• Journal of Microbiology and Biotechnology
• /
• v.24 no.6
• /
• pp.781-787
• /
• 2014

2-Ketogluconic acid production by Klebsiella pneumoniae is a pH-dependent process, strictly proceeding under acidic conditions. Unfortunately, cell growth is inhibited by acidic conditions, resulting in low productivity of 2-ketogluconic acid. To overcome this deficiency, a two-stage fermentation strategy was exploited in the current study. During the first stage, the culture was maintained at neutral pH, favoring cell growth. During the second stage, the culture pH was switched to acidic conditions favoring 2-ketogluconic acid accumulation. Culture parameters, including switching time, dissolved oxygen levels, pH, and temperature were optimized for the fed-batch fermentation. Characteristics of glucose dehydrogenase and gluconate dehydrogenase were revealed in vitro, and the optimal pHs of the two enzymes coincided with the optimum culture pH. Under optimum conditions, a total of 186 g/l 2-ketogluconic acid was produced at 26 h, and the conversion ratio was 0.98 mol/mol. This fermentation strategy has successfully overcome the mismatch between optimum parameters required for cell growth and 2-ketogluconic acid accumulation, and this result has the highest productivity and conversion ratio of 2-ketogluconic and produced by microorganism.

• Korean Journal of Soil Science and Fertilizer
• /
• v.35 no.1
• /
• pp.59-67
• /
• 2002

A phosphate solubilizing bacterium. strain 60-2G, possessing a strong ability to solubilize insoluble phosphate was isolated from the rhizosphere of grass. On the basis of GC-FAME profile, carbon utilization pattern, and the DNA sequence of a conserved partial 16S rRNA gene, the 60-2G was identified as Enterobacter intermedium. The analysis by HPLC revealed that the strain 60-2G produced mainly gluconic and 2-ketogluconic acids with small amounts of lactic acid in broth culture medium containing hydroxyapatite. During the incubation period of the strain 60-2G in broth culture, pH of the medium decreased upto 3.8 while the soluble phosphate concentration increased. The reversed correlation between pH and soluble phosphate concentration indicated that the solubility of P was due to the produced organic acids. The sequence homology of the deduced amino acids suggested that E. intermedium 60-2G synthesized PQQ which is essential for the oxidation of glucose by glucose dehydrogenase.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.6
• /
• pp.955-959
• /
• 2003

Rhizobium tropici CIAT 899 is capable of synthesizing inactive apo-glucose dehydrogenase (GDH). To become an active holo enzyme, the GDH requires a cofactor, PQQ. When R. tropici CIAT 899 was grown in a broth culture medium containing hydroxyapatite and pyrrolo quinoline quinone (PQQ), pH decreased while the concentration of soluble P increased. The solubilization of hydroxyapatite was associated with the production of gluconic acid and 2-ketogluconic acids. The organic acid production and P solubilization were greatly enhanced when the bacterium was grown with air supply. Effect of R. tropici CIAT 899 with (CI＋PQQ) and without PQQ (CI) on the common bean growth was examined. Shoot and root weight, and N and P contents in CI＋PQQ treatment, were significantly higher than those in control and CI treatment. Nodule weight and acetylene reducing activities were also significantly higher in CI＋PQQ treatment than in other treatments.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.34 no.8
• /
• pp.1251-1257
• /
• 2005

This study was carried out to investigate characteristics of acetic acid bacteria for persimmon vinegar fermen-tation. Acetic acid bacteria were isolated from statically fermented vinegar. Microscopically the cells appeared as non-motile and non-flagellated, preferentially occuring in pairs. Isolated strains were able to grow in the presence of acetic acid, ethanol, and glucose. Four of them produced 2-ketogluconic acid but did not produce 5-ketogluconic acid. The four strains isolated from statically fermented vinegar were considered to be grouped into Acetobacter. The strains were inoculated into persimmon juice for persimmon vinegar fermentation. They produced acetic acid in the range of 5.25$\∼$5.68$\%$.

• Microbiology and Biotechnology Letters
• /
• v.22 no.6
• /
• pp.571-576
• /
• 1994

One strain of cellulose-producing Acetobacter was isolated from the traditionally fermen- ted grape vinegar in Korea. The isolated strain, designated as KI strain was identified as the Acetobacter xylinum with respect to physiological and biochemical characteristics. KI produced acetic acid from ethanol, and then decomposed acetate to CO$_{2}$ and H$_{2}$O. When the isolated strain was cultivated statically in broth culture, a thick cellulose pellicle was formed. KI was tolerance of 8% ethanol and 30% glucose, and the isolate was positive in ketogenesis from glycerol, $\gamma$-pyrone from glucose and fructose, and 2-ketogluconic acid from glucose. KI strain possessed straight-chain C$_{18:1}$, C$_{16:0}$, and C$_{14:0}$ fatty acid, and contained ubiquinone Q$_{9}$ and Q$_{10}$ as isoprenoid quinone. DNA base composition of KI strain was 57.6% G+C.

• Korean Journal of Soil Science and Fertilizer
• /
• v.42 no.1
• /
• pp.29-36
• /
• 2009

Fourteen bacterial strains were isolated from crop rhizosphere and identified as phosphate solubilizing bacteria (PSB) by 16S rRNA analysis. Only 3 strains exhibited a strong ability to solubilize insoluble phosphate in agar medium containing a hydroxyapatite. The rates of P solubilization by isolates were ranged from 200 and $2300\;mg\;L^{-1}$, which are inversely correlated with pH in culture medium. Furthermore, HPLC analyses reveal the production of organic acid from the culture filtrates of PSB. Among these, strain Acinetobacter sp. released only gluconic acid, Pseudomonas orientalis produced gluconic acid which was subsequently converted into 2-ketogluconic acid, and Enterobacter asburiae released acetic acid and succinic acid. On the other hand, P. orientalis and E. asburiae released $372\;mg\;L^{-1}$ and $191\;mg\;L^{-1}$ of IAA into broth culture, respectively, while Acinetobacter sp. did not produce IAA. Furthermore, in vivo study showed that plant growth promoting effect by bacteria generally seemed to be increased IAA production and phosphate solubilization.

• Korean Journal of Soil Science and Fertilizer
• /
• v.38 no.1
• /
• pp.15-24
• /
• 2005

Enterobacter intermedium oxidizes glucose to gluconic acid and sequentially converts gluconic acid to 2-ketogluconic acid (2-KGA) under aerobic condition. Shaking incubation of E. intermedium in a broth medium containing 22.5 g glucose, 8.2 g gluconic acid and 40 g rock phosphate per liter resulted in $1028mg\;L^{-1}$ soluble phosphate. The culture broth was used as phosphate bio-fertilizer (PBF) in this experiment. To evaluate PBF produced by E. intermedium on lettuce growth, five treatments (PBF1/3, PBF2/3, PBF3/3, BP, and MF) were used. In MF and BP treatments, $P_2O_5$ 5.9 kg of mineral fertilizer per 10a was added, while in PBF1/3, PBF2/3, and PBF3/3 treatments, culture broth containing one third, two third, and same amount of soluble $P_2O_5$ 5.9 kg per 10a was applied, respectively. At 20, 35, and 50 days after transplanting of lettuce, plant growth components, biomass, enzyme activities and soil chemical properties were analyzed. Dehydrogenase activity and available phosphate concentration of rhizosphere in phosphate bio fertilizer treatments (PBF1/3, PBF2/3, PBF3/3) were generally higher compared to MF and BP treatments. Soil biomass in PBF3/3 treatment was significantly higher than MF and BP treatments at early growth stage. However, there was no significant difference among all treatments with time. Plant fresh weights in PBF1/3, PBF2/3, and MF treatments were better than those in BP and PBF3/3 treatments. However, in PBF2/3 treatment the highest fresh weight was discovered where alkaline phosphatase activity was generally higher than other treatments at 35 and 50 days. Enhancement of lettuce growth at 35 and 50 days in PBF2/3 treatment was associated with greater phosphate uptake in lettuce tissue. As regarding all results, PBF showed better lettuce growth compared to mineral phosphate fertilizer where PBF2/3 treatment resulted in increase of lettuce fresh weight by 23% and phosphate uptake by 50%.

• Korean Journal of Microbiology
• /
• v.31 no.2
• /
• pp.99-104
• /
• 1993

Two strains of the gram-negative acetic acid bacteria, Acetobacter sp. strain CS2- AND CS5, were isolated form the traditional raw rice wine vinegar of Haenam area. The strains oxidized ethanol to acetic acid and over-oxidized acetate and lactate to $CO^{2}$ and $H ^{2}$O. They produced 2-ketogluconic acid from glucose but did not produce .gamma.-pyrones from glucose and dihydroxyacetone from glycerol. The CS strains possessed ubiquinone-9 as a major isoprenoid quinone and contained straight-chain $C_{18 :1}$, $_C{16 : 0}$, and $C _{14 : 0}$ fatty acids. The DNA base composition of the CS2 and CS5 strains was 56.2 and 57.3 mole% G + C, respectively. The isolates were grown well on methanol, gluconate, erythritol, raffinose, dulcitol and xylitol as sole sources of carbon and energy which are different from those of other Acetobacter species and producedd acid from sucrose, glycerol, fructose, inositol, mannitol, and ribose.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.7 no.4
• /
• pp.201-205
• /
• 2002

The growth conditions of Pantoea aggicmerans, a phosphate solubilizing organism, were studied In our laboratory to determine the optimal conditions. Pantoea aggionerans showed the highest growth rate at 30$\^{C}$, pH 7.0 and 2 vvm, after 50 h cultivation. A certain relationship between pH and phosphate concentration was evident when the glucose concentration in the me dium was changed. Increasing glucose concentration increased the pH buffer action of the broth. At glucose concentrations higher than the optimum concentration of 0.2 M, the cell growth was retarded. P. agglomerans consumed glucose as a substrate to produce organic acids which caused the pH decrease in the culture medium. The phosphate concentration in the medium was increased by the presence of the organic acids, which solubilized insoluble phosphates such as hydroxyapa-tite.

• Korean Journal of Environmental Agriculture
• /
• v.20 no.1
• /
• pp.1-7
• /
• 2001

We investigated the capability of the phosphate-solubilizing fungus, Penicillium sp. GL-101, to solubilize in vitro some insoluble rock phosphate via possible mechanisms: acidification of the medium, production of chelating metabolites, redox activity, and so on. GL-101 was able to solubilize rock phosphate (mostly calcium phosphate) in a liquid potato dextrose broth(PDB) medium, as determined by spectrophotometric analyses. Acidification was the major mechanism of solubilization since the pH of cultures fell below 4.0 and in cultures containing 1.0%(w/v) loess the pH dropped from 7.0 to 3.2. More than 10 mg/mL concentrations of citric acids were detected by high-performance liquid chromatography(HPLC) in the culture supernatants. Also this fungus showed the phosphatase activity (over 1.3 unit) to contribute partially releasing phosphate from rock phosphate, when supplemented with 1.0% loess in culture broth. The chelating activity of GL-101 in culture supernatants was not present because 2-ketogluconic acid, a chelating agent for the phosphate, was produced only a basal level. Therefore, the solubilization mechanism of rock phosphate by Penicillium sp. GL-101 involves both acidification due to citric acid production and phosphatase activity.