• Title/Summary/Keyword: flask culture

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Determining the Wood (Parashorea spp.) Decaying and Metal Corroding Abilities of Eight Fungi

  • SUPRAPTI, Sihati;DJARWANTO, DJARWANTO;DEWI, Listya Mustika
    • Journal of the Korean Wood Science and Technology
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    • v.48 no.1
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    • pp.50-60
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    • 2020
  • Fungi are wood-decaying organisms, and this is an important trait that should be considered in wood utilization. When fungi attack wood, it decreases the quality of the wood. The use of metal screws has become an important part of woodworking. The ability of fungi to decay wood and damage metal screws that are embedded into wood is varied. In this study, eight fungal species were evaluated with respect to their ability to decay Parashorea smythiesii and P. tomentella wood. In addition, the effect of fungi on corroding metal screws was determined using the Kolle flask method. The evaluation showed that the fungal species Schizophyllum commune, Pycnoporus sanguineus, and Polyporus arcularius were highly capable of decaying Parashorea spp. woods. The greatest wood weight loss occurred with the heartwood of P. tomentella exposed to S. commune. Based on the classification of wood resistance against fungal attack, the two Parashorea spp. were classified as moderately resistant woods (class III). Schizophyllum commune was classified as highly capable of decaying wood that was embedded with metal screws and was highly capable of corroding metal screws placed in fungi-culture media. The greatest weight of rust powder formed because of screw corrosion was obtained from screw-embedded wood exposed to S. commune. Additionally, the most severe corrosion of metal screws that were embedded into woods was caused by the activities of P. arcularius. Moreover, the average weight loss of screw-embedded wood was greater than that of unscrewed wood.

Production of Bio-Based Isoprene by the Mevalonate Pathway Cassette in Ralstonia eutropha

  • Lee, Hyeok-Won;Park, Jung-Ho;Lee, Hee-Seok;Choi, Wonho;Seo, Sung-Hwa;Anggraini, Irika Devi;Choi, Eui-Sung;Lee, Hong-Weon
    • Journal of Microbiology and Biotechnology
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    • v.29 no.10
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    • pp.1656-1664
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    • 2019
  • Isoprene has the potential to replace some petroleum-based chemicals and can be produced through biological systems using renewable carbon sources. Ralstonia eutropha can produce value-added compounds, including intracellular polyhydroxyalkanoate (PHA) through fatty acid and lipid metabolism. In the present study, we engineered strains of R. eutropha H16 and examined the strains for isoprene production. We optimized codons of all the genes involved in isoprene synthesis by the mevalonate pathway and manipulated the promoter regions using pLac and pJ5 elements. Our results showed that isoprene productivity was higher using the J5 promoter ($1.9{\pm}0.24{\mu}g/l$) than when using the lac promoter ($1.5{\pm}0.2{\mu}g/l$). Additionally, the use of three J5 promoters was more efficient ($3.8{\pm}0.18{\mu}g/l$) for isoprene production than a one-promoter system, and could be scaled up to a 5-L batch-cultivation from a T-flask culture. Although the isoprene yield obtained in our study was insufficient to meet industrial demands, our study, for the first time, shows that R. eutropha can be modified for efficient isoprene production and lays the foundation for further optimization of the fermentation process.

Enhanced Production of C30 Carotenoid 4,4'-Diaponeurosporene by Optimizing Culture Conditions of Lactiplantibacillus plantarum subsp. plantarum KCCP11226T

  • Siziya, Inonge Noni;Yoon, Deok Jun;Kim, Mibang;Seo, Myung-Ji
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.892-901
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    • 2022
  • The rising demand for carotenoids can be met by microbial biosynthesis as a promising alternative to chemical synthesis and plant extraction. Several species of lactic acid bacteria (LAB) specifically produce C30 carotenoids and offer the added probiotic benefit of improved gut health and protection against chronic conditions. In this study, the recently characterized Lactiplantibacillus plantarum subsp. plantarum KCCP11226T produced the rare C30 carotenoid, 4,4'-diaponeurosporene, and its yield was optimized for industrial production. The one-factor-at-a-time (OFAT) method was used to screen carbon and nitrogen sources, while the abiotic stresses of temperature, pH, and salinity, were evaluated for their effects on 4,4'-diaponeurosporene production. Lactose and beef extract were ideal for optimal carotenoid production at 25℃ incubation in pH 7.0 medium with no salt. The main factors influencing 4,4'-diaponeurosporene yields, namely lactose level, beef extract concentration and initial pH, were enhanced using the Box-Behnken design under response surface methodology (RSM). Compared to commercial MRS medium, there was a 3.3-fold increase in carotenoid production in the optimized conditions of 15% lactose, 8.3% beef extract and initial pH of 6.9, producing a 4,4'-diaponeurosporene concentration of 0.033 A470/ml. To substantiate upscaling for industrial application, the optimal aeration rate in a 5 L fermentor was 0.3 vvm. This resulted in a further 3.8-fold increase in 4,4'-diaponeurosporene production, with a concentration of 0.042 A470/ml, compared to the flask-scale cultivation in commercial MRS medium. The present work confirms the optimization and scale-up feasibility of enhanced 4,4'-diaponeurosporene production by L. plantarum subsp. plantarum KCCP11226T.

Effect of carbon sources on somatic embryogenesis and cotyledon number variations in carrot (Daucus carota L.)

  • Young Jin Lee;Kyu Seog Hwang;Pil Son Choi
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.89-95
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    • 2023
  • In order to investigate the effect of carbon sources on somatic embryogenesis and cotyledon number variations in carrot, embryogenic callus were cultured in the medium supplemented with various concentrations of sucroseor glucose, and with combination of 2% sucrose and various concentrations of mannitol or sorbitol. The maximum number of somatic embryos formed per flask (1,555.70) was obtained in the medium supplemented with 2% sucrose rather than glucose alone or a combination of mannitol or sorbitol and 2% sucrose, and the number of somatic embryos was decreased with the increasing of mannitol or sorbitol concentration. The frequencies of somatic embryos with two cotyledons were 35.14% for sucrose, 19.88% for glucose, 32.55% for mannitol + 2% sucrose, and 38.59% for sorbitol + 2% sucrose, respectively, and the frequencies of abnormal somatic embryos having 3 or more cotyledons were 64.86% for sucrose, 80.12% for glucose, 67.44% for mannitol + 2% sucrose, and 61.41% for sorbitol + 2% sucrose, respectively. Particularly, the frequency of somatic embryos with two cotyledons (59.16%) was the highest in the 2% sucrose medium compared to the frequency of abnormal somatic embryogenesis with three or more cotyledons, and the frequency gradually decreased with increasing concentration of glucose, mannitol or sorbitol. According to these results, it was found that the ratio of abnormal somatic embryo was higher than the normal somatic embryo in carrot, and was shown that somatic embryogenesis and the cotyledon number was affected by the concentrations of sucrose, glucose as carbon source, and mannitol and sorbitol as osmotic agents in culture medium.

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.

Cell Growth and Lipid Production from Fed-batch Cultivation of Chlorella minutissima according to Culture Conditions (유가식 배양에서 배양조건에 따른 Chlorella minutissima의 생육 및 지질생산)

  • Oh, Sung-Ho;Han, Jae-Gun;Kim, Na-Young;Cho, Jeong-Sub;Yim, Tae-Bin;Lee, Shin-Young;Lee, Hyeon-Yong
    • KSBB Journal
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    • v.24 no.4
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    • pp.377-382
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    • 2009
  • The culture condition of growing Chlorella minutissima was optimized to produce biodiesel for fed-batch cultivation. First, under heterotrophic cultivation, the optimum level of glucose was determined to be 10 g/L for 20 days. After, three cultivation conditions were operated: autotrophic, heterotrophic, and mixotrophic growth. The lipid level and the maximum cell concentration from the fed-batch heterotrophic process were 32.0 (%, v/v) and 15.0 (g-dry wt./L) in 20 L flask, respectively. In addition, since the relatively constant specific lipid production rate was observed as 0.040 (% lipid/g-dry wt./day) at the latter period of cultivation time, the fed-batch process could maintain continuous lipid production. Fed-batch process is higher than those values from the batch process. The lipids from the fed-batch process contained over 38% of $C_{18}$, known as the suitable composition for the biodiesel application. For mixotrophic and heterotrophic growth under fed-batch condition, glucose was proved to be an appropriate carbon source for a large scale outdoor cultivation. For fed-batch cultivation, the feeding rate of seawater medium containing glucose was decided to be 0.5 L/day. The mixotrophic cultivation maintained maximum cell concentration of 24 (g-dry wt./L) and the lipid level of 43 (%, w/w). The lipid composition from this process was also proved to be suitable for the biodiesel production. The fatty acids from the mixotrophic growth contains 18% of $C_{17}$ and 49% of $C_{18}$, implying It also tells that C. minutissima is a suitable resource of biodiesel. Especially, the mixotrophic cultivation with fed-batch process might be useful for the large scale cultivation for the biodiesel production.

Biological control of Gray Mold Rot of Perilla Caused by Botrytis cinerea II. Formulation of Antagonistic Bacteria and Its Control Effect (들깨 잿빛곰팡이병의 생물학적 방제 II. 미생물농약의 제조 및 그 방제효과)

  • Moon, Byung-Ju;Kim, Choul-Soung;Song, Ju-Hee;Kim, Ju-Hee;Lee, Jae-Pil;Park, Hyean-Cheal;Shin, Dong-Bum
    • Research in Plant Disease
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    • v.8 no.3
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    • pp.184-188
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    • 2002
  • An antagonistic bacteria, Bacillus licheniformis Nl strain which effectively inhibited mycelial growth of gray mold rot pathogen, Botrytis cinerea was isolated from the rhizosphere of perilla crop. Powder soy formulation by B. lichentfomis Nl strain as a biocontrol agent was developed far the first time and estimated its control effect on perilla leaves in this study. First of all, far the mass production of antifungal metabolites of B. lichentfomis Nl strain in flask liquid culture, the most effective carbon and nitrogen source were selected as glucose and tryp-tone, respectively, For the formulation, vegetative biomass of B. licheniformis Nl strain from 5-day-old liquid culture in nutrient broth added glucose and tryptone was mixed with soy flour, rice flour glucose, FeSo$_4$~7$H_2O$, and MnCl$_2$. 4$H_2O$, and dried and pulverized. In plastic house test, powder soy formulation effectually controlled gray mold rot as the control value of 93.1 %, was more effective than chemical fungicide, benomyl showing the control value of 86.1%. Thus, development of powder soy formulation of B. lichentfomis Nl will aid large-scale application of biological control in field trials.

Ex situ acclimatization of asymbiotically germinated seedlings of endangered lady's slipper orchid(Cypripedium macranthos Sw.) (멸종위기종 복주머니란 발아특성 및 실생묘 현지외 적응성 검토)

  • Lee, Joung Kwan;Kwon, Young Hee;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Jeong, Mi Jin;Son, Sung Won;Suh, Gang Uk
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.37-37
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    • 2018
  • We use two different methods for laboratory propagation from seed of lady's slipper orchid(Cypripedium macranthos Sw.); immature seed which also called green capsule or fully mature seed about 120~130 days from pollination. In green capsule culture, the seed pods should be collected within precisely right time. The right time of seed collection could be diverse under the wether conditions or nutritional factors of the plants. In fully matured seed culture, the more complicated procedures are needed to break the dormancy of the seed; thermal or chemical treatment. The seedlings in this study were easily germinated from immature seeds in Harvais medium; 53 days after pollination(DAP) in Cypripedium pubescens, DAP 65 in C. parviflorum and C. macranthos. The germinated seedlings were transplanted to hormone free media immediately to avoid abnormal growth of seedlings. When the seedlings have roots with a minimum length of around 2-3cm and have visible dormant buds, the seedlings were removed from the flask and stored in refrigerator for vernalization. To examine the correlation of seedlings and maternal plants, the 125 seedlings of C. macranthos were transplanted in the soil bed at a distance of 20-100 cm from mother plants on April 20. The survival rate of seedlings were 92% in 20 cm distance from the ripe plants, and 56 % in 100 cm distance. The seedlings which were transplanted near mother plants showed vigorous growth in plant height, leaf width, and especially dormant buds. Considering the existence of mycorrhiza which is a symbiotic association between a fungus and the roots of a orchid vascular, the various fungus from mother plants could affect the growth of the seedlings. These results indicate the possibility of high and stable production and practical industrialization of endangered lady's slipper orchids.

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Production of Hydroxymethylfurfrual by Sesamum indicum L. Root Cultures (참깨 뿌리배양에 의한 hydroxymethylfurfrual 생산)

  • Chun, Jae-An;Lee, Jin-Woo;Yi, Young-Byung;Hong, Seong-Sig;Cho, Kang-Jin;Chung, Chung-Han
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1836-1840
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    • 2009
  • Recently, hydroxymethylfurfrual (HMF) has been highlighted as a key intermediate for the production of liquid biofuels and other valuable compounds. We used sesame roots as a biocatalyst to synthesize HMF using flask cultures. The synthesis of HMF was identified by GC-mass analysis. The highest root growth was observed in cultures with 1.0 mg/l NAA at $30^{\circ}C$, while root growth was not found in those without NAA treatment. When silver nitrate ($AgNO_3$) was added, the root growth was greatest in those treated with 0.5 mg/l $AgNO_3$ and cultured at $30^{\circ}C$. In the case of HMF synthesis, its highest yield was obtained in those treated with 0.5 mg/l NAA at $25^{\circ}C$, but low HMF was detected in those treated without naphthaleneacetic acid (NAA). The addition of $AgNO_$ to the culture medium showed a 8-10% reduction in HMF yield compared to that of the control, indicating its inhibitory effect on the synthesis of HMF. On the whole, an optimal culture temperature for HMF synthesis seemed to be between $25-30^{\circ}C$.

Bioethanol production from wood biomass hydrolysate with supercritical water treatment (초임계수 처리로 가수분해된 목질계 바이오매스를 이용한 바이오 에탄올 생산)

  • Seo, Hyeon-Beom;Han, Jae-Gun;Choi, Won-Seok;Lee, Oh-Kyu;Lee, Soo-Min;Choi, Seok-Hwan;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • KSBB Journal
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    • v.23 no.6
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    • pp.494-498
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    • 2008
  • We investigated the bioethanol production using wood biomass hydrolysate which obtained from the supercritical water (SCW) treatment. SCW-treated hydrolysate was used C-source of culture medium in shaking flask culture for bioethanol production. When the concentrated SCW-treated hydrolysate (SCW3) was used, yeast cell growth was slower compared with those in other SCW-treated hydrolysate (SCW1, SCW2). In addition, the bioethanol productions were 0.51 to 0.56 (%,w/v) when SCW1, SCW2, and SCW3 were used. Therefore, we removed the toxic phenolic compound in SCW-treated hydrolysate by pretreatments of activated charcoal and calcium hydroxide. Activated charcoal reduced more efficiently the phenolic compounds in SCW3 by 94.6%. Finally, when we pretreated SCW3 by activated charcoal and this was used for bioethanol production, 0.96 (%,w/v) bioethanol was produced and the ethanol yield based on reducing sugar reached 0.5.