The discharge of waste nutrient solution from greenhouse to natural ecosystem leads to the accumulation of excess nutrients that results in contamination or eutrophication. There is a need to recycle the waste nutrient solution in order to prevent the environmental hazards. The amount and kind of nutrients in waste nutrient solution might be enough to grow photosynthetic microorganisms. Hence in the present study, we examined the growth and mass cultivation of cyanobacteria in the waste nutrient solution with an objective of removing N and P and concomitantly, its mass cultivation. Four photosynthetic filamentous cyanobacteria (Anabaena HA101, HA701 and Nostoc HN601, HN701) isolated from composts and soils of the Chungnam province were used as culture strains. Among the isolates, Nostoc HN601 performed faster growth rate and higher N and P uptake in the BG-II ($NO_3{^-}$) medium when compared to those of other cyanobacterial strains. Finally, the selected isolate was tested under optimum conditions (airflow at the rate of $1L\;min^{-1}$. in 15 L reactor, initial pH 8) in waste nutrient solution from tomato hydroponic in green house condition. Results showed to remove 100% phosphate from the waste nutrient solution in the tomato hydroponics recorded over a period of 7 days. The growth rate of Nostoc HN601 was $16mg\;Chl-a\;L^{-1}$ in the waste nutrient solution from tomato hydroponics with optimum condition, whereas growth rate of Nostoc HN601 was only $9.8mg\;Chl-a\;L^{-1}$ in BG-11 media. Nitrogen fixing capacity of Nostoc HN601 was $20.9nmol\;C_2H_4\;mg^{-1}\;Chl-a\;h^{-1}$ in N-free BG-11. The total nitrogen and total phosphate concentration of Nostoc HN601 were 63.3 mg N gram dry weight $(GDW)^{-1}$ and $19.1mg\;P\;GDW^{-1}$ respectively. Collectively, cyanobacterial mass production using waste nutrient solution under green house condition might be suitable for recycling and cleaning of waste nutrient solution from hydroponic culture system. Biomass of cyanobacteria, cultivated in waste nutrient solution, could be used as biofertilizer.
Kim, Sun-Hee;Kim, Sung-Hoon;Lee, Jo-No;An, Sang-Wook;Kim, Kwang-Soo;Hwnag, Baik;Lee, Hyeong-Yong
Microbiology and Biotechnology Letters
/
v.26
no.5
/
pp.435-441
/
1998
Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate, ${\mu}$$\sub$max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q$\^$max/$\sub$p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between ${\mu}$ and q$\sub$p/ showed that the product formation parameters, ${\alpha}$ and ${\beta}$$\sub$p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1 ${\mu}$M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X$\sub$max/, and maximum production, P$\sub$max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.
For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.
As information technology fusion is accelerated, the researches to improve the quality and productivity of crops inside a plant factory actively progress. Advanced growth environment management technology that can provide thermal environment and air flow suited to the growth of crops and considering the characteristics inside a facility is necessary to maximize productivity inside a plant factory. Currently running plant factories are designed to rely on experience or personal judgment; hence, design and operation technology specific to plant factories are not established, inherently producing problems such as uneven crop production due to the deviation of temperature and air flow and additional increases in energy consumption after prolonged cultivation. The optimization process has to be set up in advance for the arrangement of air flow devices and operation technology using computational fluid dynamics (CFD) during the design stage of a facility for plant factories to resolve the problems. In this study, the optimum arrangement and air flow of air circulation fans were investigated to save energy while minimizing temperature deviation at each point inside a plant factory using CFD. The condition for simulation was categorized into a total of 12 types according to installation location, quantity, and air flow changes in air circulation fans. Also, the variables of boundary conditions for simulation were set in the same level. The analysis results for each case showed that an average temperature of 296.33K matching with a set temperature and average air flow velocity of 0.51m/s suiting plant growth were well-maintained under Case 4 condition wherein two sets of air circulation fans were installed at the upper part of plant cultivation beds. Further, control of air circulation fan set under Case D yielded the most excellent results from Case D-3 conditions wherein air velocity at the outlet was adjusted to 2.9m/s.
Tamburini, Elena;Vaccari, Giuseppe;Tosi, Simona;Trilli, Antonio
Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
/
2001.06a
/
pp.3104-3104
/
2001
The research described here was undertaken with the aim of monitoring, optimizing and ultimately controlling the production of heterofermentative microbes used as starters in the salami industry. The use of starter cultures in the fermented meats industry is a well-established technique used to shorten and standardize the ripening process, and to improve and control the organoleptic quality of the final product. Starter cultures are obtained by the submerged cultivation of suitable microorganisms in stirred, and sometimes aerated, fermenters where monitoring of key physiological parameters such as the concentration of biomass, substrates and metabolites suffers from the general lack of real-time measurement techniques applicable to aseptic processes. In this respect, the results of the present work are relevant to all submerged fermentation processes. Previous work on the application of on-line NIR spectroscopy to the lactic acid fermentation (Dosi et al. - Monreal NIR1995) had successfully used a system based on a measuring cell included in a circulation loop external to the fermenter. The fluid handling and sterility problems inherent in an external circulation system prompted us to explore the use of an in-line system where the NIR probe is immersed in the culture and is thus exposed to the hydrodynamic conditions of the stirred and aerated fluid. Aeration was expected to be a potential source of problems in view of the possible interference of air bubbles with the measurement device. The experimental set-up was based on an in-situ sterilizable NIR probe connected to the instrument by means of an optical fiber bundle. Preliminary work was carried out to identify and control potential interferences with the measurement, in particular the varying hydrodynamic conditions prevailing at the probe tip. We were successful in defining the operating conditions of the fermenter and the geometrical parameters of the probe (flow path, positioning, etc.) were the NIR readings were reliable and reproducible. The system thus defined was then used to construct and validate calibration curves for tile concentration of biomass, carbon source and major metabolites of two different microorganisms used as salami starters. Real-time measurement of such parameters coupled with the direct interfacing of the NIR instrument with the PC-based measurement and control system of the fermenter enabled the development of automated strategies for the interactive optimization of the starter production process.
Kim, Hee Sook;Kim, Ji-Youn;Lee, Song Min;Park, Hye-Jung;Lee, Sang-Hyeon;Jang, Jeong Su;Lee, Mun Hyon
Microbiology and Biotechnology Letters
/
v.47
no.4
/
pp.603-613
/
2019
This study was carried out to examine the antagonistic effect against phytopathogenic fungi of isolated strains from soil samples collected from Busan, Changwon, and Jeju Island: Botrytis cinerea, Colletotrichum acutatum, Corynespora cassiicola, Fusarium sp., Rhizoctonia solani, Phytophthora capsici, and Sclerotinia sclerotiorum. According to results of our studies, isolated strains showed an antagonistic effect against phytopathogenic fungi. Such an antagonistic effect against phytopathogenic fungi is seen due to the production of siderophores, antibiotic substances, and extracellular amylase, cellulase, protease, and xylanase enzyme activities. Extracellular enzymes produced by isolated strains were significant, given that they inhibited the growth of phytopathogenic fungi by causing bacteriolysis of the cell wall of plant pathogenic fungi. This is essential to break down the cell wall of plant pathogenic fungi and thus help plant growth by converting macromolecules, which cannot be used by the plant for growth, into small molecules. In addition, they are putative candidates as biological agents to promote plant growth and inhibit growth of phytopathogenic fungi through nitrogen fixation, indole-3-acetic acid production, siderophore production, and extracellular enzyme activity. Therefore, this study suggests the possibility of using Bacillus subtilis ANGa5, Bacillus aerius ANGa25, and Bacillus methylotrophicus ANGa27 as new biological agents, and it is considered that further studies are necessary to prove their effect as novel biological agents by standardization of formulation and optimization of selected effective microorganisms, determination of their preservation period, and crop cultivation tests.
Seo, Ho-Chan;Lee, Jung-Suk;Yun, Zu-Whan;Yi, Yun-Seok;Cho, Hong-Yon
Applied Biological Chemistry
/
v.41
no.6
/
pp.465-470
/
1998
For the need of bio-degradable flocculant in stage of wastewater treatment, some cultural conditions of bioflocculant production were optimized with Aeromonas hydrophila KH-54. About 260 strains of type culture and bacteria isolated from marsh, pond, activated sludge, etc were examined for their ability to flocculate kaolin particles and swine wastewater. Among them, KH-54 showed the highest flocculating activity and was identified as Aeromonas hydrophila according to the cultural, morphological and physiological properties. The maximum production of the flocculant secreted by Aeromonas hydrophila KH-54 was observed in culture medium containing 2.0% mannitol, 0.05% ammonium chloride, 0.02% potassium phosphate dibasic, 0.01% $MgSO_4{\cdot}7H_2O$ and 0.05% yeast extract at initial pH 7.0 when cultured on rotary shaker controlled at $25^{\circ}C$ and 150 rpm. Under the optimized condition, the flocculating ability reached to 770 units/ml of kaolin flocculating activity and 81% of NTU removal efficiency against swine wastewater after 4 days cultivation. The bioflocculant was also effective on various organic wastewaters other than swine wastewater, showing NTU removal rate ranging from 92% to 34%.
A strong constitutive PJH promoter from Bacillus sp. was applied to overexpress the endoxylanase gene (639 bp) in Bacillus subtilis. The expression plasmid, pJHKJ4, was designed to contain the $P_{JH}$ promoter and open reading frame of endoxylanase including its own promoter. The plasmid was introduced into B. subtilis DB431 and the resulting transformant was grown on LB glucose medium. At the end of cultivation, the endoxylanase activity in the culture supernatant reached about 140 DIm!. The enzyme in the supernatant was concentrated by ultrafiltration (MW cut-off 10 kDa and 30 kDa) and ammonium sulfate precipitation. For the concentration of the enzyme, ultrafiltration was more efficient than 70% ammonium sulfate precipitation. The stabilization of concentrated enzyme solution at $50^{\circ}C$ was examined with various stabilizers such as NaCI, glycerol, polyethylene glycol, sorbitol, and $CaCI_2$. The most effective stabilizers were found to be NaCI and $CaCI_2$.
Based on the results in this study, here we propose a systematic micropropagation process for 'Gisela 5' that is one of the important dwarfing cherry rootstocks. When the apical tips detached from newly developed shoot in spring season were cultured on the half strength MS media with 0.5 mg/L IBA and 0.5 ~ 1.0 mg/L BA, the cultures scored the highest acquisition rate at 90% for normal shoot with vigorous growth and without hyperhydricity. As next step, the young shoots maintained in vitro well multiplied on the full strength MS medium supplemented with 0.5 mg/L IBA and 0.5 mg/L BA, in which multiplication rate was approximately nine-fold. Given the half strength MS medium containing 2.0 mg/L IBA, each transplanted shoot further developed robust roots. Finally, the plantlets were easily acclimatized in the compost consisted of vermiculite, perlite, and peatmoss in the proportion of 1:1:1. We expect that the results are useful for cherry cultivation and its rootstock production.
Lee, Hyun-Ju;Son, Jae-Young;Lee, Sang-Jae;Lee, Han-Seung;Lee, Bae-Jin;Choi, In-Soon;Sohn, Jae Hak
Journal of Life Science
/
v.27
no.5
/
pp.567-574
/
2017
This study is to isolate and identify ${\gamma}$-amino butyric acid (GABA) producing lactic acid bacteria (LAB) from Makgeolii, traditional Korean rice wine and then establish the optimal culture conditions for GABA production. Sixty four LAB from Makgeolli were isolated according to the characteristics of the shape and color of the colony grown on MRS agar plate. The GABA production of the isolated strain cultured in MRS broth contained 1% MSG (mono-sodium glutamate) were determined and evaluated by TLC and HPLC analysis. Strain B-134 was selected for highest GABA production. From the analysis of 16S rRNA and glutamate decarboxylase B (gadB) gene sequences, strain B-134 was tentatively identified as a Lactobacillus plantarum subsp. plantarum B-134. Effects of culture parameters, including glutamic acid level, culture temperature, NaCl level, and pH on GABA production were investigated for culture optimization. The optimum culture condition for GABA production by B-134 were culture temperature of $37^{\circ}C$, pH of 5.7, NaCl content of 0% (w/v) and MSG content of 3% (w/v), which produced 25 mM of GABA during cultivation time of 48 hr. From these results, strain B-134 is expected to be utilized as useful microorganisms for GABA-enriched health beneficial food.
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