• Title/Summary/Keyword: filamentous growth

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Apolar growth of Neurospora crassa leads to increased secretion of extracellular proteins

  • Lee, In-Hyung;Rodney G. Walline;Michael Plamann
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.78-89
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    • 2000
  • Protein secretion in filamentous fungi has been shown to be restricted to actively growing hyphal tips. To determine whether an increase in the amount of growing surface area of a fungus can lead to an increase in the amount of protein secretion, we examined secretion in a temperature-sensitive Neurospora crassa mcb mutant that shows a loss of growth polarity when incubated at restrictive-temperature. Incubation of the mcb mutant at restrictive-temperature results in a three- to five-fold increase in the level of extracellular protein and a 20- fold increase in carboxymethyl cellulase activity relative to a wild-type strain. A mutation in the cr-l gene has been shown previously to suppress the apolar growth phenotype of the mcb mutant, and we find that the level of extracellular protein produced by a mcb; cr-l double mutant was reduced to that of the wild-type control. Immunolocalization of a secreted endoglucanase revealed that proteins are secreted mainly at hyphal tips in hyphae exhibiting polar growth and over the entire surface area of bulbous regions of hyphae that are produced following a shift of the mcb mutant to restrictive-temperature. These results support the hypothesis that secretion of extracellular protein by a filamentous fungus can be significantly increased by mutations that alter growth polarity.

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Establishment of Miniaturized Cultivation Method for Large and Rapid Screening of High-yielding Monascus Mutants, and Enhanced Production of Monacolin-K through Statistical Optimization of Production Medium (Monascus 균사체의 소규모 배양을 통한 고생산성 균주의 대규모 선별방법 확립과 통계적 생산배지 최적화를 통한 Monacolin-K 생산성 향상)

  • Lee, Mi-Jin;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.305-312
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    • 2007
  • It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.

Physiological and Molecular Characterization of Cephaleuros virescens Occurring in Mango Trees

  • Vasconcelos, Camila Vilela;Pereira, Fabiola Teodoro;Duarte, Elizabeth Amelia Alves;de Oliveira, Thiago Alves Santos;Peixoto, Nei;Carvalho, Daniel Diego Costa
    • The Plant Pathology Journal
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    • v.34 no.3
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    • pp.157-162
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    • 2018
  • The objective of this work was to accomplish the isolation, molecular identification and characterizing the physiology of the causal agent of the algal spot in mango trees. For this purpose, the pathogen growth was assessed in different culture media, with subsequent observation and measurements of the filamentous cells. The molecular identification was made using mycelium obtained from leaf lesions and pure algae colonies grown in culture medium. Descriptions based on DNA sequencing indicated that the algae is Cephaleuros virescens. The algae must be isolated primarily in liquid medium for further pricking into agar medium. The highest mycelial growth average in Petri dishes occurred when the algae were grown in Trebouxia and BBM. Trebouxia enabled larger cells in the filamentous cells when compared to other culture media.

Influence of Ammonium Phosphate on Mycelial Morphology during Submerged Cultivation of Ganoderma lucidum (영지의 액체배양에 있어서 균사체 형태에 미치는 Ammonium Phosphate의 영향)

  • Lee, Kyu-Min;Lee, Shin-Young
    • The Korean Journal of Mycology
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    • v.29 no.2
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    • pp.91-98
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    • 2001
  • The mycelial morphology during submerged cultivation of Ganoderma ludium using by air-lift fermenter system were analyzed by image processing system and the characterization of mycelial morphology were investigated. In submerged culture using medium with different ammonium phosphate concentrations, the various morphological forms of G. lucidum mycelium were observed. The filamentous forms such as non-branched long filamentous mycelium, non-branched short mycelium, branched long filamentous mycelium, branched short mycelium, entangled mycelium and clump were observed, and also, and also, the pelleted forms such as smooth pellet, rough pellet and hollow rough pellet were observed. The mycelial morphology was changed from the filamentous to the pelleted forms by addition of ammonium phosphate. The fractal dimensions of pelleted and filamentous forms were 1.05 and 1.3, respectively, while the fractal dimension of mixtures of pelleted and filamentous forms was 1.16. Therefore, the fractal dimension was found to be more effective index for the detection of the mycelial morphology and morphological change during batch cultivation. The circularity was also found to be useful for evaluating the surface growth of pelleted mycelium.

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Effect of Ammonium Phosphate on Mycelial Growth and Exopolysaccharides Production of Ganoderma lucidum in an Air-Lift Fermenter

  • Lee, Kyu-Min;Lee, Shin-Young;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.726-731
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    • 1999
  • It was discovered that ammonium phosphate in the medium played an important role in both growing mycelium and producing exopolysaccharides (EPS) from G. lucidum. In lower concentration levels of ammonium phosphate (0-3 g/l), an improved mycelial growth was observed by maintaining more filamentous morphology than in high concentrations (5-11 g/l). In addition, it was confirmed by comparing the factual dimension and frequency of the area regarding the mycelial pellets. This must be attributed to limitations of nutrient transfer by maintaining filamentous mycelium during the cultivation in a low ammonium phosphate containing medium. On the other hand, the best EPS production was observed in medium with the absence or low concentration of ammonium phosphate. The shear stress of the culture broth was greatly affected by the shear rate, as compared with that of the culture broth with high ammonium phosphate concentration. The rheological characteristics of the fermentation broth and filtrate worked well according to the Herschel-Bulkley model. It was also found that the morphological changes of the mycelium resulting from the ammonium phosphate concentration directly affected the rheological characteristics of the system and resulted in reversely affecting the EPS production levels. Based on these results, it can be concluded that delicate regulation of the ammonium phosphate concentration in the culture media should be provided in order to obtain optimal mycelial growth and/or EPS production.

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Penicillin Fermentation using a Carrier-supported Mycelial Growth (담체에 고정화된 균사체 증식을 이용한 페니실린의 발효)

  • Park, Sang K.;Kim, Jung H.;Park, Young H.
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.273-278
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    • 1985
  • A carrier-supported mycelial growth of Penicillium chrysogenum was applied to penicillin fermentation system. Among various materials tested, celite was found to be most effective for both spore adsorption and bioparticle development. Hyphal growth through pore matrices of the material showed strong anchorages and provided highly stable biofilm growths. When 5-10% celite was employed, both cell growth and penicillin production were observed to increase significantly comparing to the dispersed filamentous growth. Specific productivity of penicillin, however. was found to be kept almost constant at a value of 1,900 unit/g cell/hr. A semicontinuous fermentation in a fluidized-bed reactor. using the tarrier-supported biofilm growth, was conducted successfully although free mycelia appeared in the late phase of the fermentation made the reactor operation difficult. Control of the size of bioparticles was considered as a major operating factor to maintain the reactor productivity at a desired level.

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Ontogenetic Food Habits of Four Common Fish Species in Seagrass Meadows (해초생태계에 서식하는 4 우점어종의 성장에 따른 먹이의 변화에 관한 연구)

  • 허성희
    • 한국해양학회지
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    • v.21 no.1
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    • pp.25-33
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    • 1986
  • Ontogenetic food habits of the four most abundant fish species in seagrass neadows of Redfish Bay, Texas, were examined quantitatively during 1982-1983. The darter goby (Gobionellus bolelsoma) and pinfish (Lagodon rhomboides) were trophic generalists, which used a wide range of food items. The darter goby had relatively diverse, omnivorous diet which included amphipods, copepods, polychaetes, filamentous algae, diatoms, and detritus. This species did not show distinct ontogenetic changes in foodpreferences. Unlike the darter goby, the pinfish showed ontogenetic progression of four feeding stages. An initial feeding stage was a planktivorous stage in which copepods were int major food items, followed by a carnivorous stage in which amphipods became the major food items, an omnivorous stage in which filamentous algae, diatoms, amphipods, and polychaetes were the major food items, and finally a herbivorous stage in which seagrass pieces with attached epiphytes and their debris were the major food items. The code goby (Gobiosoma robustrm) and Gulf pipefish (Syngathus scovelli) appeared to be relatively specialized in food havits as carnivorous. Similar ontogenetic changes in food habits were observed for these two species, i.e.initially, copepods were the major food items, followed by a gradual transition to amphipods with growth.

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