriate conditions for the mass production of the Bacillus altitudinis HC7, we have investigated appropriate culture conditions and effects of various nutrient source on the bacterial growth. The appropriate initial pH and temperature were determined as pH 6.0 and 30℃, respectively. The appropriate concentration of medium elements for the growth of pathogen inhibitor bacterium(Bacillus altitudinis HC7) was determined as follows: 3.0% soluble startch, 10% soytone, 1.0% (NH4)2HPO4, 1.0 mmol KCl, and 0.5% L-asparagine.

Analysis of the Reduction Effect of Combined Treatment with UV-C and Organic Acid to Reduce Aspergillus ochraceus and Rhodotorula mucilaginosa Contamination (Aspergillus ochraceus와 Rhodotorula mucilaginosa 저감을 위한 자외선과 유기산 복합처리 효과 분석)

  • Eun-Seon Lee;Jong-Hui Kim;Bu-Min Kim;Mi-Hwa Oh
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.54-60
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    • 2024
  • This study investigated the effectiveness of using pathogens and aqueous acids to reduce the Aspergillus ochraceus and Rhodotorula mucilaginosa contamination in livestock production environments. For this study, 1 mL of each bacterial suspension (107-108 spores/mL) was inoculated on a knife surface, dried at 37℃, and used under each treatment condition. First, to investigate the effect of organic acids, acetic, lactic, and citric acids were used. Subsequently, to select the appropriate concentration, they were prepared at concentrations of 0.5, 1, 2, 3, 4, and 5%, respectively. Accordingly, to further maximize the effect of organic acid treatment, we combined the treatment with ultraviolet light. The two strains showed a significant difference (P<0.05) compared to the initial strain, with a greater than 90% decrease in the concentrations of all organic acids. Consequently, acetic and lactic acids decreased by approximately 5 and 2 log colony forming unit (CFU)/cm2, respectively, when treated with ultraviolet light (360 mJ/cm2); however, citric acid decreased by less than 1 log CFU/cm2. However, when manufactured with 4% acetic acid, a severe malodor was emitted, making it difficult for workers to use it in a production environment. Accordingly, the optimal treatment conditions for organic acid and ultraviolet light for application were selected as follows: immersion in a 4% lactic acid solution for 1 minute and then, sterilization with ultraviolet light at 360 mJ/cm2. Finally, when a pork meat sample was cut with a knife that was finally washed with lactic acid and treated with ultraviolet light, the low level of inoculum transferred from the cleaned knife to the surface of the sample was not detected. In conclusion, using this established method can prevent cross-contamination of the surface of the meat during processing.

Studies on Growth Stimulation of Lactic Starter (유산균(乳酸菌) Starter의 생육촉진(生育促進)에 관(關)한 연구(硏究))

  • Kim, Jong Woo;Lee, Young Soo
    • Korean Journal of Agricultural Science
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    • v.19 no.1
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    • pp.40-50
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    • 1992
  • This experiment was carried out to stimulate lactic starter culture for yoghurt manufacturing. A each of 1.5% of Bios 2000, CR starter medium, and Yeast extract were added to bulk medium, acidity, pH and changes in the number of lactic acid bacteria were investigated at, intervals of two hours for Lactobacillus bulgaricus and four hours for Streptococus thermophilus and Lactobacillus casei. The results obtained were summarized as follows. 1. The acidity of control arrived at 0.99% after 16 hours of incubation during the incubation of Lactobacillus bulgaricus. Whereas that of CR starter medium reached 1.00% at 12 hours of incubation. Yeast extract, 1.12% at 12 hours, and Bios 2000 reached 0.97% at 10 hours respectively, Thus, Bios 2000 showed the fastest rate of acid production. 2. When the acidity of experiment medium peaked on optimum levels. pH of control was 4.03 in 16 hours of incubation during the incubation of Lactobacillus bulgaricus. Whereas that of Bios 2000 reached 4.10 of Yeast extract reached 3.97 at 12 hours, and of CR starter medium reached 4.05 at 12 hours. 3. Lactic acid bacterial counts were $3.1{\times}10^{10}/ml$ after 16 hours of incubation during the incubation of lactobacillus bulgaricus, Whereas those of Bios 2000 reached $2.1{\times}10^{10}/ml$ at 10 hours, with the fastest stimulation of growth, The counts in CR starter medium were at $2.9{\times}10^{10}/ml$ at 12 hours, and Yeast extract were $3.8{\times}10^{10}/ml$ at 12 hours. 4. The acidity of control, CR starter medium, and Yeast extract reached 0.92% at 44 hours, and 0.96% at 32 hours, and 0.90% at 32 hours respectively, Also, that of Bios 2,000 reached 0.97% at 32 hours, which exhibited the highest, among the treatments. 5. The pH of control was 4.27 at 44 hours. that of CR starter medium was 4.33 at 40 hours and that of Yeast extract was 4.25 at 32 hours during the incubation in Streptococcus thermophilus. Besides, pH of Bios 2000 is lowest as 4.18 at 32 hours. 6. Lactic acid bacterial counts in control, CR starter medium, and Yeast extract during the incubation of Streptococcus thermophilus were $9.8{\times}10^{9}/ml$ at 44 hours,$9.5{\times}10^{8}/ml$ at 40 hours, and $9.6{\times}10^{8}/ml$ at 32 hours. And, the highest number was $2.0{\times}10^{9}/ml$ for Bios 2000 at 32 hours. 7. The acidity of control during the incubation of Lactobacillus casei reached 0.92% at 40 hours, and those of CR starter medium and Yeast extract were 0.95% at 40 hours, and 1.01% at 36 hours respectively. Also, Bios 2000 had the highest acidity as 0.94% at 32 hours. 8. The pH of control, CR starter medium and Yeast extract during the incubation Lactobacillus casei was 4.27 at 40 hours. 4.21 at 40 hours, and 4.15 at 36 hours respectively. Also, Bios 2000 showed the lowest pH, as 4.23, at 32 hours. 9. Lactic acid bacterial counts in control, CR starter medium and Yeast extract during the incubation of Lactobacillus casei were $9.4{\times}10^{7}/ml$ at 40 hours, $1.1{\times}10^{8}/ml$ at 40 hours, and $5.0{\times}10^{8}/ml$ at 36 hours respectively. And, the progress of 32 hours showed the highest number of lactic acid bacteria as$6.4{\times}10^{8}/ml$ in Bios 2000.

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Intensive Culture of the Pacific White Shrimp Litopenaeus vannamei under Limited Water Exchange I. Indoor Nursery Culture of Postlarvae (사육수 비교환 방식에 의한 흰다리새우의 고밀도 사육 I. 후기유생(postlarva)의 실내 중간육성)

  • Jang, In-Kwon;Kim, Jong-Sheek;Cho, Kook-Jin;Seo, Hyung-Chul;Cho, Yeong-Rok;Gopalakannan, Ayyaru;Kim, Bong-Lae
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.339-345
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    • 2008
  • Farming of the fleshy shrimp Fenneropenaeus chinensis which is a major cultured species in the west coast of South Korea, has been suffered :trom mass mortality due to disease epizootics including viruses. Since the Pacific white shrimp Litopenaeus vannamei was introduced to Korea in 2003, farming of this species has rapidly increased for years, occupying 62.5% of total cultured shrimp production in 2007. However the studies on L. vannamei culture methods for shrimp farming situations in Korea are very limited. Nursery culture of shrimp larvae has some advantages including increased survival, improved feed efficiencies, enhanced growth performance and reduced grow-out period. In this study, L. vannamei postlarvae (${PL_3}-{PL_{10}}$) with a density of $3,750-9,090/m^3$ were cultured in four raceways under limited water exchange condition for 35 days. Survival was the highest (93.6%) in tank stocked with $4,090/m^3$ and was the lowest in tank with $9,090/m^3$ (58.1 %). Mean body weight at harvest ranged from 0.071 to 0.108 g, and FCR was 0.59-0.70 in all tanks. Concentration of total ammonia nitrogen was increased up to 20 ppm on day 10 in all tanks and thereafter gradually decreased by the third week of culture. Nitrite-nitrogen was rapidly increased from the third week, representing bio-floc condition by developed nitrifying bacterial community. Of the present nursery system some modification of structure and consideration for commercial scale are needed in order to be implemented to shrimp farmers.

Immuno-stimulatory Activities of a High Molecular Weight Fraction from Cynanchum wilfordii Radix Obtained by Ultrafiltration (한외거르기(Ultrafiltration)에 의하여 분리된 백수오 고분자 분획물의 면역증진 활성)

  • Jang, Mi;Lim, Tae-Gyu;Hong, Hee-Do;Rhee, Young Kyoung;Kim, Kyung-Tack;Lee, Eunjung;Lee, Jeong Hoon;Lee, Yun Ji;Kim, Yeon Bok;Cho, Chang-Won
    • Korean Journal of Food Science and Technology
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    • v.48 no.3
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    • pp.268-274
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    • 2016
  • The purpose of this study was to investigate the immuno-stimulatory activity of the high-molecular-weight fraction (HMWF) of Cynanchum wilfordii (CW) extracts obtained by ultrafiltration in murine macrophage RAW 264.7 cells and to assess its immuno-stimulatory effect in mice. Ultrafiltration was performed with polyethersulfone membranes (30 kDa cutoff) in a cross-flow filtration system to obtain the HMWF of CW. The results showed that the HMWF increased the production of various cytokines such as tumor necrosis factor-${\alpha}$, interleukin-6, and nitric oxide in dose-ependent manners. In addition, HMWF treatment increased the relative spleen weight as well as splenocyte proliferation induced by concanavalin A or bacterial lipopolysaccharide in mice. Natural killer (NK) cell activity in the HMWF-treated group was significantly increased compared to that in the control group. These results suggest that the HMWF of CW can support the immune system through secretion of macrophage cytokines, thereby enhancing NK cell activity and murine splenocyte proliferation.