ε-Poly-L-lysine (ε-PL) is a homopolymer of L-lysine molecules connected between the epsilon amino and alpha carboxyl groups. This polymer is currently used as a natural preservative in food. Insufficient biomass is a major problem in ε-PL fermentation. Here, to improve cell growth and ε-PL productivity, various nitrogen-rich nutrients were supplemented into flask cultures after 16 h cultivation, marking the onset of ε-PL biosynthesis. Yeast extract, soybean powder, corn powder, and beef extract significantly improved cell growth. In terms of ε-PL productivity, yeast extract at 0.5% (w/v) gave the maximum yield (2.24 g/l), 115.4% higher than the control (1.04 g/l), followed by soybean powder (1.86 g/l) at 1% (w/v) and corn powder (1.72 g/l) at 1% (w/v). However, supplementation with beef extract inhibited ε-PL production. The optimal time for supplementation for all nutrients examined was at 16 h cultivation. The kinetics of yeast-extract-supplemented cultures showed enhanced cell growth and production duration. Thus, the most commonly used two-stage pH control fed-batch fermentation method was modified by omitting the pH 5.0-controlled period, and coupling the procedure with nutrient feeding in the pH 3.9-controlled phase. Using this process, by continuously feeding 0.5 g/h of yeast extract, soybean powder, or corn powder into cultures in a 30 L fermenter, the final ε-PL titer reached 28.2 g/l, 23.7 g/l, and 21.4 g/l, respectively, 91.8%, 61.2%, and 45.6% higher than that of the control (14.7 g/l). This describes a promising option for the mass production of ε-PL.
Kim, Ki-Hyuk;Moon, Hye-Na;Noh, Yun-Hye;Yeo, In-Kyu
Development and Reproduction
/
v.24
no.1
/
pp.19-30
/
2020
The pH of water is one of the main environmental factors exerting selective pressure on marine and freshwater organisms. Here, we focus on the influence of pH on an organism's ability to maintain homeostasis and investigate the effects of acidification on immunity-related genes and osmotic pressure during early development of the olive flounder, Paralichthys olivaceus. The aim of our study was to determine the influence of various pH levels on the fertilized eggs and larvae of P. olivaceus. Gametes of P. olivaceus were artificially introduced and the resulting fertilized eggs were incubated at pH 4.0 (low), 6.0, and 8.0 (equivalent to natural sea water; control). We found that all eggs sank from the water column at pH 4.0. After 38 h, these eggs showed slow development. Hatching occurred more slowly at pH 4.0 and 6.0 and did not occur at all at pH 4.0. Result of gene expression, caspase and galectin-1 were expressed from the blastula to pre-hatch stages, with the exception of the two-cell stage. HSP 70 was also steadily expressed at all pH levels over the five days. The osmolality of fertilized eggs differed marginally at each stage and across pH levels. So, this results demonstrates that low pH level is detrimental to P. olivaceus fertilized eggs.
In this paper, the robust superconductor flywheel energy storage system(SFESS) controller using $H_{\infty}$ control theory was designed to damp low frequency oscillation of power system. The main advantage of the $H_{\infty}$ controller is that uncertainties of power system can be included at the stage of controller design. Both disturbance attenuation and robust stability for the power system were treated simultaneously by using mixed sensitivity $H_{\infty}$ problem. The robust stability and the performance for uncertainties of power system were represented by frequency weighted transfer function. To verify control performance of proposed SFESS controller using $H_{\infty}$ control, the closed loop eigenvalue and the damping ratio in dominant oscillation mode of power system were analyzed and nonlinear simulation for one-machine infinite bus system was performed under disturbance for various operating conditions. The results showed that the proposed $H_{\infty}$ SFESS controller was more robust than conventional power system stabilizer (PSS).
To determine effects of simulated acid rain on the growth and yield of rice, soybean and sesame, and on the soil acidity, two cultivars of rice, soybean and sesame were exposed twice a week to one of four simulated rain acidities[pH 2. 6, 3. 6, 4. 6 and 5. 6(control)]. The plants were protected from ambient rain by a polyethylene film cover. No visible damage due to the simulated acid rain was observed at any stage of growth for either of the crops. In rice, heading date, plant height and yield components except ripening ratio were not significantly affected by the pH ofsimulated rain, but the ripening ratio and rough rice yield linearly decreased as the pH of simulated rain was decreased. In soybean, maturing date and yield components except 100 seed weight were not significantly influenced by the pH of simulated rain, but the 100 seed weight and yield linearly decreased as the pH of simulated rain was decreased. Seed yield and the other agronomic characteristics of sesame were not significantly affected by the pH of simulated rain. Soil pH after cropping rice, soybean and sesame decreased as pH of simulated rain was decreased.
The present study was carried out to investigate the localization and isozyme patterns of acid phosphatase and alkaline phosphatase in metacercariae and in adults of F. seoulensis by enzyme-histochemistry method and electrophoresis. Acidphosphatase showed a strong activity at pH 5 in the intestinal caecum of adults, but showed no reactions in the nonsubstrate control and in the inhibitor-treated control. Alkaline phosphatase showed a strong activity at pH 8 in the intestinal caecum and the tribocytic organ of adults, and in the intestinal caecum and in the genital anlagen of metacercariae. In non-denature PAGE, ten bands of protein fraction from the extracts of metacercariae and twenty-two bands from adults were detected. In denature PAGE, two protein bands having molecular weights of 192 kDa and 123 kDa were detected in the metacercariae, but absent from adult stage. In adults, protein fractions of 27.5 kDa, 24.5 kDa, 21.4 kDa, 18 kDa, 16 kDa and 15 kDa were detected. In non-denature PAGE, isozymes of acid phosphatase showed the most strong activity at pH 5, whereas no activity was shown at pH 2 and pH 7. One isozyme 85 kDa, 73 kDa and 62 kDa) in adults.
For general nonlinear processes, it is difficult to control with a linear model-based control method and nonlinear controls are considered. Among the numerous approaches suggested, the most rigorous approach is to use dynamic optimization. Many general engineering problems like control, scheduling, planning etc. are expressed by functional optimization problem and most of them can be changed into dynamic programming (DP) problems. However the DP problems are used in just few cases because as the size of the problem grows, the dynamic programming approach is suffered from the burden of calculation which is called as 'curse of dimensionality'. In order to avoid this problem, the Neuro-Dynamic Programming (NDP) approach is proposed by Bertsekas and Tsitsiklis (1996). To get the solution of seriously nonlinear process control, the interest in NDP approach is enlarged and NDP algorithm is applied to diverse areas such as retailing, finance, inventory management, communication networks, etc. and it has been extended to chemical engineering parts. In the NDP approach, we select the optimal control input policy to minimize the value of cost which is calculated by the sum of current stage cost and future stages cost starting from the next state. The cost value is related with a weight square sum of error and input movement. During the calculation of optimal input policy, if the approximate cost function by using simulation data is utilized with Bellman iteration, the burden of calculation can be relieved and the curse of dimensionality problem of DP can be overcome. It is very important issue how to construct the cost-to-go function which has a good approximate performance. The neural network is one of the eager learning methods and it works as a global approximator to cost-to-go function. In this algorithm, the training of neural network is important and difficult part, and it gives significant effect on the performance of control. To avoid the difficulty in neural network training, the lazy learning method like k-nearest neighbor method can be exploited. The training is unnecessary for this method but requires more computation time and greater data storage. The pH neutralization process has long been taken as a representative benchmark problem of nonlin ar chemical process control due to its nonlinearity and time-varying nature. In this study, the NDP algorithm was applied to pH neutralization process. At first, the pH neutralization process control to use NDP algorithm was performed through simulations with various approximators. The global and local approximators are used for NDP calculation. After that, the verification of NDP in real system was made by pH neutralization experiment. The control results by NDP algorithm was compared with those by the PI controller which is traditionally used, in both simulations and experiments. From the comparison of results, the control by NDP algorithm showed faster and better control performance than PI controller. In addition to that, the control by NDP algorithm showed the good results when it applied to the cases with disturbances and multiple set point changes.
This study was to examine whether Hanwoo IVM/IVF/IVC blastocyst can be successfully survived in vitro/in vivo after vitrification and one-step dilution. For vitrification, blastocysts were serially exposed in glycerol (G) and ethylene glycol(EG) mixtures[10% (v Iv) G for 5 min., 10% G plus 20% EG (v/v) for 5 min., and 25% G plus 25% EG (v/v) for 30 sec.] which is diluted in 10% FBS added D-PBS. And then they were loaded in the straw, placed in cold nitrogen vapor for 3 min. and plunged into L$N_2$(-196$^{\circ}C$). One-step dilution within the straw was done in $25^{\circ}C$ and 36$^{\circ}C$ water for about 5 min. and 3 min., respectively. Recovered embryos after one-step dilution were cultured in cumulus cell mono-layered drop for 48 h or were transferred into recipient cows. When the embryo survival in vitro was assessed to re-expanded and hatched rates at 24 hand 48 h after one-step dilution, the results of vitrified group (85.4, 43.8%) was high, although these results were significantly lower than normal development (100.0, 63.3%) of control group, respectively (P<0.001, P<0.05). When in vitro survival of vitrified groups according to developmental stage was compared, the results of fast developed embryos (expanded blastocyst and early hatching blastocyst stage) were significantly higher than those of delayed developed one (early blastocyst stage) after one-step dilution (early hatching: 88.0, 48.0%: expanded: 81.1, 45.3%; early: 66.7, 14.3%) (P<0.05). Also, in case of in vitro survival of vitrified groups according to embryo age (day 7, 8 and 9), when embryo age was younger, in vitro survival was significantly higher (day 7: 67.3, 34.5%; day 8: 76.9, 40.7%; day 9: 60.9, 23.9%)(P<0.05). Finally, when in vivo development potential of vitrified and one-step diluted Hanwoo blastocysts was examined, 4 of 8 recipient (50%) cows became confirmed pregnant. These results demonstrated that our vitrification and one-step dilution technique can be applied easily and effectively on field trial without the equipment and embryological skills required for conservative dilution and transfer.
To compare the tolerance of crops to acid rain at different growth stages a simulated acid rain (SAR) of pH 2.7 was applied to rice, soybean, and hot pepper from vegetative growth stage to harvest (Veget.-Harvest) and from reproductive growth stage to harvest (Reprod.-Harvest). Visual damages of crops by SAR were greater in the order of rice < hot pepper < soybean and greater at Veget.-Harvest than at Reprod.-Harvest treatment. Chlorophyll content of all crops was greater in the order of Veget.-Harvest < Reprod.-Harvest treatment < control, but photosynthetic activity was not affected by SAR treatments. Nitrogen concentration and uptake of rice plants at harvest were similar among SAR treatments, but those of soybean and hot pepper were greater at Veget.-Harvest treatment than at Reprod.-Harvest treatment or control. Sulfur concentration of all crops was not affected by SAR treatments, but total sulfur uptake of soybean was greater in SAR treatments than untreated control. Grain yield of rice and soybean was not affected by SAR although grain fertility, percent ripened grains, and 1,000-grain weight of rice at Veget.-Harvest treatment were lower compared with Reprod.-Harvest treatment or control. Fruit dry weight of hot pepper was greater in the order of Veget.-Harvest < Reprod.-Harvest < control due to decreased fruit number per plant and average fruit weight. At one time application of SAR at flowering stage, brown spots were observed on the spikelets of rice at below pH 2.3. Petals of soybean and hot pepper were wilted at pH below 1. 7 and 2.0, respectively, but fruit setting was not affected by the pH of the SRA.
The aim of this study was to evaluate the effect of incorporating wet sorghum distiller's grains (WSDG) as part of their diet on the lactating performance of dairy cows. Twenty-seven Holstein milking cows were selected, all in the early lactating stage, with an average weight of 550 kg, and producing an average of 30 kg of milk daily. The cows were divided into three groups according to milk yield and lactation and were fed different total mixed rations. The diets were formulated according to NRC (1989) recommendations in three rations to (1) control diet, (2) 15% WSDG diet and (3) 30% WSDG diet. The three different diets were all formulated as iso-nitrogen and iso-energetic diets. After one week adaptation period, the experimental feeding was conducted for 8 weeks. Three ruminal cannulated cows were also examined in order to investigate ruminal fermentation of the three total mixed rations. The results showed that the milk yield, as corrected to the 4.0% fat standard, had no significant difference among the control, 15% WSDG and 30% WSDG treatment groups (p>0.05). The daily dry matter intake of the control group was higher than the other groups (p<0.05). with respect to milk composition, milk fat, milk protein and total solids, there was no significant difference among the treatment groups (p>0.05). The energy efficiency of the 30% WSDG group were significantly higher than the other treatment groups (p<0.05). Ruminal pH value showed no difference among the treatment groups (p<0.05). Ammonia-nitrogen concentration in the control group was higher than the other treatment groups (p<0.05). The concentration of total ruminal volatile fatty acid was similar in all three dietary groups.
This study was conducted to examine the viability of nuclear transfer bovine embryos following embryo transfer. Donor embryos were treated with nocodazole to arrest their cell-cycle-stage at mitotic(M) phase. After releasing from nocodazole blastomeres were separated and transferred into the enucleated oocytes(BC), or cultured in medium with aphidicolin. Freshly cleaved blastomeres within 1.5h after cleavage(AC) and non-cleaved ones up to 3h after releasing from nocodazole(NC) were transferred into the enucleated oocytes. Blastocysts derived from nuclear transfer were transferred to Day 7~8 recipient cows. Some blastocysts were vitrified and thawed before embryo transfer. Developmental rates to the blastocyst stage were higher in AC(18.1%, P<0.05) than BC(8.6%) and NC(5.1%). Blastocyst development slightly enhanced with aphidicolin(1~2$\mu\textrm{g}$/ml) treatment(16.9~22.6%) compared to non treated control(11.1%). Survival rate fo vitrified nuclear transfer embryos after thawing was 75%(24/32). Twnety-three vitrified nuclear transfer embryos and 3 fresh ones were transferred to 23 recipients, 6 heads were pregnant and 1 male calf(24 kg) was born from a recipient cow recevied one vitrifiedthawed nuclear transfer embryo at 277 days after embryo transfer. This result suggests that the nuclear transfer embryos can developed to term after vitrification andembryo transfer.
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