In order to obtain a regulatory mutant strain with high cellulase activity, a newly isolated Penicillium verrculosum, strain F-3 was used as parental strain since it was proved to be an efficient cellulase producer. A number of experiments were conducted to determine the optimum conditions to in-duce mutagenesis and isolate the desirable mutant strains. Out of several restriction compounds tested, 1.5% oxgall was found to be most effective to restrict the colony size by suppressing overgrowth. Derepression of catabolites was employed as a criterion in selecting mutant strains with high cellulase productivity. Production of cellulase by Penicillium venculosum F-3 was suppressed when cultured on the media with more than 1% of glucose or glycerol. It was found that either irradiation with UV light for 19 mins or treatment with nitrosoguanidine at 200$\mu\textrm{g}$/m1 for 60 mins, induced mutagenesis at desired level, when the survival rate of the spore was 0.2% and 48%, respectively. Three mutant strains of F-3, UV-9, UV-10, and NTG-3 that had the highest cellulase productivity were finally selected, based on filter paper degradation rate, size of clearing zone on the screening plate and cellulase activity in the medium containing cellulose powder. When the mutant strains were compared with parental strain F-3, on the KC-M-W medium containing cellulose powder, the filter paper activities of UV-9, UV-10, and NTG-3 were increased by 34%, 55%, and 41%, respectively. However, the assimilation of cellobiose octaacetate by UV-9 or NTG-3 was markedly reduced. When the mutant UV-10 was grown on cellobiose octaacetate medium (CCA-4) in shaking flasks, the cellulase activities of the mutant increased by 20 to 50% compared to the parental strain. Excreation of soluble protein from the mutant also elevated up to 30%. The mutant also constitutively produced both CMCase and $\beta$-glucosidase, though at relatively low level, in the presence of glucose or cellobiose as carbon sources.
Kim, Young-Kwon;Hong, Myung-Pyo;Kim, Myung-Jin;Hong, Suk-Il;Park, Myung-Suk;Kim, Jong-Suk;Chang, Ho-Geun
Journal of the Korea Organic Resources Recycling Association
/
v.6
no.2
/
pp.95-112
/
1998
For the effective disposal of organic food wastes, we seleted 4 strains of microorganism from 186 microbial candidate via enzyme activity test, salt tolerance, food decomposition rate, stability and safety of strains. The identity of these 4 strains are as follows : Fungi is Rhizopus sp., yeasts are Galactomyces sp., Pichia sp. and Hyphopichia sp., In the 50L fermenter scale, we tested various fermenting factor for the optimization of conditions of food waste decomposition using 4 selected strains. The optimum fomenting conditions were as follows : BIO-CHIP Volume 25-30 L, BIO CHIP size 2.0-6.0mm, air flow 200-280L/min, mixing intensity 2-4rpm, temperature $30-45^{\circ}C$. In these fermenting conditions, the efficiency of decomposition(rate of weight loss of food wastes) were 93%. Also the quality of fermenting output were assayed at the basis of fertilizer, and the results were as good as general compost.
In this study, powered superplasticizer(PSP) agents to improve the slump loss rate of recycled aggregate concrete were developed. To evaluate the variation of fluidity against elapsed time and the mechanical properties, twenty four specimens whose main variables had the mixing condition of aggregates, such as natural and recycled gravels, and natural and recycled fine aggregates, were tested. The concrete slump with a liquid superplasticizer greatly decreased against the elapsed time and dropped by less than 50% of initial slump after two hours. However the concrete slump with the PSP agents hardly varied until after half an hour and maintained more than 85% of initial slump even after an hour. Also the PSP agents made the compressive, splitting tensile, and flexural strength of concrete increased and the shrinkage strain decreased. Considering the properties improvement of concrete, it can be recommended that optimum mixing amount of the PSP agents should be 5% of the amount of cement.
Park, Kyoung-Joo;Cho, Kyoung-Sook;Kim, Jeong-Bo;Lee, Min-Gyu;Lee, Byung-Hun;Hong, Young-Ki;Kim, Joong-Kyun
한국생물공학회:학술대회논문집
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2005.10a
/
pp.450-455
/
2005
To treat wastewater efficiently by a one-step process of nitrogen removal, a new strain of $N_2-producing$ bacteria from $NH{_4}{^+}$ under an aerobic condition was isolated and identified. By 16S-rDNA analysis, the isolate was identified as Enterobacter asburiae with 96% similarity. The isolate shows that the capacity of $N_2$ production under an oxic condition was approximately three times higher than that under an anoxic condition. The optimal conditions (pH, temperature and C/N ratio) of the immobilized isolate for $N_2$ production were found to be 7.0, $30^{\circ}C$ and 5, respectively. Under all the optimum reaction conditions, the removal efficiency of $COD_{Cr}$ and TN reached 56.1 and 60.9%, respectively. The removal rates of $COD_{Cr}$ and TN were highest for the first 2.5 hrs (with the removal $COD_{Cr}$ ratios of 32.1), and afterwards the rates decreased as reaction proceeded. For application of the immobilized isolate to a practical process of ammonium removal, a continuous bioreactor system exhibited a satisfactory performance at HRT of 12.1 hr, in which the effluent concentrations of $NH{_4}{^+}-N$ was measured to be 15.4 mg/L with its removal efficiency of 56.0%. The maximum removal rate of $NH{_4}{^+}-N$ reached 1.6 mg $NH{_4}{^+}-N/L/hr$ at HRT of 12.1 hr (with N loading rate of 0.08 $Kg-N/m^3-carrier/d)$. As a result, the application of the immobilized isolate appears a viable alternative to the nitrification-denitrification processes.
The Photosynthetic bacterium, Rhodopseudomonas sphaeroides strain B6 was irnmobilized on agar gel. The optimum concentration of agar for hydrogen production was 2% (w/v). Maximum rates of hydrogen production by immobilized (300 ml of gel; 2.85 rng dry cells/ml) and free cells (1l culture; 0.87 mg dry cells/ml) were 47.5 and 48.0 ml/hr/culture, respectively. However, when both cultures were fed by 10 mmoles of lactate as limited electron donor at the later period of incubation, the activity of hydrogen production by free cells was significantly decreased but, immobilized cells continued hydrogen production with almost the same initial rate. Wc examined hydrogen production by immobilized cells of strain B6 under periodic illumination for 12 hr-intervals. When the culture was periodically fed by basal medium containing 9.3 rnmoles of DL-lactate and 1.86 mmoles of L-glutamate as consumed electron donor and nitrogen source, respectively, for every one liter of hydrogen produced, hydrogen was evolved continuously with the average rate of 510 ml/day/300 ml gel (2.9 rng dry cellslml) during the incubation time for 228 hr.
Park, Kyoung-Joo;Cho, Kyoung-Sook;Kim, Jeong-Bo;Lee, Min-Gyu;Lee, Byung-Hun;Hong, Young-Ki;Kim, Joong-Kyun
Environmental Engineering Research
/
v.10
no.5
/
pp.213-226
/
2005
To treat wastewater efficiently by a one-step process of nitrogen removal, a new bacterial strain producing $N_2$ gas from ${NH_4}^+$ under an aerobic condition was isolated and identified. The cell was motile and a Gram-negative rod, and usually occurred in pairs. By 16S-rDNA analysis, the isolated strain was identified as Enterobacter asburiae with 96% similarity. The isolate showed that the capacity of $N_2$ production under an oxic condition was approximately three times higher than that under an anoxic condition. Thus, the consumption of ${NH_4}^+$ by the isolate was significantly different in the metabolism of $N_2$ production under the two different environmental conditions. The optimal conditions of the immobilized isolate for $N_2$ production were found to be pH 7.0, $30^{\circ}C$ and C/N ratio 5, respectively. Under all the optimum reaction conditions, $N_2$ production by the immobilized isolate resulted in reduction of ORP with both the consumption of DO and the drop of pH. The removal efficiencies of $COD_{Cr}$, and TN were 56.1 and 60.9%, respectively. The removal rates of $COD_{Cr}$, and TN were the highest for the first 2.5 hrs with the removal $COD_{Cr}/TN$ ratios of 32.1, and afterwards the rates decreased as reaction proceeded. For application of the immobilized isolate to a practical process of ammonium removal, a continuous operation was executed with a synthetic medium of a low C/N ratio. The continuous bioreactor system exhibited a satisfactory performance at 12.1 hrs of HRT, in which the effluent concentrations of ${NH_4}^+$-N was measured to be 15.4 mg/L with its removal efficiency of 56.0%. The maximum removal rate of ${NH_4}^+$-N reached 1.6 mg ${NH_4}^+$-N/L/hr at 12.1 hrs of HRT(with N loading rate of $0.08\;Kg-N/m^3$-carrier/d). As a result, the application of the immobilized isolate appears a viable alternative to the nitrification-denitrification processes.
Bacterial strains exhibiting fibrinolytic activity were screened from traditional Korean soybean sauce. The Fibrinolytic activities of the various isolated microorganism were further examined and the superior strain YJ11-21 was selected for further analyses. Gene sequence analysis of 16S rDNA of the YJ11-21 strain revealed Bacillus licheniformis. Optimal culture conditions were investigated in order to maximize the production of the fibrinolytic enzyme by YJ11-21. Amongst the carbon sources tested, glucose was the most effective for enzyme production and amongst the nitrogen sources tested, yeast extract was seen to be the most effective. A one percent addition of NaCl to the medium resulted in the highest fibrinolytic activity. Interestingly, a 10% addition of NaCl resulted in a high activity together with a high cell growth rate. Therefore, YJ11-21 is speculated of being a halotolerant. The optimum pH and temperature for enzyme production were a pH of 9.0 and $30^{\circ}C$, respectively.
Lee Jung-Bok;Kim Dong-Geol;Choi Chung-Sig;Sohn Ho-Yong;Kim Jang-Eok;Kwon Gi-Seok
Microbiology and Biotechnology Letters
/
v.34
no.2
/
pp.174-179
/
2006
Bacterium capable of using biphenyl as a sole source of carbon and energy were isolated from soil, and based on the results of 16S rDNA sequence, strain BK10 identified as a Sphingobium yanoiktiyae. The optimum cultural conditions were as follows; $NH_4NO_3$ 1g, $K_2HPO_4$ 1g, $MgSO_4{\cdot}7H_2O$ 0.5g, $CaCO_3$ 0.2 g per 1 liter of distilled water. The Sphingobium yanoikuyae BK10 strain was completely utilized biphenyl in mineral salt media containing biphenyl at concentration 500 $\mu$g/ml of biphenyl as a sole carbon and energy source within 48 hours. Optimumal pH and temperature for biphenyl degradation and cell growth of strains were 6.0$\sim$8.0 and 20$\sim$50$^{\circ}C$, respectively. Especially, at 30$^{\circ}C$, cell-growth were higher than other temperature. Cell grown on biphenyl has been shown to have a higher removal rate for biphenyl than grown on sucrose. This study shows that Sphingobium yanoikuyae BK10 strain had a high biodegradation capability of biphenyl and can be simulate a candidate compounds the bioremediation of PCBs (Polychlorinated biphenyl) contaminant soil and water.
Hiroyuki Kokawa;Masayuki Shimada;Wang, Zhan-Jie;Yutaka S. Sato
Proceedings of the KWS Conference
/
2002.10a
/
pp.250-254
/
2002
Intergranular corrosion of austenitic stainless steels is a conventional and momentous problem during welding and high temperature use. One of the major reasons for such intergranular corrosion is so-called sensitization, i.e., chromium depletion due to chromium carbide precipitation at grain boundaries. Conventional methods for preventing sensitization of austenitic stainless steels include reduction of carbon content in the material, stabilization of carbon atoms as non-chromium carbides by the addition of titanium, niobium or zirconium, local solution-heat-treatment by laser beam, etc. These methods, however, are not without drawbacks. Recent grain boundary structure studies have demonstrated that grain boundary phenomena strongly depend on the crystallographic nature and atomic structure of the grain boundary, and that grain boundaries with coincidence site lattices are immune to intergranular corrosion. The concept of "grain boundary design and control", which involves a desirable grain boundary character distribution, has been developed as grain boundary engineering. The feasibility of grain boundary engineering has been demonstrated mainly by thermomechanical treatments. In the present study, a thermomechanical treatment was tried to improve the resistance to the sensitization by grain boundary engineering. A type 304 austenitic stainless steel was pre-strained and heat-treated, and then sensitized, varying the parameters (pre-strain, temperature, time, etc.) during the thermomechanical treatment. The grain boundary character distribution was examined by orientation imaging microscopy. The intergranular corrosion resistance was evaluated by electrochemical potentiokinetic reactivation and ferric sulfate-sulfuric acid tests. The sensitivity to intergranular corrosion was reduced by the thermomechanical treatment and indicated a minimum at a small roll-reduction. The frequency of coincidence-site-lattice boundaries indicated a maximum at a small strain. The ferric sulfate-sulfuric acid test showed much smaller corrosion rate in the thermomechanically-treated specimen than in the base material. An excellent intergranular corrosion resistance was obtained by a small strain annealing at a relatively low temperature for long time. The optimum parameters created a uniform distribution of a high frequency of coincidence site lattice boundaries in the specimen where corrosive random boundaries were isolated. The results suggest that the thermomechanical treatment can introduce low energy segments in the grain boundary network by annealing twins and can arrest the percolation of intergranular corrosion from the surface.
Two strains of pure lactic acid bacteria capable of forming both acid and slime were isolated from the kefir made of goat milk. The isolated strains observed by morphological and physiological properties, and their 16S rDNA partial sequence were identified as Streptococcus salivarius subsp. thermophilus(LFG-1) and Lactococcus lactis subsp. lacits(LFG-2) with over 99% homology. The optimum temperature of Str. salivarius subs. thermophilus LFG-1 for growth was $40-45^{\circ}C$, and its generation time was 40.6 minutes. The final pH of cultured broth by Str. salivarius subsp. thermophilus LFG-1 and the commercial strain Str. thermophilus Body-1 for 24hr at $37^{\circ}C$ were 4.30 and 4.55, respectively. The coagulative activity of Str. salivarius subsp. thermophilus LFG-1 was almost as strong as that of commercial strain Str. thermophilus Body-1. However, the LFG-2 strain showed lower coagulative activity than Str. thermophilus Body-1. The survival rate of lactic acid bacteria were between 22-29% in 0.3% bile extract. At pH 1.0 all of the bacteria were killed, and most of lactic acid bacteria died against pH 3.0. However, all lactic acid bacteria survived well at pH 4.5.
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