• Title/Summary/Keyword: Spore Production

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Kinetics of Enriched Chitinase as Extracellular Metabolite in Beauveria bassiana

  • Mondal, Subhoshmita;Datta, Siddhartha;Mukherjee, Alakananda;Bhattacharya, Pinaki
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.96-104
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    • 2019
  • Beauveria bassiana, one of the most common entomopathogenic fungi, has been isolated, pre defined and characterized in-house from soil of tea cultivation area. Experiments have been performed to verify the presence of chitinase as intracellular metabolite and its release as extracellular product rendering the spores with biopesticide activity. Although there are many responsible enzymes for the pest killer action of B. bassiana, binding property of chitinase depending on presence as well as absence of serine supplemented in the media has been studied with respect to the production and kinetics. A programmed investigation conclusively indicates that the isolated spore (hyphae) of B. bassiana has been metabolically enriched with the enzyme chitinase in presence of an externally added amino acid serine with its inhibitory kinetics.

Development of a Culture Medium for Growth and Sporulation of Bacillus polyfermenticus SCD (프로바이오틱 비스루트균의 아포생산을 위한 최적배지 개발)

  • Lee, Kwang-Ho;Park, Kyu-Yong;Kim, Seong-Mi;Kim, Won-Seok;Paik, Hyun-Dong
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.263-268
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    • 2002
  • Bacillus polyfermenticus SCD, which is commonly called a 'Bisroot' strain, has been appropriately used for the treatment of long-term intestinal disorders, since the live strains, in the form of active endospores, can successfully reach the target intestine. Goal of this study was to develop an industrial medium for growth and sporulation of B. polyfermenticus SCD. From the results of effect of mixed carbon sources on growth and sporulation of B. polyfermenticus SCD, glucose 2% and starch 2% was particularly found to be the most effective for the maximum number of spore production, resulting in spore cells of $4.3{\times}10^9\;spores/mL$ with a sporulation yield of 91%. For the effect of nitrogen sources, the maximum spore cells of $5.7{\times}10^9\;spores/mL$ of B. polyfermenticus SCD with a sporulation yield of 97% was obtained when B. polyfermenticus SCD was cultivated in an optimum nitrogen source medium containing 5% soybean flour. A medium involving proper phosphate salt yielded the maximum number of a spore cells of $6.0{\times}10^9\;spores/mL$ with a sporulation yield of 95%. Finally, the efficacy of an industrial medium (KH5 medium) on growth and sporulation of B. polyfermenticus SCD was investigated in jar fermenter. The higher number of viable cells $(3.3{\times}10^{10}\;cells/mL)$ and spore cells $(3.0{\times}10^{10}\;spores/mL)$ were obtained in 5 L fermenter when compared with a 500 mL baffle flask cultivation. Thus, KH5 medium developed in this study shows promise as an industrial medium because of higher cells and sporulation yield.

Isolation and Characterization of Airborne Mushroom Damaging Trichoderma spp. from Indoor Air of Cultivation Houses Used for Oak Wood Mushroom Production Using Sawdust Media

  • Kim, Jun Young;Kwon, Hyuk Woo;Lee, Dong Hyeung;Ko, Han Kyu;Kim, Seong Hwan
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.674-683
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    • 2019
  • Some species of the Trichoderma genus are reported as the major problem in oak wood mushroom production in Korea. In spite of economic loss by the fungi, scientific information on airborne Trichoderma species is not much available. To generate information for disease management development we analyzed airborne Trichoderma. A total of 1,063 fungal isolates were purely obtained from indoor air sampling of cultivation houses used for oak wood mushroom using sawdust media. Among the obtained isolates, 248 isolates were identified as Trichoderma fungi including T. harzianum, T. atroviride, T. citrinoviride, and T. pseudokoningii, by morphological and molecular analysis. T. harzianum was dominant among the four identified species. All the four Trichoderma species grew fast on solid nutrient media tested (potato dextrose agar [PDA], malt extract agar [MEA], Czapek's Dox + yeast extract agar [CYA] and cornmeal dextrose agar). Compact mycelia growth and mass spore production were better on PDA and CYA. In addition, T. harzianum and T. citrinoviride formed greenish and yellowish mycelium and spores on PDA and CYA. Greenish and yellowish pigment was saturated into PDA only by T. pseudokoningii. These four Trichoderma species could produce extracellular enzymes of sawdust substrate degradation such as β-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease. Their mycelia inhibited the growth of oak wood mushroom mycelia of two tested cultivars on dual culture assay. Among of eleven antifungal agents tested, benomyl was the best to inhibit the growth of the four Trichoderma species. Our results demonstrate that the airborne Trichoderma fungi need to be properly managed in the cultivation houses for safe mushroom production.

Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

Production and Characterization of Antifungal Chitinase of Bacillus licheniformis Isolated from Yellow Loess (황토로부터 분리한 Bacillus licheniformis의 항진균 chitinase 생산과 효소 특성)

  • Han, Gui Hwan;Bong, Ki Moon;Kim, Jong Min;Kim, Pyoung Il;Kim, Si Wouk
    • KSBB Journal
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    • v.29 no.3
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    • pp.131-138
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    • 2014
  • In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.

Studies on the Production of Polysacrhacides by Agrobacterium radiobacter (Agrobacterium radiobacter에 의(依)한 다당류(多糖類) 생산(生産)에 관(關)한 연구(硏究))

  • Lee, Ke-Ho;Shin, Hyun-Sung
    • Applied Biological Chemistry
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    • v.26 no.2
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    • pp.110-118
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    • 1983
  • A slimy non-spore-forming bacterium strain SAF-C isolated from bean stem and root was motile with flagella and identified to one of Agrobacterium radiobacter. Studies were made on the conditions necessary for maximal production of this acidic succinoglucan polysaccharide by this strain in shaken culture. Much production was observed with yeast extract,$(NH_4)_2\;HPO_4$, distillers' dried solubles(D.D.S.), as nitrogen source in the medium composed of 4% glucose, 0.5% nitrogen source, 0.3% $CaCO_3$. The yield was greatest with yeast extract and decreased in order with the above nitrogen source from 22.9% to 9.6 percent. A polysaccharide was produced in a yield of about 25% in a medium composed of 3% glucose, 0.4% D.D.S., 0.5% $K_2\;HPO_4$, 0.01% $MgSO_4{\cdot}7H_2O$.

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Study on Marteilioides chungmuensis Comps et al., 1986 Parasite of the Pacific Oyster, Crassostrea gigas Thunberg (참굴의 난(卵)에 기생(寄生)하는 Marteilioids chungmuensis Comps et al., 1986에 관하여)

  • Park, Mi-Seon;Chun, Seh-Kyu
    • Journal of fish pathology
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    • v.2 no.2
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    • pp.53-70
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    • 1989
  • An ovarian parasite, Marteilioides chungmuensis of the Pacific oyster, Crassostrea gigas has been observed on several occasions in the Pacific sector of production of this oyster species(Matsuzato et al., 1977 ; Chun, 1979). This study was carried out on the specimens collected at Hwado, Och'$\check{o}$n, and Sinchang respectively located the southern, western, and eastern coasts of Korean Peninsula from 1986 through 1988 to investigate M. chungmuensis to the Pacific oyster. Uitrastructural studies were also carried out on infected oysters, to allow detailed examination of the structure and consepuently the systematic position of this parasite. Infection rates of M. chungmuensis at Hwado and Och'$\check{o}$n oyster farms were 5.3% and 4.2% each in 1986, 6.7% and 2.8% each in 1987, but they were not found at Sinchang oyster habitat. M,. chungmuensis-infected oysters were found from June to November at Hwado and from June to October at Och'$\check{o}$n. Twenty five of three hundred oysters transplanted from Sinchang to Hwado were found infected with M. chungmuensis. Some abnormal eggs infected with M. chungmuensis are liberated through the gill together with normal mature eggs on the spawning and the rest remain necrotized after spawning season. The earliest known stages consist of a stem cell or primary cell, including a secondary cell in which ovoid haplosporosomes are found. During sporulation, 2 or 3 secondary are produced by exogenous budding from the first secondary cell and, each secondary cell evolves into a sporont upon the tertiary cell differentiation (enodogenous budding) ; then, haplosporosomes are formed in the young sporont. Internal cleavages involve the differentiation of one tricellular spore per sporont. The outermost spore cell contains membrane-bounded osmiophilic bodies : the middle and the inner, most spore cells contain high density cytoplasmic ribosomes. The mechanism of spore formation from the stem cell of M. chungmuensis is the simplest of the class Paramyxea known up to now.

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Metabolic engineering of the genus Clostridium for butanol production (Clostridium 속 미생물 대사공학을 통한 butanol 생산)

  • Woo, Ji Eun;Kim, Minji;Noh, Hyeon Ji;Hwang, NuRi;Kim, Jin-Hyo;Lee, Sang Yup;Jang, Yu-Sin
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.391-397
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    • 2016
  • Clostridium is a genus of Gram-positive, rod shape, spore-forming obligate anaerobe. Recently, Clostridium has been attracted as a host for bio-based chemical production, due to its diversity of substrate utilization and the production ability for metabolites which can be used as a building block for chemical production. Especially, butanol produced from Clostridium has been considered as an alternative fuel. As a transportation fuel, butanol has a higher energy density and lower hygroscopicity compared to ethanol, the first generation biofuel. Recently, metabolic engineering of Clostridium has been massively conducted for butanol production. In this study, the metabolic engineering strategy of Clostridium for butanol production has been reviewed with a brief perspective.

Novel Approaches to Clubroot Management in Western Canada

  • Hwang, Sheau-Fang;Strelkov, Stephen E.
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.49-49
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    • 2015
  • Over the past decade, clubroot has emerged as a major constraint to canola (Brassica napus) production in central Alberta, Canada. The number of fields with confirmed P. brassicae infestations in Alberta has increased steadily from 12 in 2003 to nearly 2,000 in 2014. Management of clubroot on canola has focused on sanitization of field equipment, soil amendments to reduce viable pathogen populations, long rotations out of susceptible crops and cropping of resistant cultivars. Clubroot resistance is the most effective and economical method of disease mitigation, but the recent identification of isolated P. brassicae populations with novel virulence phenotypes capable of overcoming resistance in most canola cultivars highlights the variable nature and adaptability of the pathogen. Recent studies have shown slight reductions in pathogen populations through crop rotations, but much more substantial reductions in spore populations in heavily infested areas near field entrances using fumigants such as Vapam (metam-sodium) or Basamid (dazomet). Greenhouse trials showed that seedling emergence, plant height and root weight increased, while primary and secondary infection and disease severity decreased with increased Basamid dosage. However, field trials showed some phytotoxicity. Application of Vapam at rates of 0.4 to $1.6mL\;L^{-1}$ soil resulted in 12-16 fold reductions in clubroot severity and primary and secondary infection. Vapam also was effective in reducing clubroot severity and improving canola seed yield under field conditions. These studies underscore the need for good resistance stewardship and for the integration of multiple products and practices for successful management of clubroot on canola.

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Isolation and Identification of Alkalophilic Microorganism and its Mutant Growing at Neutral pH (호알칼리성 미생물의 분리, 동정 및 중성에서 생육 가능한 변이주의 분리)

  • 심창환;신원철;유주현
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.543-547
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    • 1991
  • An alkalophilic microorganism, SH-8, was isolated from soil samples. It was a Grampositive, catalase-positive, spore-forming and motile rod which was capable of growth in aerobic condition at the initial pH 9.0 or above up to 11.0 and between 15 and $42^{\circ}C$ The characteristics of this strain resembled those of the Bacillus group of bacteria. The mutant, Bacillus sp. SH- 8M, was selected from Bacillus sp. SH-8 by U.V. mutagenesis and was able to grow at pH 6.9 up to 11.0. These two strains will be suitable for the comparative study on growth and enzyme production at various pH conditions.

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