The main objective of this topic is to establish strategies and to plan biotechnology researches which are related to the agricultural improvements especially focusing on the crop breeding in Korea. From 1960's to 1980's government policy had been emphasized to develope high yielding cultivars for the self sufficient supply of the staple food crops. As a result, considerable increase of rice production has been made with accumulating technology and man's powers. Recently genetically modified crops harboring useful characteristics have been developed using biotechnology and released in the developed countries. National research institutes and private companies have been developed biotechnology researches to establish competitive capabilities, however they have not been successfully used in commercialization. Therefore it is necessary to promote the practical. application by connecting molecular technology with conventional breeding. Proposed research projects are; (1) basic researches including plant genome studies, (2) developing new cultivars through gene transformation, (3) screening and producing antioxidants, secondary metabolite substances and edible vaccines. To set a government policy, both domestic and international research trends were reviewed and possibility of success based on the economic view point were discussed. The intellectual property and preservation of environment play a key role to decide the research priority. It is also necessary for us to make one step system for the distribution of research resources such as microorganisms, genes cloned, plant seeds and research informations for promoting research activities.
You, Young-Hyun;Park, Jong Myong;Kang, Sang-Mo;Park, Jong-Han;Lee, In-Jung;Kim, Jong-Guk
The Korean Journal of Mycology
/
v.43
no.3
/
pp.200-205
/
2015
Perilla frutescens var. japonica Hara was collected from farmland in Seongju-gun. Fifteen endophytic fungal strains with different colony morphologies were isolated from roots of P. frutescens. Waito-c rice seedlings were treated with the concentrated culture filtrates (CF) of endophytic fungi for observation of their plant growth-promoting activities. In the results, the CF of Y2H001 fungal strain promoted the growth of the waito-c rice seedlings. The phylogenetic tree of Y2H001 strain was analyzed by the combined sequences of the partial internal transcribed spacer region (ITS) and partial betatubulin gene. Molecular and morphological studies identified the Y2H001 strain as belonging to Aspergillus flavus. In gas chromatography mass spectrometry (GC/MS) analysis of the CF of Y2H001 strain, gibberellic acid (GA) was detected and quantified. Therefore, we describe Y2H001 strain as a new $GA_3$-producing A. flavus based on morphological, molecular characteristics and analysis of secondary metabolite.
The large-scale production of antibiotics by filamentous mycelial organism requires and adequate supply of dissolved oxygen. In terms of productivity, it means that oxygen transfer is the rate-limiting step. Therefore, the oxygen transfer coefficients(K$\_$L/A) were determined in a broth involving a filamentous mycelial organism such as Streptomyces avermitilis for use in fermentations. To determine (K$\_$L/A) inn a stirred vessel, a great deal of effort is required to provide all the cells with a sufficient oxygen supply. To overcome the oxygen limitation in a batch culture, a fed-batch culture was applied to control the growth rate by an intermittent supply of nutrients. Thus, it was possible to maintain a suitable dissolved oxygen concentration at a low agitation rate. The optimal agitation speed was 350 rpm at low cell concentrations (below 7 g/L) by considering the efficiency of agitation and shear stress. The (K$\_$L/A) was found to decrease from 64.26 to 29.21h.$\^$-1/ when the biomass concentration was increased from 9.82 to 12.06 g/L. In addition, and increase in viscosity was also observed during the growth phase. By comparing the (K$\_$L/A) values for the various agitation and aeration rates, it was found that the effect of an increase in (K$\_$L/A) by aeration was reduced dramatically at high biomass concentrations. However, this effect was not observed when altering the agitation rate. This suggests that controlling the dissolved oxygen concentration by altering the agitation rate was more efficient than increase the aeration rate.
The biosynthesis of Lovastatin, a cholesterol lowering agent formed by the filamentous fungus, cerulenin-resistant Aspergillus terreus mutant was studied in shake flasks and bioreactors. The lovastatin production could be improved by fed-batch under the limited condition of carbon source. The relationship between the fungal morphology and the lovastatin production was also examined during the fed-batch cultures. The fed-batch studies in shake flasks were carried out to find the optimum glucose feeding method, and the pulsed feeding of glucose from 3 days onward at 24 hours intervals was found to be optimal to increase the lovastatin production and reduce the average pellet size. When the pH was controlled at around 5.8 during the whole fermentation period, the lovastatin concentration reached 384 mg/L, which is much higher than the values obtained pH-uncontrolled and pH 7.4. The optimal glucose feeding strategies was found that 30 g/L of glucose was added initially in batch mode, and then fed-batch was conducted by continuous addition of glucose solution(180 g/L) from 72 to 240 hr at a rate of 1.2 mL/hr at $28^{\circ}C$, pH 5.8, 400 rpm, and 1.0 vvm. The lovastatin concentration of 547 mg/L was obtained in 168 hr. It was about 1.5 times higher than the value of the batch fermentation.
Journal of the Korean Society of Food Science and Nutrition
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v.37
no.5
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pp.666-670
/
2008
Lovastatin (Mevinolin, Monacolin K) is a well-known drug for the therapy of hypercholesterolemia. It is an important fungal secondary metabolite as it inhibits 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase, EC 1.1.1.34) which catalyzes a major rate-limiting step in the biosynthesis of cholesterol. Both soybeans and black soybeans with Aspergillus terreus ATCC 20542 mutant were used as the seed culture for the mass production of lovastatin. The production of lovastatin in soybean seed culture of Asp. terreus was twofold compared to that of black soybean seed culture. The effect of two different vessels (petri dish and Erlenmeyer flask) on lovastatin production was also studied. The production of lovastatin on petri dish was tenfold to that of Erlenmeyer flask. Furthermore, the most lovastatin production on rice bran was achieved when the soybean seed culture was treated by heat shock at $30^{\circ}C$ for 1 hour, representing 82% of HMG-CoA reductase inhibition in the koji extract. We estimated that the heat treated soybean seed culture could be a new method for the mass production of lovastatin.
An actinomycetes isolate G-0299 obtained from a forest soil showed strong phytotoxic activity to Digitaria ciliaris. For the foliar application study, the culture filtrate of the isolate G-0299 showed strong herbicidal activity only to D. ciliaris among the 12 monocot and 5 dicot weed species. And herbicidal activity at a concentration of 500, 250, 125 and $62.5{\mu}gmL^{-1}$ of culture filtrate was 100%, 98%, 70% and 40%, respectively. Phytotoxic symptoms of the culture filtrate by foliar application were desiccation and burn-down or bleaching of leaves and finally plant death. And then the herbicidal activity was exhibited only under the light condition. Also, chlorophyll loss of D. ciliaris leaf tissues in the light condition was much higher than in the dark condition and then chlorophyll content decreased 82%, 5%, respectively. In conclusion, our results suggest that soil bacteria, isolate G-0299 could be a good candidate for new bio-herbicide and provide a new lead molecule for a more unique herbicide.
Kim, Chang Joon;Park, Sun Ok;Chang, Yong Keun;Chun, Gie-Taek;Lee, Jong-Dae;Kim, Sangyong
Clean Technology
/
v.4
no.1
/
pp.45-59
/
1998
Continuous immobilized-cell culture was carried out for the production of kasugamycin, a secondary metabolite by a filamentous bacteria, Streptomyces kasugaensis, with an intention of reducing waste generation. A sporulation medium was developed for production of bulk amounts of spores, and the spores were entrapped into celite biosupports for immobilization. It was possible to effectively keep the immobilized-cells inside the reactor during the continuous culture by an efficient immobilized cell separator of decantor type on the outlet of the fermentor. Using this continuous immobilized-cell fermentor system, we investigated the effects of feed substrate and phosphate concentrations on kasugamycin production and chemical oxygen demand(COD). Comparing with the conventional suspended-cell batch culture, the kasugamycin productivity was observed to increase by 2.5 times, whereas COD per unit kasugamycin production decreased by 2.3 times in the continuous immobilized-cell culture. Based on these results, the continuous immobilized-cell system was considered to be a cleaner bioprocess than the conventional batch suspended-cell system.
Certain Fungi Including Aspergillus flavus produce low molecular secondary metabolite that is toxic to human and animals, which have been termed mycotoxin. Given the proper humidity and temperature like summer in Korea, are capable of growing of those hazard fungi and elaborating mycotoxin on almost any organic substrate such as traditional foodstuffs and their raw materials including rice, barley, corn, meju, doenjang and gochujang etc. Until now, some people have examined to isolate various fungi such as Aspergillus sp., Penicillium sp. and Fusarium sp. from traditional foodstuffs and raw materials, and have screened various mycotoxin producing strains. Some mycotoxin contamination such as aflatoxin, ochratoxin, deoxynivalenol (DON) and zearalenone etc. also have been confirmed from similar above samples. But these data are different each other and inconsistent in experimental conditions and methods. Especially, almost experiments have been finished for one time. So more consistent experimental method and data are necessary to evaluate objectiely the safety of traditional foodstuffs subjected to the mycotoxin. For this purpose, we have to apply a new advanced technology to develop more simple and rapid methods for determination of mycotoxin and also have to concentrate our efforts on activation of research and accumulation of technology nth sustaining investment of financial support and enlargement of research installation. With those harmonious efforts, it should be possible to examine continuously nd systematically the mycotoxin contamination in our traditional foodstuffs and to assure the safety of them. Then we can maintain and develop the better traditional foodstuffs suited to internationalization.
Avermectin (AVM) $B_{1a}$ produced by Streptomyces avermitilis via polyketide pathway is a secondary metabolite with powerful anthelmintic and insecticidal activities, thus being used as an efficient agent in the field of agriculture and animal health. It has been reported that a precursor for AVM $B_{1a}$ biosynthesis was isoleucine and the biosynthetic pathway of AVM $B_{1a}$ was closely similar to that of fatty acid. Based on understanding of the biosynthetic pathway of AVM $B_{1a}$, we intended to screen various mutants resistant against O-methyl threonine (OMT), an isoleucine-anti metabolite, and/or mutants resistant against p-fluoro phenoxy acetic acid (pFAC), an inhibitor of fatty acid biosynthesis. It was inferred that these mutants could produce AVM $B_{1a}$ more efficiently, due to the acquired capability of not only overproducing isoleucine intracellularly but also channelling metabolized carbon-sources into the polyketide pathway, thus leading to enhanced biosynthesis of AVM $B_{1a}$. The resulting mutant (PFA-1 strain) resistant against 100 ppm of pFAC was able to produce approximately 42 fold higher amount of AVM $B_{1a}$ compared to the parallel mother strain (4,200 vs. 100 units/l). In addition, through the process of continuous strain improvement program carried out by gradually increasing the OMT concentration, it was possible to obtain a more attractive mutant with greater AVM $B_{1a}$ production capacity (9,000 units/l). Notable was that significantly higher producer (12,000 units/l) could be selected through further screening of the resistant mutants, this time, to even higher concentration of PFAC. Meanwhile, through the analysis of AVM Bla production histograms (i.e., number of strains according to their AVM $B_{1a}$ biosynthetic ability) for the earlier strains in comparison with the high producers having the characteristics of resistance to OMT and pFAC, it was found that production stability of the high-yielding producers were remarkably improved, as demonstrated by the fact that larger proportion of the mutated strains had greater capability of AVM $B_{1a}$ biosynthesis ($71\%$ in the range between 5,000 and 7,000 units/L; $47\%$ in the range between 6,000 and 7,000 units/l). Based on these consequences, it was concluded that the rational screening strategy based on the understanding of the biosynthetic pathway of AVM $B_{1a}$ was very effective in obtaining high-yielding mutants with the features of enhanced production stability.
Aim of this study was to investigate the effects of different nutrient solutions and various light qualities generated by LED on the growth and glucosinolates contents of watercress (Nasturtium officinale) grown under hydroponics for 3 weeks. The seeds of watercress were sown on crushed rockwool media and raised them for two weeks. They were transplanted in a semi-DFT (deep flow technique) hydroponics system. A controlled-environment room was maintained at $20{\pm}1^{\circ}C$ and $16{\pm}1^{\circ}C$ temperatures and $65{\pm}10%$ and $75{\pm}10%$ relative humidity (day and night, respectively), with a provided photosynthetic photon flux density (PPFD) of $180{\pm}10{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ and a photoperiod of 16/8h. To find out the best kinds of nutrient solutions for growing watercress, Otsuka House 1A (OTS), Horticultural Experiment Station in Korea (HES), and Netherland's Proefstaion voor Bloemisterij en Gasgroente (PBG) were adapted with initial EC of $1.0-1.3dS{\cdot}m^{-1}$ and pH of 6.2, irradiating PPFD with fluorescent lamps (Ex-1). Either monochromatic (W10 and R10) or mixed LEDs (R5B1, R3B1, R2B1G1, and W2B1G1) were irradiated with a differing ratio of each LED's PPFD to understanding light quality on the growth and glucosinolates contents of watercress (Ex-2). Although significant difference in the shoot growth of watercress was not found among three nutrient solutions treatments, but the root fresh weight increased by 13.7% and 55.1% in PBG and OTS compared to HES, respectively. OTS increased the gluconasturtiin content by 96% and 65% compared to PBG and HES. Compared with the white light (W10), the red light (R10) showed a 101.3% increase in the shoot length of watercress. Increasing blue light portion positively affected plant growth. The content of total glucosinolates in watercress was increased by 144.5% and 70% per unit dry weight in R3B1 treatment compared with R2B1G1 and W10 treatments, respectively. The growth and total glucosinolates contents of the watercress were highest under R3B1 among six light qualities.
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