• Title/Summary/Keyword: Liquid cultures

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Griseofulvin from Xylaria sp. Strain F0010, an Endophytic Fungus of Abies holophylla and its Antifungal Activity Against Plant Pathogenic Fungi

  • PARK, JOONG-HYEOP;CHOI, GYUNG-JA;LEE, SEON-WOO;LEE, HYANG-BURM;KIM, KYOUNG-MO;JUNG, HACK-SUNG;JANG, KYOUNG-SOO;CHO, KWANG-YUN;KIM, JIN-CHEOL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.112-117
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    • 2005
  • Abstract Griseofulvin has been used as an antifungal antibiotic for the treatment of mycotic diseases of humans and veterinary animals. The purpose of this work was to identify a griseofulvin-producing endophytic fungus from Abies holophylla and evaluate its in vivo antifungal activity against plant pathogenic fungi. Based on nuclear ribosomal ITS1-5.8SITS2 sequence analysis, the fungus was identified and labeled as Xylaria sp. F0010. Two antifungal substances were purified from liquid cultures of Xylaria sp. F0010, and their chemical identities were determined to be griseofulvin and dechlorogriseofulvin through mass and NMR spectral analyses. Compared to dechlorogriseofulvin, griseofulvin showed high in vivo and in vitro antifungal activity, and effectively controlled the development of rice blast (Magnaporthe grisea), rice sheath blight (Corticium sasaki), wheat leaf rust (Puccinia recondita), and barley powdery mildew (Blumeria graminis f. sp. hordei), at doses of 50 to 150 ${\mu}$g/ml, depending on the disease. This is the first report on the production of griseofulvin and dechlorogriseofulvin by Xylaria species.

Genetic and biochemical evidence for redundant pathways leading to mycosporine-like amino acid biosynthesis in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024

  • Geraldes, Vanessa;de Medeiros, Livia Soman;Lima, Stella T.;Alvarenga, Danillo Oliveira;Gacesa, Ranko;Long, Paul F.;Fiore, Marli Fatima;Pinto, Ernani
    • ALGAE
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    • v.35 no.2
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    • pp.177-187
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    • 2020
  • Cyanobacteria have been widely reported to produce a variety of UV-absorbing mycosporine-like amino acids (MAAs). Herein, we reported production of the unusual MAA, mycosporine-glycine-alanine (MGA) in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024 using a newly developed UHPLC-DAD-MS/HRMS (ultra-high performance liquid chromatography-diode array detection-high resolution tandem mass spectrometry) method. MGA had previously been first identified in a red-algae, but S. torques-reginae strain ITEP-024 is the first cyanobacteria to be reported as an MGA producer. Herein, the chemical structure of MGA is fully elucidated from one-dimensional / two-dimensional nuclear magnetic resonance and HRMS data analyses. MAAs are unusually produced constitutively in S. torques-reginae ITEP-024, and this production was further enhanced following UV-irradiance. It has been proposed that MAA biosynthesis proceeds in cyanobacteria from the pentose phosphate pathway intermediate sedoheptulose 7-phosphate. Annotation of a gene cluster encoded in the genome sequence of S. torques-reginae ITEP-024 supports these gene products could catalyse the biosynthesis of MAAs. However, addition of glyphosate to cultures of S. torques-reginae ITEP-024 abolished constitutive and ultra-violet radiation induced production of MGA, shinorine and porphyra-334. This finding supports involvement of the shikimic acid pathway in the biosynthesis of MAAs by this species.

Induction of the T7 Promoter Using Lactose for Production of Recombinant Plasminogen Kringle 1-3 in Escherichia coli

  • Lim, Hyung-Kwon;Lee, Shi-Uk;Chung, Soo-Il;Jung, Kyung-Hwan;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.225-230
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    • 2004
  • A plasminogen kringle domain 1 to 3, rKl-3, was expressed in Escherichia coli under the control of T7 promoter. For the cost-effective production of rKl-3, the induction process was analyzed and optimized. Induction characteristics with lactose were analyzed in terms of induction time and inducer concentration in various culture conditions including batch and high-cell-density fed-batch cultures. In the fed-batch culture, the induction around 6 h after initiation of the DO-stat fed-batch culture resulted in the highest expression level of rKI-3 among the induction points examined. The highest demand of oxygen at this point was crucial for the maximum expression level of rKI-3. As the lactose concentration increased, the expression level also increased, though the expression level showed a plateau above a concentration of 14 mM of lactose. Lactose acted less specifically than IPTG since most of it was hydrolyzed to glucose and galactose. However, using lactose, the cell growth and the maximum expression level of rKl-3 increased by 20% and 24%, respectively, compared with those using IPTG in the fed-batch culture. The lactose seemed to be hydrolyzed by intracellular and extracellular $\beta$-galactosidase liberated by cell lysis at the same time. Residual concentration of glucose was maintained to a a limit of detection by high performance liquid chromatography, and galactose was not consumed by the host strain Escherichia coli BL2l(DE3).

Macrophage Stimulating Activity of Exo-Biopolymer from Submerged Culture of Lentinus edodes with Rice Bran

  • Yu, Kwang-Won;Shin, Kwang-Soon;Choi, Yang-Mun;Suh, Hyung-Joo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.658-664
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    • 2004
  • To find a new utilization of rice bran, nine higher fungi were examined for the production of exo-biopolymer with macrophage stimulating activity from rice bran. Among the exo-biopolymers produced from submerged cultures, Lentinus edodes showed the highest activity, followed by Grifola frondosa, Schizophyllum commune, and Coriolus versicolor. L. edodes also had the most potent macrophage stimulating activity in a liquid culture rather than in a solid culture. In order to improve rice bran utilization and the yield of exo-biopolymer with macrophage stimulating activity, the treatment of Rapidase effectively increased the macrophage stimulating activity (about 30% increase), whereas the other enzymes (Econase, Viscozyme, Ultraflo, Celluclast, and Thermylase) treatments did not increase the macrophage stimulating activity. Exo-biopolymer with macrophage stimulating activity from L. edodes contained mainly neutral sugars (58.7%) with considerable amounts of uronic acid (32.2%) and a small amount of proteins (9.1%). Component sugars of exo-biopolymer consisted of mainly arabinose, galactose, glucose, mannose, and xylose (0.95:0.81:0.96:1.00:0.39, respectively). When the exo-biopolymer was treated with $NaIO_4, NaClO_2$, and pronase, the $NaClO_2$ treatment and pronase digestion had little effect, whereas $NaIO_4$ oxidation significantly decreased the macrophage stimulating activity (47.6% reduction at $100\mug/ml$). Therefore, the carbohydrate moiety in exo-biopolymer from L. edodes plays an important role in the expression of the macrophage stimulating activity.

Antibacterial and Antifungal Activities of a Naphthoquinone Derivative Isolated from the Fruits of Catalpa ovata G.$D_{ON}$

  • Kuk, Ju-Hee;Ma, Seung-Jin;Moon, Jae-Hak;Kim, Kil-Yong;Choi, Sang-Ho;Park, Keun-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.858-863
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    • 2002
  • An antimicrobial compound was isolated from the MeOH extract of Catalpa ovata G.$D_{ON}$ fruits, and its structure was Identified as 4,9-dihydroxy-2,2-dimethyl-3,4-Uihydronaphtho[2,3-b]pyran-5,10-dione (HMNP). The antimicrobial activity of the Un was determined by measuring the dose-response inhibiton of microbial growth in liquid cultures and then compared with that of lapachol, a well known antimicrobial 1,4-naphthoquinone. The antimicrobial activity of the HMNP was more effective than that of lapachol over a wide range of test organisms. Gram-positive bacteria, yeast, and fungi ($IC_{50}$ $20-75\muM$) were found to be more sensitive to the HMNP than Cram-negative bacteria ($IC_{50}$ > $100\muM$). The HMNP also inhibited germination of spores of many fungi. The morphological defurmation of the fungal spores was induced by the treatment of HMNP, as illustrated by Scanning Electron Microscopy (SEM).

Effects of Heavy Metal and pH on Bacterial Growth Isolated from the Contaminated Smelter Soil (제련소 인근 토양에서 분리한 박테리아 생장에 미치는 중금속 및 pH 영향)

  • Keum, Mi-Jung;Yoon, Min-Ho;Nam, In-Hyun
    • Journal of Soil and Groundwater Environment
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    • v.20 no.4
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    • pp.113-121
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    • 2015
  • The contaminated soil at abandoned smelter areas present challenge for remediation, as the degraded materials are typically deficient in nutrients, and rich in toxic heavy metals and metalloids. Bioremediation technique is to isolate new strains of microorganisms and develop successful protocols for reducing metal toxicity with heavy metal tolerant species. The present study collected metal contaminated soil and characterized for pH and EC values, and heavy metal contents. The pH value was 5.80, representing slightly acidic soil, and EC value was 13.47 mS/m. ICP-AES analytical results showed that the collected soil samples were highly contaminated with various heavy metals and metalloids such as lead (183.0 mg/kg), copper (98.6 mg/kg), zinc (91.6 mg/kg), and arsenic (48.1 mg/kg), respectively. In this study, a bacterial strain, Bacillus cereus KM-15, capable of adsorbing the heavy metals was isolated from the contaminated soils by selective enrichment and characterized to apply for the bioremediation. The effects of heavy metal on the growth of the Bacillus cereus KM-15 was determined in liquid cultures. The results showed that 100 mg/L arsenic, lead, and zinc did not affect the growth of KM-15, while the bacterial growth was strongly inhibited by copper at the same concentration. Further, the ability of the bacteria to adsorb heavy metals was evaluated.

Bioremediation of Heavy Metal Contaminated Mine Wastes using Urease Based Plant Extract (요소분해효소 기반 식물추출액을 이용한 광산폐기물 내 중금속 오염 저감)

  • Roh, Seung-Bum;Park, Min-Jeong;Chon, Chul-Min;Kim, Jae-Gon;Song, Hocheol;Yoon, Min-Ho;Nam, In-Hyun
    • Journal of Soil and Groundwater Environment
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    • v.20 no.1
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    • pp.56-64
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    • 2015
  • Acid mine drainage occurrence is a serious environmental problem by mining industry, it usually contains high levels of metal ions, such as iron, copper, zinc, aluminum, and manganese, as well as metalloids of which arsenic is generally of the greatest concern. An indigenous plant extract was used to produce calcium carbonate from Canavalia ensiformis as effective biomaterial, and its ability to form the calcium carbonate under stable conditions was compared to that of purified urease. X-ray diffraction and scanning electron microscopy were employed to elucidate the mechanism of calcium carbonate formation from the crude plant extracts. The results revealed that urease in the plant extracts catalyzed the hydrolysis of urea in liquid state cultures and decreased heavy metal amounts in the contaminated soil. The heavy metal amounts were decreased in the leachate from the treated mine soil; 31.7% of As, 65.8% of Mn, 50.6% of Zn, 51.6% of Pb, 45.1% of Cr, and 49.7% of Cu, respectively. The procedure described herein is a simple and beneficial method of calcium carbonate biomineralization without cultivation of microorganisms or further purification of crude extracts. This study suggests that crude plant extracts of Canavalia ensiformis have the potential to be used in place of purified forms of the enzyme during remediation of heavy metal contaminated soil.

Comparisons of Physical Properties of Bacterial Celluloses Produced in Different Culture Conditions Using Saccharified Food Wastes

  • Moon Seung-Hyeon;Park Ji-Min;Chun Hwa-Youn;Kim Seong-Jun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.1
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    • pp.26-31
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    • 2006
  • The saccharogenic liquid (SFW) obtained by the enzymatic saccharification of food wastes was used as a medium for production of bacterial cellulose (BC). The enzymatic saccharification of food wastes was carried out by the cultivation supernatant of Trichoderma harziaum FJ1 culture. Acetobacter xylinum KJ1 was employed for the BC production culture. The physical properties, such as polymerization, crystallinity, Young's modulus, and tensile strength, of BCs produced by three culture methods: the static cultures using HS (Hestrin-Schramm) as a reference medium (A) or the SFW medium (B), the shaking culture (C) or the air circulation culture (D) using the SFW medium, were investigated. The degrees of polymerization of BCs produced under the different culture conditions (A-D) showed 11000, 9500, 8500, and 9200, respectively. Young's modulus was 4.15, 5.0, 4.0, and 4.6 GPa, respectively. Tensile strength was 124, 200, 80, and 184 MPa, respectively. All of the BC had a form of cellulose I representing pure cellulose. In the case of the shaking culture, the degree of crystallinity was 51.2%, the lowest degree. Under the other culturing conditions, the trend should remain in the range of 89.7-84%. Overall, the physical properties of BC produced from SFW were similar to those of BC from HS medium, a commercial complex medium, and BC production by the air circulation culture mode brought more favorable results in terms of the physical properties and its ease of scale-up. Therefore, it is expected that a new BC production method, like air circulation culture using SFW, would contribute greatly to BC-related manufacturing.

Biodegradation of the Commercial Phenoxy Herbicide 2,4-D by Microbial Consortium (미생물 컨소시엄에 의한 시판 페녹시계 제초제 2,4-D의 생물분해)

  • 오계헌;김용석
    • KSBB Journal
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    • v.9 no.5
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    • pp.469-474
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    • 1994
  • The purpose of the work was to evaluate the feasibility of a biological treatment process for the phenoxy alkanoic herbicide 2,4-D(2,4-dichlorophenoxyacetic acid) as a commercial pesticide. The phenoxy herbicide was 2,4-D amine salts which contained 40%(vol/vol) 2,4-D and 60%(vol/vol) solvent. A microbial consortium has been derived by enrichment with 2,4-D. The consortium utilized 2,4-D as the sole source of carbon and energy. Optimal pH on the 2,4-D degradation was 7.0 in this experiment. As concentrations of 2,4-D were increased, the degradation by microbial consontium became inhibited. The amendment with yeast extract and ascorbic acid accelerated the degradation of 2,4-D. High performance liquid chromatography methodology was used to measure 2,4-D and it also resolved 2,4-DCP(2,4-dichlorophenol), the corresponding phenol as intermediate. Gas chromatography-mass spectrometry was used for preliminary identification of the intermediate 2,4-DCP. UV scans of spent cultures showed that the maximum absorption of 2,4-D at the wavelength of 283 nm was decreased toward the end of incubation, but the consortium displayed no detectable spectral changes or peak shifts in the UV absorbance.

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Metabolic Pathways of Hydrogen Production in Fermentative Acidogenic Microflora

  • Zhang, Liguo;Li, Jianzheng;Ban, Qiaoying;He, Junguo;Jha, Ajay Kumar
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.668-673
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    • 2012
  • Biohydrogen production from organic wastewater by anaerobically activated sludge fermentation has already been extensively investigated, and it is known that hydrogen can be produced by glucose fermentation through three metabolic pathways, including the oxidative decarboxylation of pyruvic acid to acetyl-CoA, oxidation of NADH to $NAD^+$, and acetogenesis by hydrogen-producing acetogens. However, the exact or dominant pathways of hydrogen production in the anaerobically activated sludge fermentation process have not yet been identified. Thus, a continuous stirred-tank reactor (CSTR) was introduced and a specifically acclimated acidogenic fermentative microflora obtained under certain operation conditions. The hydrogen production activity and potential hydrogen-producing pathways in the acidogenic fermentative microflora were then investigated using batch cultures in Erlenmeyer flasks with a working volume of 500 ml. Based on an initial glucose concentration of 10 g/l, pH 6.0, and a biomass of 1.01 g/l of a mixed liquid volatile suspended solid (MLVSS), 247.7 ml of hydrogen was obtained after a 68 h cultivation period at $35{\pm}1^{\circ}C$. Further tests indicated that 69% of the hydrogen was produced from the oxidative decarboxylation of pyruvic acid, whereas the remaining 31% was from the oxidation of NADH to $NAD^+$. There were no hydrogen-producing acetogens or they were unable to work effectively in the anaerobically activated sludge with a hydraulic retention time (HRT) of less than 8 h.