Various environmental factors such as pH, temperature and initial glucose concentration were investigated for enhancing cell growth in fermentations of Phellinus linteus WI-OOl, a producer of polysaccarides with potent anticancer activities. Optimal pH and temperature were around 5.5 and $28^{\circ}C$, respectively. Relatively little variation of pH was observed ranging between 5.5 and 6.5 during the whole fermentation period. Maximum cell concentration and specific growth rate were investigated in the media containing initial glucose concentrations of 0.5%, 1 %, 2%, 3% and 4%. High initial glucose concentration enhanced biomass production but showed negative effect on specific growth rate. In bioreactor experiments with various feeding strategies, increases of 28% and 42% in final cell concentration were obtaind as compared to conventional batch process, by adopting pulse and continuous supplement of 2% glucose solution, respectively.
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.
Lui, Zhen-Jia;Chu, Hung-Hsin;Wu, Yun-Chu;Yang, Shyi-Kuen
Asian-Australasian Journal of Animal Sciences
/
v.27
no.6
/
pp.841-846
/
2014
The present study was conducted to determine whether the two-step time-restricted feeding improves the fattening traits of one-step time-restricted feeding in geese. Thirty-six 8-wk-old geese were allotted into one of three groups. Group R1 (the 1-step restricted feeding group) was allowed access to feed for 2 h in the morning from 8 wk to 14 wk of age. Group R2 (the 2-step restricted feeding group) was treated as Group R1, but was additionally fed for 2 h in the afternoon from 12 wk to 14 wk of age. Group C (the control group) was fed ad libitum from 8 wk to 14 wk of age. Feed intake and body weight (BW) were recorded daily and weekly, respectively. At 14 wk of age, the blood samples were collected to determine the fasting plasma levels of glucose, triacylglycerols and uric acid before sacrifice. The results showed that daily feed intake (DFI) was lower, feed efficiency (FE) was higher in both Groups R1 and R2 than in Group C, and daily gain (DG) in Group R2 was higher than in Group R1 during the whole experimental period (p<0.05). Group R1 exhibited lower abdominal and visceral fat weights in carcass than did Group C (p<0.05), and Group R2 was in intermediate. The fasting plasma glucose levels in Group C were higher, and triacylglycerol levels in Group R1 were higher, compared with the other groups (p<0.05). It is concluded that time-restricted feeding in the fattening period not only increases FE but reduces DFI, and the additional meal during the late fattening period improves the DG without the expense of FE in geese.
Xylitol production from xylose and glucose was investigated using Candida tropicalis KFCC-10960. As glucose concentration in xylose medium was increased, ethanol production increased. However, xylitol production was maximum at glucose concentration of 10 g/l. The concentrated cells grown on xylose or glucose were inoculated in xylose medium. The specific activities of xylose reductase and xylitol dehydrogenase, and xylitol production in concentrated cells grown on glucose were the same as those in concentrated cells grown on xylose. The results suggested that cells grown on glucose had the same xylitol producing activity as those grown on xylose. By feeding glucose in xylose medium, cell growth was achieved from glucose and xylitol production was obtained from xylose. By using this technique, a final xylitol concentration of 261 g/l was achieved from 300 g/l xylose in 41 hours which corresponded to a xylitol yield from xylose of 87% and a xylitol productivity of 6.37 g/1-h.
A glucose clamp technique was used to compare dietary starch (ST), starch plus sucrose (ST+SU) and sucrose (SU) with regard to the effect on tissue responsiveness and sensitivity to insulin in intact adult male goats. The goats were fed diets containing 1.2 times of ME and CP for maintenance requirements twice daily for 21 d. Of the energy intake, 30% was offered with ST, ST+SU or SU for the respective diets, and 70% as alfalfa hay, ground corn and ground soybean meal at the respective weight ratio of 1, 1, and 0.3 for all diets. Tissue responsiveness and sensitivity to insulin were evaluated using a hyperinsulinemic euglycemic clamp technique with four levels of insulin infusion beyond 13 h after feeding. The concentrations of plasma metabolites and insulin were also determined at 3, 6 and 13 h after feeding to evaluate the effects of different carbohydrates on metabolic states in the body. Plasma glucose concentration was higher (p = 0.01) for SU diet than for ST and ST+SU diets. Increasing SU intake decreased (p<0.01) plasma acetate concentration across the time. At 3 h but not 6 and 13 h after feeding, high lactate (p = 0.01), and non-significant high propionate (p = 0.14) and low urea nitrogen (p = 0.19) concentrations were observed in plasma on SU compared with ST and ST+SU diets. Plasma insulin concentration was not different (p = 0.44) between ST and SU fed animals. In the glucose clamp experiment, considering the effects on the maximal glucose infusion rate (tissue responsiveness to insulin, p = 0.54) and the plasma insulin concentration at half-maximal glucose infusion rate (insulin sensitivity, p = 0.54), SU was not different from ST. It is concluded that SU may not be greatly different from ST with regard to the effect on tissue responsiveness and sensitivity to insulin in adult goats when fed twice daily as part of a high-concentrate diet. The possible greater effects of SU on plasma metabolites concentrations at 3 h than at 6 and 13 h after feeding suggest that a lack of persistency of SU effects during the postfeeding period may be associated with the poor response to SU in insulin action.
Fed-batch fermentation was used to produce the high concentrations of poly-$\beta $-hydroxybutyrate (PHB) and poly-$\beta $-(hydroxybutyrate-co-hydroxyvalerate) (PHB/V). Specific growth rate ($\mu $), yield of cell from glucose (Y$_{x/s}$) were calculated from the two samples in 3 to 5 hours of interval and they were reflected on the determination of glucose feeding rate to maintain the glucose concentration at around 10 g/l in the culture broth. PHB was accumulated after the nitrogen became limited at 60 g/l of dry cell weight by changing ammonia water to 4N-NaOH solution. As results, the final dry cell weight (DCW) of 170 g/l, PHB of 115 g/l were obtained in 50 hours and the overall productivity was 2.4 g/l$\cdot $h. After PHB accumulation, cosubstrate of glucose and propionic acid (PA) was fed to accumulate PHB/V. But, PA feeding rate was decreased from 3 g/l$\cdot $h to 1 g/l$\cdot $h to prevent PA from accumulating to high level in the broth, which is very inhibitory to the cells. As results, DCW, PHB and PHV were 147.5 g/l, 90 g/l and 8 mole % of hydroxyvalerate, respectively.
Effect of glucose addition to xylose medium on xylitol production was investigated by using Candida parapsilosis ATCC 21019 mutant. With increasing the ratio of glucose to xylose in total amount of 50 g/l as glucose and/or xylose, xylitol production was decreased but ethanol and glycerol production were increased. Ethanol and glycerol concentration were maxmum in 10 g/l of xylose and 40 g/l of glucose medium as 21.5 g/l and 3.6 g/l, respecti- vely. No xylitol was formed in glucose medium without xylose because xylitol could be not produced from glucose. With increasing the ratio of glucose to xylose, the activity of xylose reductase which converted xylose to xylitol were decreased. The activities of xylitol dehydrogeiiase which converted xylitol to xylulose and then cell materials were found to be constant regardless of the ratio of glucose to xylose. This results indicated that glucose addition to xylose medium on cell growth was not affected. In order to prevent the inhibitory effect of glucose on xylitol production, glucose in a fermentor was fed with low concentration and then ethanol and glycerol was critically decreased and the xylitol yield from xylose of the culture with glucose feeding was recovered the almost same as that with only 50 g/l of xylose. However, the xylitol yield from total sugars (xylose and glucose) was decreased and glucose was not contributed to xylitol production. Therefore, the fermentation at high concentration of xylose without glucose was carried out. A final xylitol concentration of 242 g/l which corresponding 80.7% of xylitol yield was obtained from 300 g/l of xylose for 273 hours.
Kim, Myoung-Dong;Lee, Woo-Jong;Park, Kwan-Hwa;Rhee, Ki-Hyeong;Seo, Jin-Ho
Journal of Microbiology and Biotechnology
/
v.12
no.2
/
pp.273-278
/
2002
Two-step fed-batch fermentations were carried out to overproduce Bacillus licheniformis maltogenic amylase (BLMA) in recombinant Escherichia coli. The first step was to increase the cell mass by controlling the feeding of a glucose solution, while the second step was designed to improve the amylase expression efficiency by supplementing organic nitrogen sources. The linear gradient feeding method was successfully adopted to maintain the glucose concentration below 0.2 g/l during the fed-batch mode, as effectively minimizing acetic acid formation. When the dissolved oxygen (DO) level became limiting, an accumulation of acetic acid and drastic decrease in specific BLMA productivity were observed. Glucose and organic nitrogen sources consisting of yeast extract and casein hydrolysate were simultaneously supplied in the pH-stat mode to further increase the specific BLMA expression efficiency. An organic nitrogen source consisting of 200 g/1 yeast extract and 100 g/1 casein hydrolysate was found to be the best among the various combinations tested. The feeding of an organic nitrogen source in the second-step fed-batch period was highly beneficial in enhancing the BLMA production. The optimized two-step fed-batch culture resulted in 78 g/l maximum dry cell mass and 443 U/ml maximum BLMA activity, corresponding to 1.5-fold increase in the dry cell mass and 3.7-fold enhancement in BLMA production, compared with the simple fed-batch fermentation.
Ralstonia eutropha NCIMB 11599 and ATCC 17699 were grown, and their productions of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] compared. In flask cultures of R. eutropha NCIMB 11599, cell concentration, P(3HB-co-4HB) concentration and polymer content decreased considerably with increases in the ${\gamma}-butyrolactone$ concentration, and the 4HB fraction was also very low (maximum 1.74 mol%). In fed-batch cultures of R. eutropha NCIMB 11599, glucose and ${\gamma}-butyrolactone$ were fed as the carbon sources, under a phosphate limitation strategy. When glucose was fed as the sole carbon source, with its concentration controlled using an on-line glucose analyzer, 86% of the P(3HB) homopolymer was obtained from 201g/L of cells. In a two-stage fed-batch culture, where the cell concentration was increased to 104g/L, with glucose fed in the first step and constant feeding of ${\gamma}-butyrolactone$, at 6g/h, in the second, final cell concentration at 67h was 106g/L, with a polymer content of 82%, while the 4HB fraction was only 0.7mol%. When the same feeding strategy was applied to the fedbatch culture of R. eutropha ATCC 17699, where the cell concentration was increased to 42 g/L, by feeding fructose in the first step and ${\gamma}-butyrolactone$ (1.5g/h) in the second, the final cell concentration, polymer content and 4HB fraction at 74h were 51g/L, 35% and 32 mol%, respectively. In summary, R. eutropha ATCC 17699 was better than R. eutropha NCIMB 11599 in terms of P(3HB-co-4HB) production with various 4HB fractions.
Park, Kum-Ju;Hong, Seong-Eui;Do, Myoung-Sool;Hyun, Chang-Kee
Korean Journal of Pharmacognosy
/
v.33
no.1
s.128
/
pp.21-28
/
2002
Antidiabetic effects of the acid hydrolysate of silk fibroin were investigated by oral administration to animal models for diabetes mellitus, Fibroin protein was extracted from cocoon and digested to peptides of low-molecular weight range (mainly below 3,000) and amino acids by acid hydrolysis, Feeding of the fibroin hydrolysate resulted in a significant recovering effect on reduction of body weight gain and a lowering effect on blood glucose gain in streptozotocin-induced diabetic Sprague Dawley rats (STZ rats) which were used as an insulin-dependent diabetic animal model. But the body weight and blood glucose level in C57BL/KsJ-db/db mice (db/db mice), an non-insulin-dependent diabetic animal model, were not changed significantly by the feeding, On the other hand, plasma leptin levels increased according to increased feeding amount of the hydrolysate in STZ rats and db/db mice in common, It was concluded from the results that the fibroin hydrolysate might stimulate the insulin secretion by recovering or activating pancreatic ${\beta}$ cells and result in the increased plasma leptin level. It was also deduced that the antidiabetic improvements in body weight and blood glucose gain in STZ were thought to be due to the increased insulin secretion, but in db/db mice of which the diabetic symptoms were caused by insulin resistance, the stimulated secretion of insulin was unlikely to be able to change body weight and blood glucose level significantly.
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