• Title/Summary/Keyword: Fermentation Mycelial growth

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Effects of Dissolved Oxygen Level on Avermectin $B_{1a}$ Production by Streptomyces avermitilis in Computer-Controlled Bioreactor Cultures

  • Song, Sung-Ki;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1690-1698
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    • 2006
  • In order to investigate the effect of dissolved oxygen (DO) level on AVM $B_{1a}$ production by a high yielding mutant of Streptomyces avermitilis, five sets of bioreactor cultures were performed under variously controlled DO levels. Using an online computer control system, the agitation speed and aeration rate were automatically controlled in an adaptive manner, responding timely to the oxygen requirement of the producer microorganism. In the two cultures of DO limitation, the onset of AVM $B_{1a}$ biosynthesis was observed to casually coincide with the fermentation time when oxygen-limited conditions were overcome by the producing microorganism. In contrast, this phenomenon did not occur in the parallel fermentations with DO levels controlled at around 30% and 40% throughout the entire fermentation period, showing an almost growth-associated mode of AVM $B_{1a}$ production: AVM $B_{1a}$ biosynthesis under the environments of high DO levels started much earlier than the corresponding oxygen-limited cultures, leading to a significant enhancement of AVM $B_{1a}$ production during the exponential stage. Consequently, approximately 6-fold and 9-fold increases in the final AVM $B_{1a}$ production were obtained in 30% and 40% DO-controlled fermentations, respectively, especially when compared with the culture of severe DO limitation (the culture with 0% DO level during the exponential phase). The production yield ($Y_{p/x}$), volumetric production rate (Qp), and specific production rate (${\bar{q}}_p$) of the 40% DO-controlled culture were observed to be 14%, 15%, and 15% higher, respectively, than those of the parallel cultures that were performed under an excessive agitation speed (350 rpm) and aeration rate (1 vvm) to maintain sufficiently high DO levels throughout the entire fermentation period. These results suggest that high shear damage of the high-yielding strain due to an excessive agitation speed is the primary reason for the reduction of the AVM $B_{1a}$ biosynthetic capability of the producer. As for the cell growth, exponential growth patterns during the initial 3 days were observed in the fermentations of sufficient DO levels, whereas almost linear patterns of cell growth were observed in the other two cultures of DO limitation during the identical period, resulting in apparently lower amounts of DCW. These results led us to conclude that maintenance of optimum DO levels, but not too high to cause potential shear damage on the producer, was crucial not only for the cell growth, but also for the enhanced production of AVM $B_{1a}$ by the filamentous mycelial cells of Streptomyces avermitilis.

Culture Conditions and Additives Affecting to the Mycelial Pellet Size of Penicillium sp. GL-101 in the Submerged Culture (Penicillium sp. GL-101의 액침배양중 Mycelial Pellet 크기에 영향을 주는 배양조건 및 첨가물)

  • Lee, Dong-Gyu;Ha, Chul-Gyu;Lee, Tae-Geun;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.42 no.3
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    • pp.188-192
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    • 1999
  • In order to minimize the mycelial pellet formation, one of the critical obstacles during the fermentation processes of filamentous fungi, an investigation was focused on the culture conditions(media and initial inoculum) and additives(soils, surfactants and polyethylene glycol 200) when a high phosphate-dissolving fungus, Penicillium sp. GL-101, was cultured in liquid media. Culturing the strain in PDB, SDB and YPD media, their pellet sizes decreased to the order of YPD > SDB > PDB. And at the high concentrations of the initial inoculum in the range from $1{\times}10^3\;to\;1{\times}10^6$ conidia/ml, the small sizes of pellet were formed in the PDB media. For the initial inoculum between $1{\times}10^7\;and\;1{\times}10^8$ conidia/ml, however, an amorphous pellet or loose aggregate was formed. The addition of soils, zeolite and diatomite, up to 1.0% decreased the pellet sizes to 3/4 and 1/2, respectively, but the pellet was increased to 2.5 times by the addition of bentonite. Surfactants also affected on the size of pellet; the addition of Triton X-100 and Tween 80 up to 1.0% decreased the pellet sizes maximally to 1/10 and 1/4, respectively, while SDS completely inhibited the fungal growth. Among the four additives tsted, polyethylene glycol 200 was the most effectively reduced the pellet sizes to $0.2{\pm}0.1$mm that resulted in about 25- fold reduction compared to the control.

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Isolation and Characterization of Growth Stimulating Thermophilic Fungi on Oyster Mushroom from Oyster Mushroom (Pleurotus ostreatus) Compost (느타리버섯 배지로부터 느타리버섯 균사의 성장을 돕는 고온성 곰팡이의 분리 및 동정)

  • 이호용;현성희
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.33-39
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    • 2000
  • Some of thermophilic fungi which has growth-promoting effect on Pleurotus ostreatus were isolated from compost during high temperature fermentation process. The temperature optima of 7 isolated thermophilic fungi were $50^{\circ}C$ on PDA media. Isolated strains S-1 and S-2 have the best mycelial growing rate, so these isolates were expected as excellent thermophilic fungi for high temperature composting and mycelial growing of oyster mushroom. In liquid culture, the optimal pH of thermophilic fungi observed variously, pH 7.0-10.0 but most of thermophilic fungi grow well in pH 8.0-pH 9.0 and the final pH of media after cultured was done pH 5.5-6.0. In liquid culture of thermophilic fungi on the optimal condition, S-2 have the best mycelial growing rate. The growing rate of thermophilic fungi S-1, S-2, S-5, and S-10 on lignocellulosic substrates was good but Humicola grisea var. thermoidea, well know thermophilic fungi which has growth-promoting effect on Agaricus bisporus, was poor and which was well grown on PDA at $50^{\circ}C$, pH 7.0. Isolated strain S-1 was identified as Trichophyton sp. and other 6 strains were identified as Sepedonium sp. by morphological characteristics.

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Taxonomy and Fermentation of Kitasatosporia kimorexae Producing New Thiopeptide Antibiotics, Kimorexins

  • Yeo, Woon-Hyung;Kim, Si-Kwan;Kim, Sang-Seock;Yu, Seung-Hun
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.354-359
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    • 1994
  • An isolate, 90-GT-302, was found to produce antibiotics inducing typical mycelial swelling in Magnaporthe grisea and Fusarium solani. This isolate formed yellow substrate and white rectiflexbiles aerial mycelia in the early stages of growth. The aerial mycelium gradually changed its color to white and finally formed a gray spore mass. Analysis of the cell wall acid hydrolysate revealed the presence of LL- and meso-diaminopimelic acids, glycine, and galactose, which indicated cell wall type X. This result placed our isolate in genus Kitasatosporia. A comparison of isolate 9O-GT-302 with reference strains of Kitasatosporia spp., which not only demonstrated several differences in their physiological properties but also novelty of the active compounds produced by this isolate, led us to designate the isolate as Kitasatosporia kimorexae.

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The effect of antagonists produced by Paenibacillus polymyxa CK-1 on the growth of Trichoderma sp. (Paenibacillus polymyxa CK-1이 생산한 길항물질이 Trichoderma sp. 생육에 미치는 영향)

  • Lee, Sang-Won;Choi, Jin-Sang;Kim, Chul-Ho
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.201-208
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    • 2014
  • The separation of the bacteria inhibiting Trichoderma sp. mold, the strain causing blue mold disease that occurs frequently when cultivating mushroom while carrying out the efficient fermentation of mushroom medium, from the growth was done. In about 200 strains isolated primarily from fungus garden samples, 6 strains were secondly isolated, which had fast growth rates and a clear zone on the plate medium of SM, AM, and CM. Among the 6 strains isolated, the C-1 strain showed high enzymatic activity of cellulase, amylase, and protease, and strong antibacterial activity for the T. virens and T. harzianum, selected finally. The selected C-1 strain was identified as Paenibacillus polymyxaby the result of the identification by Bergey's Manual of Systematic Bacteriology and the analysis of the nucleotide sequence of 16S rRNA, and named as P. polymyxa CK-1. In reviewing the growth conditions of the P. polymyxa CK-1 strain, the optimum cultivation temperature was $45^{\circ}C$, and the optimum pH for growth was in the range of 6.0~7.0. Appropriate incubation time of P. polymyxa CK-1 for the growth inhibition of the fungus T. virens and T. harzianum was 22 to 36 hours. And the fungal growth was not observed, even when leaving two molds inoculated on each petri dishes, which were treated with 24 hour culture solution of P. polymyxa CK-1 strain for 10 days. As a result of studying the thermal stability of the antagonists produced by the P. polymyxa CK-1 strain, no mycelial growth of the two fungi was observed in the test group treated for 20 minutes at $60^{\circ}C$ and $100^{\circ}C$, but mycelial growth was slightly observed in the test group treated for 20 minutes at $121^{\circ}C$. As aresult of reviewing the impact of the P. polymyxa CK-1 culture medium on mushroom mycelial growth, it showed no effect on a variety of mushroom mycelial growth including enoki mushroom and shiitake mushroom.

Development of Miniaturized Culture Systems for Large Screening of Mycelial Fungal Cells of Aspergillus terreus Producing Itaconic Acid

  • Shin, Woo-Shik;Lee, Dohoon;Kim, Sangyong;Jeong, Yong-Seob;Chun, Gie-Taek
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.101-111
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    • 2017
  • The task of improving a fungal strain is highly time-consuming due to the requirement of a large number of flasks in order to obtain a library with enough diversity. In addition, fermentations (particularly those for fungal cells) are typically performed in high-volume (100-250 ml) shake-flasks. In this study, for large and rapid screening of itaconic acid (IA) high-yielding mutants of Aspergillus terreus, a miniaturized culture method was developed using 12-well and 24-well microtiter plates (MTPs, working volume = 1-2 ml). These miniaturized MTP fermentations were successful, only when highly filamentous forms were induced in the growth cultures. Under these conditions, loose-pelleted morphologies of optimum sizes (less than 0.5 mm in diameter) were casually induced in the MTP production cultures, which turned out to be the prerequisite for the active IA biosynthesis by the mutated strains in the miniaturized fermentations. Another crucial factor for successful MTP fermentation was to supply an optimal amount of dissolved oxygen into the fermentation broth through increasing the agitation speed (240 rpm) and reducing the working volume (1 ml) of each 24-well microtiter plate. Notably, almost identical fermentation physiologies resulted in the 250 ml shake-flasks, as well as in the 12-well and 24-well MTP cultures conducted under the respective optimum conditions, as expressed in terms of the distribution of IA productivity of each mutant. These results reveal that MTP cultures could be considered as viable alternatives for the labor-intensive shake-flask fermentations even for filamentous fungal cells, leading to the rapid development of IA high-yield mutant strains.

Impeller Types and Feeding Modes Influence the Morphology and Protein Expression in the Submerged Culture of Aspergillus oryzae

  • Heo, Joo-Hyung;Vladimir Ananin;Park, Jeong-Seok;Lee, Chung-Ryul;Moon, Jun-Ok;Ohsuk Kwon;Kang, Hyun-Ah;Kim, Chul-Ho;Rhee, Sang-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.3
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    • pp.184-190
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    • 2004
  • The influences of impeller types on morphology and protein expression were investigated in a submerged culture of Aspergillus oryzae. The impeller types strongly affected mycelial morphology and protein production in batch and fed-batch fermentations. Cells that were cultured by propeller agitation grew in the form of a pellet, whereas cells that were cultured by turbine agitation grew in a freely dispersed-hyphal manner and in a clumped form. Pellet-grown cells showed high levels of protein production for both the intracellularly heterologous protein (${\beta}$-glucuronidase) and the extracellularly homologous protein (${\alpha}$-amylase). The feeding mode of the carbon source also influenced the morphological distribution and protein expression in fed-batch fermentation of A. oryzae. Pulsed-feeding mainly showed high protein expression and homogeneous distribution of pellet whereas continuous feeding resulted in less protein expression and heterogeneous distribution with pellet and dispersed-hyphae. The pellet growth with propeller agitation paralleling with the pulsed-feeding of carbon source showed a high level of protein production in the submerged fed-batch fermentation of recombinant A. oryzae.

Analysis of the behavior of microorganisms isolated from the medium during cultivation of Agaricus bisporus (button mushroom) (양송이 재배 중 배지에서 분리한 미생물의 상호작용 분석)

  • Min, Gyeong-Jin;Park, Hae-sung;Lee, Eun-Ji;Yu, Byeong-kee;Lee, Chan-Jung
    • Journal of Mushroom
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    • v.19 no.2
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    • pp.103-108
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    • 2021
  • This experiment investigates the characteristics of microorganisms isolated from a medium during cultivation process and reveals the relationship between these microorganisms and the growth of Agaricus bisporus. The domestically grown strains of Agaricus bisporus displayed a higher inhibition growth rate against microorganisms isolated from straw, chicken manure, and medium than imported strains. As for inhibition of mycelial growth among mushroom cultivars of the microorganisms separated by each fermentation step from the mushroom medium, the domestic cultivar, 'Saedo,' grew more vigorously among other cultivars. As the fermentation progressed, it was confirmed that inhibitation of microorganisms against Agaricus bisporus was weakened. A total of 21 strains of microorganisms that promote mushroom growth were isolated in the 4th turning process, and the microorganisms isolated from the mushroom medium affect the growth and as yield of the mushroom through secretory substances.

Utilization and Application of Microorganisms in Treating Food Processing Wastes -Recovery of Mycelial Proteins- (식품가공공장 폐수의 미생물학적 처리 및 응용 -미생물 균체단백질 회수-)

  • Cho, Sung-Hwan;Choi, Jong-Duck;Lee, Sang-Yeol;Ki, Woo-Kyung;Kim, Ze-Uook
    • Applied Biological Chemistry
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    • v.32 no.4
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    • pp.424-434
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    • 1989
  • The rationale for the use of fungi in treating waste streams from food processing plants I~as been that of incorporating the dissolved and suspending nutrients into a macroscopic organism which can be filtered out readily. In order for a process using fungi to meet these objectives we examined a strain of fungi, Aspergillus fumigatus, which grew well on a variety of polysaccharide-containing materials and showed both efficient BOD removal and high quality protein recovery. In this experiment the fungal choice was based on the laboratory screening studies where the criteria used was BOD and COD reduction, growth response, mycelial yield, and the ability to compete with the natural flora. In the fermentation system used far the continuous culture of Aspergillus fumigatus the best combination of operating variables, inoculum ratio, temperature, initial pH, fermentation time and agitation rate was 5%(v/v), $35{\sim}40^{circ}C,\;pH\;4.5{\sim}5.0$, 2days and 150rpm, respectively. The fungus reduced BOD and COD to 94.0 and 90.4%, respectively and 3.15g of dry mycelium per liter of alcohol waste was harvested during 48hr of incubation time. The protein efficiency ratios for the control diet and the experimental diet containing the fungal protein were $3.42{\pm}0.15$ and $3.40{\pm}0.43$, respectively.

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Mycelial Growth Using the Natural Product and Angiotensin Converting Enzyme Inhibition Activity of Pleurotus eryngii (천연물을 이용한 큰느타리 균사배양 및 Angiotensin Converting Enzyme 저해활성)

  • Kang, Tae-Su;Jeong, Heon-Sang;Lee, Myong-Yul;Park, Hee-Joeng;Jho, Taek-Sang;Ji, Seung-Taek;Shin, Myung-Keun
    • The Korean Journal of Mycology
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    • v.31 no.3
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    • pp.175-180
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    • 2003
  • To develop the health/functional food materials, we investigated the cultural condition of mycelial growth on the solid state fermentation using the brown rise, Acanthopanax sp. and Artemisia sp., and also evaluated inhibitory activity of angiotensin converting enzyme (ACE) of hot water extracts from cultured media of Pleurotus eryngii. As the amount of Acanthopanax nnd Artemisia In the cultural media increased, the mycelial growth rate decreased. Especially, addition of Aeantopanax showed marked effect than Artemisia. Moisture contents in three kinds of cultured media were in the range of $10.9{\sim}12.0%$. Crude protein fat and crude fiber content were the highest value in cultured brown rice medium, whereas the mineral contents (Ca, K and P) were higher in the Acanthopanax supplemented (5%) medium than the other media, The extraction yield of the Artemisia supplemented (5%) medium was the highest value of 4.80%, and the pH of hot water extract from cultured brown rice medium showed the lowest value of 6.1. Lightness (L) values in three kinds of extracts from cultured media were in the range of $85.8{\sim}87.1$. Redness (a) value was the highest In the brown rice and Acanthopanax supplemented media, however cultured Artemisia supplemented medium showed the highest value in yellowness (b). In comparison of sugar components analyzed by the thin layer chromatography with three kinds of samples, two spots were detected to be glucose and maltose, respectively. The ACE inhibitory activity of hot water extract from the cultured Acanthopanax supplemented medium showed the highest value at the concentration of $0.2{\sim}1.0\;mg/ml$. These results suggest that the Pleurotus eryngii grew in natural media using brown rice and Acanthopanax can be supplemented to the brown rice medium to enhance its ACE inhibitory activity as health/functional food materials.