• Title/Summary/Keyword: mycelial culture

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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 characteristics and genetic relationship of morel mushroom (Morchella spp.) isolates from Korea and other countries (곰보버섯 (Morchella spp.) 수집균주의 배양적특성 및 유전적 유연관계)

  • Min, Gyeong-Jin;Park, Hye-sung;Lee, Eun-ji;Lee, Chan-Jung
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.100-106
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    • 2020
  • Eight morel mushroom species were collected from Korea and other countries. The culture characteristics, genetic relationships, and beta-glucan content of the strains were analyzed. The mycelia of Morchella species exhibited optimal growth when cultured in dark at 25 ℃ in media with pH 7. The mycelia had a distinctive mycelial scent and characteristically changed color, being white initially, and then turning dark yellow to dark brown as it grew. The mycelia were classified into five types based on morphology. The isolates were identified as Morchella conica, two M. sextelata, M. importuna, M. esculenta, and three M. crassipes, based on ITS-rDNA sequences. PCR polymorphisms were variably produced within Morchella spp. using Universal Fungal Fingerprinting Primers (UFPF) and classified into four groups at the intra and inter species level. The strains, KMCC04971 and KMCC04407, showed the same banding pattern as M. conica and M. sextelata, respectively; however, these results were different from those of ITS analysis. Glucan content analysis by strain showed that the KMCC 04973 strain of M. importuna had the highest alpha- and beta-glucan content, at 16.4 g and 33.1 g per 100 g, respectively.

Isolation and Selection of Antagonistic Microbes for Biological Control of Zoysiagrass Large Patch Disease (한국잔디 갈색퍼짐병의 생물학적 방제를 위한 길항미생물의 분리 및 선발)

  • Ma, Ki-Yoon;Kwark, Soo Nyeon;Lee, Geung-Joo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.657-665
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    • 2013
  • A large patch disease caused by Rhizoctonia solani AG2-2 (IV) is a serious problem in Korean lawngrass (Zoysia japonica) sites including golf courses and sports fields in Korea. Antagonistic microorganisms against R. solani AG2-2 (IV) were isolated from various forest and crop soil sources in Southern Korea. Among the 61 isolates, I-009, FRIN-001-1, and YPIN-022 strains showing dramatic inhibition of the mycelial growth of R. solani AG2-2 (IV) in the pairing culture were selected as the most potential antagonistic microorganisms for this study. Based on the 16s RNA sequence comparison, I-009 and FRIN-001-1 isolates were identified as Bacillus spp., while YPIN-022 isolate belongs to the genus Pseudomonas. The greater inhibition (clear) zone between two edges of the selected and pathogenic microbes ranged from 11 to 15 mm in three selections, but the others averaged to 7 mm out of 30 mm distance. In another antifungal test using culture filtrate, those three isolates represented a range of 51.7 to 63.5% suppression potential. The selected isolates also inhibited significantly the stem-segment colonization by R. solani AG2-2 (IV) in vivo test by 28.1%, 43.0%, and 23.7% when inoculated with I-009, FRIN-001-1, and YPIN-022, respectively. The highest antagonistic activity for the large patch disease was demonstrated by the isolate FRIN-001-1, which will be useful for developing a bio-pesticide against Rhizoctonia.

Optimization of γ-Aminobutyric Acid (GABA) Production Using Immobilized Lactobacillus plantarum K154 in Submerged Culture of Ceriporia lacerata (Ceriporia lacerata 배양액과 고정화 Lactobacillus plantarum K154를 이용한 감마아미노뷰티르산 생산 최적화)

  • Lee, Eun-Ji;Lee, Sam-Pin
    • Korean Journal of Food Science and Technology
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    • v.47 no.4
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    • pp.438-445
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    • 2015
  • The production of GABA was optimized by co-cultivation of immobilized Lactobacillus plantarum K154 (ILK) with Ceriporia lacerata cultures. The mycelial culture of C. lacerata was performed in a defined medium containing 3% glucose, 3% soybean flour, and 0.15% $MgSO_4$ in a submerged condition for 7 days at $25^{\circ}C$, resulting in the production of 29.7 g/L mycelia, 3.1 g/L exopolysaccharides, 2% (w/w) ${\beta}$-glucan, 68.96 unit/mL protease, and 10.37 unit/mL ${\alpha}$-amylase. ILK in C. lacerata culture showed viable cell counts of $3.13{\time}10^9CFU/mL$ for immobilized cells and $1.48{\time}10^8CFU/mL$ for free cells after 1 day. GABA production in the free and immobilized cells was 9.96 mg/mL and 6.30 mg/mL, respectively, after 7 days. A recycling test of ILK in the co-fermentation was consequently performed five times at $30^{\circ}C$ for 15 days, resulting in the highest production of GABA. GABA could also be efficiently overproduced by co-cultivation with the produced polysaccharides, ${\beta}$-glucan, peptides, and probiotics.

Batch Kinetics of Exo-polysaccharide Production by Submerged Cultivation of Ganoderma lucidum (영지의 액체배양에 의한 세포외 다당 생산의 동력학적 특성)

  • Lee, Shin-Young;Lee, Hak-Su;Park, Heung-Cho
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.304-311
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    • 1999
  • Batch kinetics during the exo-polysaccharide (EPS) fermentation of Ganoderma lucidum was investigated as a function of different substrates (glucose and starch), substrate concentration $(1{\sim}7%,\;w/v)$ and subculture (3 times). Logistic model for mycelial growth fitted the experimental data better than Monod and two thirds power model. The Luedeking-Pirt equation was adequate to fit the kinetic data of product formation and substrate consumption. The EPS production was strongly non-growth associated, although it was mixed type. The product formation and sustrate consumption by growth associated mechanism decreased as the concentration of glucose increased, while those of the non-growth associated mechanism increased. However, starch medium increased the growth associated and non-growth associated substrate consumption indicating higher availability of substrate. Also, batch culture in starch medium showed the higher specific growth rate and stability during subculture than those in glucose medium. In conclusion, the enhanced EPS production and stability in the subculture was found to be remarkably improved by use of starch as sole carbon source in medium. The maximum mycelium dry weight and EPS production of 9.463 and 10.410 g/l, respectively, were obtained after shake culture of 7 days at $30^{\circ}C$ from the media containing 7% starch.

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Antimicrobial Activity of the Coriolus versicolor Liquid Culture Extracts Against Antibiotic Resistant Bacteria and Purification of Active Substance (구름버섯 균사체 배양 추출물의 복합내성 세균에 대한 항균활성 및 활성물질의 정제)

  • Lee, Jung-Sun;Kim, Taeg;Lee, Yoon-Hi;Jin, Cheng-Min;Kim, Hyun-Guell;Kim, Woo-Jung;Oh, Duek-Chul;Park, Yong-Il
    • The Korean Journal of Mycology
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    • v.34 no.2
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    • pp.92-97
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    • 2006
  • The liquid culture extract of Coriolus versicolor was prepared by directly boiling the whole culture broth 7 days after incubation in 12% citrus extract medium. After removal of mycelial debris through filtration, this extract was further extracted with equal volume of ethyl acetate (1 : 1, v/v). The ethyl acetate extracts showed significant antibacterial activities against Stapylococcus aureus CCARM3230 and Psudomonas aeruginosa CCARM2171, which are resistant to several antibiotics. The most active fraction was eluted from a silica gel column with a mixture of dichloromethane and methanol (9 : 1, v/v) and the purity of this active substance was confirmed by HPLC analysis. The results suggest that the purified active substance could be a good source for the development of a new antimicrobial agent, especially for the treatment of antibiotic resistant bacteria.

High density culture of Bacillus subtilisBSM320 in aqueous extract of composted spent mushroom substrate of Lentinula edodes and biological control of green mold disease (표고수확후배지 퇴비 물 추출물에서 Bacillus subtilis BSM 320의 고밀도 배양 및 표고 푸른곰팡이병의 생물학적 방제)

  • Ja-Yoon Kim;Se-Hyun Park;Seong-Joon Park;Ju-Hyeong Hwang Bo;Hee-Wan Kang
    • Journal of Mushroom
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    • v.21 no.3
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    • pp.140-144
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    • 2023
  • The objective of this study was to achieve biological control of green mold disease in Pyogo mushrooms using antagonistic microorganisms. Bacillus subtilis BSM320 cells inhibited mycelial growth by 48-60% against three Trichodermaisolates including T. hazianumisolated from the substrates of Lentinula edodes, showing their antifungal activity.The bacteria were cultured to a high density of 4.2 × 109±113.7 cfu/mlin aqueous extract of composted spent mushroom substrates of L. edodes containing 1% glucose and showed a higher growth rate than that observed when using the commercial medium, Luria-Bertani broth. The bacterial culture showed a 75% protective effect without damaging the mushroom fruiting bodies. These results suggest that B. subtilis BSM320culture is suitable for biological control of green mold disease during mushroom cultivation.

Cultural Characteristics and Artificial Cultivation of Edible Mushroom, Clitocybe maxima (흰깔대기버섯의 배양적 특성 및 인공재배에 관한 연구)

  • Kim, M.K.;Kim, D.U.;Kang, H.W.;Seo, G.S.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.20 no.1
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    • pp.35-47
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    • 2018
  • A edible mushroom, Clitocybe maxima (Lentinus giganteusis) commercially cultivated in China and Taiwan. However, the researches of cultivation and cultural characteristics were not reported in Korea. In this study, we conducted on cultural characteristics and artificial cultivation of C. maxima. Six isolates were collected from China(3 isolates, commercial strain), Taiwan(1 isolate, commercial strain) and Korea(2 isolates, wild type). C. maxima and L. giganteus collected in China and Taiwan, respectively, are the same in China and are estimated to be of the same species as cultured characteristics. The mycelial growth of the collected strains was not significantly different in agar medium but it showed the best growth in YPMG in liquid culture. Optimum temperature for mycelial growth and induction of fruit body were 25℃ and 30℃, respectively. In order to artificial cultivation of C. maxima, cultural characteristics and artificial cultivation were carried out using agricultural by-products and forestry by-products materials. Mycelial growth was suitable in rice straw, cottonwood sawdust, corncob and rice seed medium, and it was selected as a cultivation medium. The suitable medium for artificial cultivation of C. maxima was selected to mixed medium 2(compounding ratio(v/v): 55% of hardwood sawdust, 5% of cottonseed pellets, 10% of cottonseed, 15% of beet pulp, 15% of swollen rice husks). It took about 30 days to be able to harvest, it was faster than oyster mushrooms. The cultivation period was about 30days. A isolate, CMA-002 was not initiation to fruit body primordiuma on the used cultivation substrate. Other 5 isolates were initiate and development to fruit body on the substrate used in this study. The strain CMA-003 was initiated to be fruiting body by 8~10 days after induction of fruiting body in all of the substrates. Isolate CMA-003 was generate to a bundle fruit body. Other isolates, however, were form fruit body individually. The CMA-003 strain was likely highly recommendable strains for farming. The optimum conditions for the induction and growth of C. maxima fruit body were 25~30℃, 8 hr illumination per day with white fluorescent lamp, 90~95% relative humidity, and 1,500 ppm of CO2 concentration in a cultivation room.

Control of Phythophthora capsici and residual characteristics by drenching of pesticides on tomato in hydroponic culture system (약제 관주처리에 의한 양액재배 토마토의 역병 방제 및 농약잔류 특성)

  • Ihm, Yang-Bin;Lee, Jung-Sup;Kyung, Kee-Sung;Kim, Chan-Sub;Oh, Kyeong-Seok;Jin, Yong-Duk;Lee, Byung-Moo
    • The Korean Journal of Pesticide Science
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    • v.6 no.4
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    • pp.287-292
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    • 2002
  • To establish effective and safe control method against Phytophthora root rot caused by Phytophthora capsici on tomato in hydroponic culture, three pesticides, oxadixyl copper hydroxide 8% WP, metalaxyl copper oxychloride 15% WP, and dimethomorph. dithianon 38% WP at 4 concentration levels were tested on potato dextrose agar medium inoculated with Phytophthora capsici. All pesticides inhibited mycelial growth, but two pesticides of them, metalaxyl copper oxychloride WP and dimethomorph. dithianon WP, were selected as effective pesticides for the efficacy test in a hydroponic culture. Forty days after transplanting of tomato seedlings, 4 ml of sporangia of P. capsici (about 25 sporangi/ml) per plot was inoculated around tomato plant root, and then 5 days after inoculation, the pesticides diluted at 5,000 times were drenched 1, 2 or 3 times per plot on the culture cube at 15 days interval. Fifteen days after drenching, tomato fruits and hydroponic culture solution were sampled for the analysis of pesticide residues. Dimethomorph was detected 0.001 and 0.003 mg/kg in tomato of the plots sprayed 2 and 3 times with dimethomorph dithianon WP of which detection levels were far below compared with 1.0 mg/kg of the Korean MRL of dimethomorph on tomato. Incidences of Phytophthora root rot were $30.5{\sim}50%$ in the plots drenched at 1 or 2 times with metalaxyl.copper oxychloride WP, and $16.7{\sim}25%$ in the plots treated with dimethomorph dithianon WP. However, there was no incidence of Phytophthora root rot in the plots treated at 3 times with both of pesticides, showing no phytotoxic effect. Based on the results, the drenching of these pesticides on the culture cube could be recommended as a very safe and effective control method for Phytophthora root rot in tomato.

Degradation of Three Aromatic Dyes by White Rot Fungi and the Production of Ligninolytic Enzymes

  • Jayasinghe, Chandana;Imtiaj, Ahmed;Lee, Geon-Woo;Im, Kyung-Hoan;Hur, Hyun;Lee, Min-Woong;Yang, Hee-Sun;Lee, Tae-Soo
    • Mycobiology
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    • v.36 no.2
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    • pp.114-120
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    • 2008
  • This study was conducted to evaluate the degradation of aromatic dyes and the production of ligninolytic enzymes by 10 white rot fungi. The results of this study revealed that Pycnoporus cinnabarinus, Pleurotus pulmonarius, Ganoderma lucidum, Trametes suaveolens, Stereum ostrea and Fomes fomentarius have the ability to efficiently degrade congo red on solid media. However, malachite green inhibited the mycelial growth of these organisms. Therefore, they did not effectively decolorize malachite green on solid media. However, P. cinnabarinus and P. pulmonarius were able to effectively decolorize malachite green on solid media. T. suaveolens and F. rosea decolorized methylene blue more effectively than any of the other fungi evaluated in this study. In liquid culture, G. lucidum, P. cinnabarinus, Naematoloma fasciculare and Pycnoporus coccineus were found to have a greater ability to decolorize congo red. In addition, P. cinnabarinus, G lucidum and T. suaveolens decolorized methylene blue in liquid media more effectively than any of the other organisms evaluated in this study. Only F. fomentarius was able to decolorize malachite green in liquid media, and its ability to do so was limited. To investigate the production of ligninolytic enzymes in media containing aromatic compounds, fungi were cultured in naphthalene supple mented liquid media. P. coccineus, Coriolus versicolor and P. cinnabarinus were found to produce a large amount of laccase when grown in medium that contained napthalene.