• Title/Summary/Keyword: Bacterial mixture

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Effect of biocide addition on plantlet growth and contamination occurrence during the in vitro culture of blueberry

  • Huh, Yoon Sun;Lee, Joung Kwan;Kim, Ik Jei;Kang, Bo Goo;Lee, Ki Yeol
    • Journal of Plant Biotechnology
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    • v.42 no.2
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    • pp.111-116
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    • 2015
  • Interest and great demand for blueberry (Vaccinium corymbosum) have increased, as V. corymbosum is now one of the most economically important crops in Korea. It is expected that blueberry production and the area planted for cultivation will increase consistently in the years ahead because of high profitability and the consumer's demand for healthy ingredients. Effective mass production of blueberry is urgently needed for commercial cultivation establishment, but a main limitation is lack of a propagation system that produces a disease-free plant material for commercial plantation. A large amount of research has focused entirely on developing tissue culture techniques for blueberry propagation. However, controlling fungal and bacterial contamination of woody plant material is extremely difficult. Our study was conducted to investigate the effect of biocide addition during the in vitro culture of blueberry on plantlet growth and contamination occurrence. Four biocides, including Plant Preservative Mixture ($PPM^{TM}$), vancomycin, nystatin and penicillin G, were used in varying concentrations during the in vitro propagation of blueberry. When nystatin was added into the medium at low concentrations, the overall growth of blueberry plantlets was retarded. Addition of vancomycin and penicillin G in high concentrations decreased contamination but induced plantlet mortality. On the other hand, when 1ml/L $PPM^{TM}$ was added, the growth characteristics of blueberry plantlets did not significantly differ from non-treatment (control), and the contamination occurrence rate was very low. From these results, we found that the addition of the appropriate biocide could provide an effective method to reduce contamination in the culture process, thereby raising in vitro production efficiency.

Characterization of a Toxin isolated from Bacillus thuringiensis var. alesti and Its Toxicity to the Silkworm, Bombyx mori (Bacillus thuringiensis var. alesti로부터 분리된 독소의 성상과 누에에 대한 독성)

  • 이영근;조용섭;임종성
    • Journal of Sericultural and Entomological Science
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    • v.17 no.1
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    • pp.55-62
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    • 1975
  • In the studies of characterization of crystal prototoxin isolated from Bacillus thuringinensis var. alesti and its bioassay to the silkworm larvae, Bombyx mori L., the following results were obtained. 1. The complete lysis of bacteria grown on the nutrient agar at 30$^{\circ}C$ in an incubator took 30 days, but the period could be reduced by a half in a devised broth media in a fermentation jar. 2. The protein toxin extracted directly from a mixture of crystals and spores without separation of crystals and spores was pure as same as the protein toxin extracted from only crysals separated. 3. By SLS polyacrylamide gel electrophoresis of prototoxin released from crystals in alkali (pH 9.5) three different proteins in molecular weights of 120,000, 87,000, 74,000 were separated. 4. In the bioassay of the toxin to the silkworm larvae, LD$\sub$50/ per gram of the larvae in the 4th instar was 0.080 $\mu\textrm{g}$.

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High-Level Production of Low-Branched Levan from Pseudomonas aurantiaca S-4380 for the Production of $di-\beta-D-Fructofuranose$ Dianhydride IV

  • JANG KI-HYO;JANG EUN-KYUNG;KIM SEUNG-HWAN;KIM IN-HWAN;KANG SOON AH;KOH ISSAC;PARK YOUNG-IL;KIM YOUNG-JUN;HA SANG-DO;KIM CHUL HO
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.102-108
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    • 2006
  • The IscA gene, encoding a levansucrase of 424 amino acids (aa) residues, was cloned from the genomic DNA of Pseudomonas aurantiaca S-4380, and overexpressed in Escherichia coli. The recombinant levansucrase overexpressed in E. coli was then used to produce levan from sucrose. Levan crystals with 98% purity could be obtained from the reaction mixture with $62\%$ yield using an alcohol precipitation method. The molecular weight of the levan was $7\times10^5$ daltons. Methylation studies showed that the levan was branched: main linkage C-2,6; branched linkage C-2,1; and degree of branching $6\%$. Three bacterial levans from different strains were incubated with levan fructotransferase (LFTase) from Arthrobacter ureafaciens K2032, which produced $di-\beta-D-fructofuranose$ dianhydride IV (DFA IV); final conversion yields from the levans to DFA IV were $39\%$ in Zymomonas mobilis, $53\%$ in Serratia levanicum, and $59\%$ in P. aurantiaca S-4380 levansucrase. The levan from P. aurantiaca S-4380 levansucrase gave the highest conversion yield of levan to DFAIV so far reported.

Cytotoxicity of Shikonin Metabolites with Biotransformation of Human Intestinal Bacteria

  • Min, Byung-Sun;Meselhy, Meselhy-R.;Hattori, Masao;Kim, Hwan-Mook;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.514-517
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    • 2000
  • Abstracts Six shikonin metabolites were obtained from human intestinal bacteria, Bacteriodes fragilis subsp. thetaotus. following biotransformation. The transformation of shikonin (1) was performed anaerobically for 3 day at $37^{\circ}C$ in thc bacterial suspension of B. Fagilis which was cultured overnight in GAM broth. The incubation mixture \vas extracted with EtGAc Lo give a dark-brown residue. The residue was apphed to a silica gel column, which was eluted successively with hexane (Fr. A), $CHCl_3$ (Fr. B), and $CHCl_3$:MeOH (9:I) (Fr. C). Six metabolites. Fr.A (2 and 3), Fr. B (6 and 7), and Fr. C (4 and 5) were isolated by repeated silica gel column chromatography, preparatlVe TLC, followed by Sephadex LH-20. In vitro cytotoxicities were tested against human tumor cell lines; PC-3 (prostate), ACHN (renal), A549 (lung), SW620 (colon), KS62 (leukemia), and Du145 (prostate). The shikonin metabolites 2. 4, 5, and 6 showed weaker cytotoxicity than the parenL shikonin (1). whereas shikonin monomenc metabolite 3 ($ED_{50}{\;}O.44-{\;}1.22{\;}\mu\textrm{g}/ml$) and dimeric metabolite 7 ($ED_{50}{\;}O.48-{\;}2.35{\;}\mu\textrm{g}/ml$) exhibited stronger activities compared with adriamycin, which was used as the positive control.ontrol.

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Bioremediation of Diesel-Contaminated Soil by Bacterial Cells Transported by Electrokinetics

  • LEE, HYO-SANG;KISAY LEE
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1038-1045
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    • 2001
  • The electrokinetic technology was applied in bioremediation for the purpose of supplying a Pseudomonas strain capable of degrading diesel to contaminated soil bed, and their biodegradation of diesel was carried out after a desired cell distribution was obtained. Electrokinetic injection of the strain was made possible because the cells acted as negatively charged particles at neutral pH, and thus the cells were transported with a precise directionality through the soil mostly by the mechanism of electrophoresis and in part by electroosmosis. A severe pH change in the soil bed was formed due to the penetration of electrolysis products, which was harmful to the cell viability and cell transport. To achieve a desirable cell transport and distribution, the control of pH in soil bed by a recirculating buffer solution in electrode chambers was essential during the appliation of an electric field. The judicious selections of electrolyte concentration and conductivity were also important for achieving an efficient electrokinetic cell transport since a higher electrolyte concentration favored the maintenance of pH stability in soil bed, but lowered electrophoretic mobility on the other hand. With electrolyte solution of pH 7 phosphate buffer, a 0.05 M concentration showed a better cell transport buffer, a 0.05 M concentration showed a better cell transport than 0.02 M and 0.08 M. The cell under pH 8 were obtained, compared to the cells under pH 7 or pH 9 in a given time period Up to $60\%$ of diesel was degraded in 8 days by the Pseudomonas cell, which were distributed electrokinetically under the conditions of pH 8 ($1,800{\mu}S/cm$, a mixture of phosphate and ammonia buffers) and 40 mA in a soil bed of 15 cm length.

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Molecular Cloning of Chitosanase Gene and Quantitative Production of Chitosan Oligomer (키토사네이즈 유전자의 클로닝과 키토산 올리고머의 정량적 생산)

  • 박유미;장혜란;허태린;김사열
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.16-21
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    • 2004
  • Six bacterial strains which formed large halo on chitosan-containing agar plate were isolated from beach mud and crabs at South coast of Korean peninsula. They were designated as Bacillus cereus KNUC51, B. cereus KNUC52, B. cereus KNUC53, B. cereus KNUC54, B. cereus KNUC55, and Paenibacillus favisporus KNUC56 by analysing their morphologies and 16S rDNA sequences. Chitosanase activities of all isolates were similar to that of B. subtilis 168. To enhance the activity of chitosanase, a powerful mutagen, MNNG was treated for P favisporus KNUC56. Three mutants showed higher activity of chitosanase than that of the original strain. The DNA fragments containing chitosanase gene from B. cereus sources were cloned, sequenced, and their deduced amino acid sequence analysis showed over 93% homologies with that of the known B. cereus ATCC14579. Extracellular sample from the isolates was incubated in proper reaction mixture including chitosan for 5 minutes at $37^{\circ}C$ to produce 3-10 chitosan oligomers which has been known to be active for clinical agents and agronomical agents.

Quality Characteristics of Sulgidduk added with Cirsium setidens Nakai (곤드레 첨가 설기떡의 품질특성)

  • Park, Sung Jin;Rha, Young-Ah
    • Culinary science and hospitality research
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    • v.22 no.7
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    • pp.1-10
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    • 2016
  • The purpose of this study was to determine the desirable mixture ratio of Cirsium setidens Nakai powder to rice flour for the preparation of Sulgidduk. The moisture of samples raged from 36.51% to 40.67%. The addition of Cirsium setidens Nakai powder in the Sulgidduk tended to decrease the lightness (L) in Hunter color value, but to increase the redness (a) and yellowness (b). With increasing Cirsium setidens Nakai powder level, the hardness, springness and chewiness of the samples increased and their cohesiveness decreased. In sensory evaluation, the addition of 5% Cirsium setidens Nakai powder showed the best score in taste and overall preference. Over 3 days of storage, Sulgidduk showed a slightly high total bacterial count compared to control. Therefore, these results suggested that the addition of 5% Cirsium setidens Nakai powder could be applied for making Cirsium setidens Nakai powder Sulgidduk.

Development of Regional Noodles Using Agricultural and Fishery Products of Cheju Island (제주특산물을 이용한 향토국수의 개발)

  • 황인주;오영주
    • Korean journal of food and cookery science
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    • v.12 no.3
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    • pp.361-366
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    • 1996
  • Acceptable fish noodles of better nutritional and sensory values than conventional noodle made only wheat flour were prepared by mixing wheat flour and ground tile fish (Branchioste gus japonicus). In oder to mask a distinctive fish odor, various herbs and spices, such as ginger (Zingiber officinale), curry (Chalcas koenigii), nutmeg (Myristica Pagrans), garlic (Ailium sativum), black pepper (Piper nigrum), lemon (Cirtus limon) or sodachi (Citrus sudachi) were added to the basic tile fish noodle. Also, for the purpose of enhancing eating qual ify and nutritional value of basic noodle were combined some agricultural or fishery products of Cheju island, citrus fruits; danyooja (Citrus danyooga), hagul (C. natfudaidai), medical plants; angelica utilis (Angelica keiskei), ginseng (Panax ginseng), cactus (Opuntia dillenii), vegetable; carrot (Daucus carota), dropwort (Oenanthe jnvanica), seaweeds; fusiforme (Hizkia fusiforme), gulfweed (Sargaceae hlvelium). The optimal mixing ratio for preparing the basic tile fish noodle amounted to ground tile fish 135 g: tile fish stock 139 $m\ell$: wheat flour 450 g: salts 10 g. The mixture of curry powder (2.5 g) and nutmeg powder (2.5 g) was proved to be the most effective combinations for masking unfavorable fish odor. The optimal amounts of materials to be added to the prepared basic noodle were 25 g citrus zest and 80m1 citrus juice for citrus fruits noodles, and ca. 140 g puree for noodles from medical plants, vegetables and seaweeds, respectively. The preference score obtained from consumer preference test, on a 9-point scales, were in oder of i) danyooja > carrot, angelica, ginseng > fusiforme, ii) hagul > dropwort, cactus > gulfweed. The shelf-life of tile fish noodle based on bacterial counting was estimated to be 7-days at 5$^{\circ}C$.

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Preparation Of levan Oligosaccharides by Acid Hydrolysis and It Application in Growth of lactic Acid-producing Bacteria (산가수 분해법에 의한 레반 올리고당의 제조 및 유산군 생육촉진 효과 연구)

  • 강태호;정성제;강순아;강기효;장은경;김승환;김철호;이상기;전억한
    • KSBB Journal
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    • v.17 no.2
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    • pp.137-141
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    • 2002
  • Levan oligosaccoharides and low molecular weight levin were produced from levin by acid hydrolysis and following column chromatography. Levan hydrolysis was progressed proportionally as increased incubation time. In terms of levan hydrolysis reaction, no differences were found from the sources of levan. Optimum hydrolysis conditions for the formation of levan oligosaccharides were, 0.38 M H$_2$S0$_4$; and incubation at 95$\^{C}$ for 4 min. The purified products were determined as the mixture of oligosaccharides (degree of polymerization (DP) of 3-6), Two of lactic acid-producing bacteria, Lactobacillus plantarum KCTC 3104 and Pediococcus pentosaceus KCTC 3507, were studied in vitro for their ability to metabolize levin oligosaccharides. Apparently, the growth of both cells were increased by levin oligosaccharide diet, compared with those of levan diets, suggesting that levan oligosaccharides may be beneficial in selectively growth of lactic acid-producing bacteria.

Effects of Methylcellulose on Cellulolytic Bacteria Attachment and Rice Straw Degradation in the In vitro Rumen Fermentation

  • Sung, Ha Guyn;Kim, Min Ji;Upadhaya, Santi Devi;Ha, Jong K.;Lee, Sung Sill
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.9
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    • pp.1276-1281
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    • 2013
  • An in vitro experiment was conducted to evaluate the effect of methylcellulose on the attachment of major cellulolytic bacteria on rice straw and its digestibility. Rice straw was incubated with ruminal mixture with or without 0.1% methylcellulose (MC). The attachment of F. succinogenes, R. flavefaciens and R. albus populations on rice straw was measured using real-time PCR with specific primer sets. Methylcellulose at the level of 0.1% decreased the attachment of all three major cellulolytic bacteria. In particular, MC treatment reduced (p<0.05) attachment of F. succinogenes on rice straw after 10 min of incubation while a significant reduction (p<0.05) in attachment was not observed until 4 h incubation in the case of R. flavefaciens and R. albus. This result indicated F. succinogenes responded to MC more sensitively and earlier than R. flavefaciens and R. albus. Dry matter digestibility of rice straw was subsequently inhibited by 0.1% MC, and there was a significant difference between control and MC treatment (p<0.05). Incubated cultures containing MC had higher pH and lower gas production than controls. Current data clearly indicated that the attachment of F. succinogenes, R. flavefaciens and R. albus on rice straw was inhibited by MC, which apparently reduced rice straw digestion.