• Journal of Microbiology and Biotechnology
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• v.23 no.2
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• pp.211-217
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• 2013

Corynebacterium glutamicum is one of the well-studied industrial strain that is used for the production of nucleotides and amino acids. Recently, it has also been studied as a possible producer of organic acids such as succinic acid, based on its ability to produce organic acids under an oxygen deprivation condition. In this study, we conducted the optimization of medium components for improved succinate production from C. glutamicum under an oxygen deprivation condition by Plackett-Burman design and applied a response surface methodology. A Plackett-Burman design for ten factors such as glucose, ammonium sulfate, magnesium sulfate, potassium phosphate ($K_2HPO_4$ and $KH_2PO_4$), iron sulfate, manganese sulfate, biotin, thiamine, and sodium bicarbonate was applied to evaluate the effects on succinate production. Glucose, ammonium sulfate, magnesium sulfate, and dipotassium phosphate were found to have significant influence on succinate production, and the optimal concentrations of these four factors were sequentially investigated by the response surface methodology using a Box-Behnken design. The optimal medium components obtained for achieving maximum concentration of succinic acid were as follows: glucose 10 g/l, magnesium sulfate 0.5 g/l, dipotassium phosphate ($K_2HPO_4$) 0.75 g/l, potassium dihydrogen phosphate ($KH_2PO_4$) 0.5 g/l, iron sulfate 6 mg/l, manganese sulfate 4.2 mg/l, biotin 0.2 mg/l, thiamine 0.2 mg/l, and sodium bicarbonate 100 mM. The parameters that differed from a normal BT medium were glucose changed from 40 g/l to 10 g/l, dipotassium phosphate ($K_2HPO_4$) 0.5 g/l changed to 0.75 g/l, and ammonium sulfate ($(NH_4)_2SO_4$) 7 g/l changed to 0 g/l. Under these conditions, the final succinic acid concentration was 16.3 mM, which is about 1.46 fold higher than the original medium (11.1 mM) at 24 h. This work showed the improvement of succinate production by a simple change of media components deduced from sequential optimization.

• Korean Journal of Food Science and Technology
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• v.4 no.3
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• pp.200-205
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• 1972

Effects of several different petroleum fractions (LGO, HGO, VGO, Diesel oil, SP(E), HGO-wax, L/M-wax), stepwise addition of calculated amounts of HGO at defined intervals, recycling of spent media on cell growth of Candida tropicalis KIST 351 were studied using $2.5{\ell}$ fermenter by batch process. In addition, continuous cultivation of the yeast was also performed in the light of biomass production using $28{\ell}$ fermenter with LGO. 1) Cell concentration, yield on the basis of gas oil and n-paraffin with the petroleum fractions were in the range of $11{\sim}15g/{\ell}$, $10{\sim}12%$ and $77{\sim}82%$, respectively. 2) By stepwise addition of the gas oil, cell concentration and yield on the oil were increased up to 18.9 g/land 13%, respectively. 3) Spent medium slowed emulsifying ability of hydrocarbon and stimulating effect on the cell growth. Without additional supplementation of $Mg^{++}$ up to 20% of spent medium could be reused, while by adding of the $Mg^{++}$, 50% of medium could be recycled. 4) Optimum condition of continuous cultivation for biomass production was attained at the dilution rate of $D=0.1{\sim}0.125\;hr^{-1}$. Maximum yield coefficient on consumed n-paraffin was 0.94 at $D=0.1\;hr^{-1}$, however, 24% of supplied n-paraffin in the media was not utilized at this dilution rate.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.2
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• pp.200-207
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• 2007

In vitro rumen incubation studies were conducted to determine effects of initial pH on bacterial attachment and fiber digestion. Ruminal fluid pH was adjusted to 5.7, 6.2 and 6.7, and three major fibrolytic bacteria attached to rice straw in the mixed culture were quantified with real-time PCR. The numbers of attached and unattached Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminocococcus albus were lower (p<0.05) at initial pH of 5.7 without significant difference between those at higher initial pH. Lowering incubation media pH to 5.7 also increased bacterial numbers detached from substrate regardless of bacterial species. Dry matter digestibility, gas accumulation and total VFA production were pH-dependent. Unlike bacterial attachment, maintaining an initial pH of 6.7 increased digestion over initial pH of 6.2. After 48 h in vitro rumen fermentation, average increases in DM digestion, gas accumulation, and total VFA production at initial pH of 6.2 and 6.7 were 2.8 and 4.4, 2.0 and 3.0, and 1.2 and 1.6 times those at initial pH of 5.7, respectively. The lag time to reach above 2% DM digestibility at low initial pH was taken more times (8 h) than at high and middle initial pH (4 h). Current data clearly indicate that ruminal pH is one of the important determinants of fiber digestion, which is modulated via the effect on bacterial attachment to fiber substrates.

• KSBB Journal
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• v.15 no.3
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• pp.268-273
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• 2000

A microbiological hydrogen production process was optimized. Anaerobic photosynthetic bacteria like Rhodospirillum rubrum which is known to produce hydrogen from carbon monoxide efficiently and remove sulfur was used. To evaluate the potenital of this microorganism the optimization of media fermentation condition light intensity and light requirement for CO conversionwas tried in batch cultures and the continuous fermenter was also applied for this process. The gas residence time on CO conversion was sought out to get high conversion of carbon monoxide to hydrogen. Through this study the possibility of microbial synthtics gas concersion process was proposed.

• Microbiology and Biotechnology Letters
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• v.27 no.1
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• pp.46-53
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• 1999

The purpose of this study was to optimize the conditions of continuous mixed culture of C.butyricum and R. spaeroides K-7, which were able to produce hydrogen using biomass-dreived substrate. To investigate the possibility of continuous culture, semi-continuous culture was carried out for 20 days. In semi-continuous culture using the reactor system, the replacement rate of fresh medium was 30% of total medium volume for the highest hydrogen evolution. In continuous culture, the optimum dilution rate was determined to be 0.05$h^{-1}$. The continuous culture produced 3.1 times as compared with the hydrogen on batch culture. On the other hand, the continuous mixed culture produced 1.3~2.1 times as much as hydrogen of the continuous monoculture of C. butyricum. When 10g of glucose in the media (1l) was supplied as a carbon source on continuous culture, mixed culture of C. butyricum and R. sphaeroides K-7 increased hydrogen evolution rate. Because considerable amount of glutamate was contained in waste water of glutamate fermentation, utilization of glutamate was examined in mixed culture. As a result of examination, production of hydorgen was slightly inhibited by high concentration of glutamate, more than 20mM, on continuous monoculture of R. sphaeroides K-7. On the other hand, both on continuous monoculture of C. butyricum and on mixed culture of C. butyricum and R. sphaeroides K-7, production of hydrogen was not inhibited by high concentration of glutamate such as 100mM. Hence this suggests that high concentration of waste water can be used as good substrate for hydrogen production on monoculture of C. butyricum and mixed culture of C. butyricum and R. sphaeroides K-7.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.815-822
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• 2005

In order to extend the shelf-life of packaged or coated foods, an antibacterial edible film was developed from soybean meal that had been fermented with Bacillus subtilis under the optimum condition of pH 7.0-7.5 and $33^{\circ}C$ for 33 h. The water vapor permeability of the fermented film ($86.0 mg/cm^2{\cdot}h$) was higher than those of normal soybean films ($66.9 mg/cm^2{\cdot}h$). Protein solubility of the fermented film was also higher than ordinary soy protein film at the pH range of 3 -10. The fermented soybean film had higher tensile strength and lower $\%$ elongation (elongation rate) than the ordinary soybean film, mainly because partial hydrolysis of proteins in the soybean film occurred during fermentation. Antimicrobial properties of the fermented film on foodstuffs were measured by placing the films on surime, jerked beef, and mashed sausage media; containing $10^2-10^3$ CFU/plate of foodborne pathogenic bacteria, and showed significantly higher inhibitory effects on the growths of all the indicating bacteria. The film could be used as a packaging material in the food industry. However, before direct application of the fermented film to the commercial food industry, its poor mechanical and antibacterial properties need to be improved.

• The Journal of the Korean Society for Microbiology
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• v.21 no.3
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• pp.345-353
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• 1986

Authors studies on the isolation of non-sucrose fermentation Vibrio species such as V. parahaemolyticus, V. vulnificus and V. damsela from sea water, shellfishes and various algae at Ulleung island in the east of Korea on summer in 1985. Authors carried out test for isolated strains to biochemical characteristics, halophilism, hemolysis and serological reaction. The results obtained were as follows: 1. Strains isolated from total specimens were 25 strains of V. parahaemolyticus, 2 strains of V. vulnificus and 1 strain of V. damsela, respectively. 2. Isolated frequency of Vibrio species from sea water and marine products was lower than on specimens in the southern sea of Korea from 1980 to 1985. 3. On hemolysis reaction on blood agar media using human and rabbit erythrocytes, 14 strains among 25 strains of V. parahaemolyticus isolated were positive, and V. vulnificus and V. damsela were positive, respectively. 4. The distributions of serotypes of V. parahaemolyticus isolated were from O1 group to O10 group, and from K 10 to K 69. The results of K serotypes were as follows ; serotype K 10 and K 33 were 2 strains, serotype K 59 and K 69 were 1 strain, respectively. And 2 strains of V. vulnificus isolated were not agglutinated by antiserum of V. vulnificus. Therefore, the probability is that 2 strains of V. vulnificus isolated were other serotypes distributed on sea weater and marine products of Ulleung island of Korea.

• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1093-1100
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• 2015

Acetate and lactate in growth media are detrimental to the production of Thermus maltogenic amylase (ThMA), a heterologous protein, as well as to the growth of recombinant Escherichia coli. Only 50 mM of acetate or 10 mM of lactate reduced 90% of specific ThMA activity. In this study, mutant E. coli strains blocked in the ackA-pta or ackA-pta and ldh pathways were created, characterized, and assessed for their culture performace in 300 L-scale fermentation. The ackApta and ldh double-mutant strain formed significantly less lactate and acetate, and produced a concomitant increase in the excretion of pyruvate (17.8 mM) under anaerobic conditions. The ackA-pta mutant strain accumulated significant acetate but had an approximately 2-fold increase in the formation of lactate. The ackA-pta and ldh double-mutant strain had superior overall performance in large-scale culture under suboptimal conditions, giving 67% higher cell density and 66% higher ThMA activity compared with those of the control strain. The doublemutant strain also achieved a 179% improvement in volumetric ThMA production.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.427-432
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• 1986

The regulation of three ammonia assimilatory enzymes, GDH (glutamate dehydrogenase), GS (glutamine synthetase) and GOGAT (glutamate synthase), has been examined in C. glutamicum. Three kinds of arginine auxotrophs blocked in each step of arginine biosynthetic pathway from glutamate were selected as arg 5, arg 6, arg 8. Histidine and tryptophan auxotrophs were also selected because histidine and tryptophan repressed GS biosynthesis in E. coli. These strains were cultured on the media containing nitrogen-excess and limited conditions, to compare the specific activities of ${\alpha}$-ketoglutarate dehydrogenase(${\alpha}-KGDH$), GDH, GS, GOGAT from the cell-free extracts. These results showed that enzyme levels of ${\alpha}-KGDH$ and GDH from 3 kinds of arginine auxotrophs, histidine and tryptophan auxotrophs in nitrogen-excess condition and those of GS and GOGAT in nitrogen limited condition were increased compared with opposite condition. The tryptophan and histidine auxotrophs showed higher level of glutamate and glutamine than parental strains and other mutants. it is assumed that the higher levels of ${\alpha-KGDH}$ and GDH from mutants in nitrogen-excess condition promoted the accumulation of glutamate and glutamine in fermentation broth. The inhibition of GS activities by ADP suggested that GS is regulated by energy charge in C. glutamicum. The results with histidine, tryptophan, glycine, alanine, serine and GMP implied that a system of feedback inhibition were effective. The GDH, GS and GOGAT biosynthesis in culture broth was markedly repressed by the nature and kinds of available nitrogen sources such as tryptophan, proline, glycine, alanine, serine and tyrosine.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1221-1225
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• 2007

A ${\beta}$-ionone-resistant mutant strain isolated from the red yeast Xanthophyllomyces dendrorhous KCTC 7704 was used for batch and continuous fermentation kinetic studies with glucose media in a 2.5-1 jar fermentor at $22^{\circ}C$ and pH 4.5. The kinetic pattern of growth and carotenoid concentration in the batch fermentations exhibited a so-called mixed-growth-associated product formation, possibly due to the fact that the content of intracellular carotenoids depends on the degree of physical maturation toward adulthood. To determine the maximum specific growth rate constant (${\mu}_m$) and Monod constant ($K_s$) for the mutant, glucose-limited continuous culture studies were performed at different dilution rates within a range of $0.02-0.10\;h^{-1}$. A reciprocal plot of the steady-state data (viz., reciprocal of glucose concentration versus residence time) obtained from continuous culture experiments was used to estimate a ${\mu}_m$ of $0.15\;h^{-1}$ and $k_s$ of 1.19 g/l. The carotenoid content related to the residence time appeared to assume a typical form of saturation kinetics. The maximum carotenoid content ($X_m$) for the mutant was estimated to be $1.04\;{\mu}g/mg$ dry cell weight, and the Lee constant ($k_m$), which was tentatively defined in this work, was found to be 3.0 h.