• Journal of Microbiology and Biotechnology
• /
• v.16 no.3
• /
• pp.374-380
• /
• 2006

In this study, glucose, sucrose, and dextrin were found to be better carbon sources for the production of pullulan by Aureobasidium pullulans HP-2001. Maximal production of pullulan with 200 g/l sucrose as a carbon source was 54.2 g/l. The highest yield of pullulan from sucrose was 0.40, when the sugar concentration was 100 g/1. Optimal conditions for the continuous production of pullulan by A. pullulans HP-2001 in a 7-1 bioreactor were determined by studying the effects of composition of feed solution, dilution rate, and concentration of sucrose in the feed solution. Pullulan concentration and productivity with 100 g/l glucose and 2.5 g/l yeast extract were 38.1 g/l and 0.53 g/l h for 72 h, respectively, in a batch culture of A. pullulans HP-2001. When the substituted medium contained 100 g/l sucrose, 2.5 g/l yeast extract, and mineral salts, which is the same composition as the medium for the production of pullulan, the pullulan concentration and productivity were 74.9 g/l and 0.55 g/l h for 120 h, respectively. The production of pullulan at the steady state increased with a dilution rate up to 0.015/h, and its concentration was 78.4 g/l with a weight average molecular weight ($M_w$) of $4.0{\times}10^5$. Unlike a batch culture, however, the decline of the $M_w$ and the number average molecular weight ($M_n$) of pullulan was not found in the continuous culture of A. pullulans HP-2001. When the concentration of sucrose in the feed solution was 200 g/l, 113.5 g/l of pullulan was obtained at the steady state. The steady state was maintained longer in the continuous culture fed with the feed solution containing 200 g/l sucrose than when fed with the feed solutions containing either 100 or 150 g/l sucrose.

• Microbiology and Biotechnology Letters
• /
• v.45 no.2
• /
• pp.149-154
• /
• 2017

To increase the efficiency of esterase production by Bacillus, high cell-density culture of recombinant Escherichia coli through fed batch fermentation was tested. Cells were cultured to $OD_{600}$ of 76 (35.8 g/l DCW) with dissolved oxygen level controlled to least above 30% air saturation by supplying pure oxygen. Cells were cultured to an $OD_{600}$ of 90 (42.4 g/l DCW) with glucose feeding controlled to at least 1 g/l. However, the cells reached stationary phase at the late stage of culture, despite glucose being supplied. Cells were cultured to an $OD_{600}$ of 185 (87.3 g/l DCW) by supplying additional medium with fortified yeast extract. To increase the productivity of the recombinant protein, cell growth and esterase productivity based on induction time were evaluated. Late exponential phase induction for esterase production in fed batch fermentation resulted in maximum optical density $OD_{600}$ of 190 (89 g/l DCW) and maximum esterase activity of 1745 U/l, corresponding to a 5.8-fold enhancement in esterase production, compared to the early exponential phase induction. In this study, we established fermentation methods for achieving maximum production of Bacillus-derived esterase by optimizing IPTG induction time in high-cell density culture by supplying pure oxygen and a nitrogen source.

• Korean Journal of Microbiology
• /
• v.37 no.1
• /
• pp.92-99
• /
• 2001

High-cell-density cultivation of Ralstonia eutopha KHB 8862 by fed-batch fermentation in a 200 l pilot plant was carried out for the mass production of poly(3-hydroxybutyrate) (PHB). After 80 h of cultivation, the dry cell weight (DCW), PHB concentration, and PHB yield from fructose syrup reached 168 g/l, 74%DCW, and 0.27 (w/w), respectively, resulting in a productivity of 1.6 g of PHB/L/h. Based on these results, the PHB production cost from bacterial fermentation was analyzed and economic evaluation was performed. In the case of new investment being implemented or not, the production cost of PHB was US$ 3.15/kg and US$ 2.41/kg, respectively. PHB productivity and PHB yield on a carbon substrate were both important factors to be optimized. The increase of PHB yield on a carbon sources significantly decreased the PHB production cost but the increase in productivity had a relatively slight effect on the decrease in PHB production cost because the cost of carbon sources (37%) for PHB was larger in proportion to total cost than the depreciation cost (17%). These results suggest that the increased PHB yield from carbon sources and the development of new cheaper substrates would be more effective in decreasing PHB production cost than the increase in productivity. It was demonstrated that PHB is not in competition with consumable plastics such as PET in present market. Therefore, it is essential to lower production cost to be used as a bulk product and desirable to develop new application fields for PHB such as biomedical and cosmeceuticals.

• Food Science and Preservation
• /
• v.22 no.1
• /
• pp.100-107
• /
• 2015

This study was conducted to develop a high-acidity vinegar production (over TA 10%) technology using grape juice to reduce the importation of high-acidity vinegar. The manufacturing condition's effect on the quality of high-acidity vinegar using pure grape juice without the addition of other nutrients for fermentation was investigated. Twelve percent acidity in vinegar was obtained from grape wine with 6% alcohol content. The acetic-acid yield from grape wine decreased when the wine's initial alcohol content was high, which extended the induction time. The pH value was similar in all the treatment groups. The sugar content of the 1st-stage fermentation (1st AAF) was proportional to the initial alcohol content whereas in the 2nd-stage fermentation (2nd AAF), the sugar content was highest in the 6%-alcohol treatment. The major organic acids of the high-acidity grape vinegar included tartaric acid, malic acid, and citric acid. The acid content of the high-acidity initial alcohol group was higher than that of the low-acidity initial alcohol group due to the alcohol content added by the fed-batch and acetic-acid yield difference. The ethyl alcohol content was 364~6,091 ppm (the main alcohol while the others had only traces in all the groups). In conclusion, it was possible to manufacture 12% high-acidity vinegar without the addition of an external nutrient source to grape wine containing 6% initial alcohol content. Finally, a complementary study will be required to shorten the fermentation period through the fed-batch-style addition of alcohol for the purpose of industrialization.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.1
• /
• pp.146-152
• /
• 2010

Streptomyces clavuligerus NRRL3585 produces a clinically important $\beta$-lactamase inhibitor, clavulanic acid (CA). In order to increase the production of CA, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene (gap) was deleted in S. clavuligerus NRRL3585 to overcome the limited glyceraldehyde-3-phosphate pool; the replicative and integrative expressions of ccaR (specific regulator of the CA biosynthetic operon) and claR (Lys-type transcriptional activator) genes were transformed together into a deletion mutant to improve clavulanic acid production. We constructed two recombinant plasmids to enhance the production of CA in the gap1 deletion mutant of S. clavuligerus NRRL3585: pHN11 was constructed for overexpression of ccaR-claR, whereas pHN12 was constructed for their chromosomal integration. Both pHN11 and pHN12 transformants enhanced the production of CA by 2.59-fold and 5.85-fold, respectively, compared with the gap1 deletion mutant. For further enhancement of CA, we fed the pHN11 and pHN12 transformants ornithine and glycerol. Compared with the gap1 deletion mutant, ornithine increased CA production by 3.24- and 6.51-fold in the pHN11 and pHN12 transformants, respectively, glycerol increased CA by 2.96- and 6.21-fold, respectively, and ornithine and glycerol together increased CA by 3.72- and 7.02-fold, respectively.

• KSBB Journal
• /
• v.20 no.6
• /
• pp.443-446
• /
• 2005

Hydrogen fermentation from food waste was attempted at different hydraulic retention time(HRT, 18-42 h). A continuous reactor fed with ground, alkali-treated and diluted food waste(average VS 4.4%) exhibited stable hydrogen production during 126 days. Hydrogen production depended on HRT, resulting in the maximum values of 25.8 mL $H_2/g\;VS_{added}$, 0.36 mol $H_2/mol\;hexose_{added}$ and 0.91 L $H_2/L/d$ at HRT 30 h. n-Butyrate and isopropanol production increased with hydrogen production increased, while acetate production decreased. The fermentation efficiency ranged from 53.3 to 65.7%, which implied that hydrogen fermentation would substitute conventional acidogenesis of food waste.

• Asian-Australasian Journal of Animal Sciences
• /
• v.31 no.10
• /
• pp.1598-1603
• /
• 2018

Objective: The goal of this study was to evaluate kenaf as a roughage source in vitro and its effects on meat quality of Hanwoo (Korean native) cattle. Methods: Three roughage materials, rice straw silage, ryegrass silage, and kenaf silage, were tested in a batch culture and feeding trial. Rumen fermentation parameters, including gas, pH, volatile fatty acid (VFA), and ammonia were analyzed. In the feeding trial, Hanwoo steers ($373.5{\pm}5.1kg$, n = 36, 11 month of age) were divided into three feeding groups (n = 12 each). Animals were fed with each silage and concentrate until the fattening stage. Results: Crude protein, ether extract, and non-structural carbohydrates were greater in kenaf silage. Total gas production was higher in ryegrass silage, followed by kenaf silage and rice straw silage (p<0.05). Total VFA and individual VFA (acetate, propionate, and n-butyrate) were greater in kenaf silage than rice straw silage (p<0.05). In vitro dry matter digestibility showed a similar trend to that of total gas and VFA production; it was higher in ryegrass silage and lower in rice straw (p<0.05). Throughout the feeding trial, the rice straw silage group showed significantly greater average daily gain than did the others (p<0.05). The feed conversion ratio in the group fed kenaf silage was significantly greater than that of others (p<0.05). No significant differences were observed in yield or quality traits, including carcass weight, ribeye area, backfat thickness, and scores for marbling, meat color, and fat color (p>0.05). Conclusion: The results indicated that no negative effects on growth performance and carcass characteristics occurred across treatments. Therefore, kenaf could be substituted for rice straw, which is most widely used as a roughage source in Korea.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.10
• /
• pp.1511-1520
• /
• 2019

Objective: The present study was conducted to select a plant oil without inhibitory effects on rumen fermentation and microbes, and to determine the optimal supplementation level of the selected oil in a series of in vitro studies for dietary application. Then, the selected oil was evaluated in a feeding study using Thai crossbred beef cattle by monitoring growth, carcass, blood and rumen characteristics. Methods: Rumen fluid was incubated with substrates containing one of three different types of plant oil (coconut oil, palm oil, and soybean oil) widely available in Thailand. The effects of each oil on rumen fermentation and microbes were monitored and the oil without a negative influence on rumen parameters was selected. Then, the dose-response of rumen parameters to various levels of the selected palm oil was monitored to determine a suitable supplementation level. Finally, an 8-month feeding experiment with the diet supplemented with palm oil was carried out using 12 Thai crossbred beef cattle to monitor growth, carcass, rumen and blood profiles. Results: Batch culture studies revealed that coconut and soybean oils inhibited the most potent rumen cellulolytic bacterium Fibrobacter succinogenes, while palm oil had no such negative effect on this and on rumen fermentation products at 5% or higher supplementation level. Cattle fed the diet supplemented with 2.5% palm oil showed improved feed conversion ratio (FCR) without any adverse effects on rumen fermentation. Palm oil-supplemented diet increased blood cholesterol levels, suggesting a higher energy status of the experimental cattle. Conclusion: Palm oil had no negative effects on rumen fermentation and microbes when supplemented at levels up to 5% in vitro. Thai crossbred cattle fed the palm oil-supplemented diet showed improved FCR without apparent changes of rumen and carcass characteristics, but with elevated blood cholesterol levels. Therefore, palm oil can be used as a beneficial energy source.

• KSBB Journal
• /
• v.7 no.4
• /
• pp.296-301
• /
• 1992

A Pseudomonas fluorescens was selected from mushrooms and studied in both batch and fed-batch cultures in order to get maximal biomass concentration. P. fluorescens is an aerobic bacterium and antagonistic to Pseudomonas tolaasii which causes blotch disease on the mushroom cap. P fluarescens and P. tolaasii were identified by Gram staining, gelatin liquefaction, oxidase test, etc. and were characterized by pigment production, temperature sensitivity, salt tolerance and rapid pitting test, etc., Celts of P. fluorescens well in medium containing 30g/L of glucose, whereas the growth was inhibited at the glucose levels at higher than 30g/L. The highest values of specific growth rate and productivity were obtained when using 10g/1 of yeast extract. Optimum concentrations of $NH_4Cl$ and ${(NH_4Cl)}_2SO_4$ for culture were found to be 1.0g/L and 0.1g/L respectively. Optimum concentration of $MgSO_4{\cdot}7H_2O$ used as a sulfursource was 1.0g/L. It was also found that the cell concentrations reached the maximum level when grown on the medium containing 1.0g/L of $KH_2PO_4$ and 0.1g/L of $CaCl_2$. Also, the optimum culture conditions were $30^{\circ}C$ and pH 6.0. Cultivation of P. fluarescens at high dissolved oxygen (DO) concentration led to a decrease of bacterial productivity in batch culture. Maximum productivity was achieved at 40% DO concentration.

• Microbiology and Biotechnology Letters
• /
• v.31 no.3
• /
• pp.211-215
• /
• 2003

When Saccharomyces cerevisiae MTY62, a high-RNA content yeast, was cultivated by fed-batch mode feeding molasses and com steep liquor, the cell density less than 45g-DCW/L and the RNA content less than 140mg/g-cell were obtained, indicating that unknown compounds inhibiting the cell growth and RNA accumulation are contained in the molasses. Therefore, in order to obtain higher levels of cell density and RNA content, $Ca^{++}$, $Cu^{++}$and $K^{+}$ ions in molasses were removed by pretreatments of molasses with various agents such as IonClear BigBead, $Na_2$$HPO_4$, $H_2$$SO_4$, citric acid, $K_2$$HPO_4$, and EDT A. Among them, IonClear BigBead, $Na_2$$HPO_4$, and EDTA gave the highest $Ca^{++}$ removal efficiency of about 60-90%. In the batch culture with pretreated molasses, the cell concentration of 18.6g-DCW/I and RNA concentration of 3127 mg/I, maximum specific growth rate of 0.459$h^{-1}$ , and specific consumption rate of reducing sugar of 1.28g-sugar/g-cell-h were obtained, which are about 10%, 17%,47%, and 36% higher levels, respectively, over the batch culture with untreated molasses.