Eight strains producing bacterial cellulose (BC) were isolated from rotten fruits and traditionally fermented vinegars. One of the isolated strains from the rotten grape in Gwangju, Korea, maintained a relatively stable BC production in shaking cultures. This isolated strain proved to be Acetobacter xylinum, based on several biochemical and morphological tests. It was shown that the slant-baffled flask was more efficient than the conventional flask for the BC production in shaking cultures. To determine the most suitable carbon and nitrogen sources for the production of BC, various compounds were examined. Fructose was found to be the most effective carbon source with an optimal concentration of 2%. Mixed carbon source (glucose:fructose=1:3) was also better than glucose or fructose alone. Optimal nitrogen source, when basal medium was used, was 10% (v/v) com steep liquor (CSL). When com steep liquor was used with a mixed carbon source (glucose:fructose=1 :3),4% CSL exhibited the best BC production. Based on these results, a defined medium was developed for the BC production by Acetobacter xylinum KJ-1. When this medium was used under optimal culture conditions, the BC production was 7.2 g/1, which was approximately 3 times higher than that with the traditional HS medium.
The morphology of filamentous fungi closely correlates with the productivity in submerged culture. Using itaconic acid (IA) production by Aspergillus terreus as a research model, the quantitative relationship between the growth form of A. terreus and IA production was investigated. IA fermentation was scaled up from shake flasks to a 7 L stirred tank bioreactor based on the quantitative relationship. Our results demonstrated the following: (1) Three morphologies of A. terreus were formed by changing the inoculum level and shape of the flask. (2) Investigation of the effects of the three morphologies on broth rheology and IA production revealed the higher yield of IA on dry cell weight (DCW, IA/DCW) and yield of glucose on DCW (consumed glucose/DCW) were achieved during clump growth of A. terreus. (3) By varying the $KH_2PO_4$ concentration and culture temperature, the relationships between clump diameter and IA production were established, demonstrating that the yield of IA on DCW ($R^2$ = 0.9809) and yield of glucose on DCW ($R^2$ = 0.9421) were closely correlated with clump diameter. The optimum clump diameter range for higher IA production was 0.40-0.50 mm. (4) When the clump diameter was controlled at 0.45 mm by manipulating the mechanical stress in a 7 L fermentor, the yield of IA on DCW and yield of glucose on DCW were increased by 25.1% and 16.3%, respectively. The results presented in this study provide a potential approach for further enhancement of metabolite production by filamentous fungi.
Park, SungJin;Choi, Seunghoon;Park, MinJoo;Lee, O-Mi;Son, Hong-Joo
Journal of Environmental Science International
/
v.29
no.8
/
pp.819-826
/
2020
Production of Bacterial Cellulose (BC) by Gluconacetobacter sp. A5 was studied in shaken culture using different cost-effective carbon sources and its structural and mechanical properties were evaluated. Glycerol showed the highest level (7.26 g/l) of BC production, which was about three times higher than the yield in glucose medium. BC production depended not only on the decrease in pH, but also on the ability of Gluconacetobacter sp. A5 to synthesize glucose from different carbon sources and then polymerize it into BC. All BC produced from different carbon sources exhibited a three-dimensional reticulated structure consisting of ultrafine cellulose fibriles. Carbon sources did not significantly change the microfibrile structure of the resulting BC. BC produced from glucose medium had the lowest water-holding capacity, while BC from molasses medium had the highest. XRD data revealed that all BC were cellulose type I, the same as typical native cellulose. The crystalline strength of BC produced in glucose medium was the highest, and that in molasses medium was the lowest. Our results suggest that glycerol could be a potential low-cost substrate for BC production, leading to the reduction in the production cost, and also to produce BC with different mechanical properties by selecting appropriate carbon source.
Diabetes mellitus is a complex metabolic derangement with hyperglycaemia being the most characteristic symptom of diabetes. Hyperglycaemia can be caused by an increase in the rate of glucose production by the liver or by a decrease in the rate of glucose use by peripheral tissues. Impaired glucose transport is one of the major factors contributing to insulin resistance in type 2 diabetic patients. The ability of insulin to mediate tissue glucose uptake is a critical step in maintaining glucose homeostasis and in clearing the post-prandial glucose load. Glucose transport is mediated by specific carriers called glucose transporters (GLUTs). In this article, the functional importance and molecular mechanisms of insulin-induced glucose transport and development of hypoglycaemic agents which increase glucose transport are reviewed.
The maintenance of normal blood glucose levels at rest and during exercise is critical. The maintenance of blood glucose homeostasis depends on the coordination and integration of several physiological systems, including the sympathetic nervous system and the endocrine system. During prolonged exercise increased demand for glucose by contracting muscle causes to increase glucose uptake to working skeletal muscle. Increase in glucose uptake by working skeletal muscle during prolonged exercise is due to an increase in the translocation of insulin and contraction sensitive glucose transporter-4 (GLUT4) proteins to the plasma membrane. However, normal blood glucose level can be maintained by the augmentation of glucose production and release through the stimulation of liver glycogen breakdown, and the stimulation of the synthesis of glucose from other substances, and by the mobilization of other fuels that may serve as alternatives. Both feedback and feedforward mechanisms allow glycemia to be controlled during exercise. This review focuses on factors that control blood glucose homeostasis during prolonged exercise.
In order to control glucose concentration during fed-batch culture for antibiotic production, we applied so called “software sensor” which estimates unmeasured variable of interest from measured process variables using software. All data for analysis were collected from industrial scale cultures in a pharmaceutical company. First, we constructed an estimation model for glucose feed rate to keep glucose concentration at target value. In actual fed-batch culture, glucose concentration was kept at relatively high and measured once a day, and the glucose feed rate until the next measurement time was determined by an expert worker based on the actual consumption rate. Fuzzy neural network (FNN) was applied to construct the estimation model. From the simulation results using this model, the average error for glucose concentration was 0.88 g/L. The FNN model was also applied for a special culture to keep glucose concentration at low level. Selecting the optimal input variables, it was possible to simulate the culture with a low glucose concentration from the data sets of relatively high glucose concentration. Next, a simulation model to estimate time course of glucose concentration during one day was constructed using the on-line measurable process variables, since glucose concentration was only measured off-line once a day. Here, the recursive fuzzy neural network (RFNN) was applied for the simulation model. As the result of the simulation, average error of RFNN model was 0.91 g/L and this model was found to be useful to supervise the fed-batch culture.
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.
The metabolic patterns of C-1 and C-6-carbon atoms of glucose were observed in the tissue homogenates of the Ehrlich ascites tumor tissue which was incubated for 3 hours in the Dubnuff metabolic shaking incubator. $C^{14}-1-and\;C^{14}-6-glucose$ were used as tracers. The glucose media in which tissue homogenate was incubated was kept at a concentration of 200mg% glucose of carrier and appropriate amount of $C^{14}-1-or\;C^{14}-6-tracer$. At the end of 3 hour incubation, respiratory $CO_2$ samples trapped by alkaline which is placed in the tenter well of incubation flask were analyzed for the total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate, pyruvate and glycogen and calculations were made on the glucose consumption rate, pyruvate and lactate accumulation rates. The following results were obtained. Data obtained in each group are as follows: 1. In the tissue homogenate, which was incubated with $C^{14}-1-glucose as a substrate, total $CO_2$ production rate averaged $19.0{\pm}5.0{\mu}M/hr/gm$ and the mean specific activity of respiratory $CO_2$ was $840{\pm}296\;cpm/mgC.$ Relative specific activity (RSA) which means the fraction of $CO_2$ derived from medium $C^{14}-1-glucose$ to total $CO_2$ production rate was calculated by ratio of SA of respiratory $CO_2$ and medium $C^{14}-1-glucose.$ RSA was $14.3{\pm}5.0%,$ Accordingly actual $CO_2$ production rate from medium $C^{14}-1-glucose$ showed a mean value of $2.79{\pm}1.35\;{\mu}m$ of which amount was equivalent to the mean value of total glucose consumption rate $(RGDco_2)$, namely, $5.1{\pm}1.3%.$ Lactate and pyruvate appearance rates averaged $7.13{\pm}1.26\;and\;0.21{\pm}0.02{\mu}M/hr/gm,$ respectively. Assuming that these 3 carbon compounds appeared in the medium were derived from glucose, calculations were made that relative glucose disappearance rate into lactate $(RGD_L)$ was $38.0{\pm}5.4%\;and\;RGD_P$ was $1.23{\pm}0.03%.$ Therefore, about 43.3% of the total glucose consumed were accounted for by conversion into the respiratory $CO_2$, lactate and pyruvate. 2. In the second group, which was incubated with $C^{14}-1-glucose$ as a substrate, glucose consumption rate, lactate and pyruvate appearance rates showed almost the same order as the values of the $C^{14}-1-glucose$ substrate group. However, RSA was remarkably decreased showing a mean value of $1.02{\pm}0.13%.$ This fact means that the C-6 carbon of glucose take the minor part in the oxidative metabolism of glucose. The glycogen level in both substrate tissue homogenate showed less than 0.3% of tissue weight. These low value suggested that there was an inhibition of carbohydrate synthesis in the Ehrlich ascites tumor tissue. 3. The catabolic pathway of glucose in the tumor tissue were analyzed on the basis of Bloom's principle from the values of RSA. It was found that in the tumor tissue more than 90% of $CO_2$ derived from glucose were oxidized via the alternate pathway other than principal EMP-TCA cycle such as hexose monophosphate pathway (HMP). From the data described above, it was assumed that in the Ehrlich tumor tissue anaerobic glycolysis proceeds normally although, the oxidation of products of anaerobic glycolysis via the TCA cycle is inhibited resulting in the accumulation of lactate and almost all of oxidative energy from glucose is released by oxidative pathway such as HMP.
Kim, Mi-Sun;Moon, Kwang-Woong;Lee, In-Gu;Lee, Tae-Jin;Sung, Chang-Keun
Microbiology and Biotechnology Letters
/
v.27
no.1
/
pp.62-69
/
1999
Clostridium butyricum NCIB 9576 evolved hydrogen gas and produced various organic acids from glucose, lactose, starch, and glycerol. Total amount of hydrogen gas produced from 1 and 2% glucose were 630 and 950ml $H_2$/l-broth, respectively, for the first 24 hrs of incubation and the maximum hydrogen production rates were 42 and 94ml $H_2$/hr/1-broth, respectively. Teh initial pH 6.8 decreased to 4.2~4.5 during the first 12~16 hrs of fermentation when the pH was not controlled, resulting in ceasing the cell growth and hydrogen evolution and in degradation of 82 and 40% glucose after 24hrs of incubation from 1 and 2% glucose, respectively. When pH was controlled to 5.5, glucose was consumed completely and resulted in increasing hydrogen production approximately 38~50% compared to the experiments without the pH control. C. butyricum NCIB 9576 produced hydrogen gas approximately 644, 1,700 and 3,080 ml $H_2$/l-broth with 0.5, 1 and 2% lactose, respectively and the maximum hydrogen production rates were 41, 141 and 179ml $H_2$/hr/l-broth, respectively. All of the lactose added was degraded completely during fermentation even though pH was not controlled. C. butyricum NCIB 9576 produced 183 and 709ml $H_2$/l-broth with 0.1 and 0.5% starch for 48 hrs, respectively, when pH was not controlled. The maximum rates of hydrogen gas production were 43 and 186ml $H_2$/l-broth, respectively and 80~100% of starch added was fermented. Approximately 107ml $H_2$/l-broth was produced using 1% glycerol by C. butyricum NCIB 9576 and the pH was maintained higher than 6.1 during fermentation without pH control. The degradation of glucose, lactose, starch and glycerol by C. butyricum NCIB 9576 were affected by the pH of fermentation broth and the organic acids released during fermentation. The pH of feremtntation broth dropped to 4.2~4.6 after 12~14 hrs incubation when glucose was used as a substrate while pHs were maintained above pH 5 under the same experimental conditions when lactose, starch and glycerol were used. The organic solvents and acids produced during glucose fermentation were mainly ethanol, butyrate, acetate and a little of propionate, while butyrate was the main organic acids during the lactose, starch, and glycerol fermentation by C. butyricum NCIB 9576.
The effects of carbon sources for exopolysaccharide production during batch cultivation of an Enterobacter sp. isolated from the composter were investigated. The highest amount of exopolysaccharide was obtained when lactose was used as carbon source. Lactose in medium was converted into glucose and galactose. Glucose was metabolized fast and was completely consumed, but about $20\%$ of lactose was accumulated as galactose. On the other hand, enzyme activity was about $350\~450$ unit with the increase of lactose concentration. Thus, it was considered that the exopolysaccharide might be produced in the course of that lactose was hydrolyzed into glucose and galactose by $\beta-galactosidase$ with respect to that enzyme activity on lactose hydrolysis was accorded to the exopolysaccharide production. When glucose and galactose were added to lactose medium, respectively, it could be considered that glucose was as a repressor and galactose was as a inducer for $\beta-galactosidase$ synthesis even though the mechanisms were not elucidated. The increase of lactose concentration was almost ineffective to the specific growth rate $(0.133\~0.151\;hr^[-1})$ but showed the difference in the biomass content. The higher carbon source concentration, the more residual sugar remained. It was assumed that the optimum lactose concentration for exopolysaccharide production was $30\~70g/L.$ On the other hand, it was considered that the nitrogen acted as growth limiting nutrients to the cell growth. In the cases of 30 and 70 g/L of the fixed carbon concentrations, the increase of the nitrogen sources concentration caused a remarkable increase within the range of $0.059\~0.225\;hr^{-1}$ and $0.141\~0.237hr^{-1}$ of the specific growth rate, respectively, while there was no significant difference in biomass.
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