• Journal of Microbiology and Biotechnology
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• v.22 no.5
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• pp.659-667
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• 2012

2,3-Butanediol (2,3-BDO) is an organic compound with a wide range of industrial applications. Although Escherichia coli is often used for the production of organic compounds, the wild-type E. coli does not contain two essential genes in the 2,3-BDO biosynthesis pathway, and cannot ferment 2,3-BDO. Therefore, a 2,3-BDO biosynthesis mutant strain of Escherichia coli was constructed and cultured. To determine the optimum culture factors for 2,3-BDO production, experiments were conducted under different culture environments ranging from strongly acidic to neutral pH. The extracellular metabolite profiles were obtained using high-performance liquid chromatography (HPLC), and the intracellular metabolite profiles were analyzed by ultra-performance liquid chromatography and quadruple time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Metabolic flux analysis (MFA) was used to integrate these profiles. The metabolite profiles showed that 2,3-BDO production favors an acidic environment (pH 5), whereas cell mass favors a neutral environment. Furthermore, when the pH of the culture fell below 5, both the cell growth and 2,3-BDO production were inhibited.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.2
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• pp.59-64
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• 2015

Lactic acid-producing bacteria such as Lactobacillus spp. function to ferment carbohydrates and produce ATP. Such Lactobacillus spp. are used for the production of commercial yogurts. Lactobacillus spp. are beneficial to the intestinal tract, and Lactobacillus acidophilus-containing yogurts have received considerable attention because of their preventive effects against early-stage cancer of the large intestine. In this study, lactic acid-producing bacteria were cultured from three different groups: commercial solid yogurt (for eating), commercial liquid yogurt (for drinking), and Lactobacillus acidophilus-containing yogurt. We first determined the optimum culture conditions for Lactobacillus spp. and then analyzed turbidity and pH in order to compare the growth abilities and lactic acid-production capacities among the groups. Finally, high-performance liquid chromatography was used to measure the lactic acid content in the culture supernatants, and the antibacterial activities against Staphylococcus aureus and Escherichia coli were compared among the three groups. The optimum culture conditions for Lactobacillus spp. were MRS medium at $25^{\circ}C$, for 24 h. The highest turbidity was found in L. acidophilus-containing yogurt, followed by liquid yogurt and solid yogurt. Similarly, the highest lactic acid production ability was found in L. acidophilus-containing yogurt, followed by liquid yogurt and solid yogurt. Culture supernatants from the three groups did not show any antibacterial activity towards S. aureus; however, supernatants derived from L. acidophilus-containing yogurt resulted in a 1.8 mm inhibitory zone against E. coli in a paper disk diffusion test. These results revealed the high level of lactic acid-production capacity and antibacterial activity in L. acidophilus-containing yogurt.

• Korean Journal of Plant Resources
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• v.21 no.1
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• pp.60-65
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• 2008

Kalopanax pictus was cultured in vitro to find out optimal condition for embryogenic cells proliferation in liquid media rapidly. Embryogenic cells were induced from leaves and petiols of Kalopanax pictus. Optimum culture medium appeared to be a 1/2MS medium supplemented with 2.0mg/L 2,4-D and 0.1mg/L BA. To find out optimal conditions, embryogenic cells were cultured some condition as different concentrations of 2,4-D, medium and sucrose. There was cultured on 1/2MS liquid medium containing different concentration of 2,4-D. When embryogenic cells were cultured on 1/2MS liquid medium supplemented with 1.0mg/L 2,4-D, cell propagation rate was higher than other concentration of 2,4-D. When embryogenic cells were cultured on different media that MS, Gambols B5, N6, White, SH medium, observed the highest multiplication rate among Gambols B5 and White medium. To find out of effect of sucrose to embryogenic cells propagation, we tested cells under different concentrations. Optimal concentration of sucrose appeared to be a basal medium added 3% sucrose. Above results suggest that optimal conditions for proliferation of embryogenic cells were established Gambols B5 and White medium added 1.0mg/L 2,4-D and 3% sucrose. There is every possibility achieving embryogenic cells proliferation via bioreactor culture system in Kalopanax pictus.

• Food Science and Preservation
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• v.24 no.7
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• pp.1034-1042
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• 2017

The quality characteristics of Yakju and survival rate of yeast were investigated by modifying the drying method for the cold adapted yeast strain Saccharomyces cerevisiae Y297 (SCY297). Viability and fermentation characteristics of the freeze-dried, air blast-dried, and liquid SCY297 cultures were compared after storing them at $25^{\circ}C$. In addition, 5% skimmed milk, ${\alpha}$-lactose, or trehalose was added as a protective agent for examining the effects of drying methods. During the 15-week storage period, the liquid and freeze-dried SCY297 cultures containing a protective agent showed a survival rate of 80%. However, the air blast-dried SCY297 culture showed 80% survival rate only in the skimmed milk supplemented group. Compared to the untreated cells, the acidity and amino acidity of Yakju prepared using freeze-dried or air blast-dried cultures of SCY297 increased by 2 fold and 5.7 fold respectively, while the alcohol content decreased by 5.07%. Compared to the untreated cells, the pH and amino acidity of Yakju prepared using the liquid culture of SCY297 increased by 1.5 fold and 2.5 fold respectively. Although the alcohol content decreased by 2.9%, decrease rate was lower than that observed for the freeze-dried and air blast-dried yeast cultures. Therefore, the results of this study showed that using a liquid starter culture was more advantageous than using the conventional solid culture.

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.357-363
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• 2011

This study focus was primarily the development of liquid starters for Aspergillus oryzae and Aspergillus niger prepared with wheat bran as a low cost culture medium. For the preparation of the liquid media wheat bran was added at rates of 0, 5, 10, 15 and 20% and the Aspergillus sp. strains were then inoculated to these prepared broths. The results indicated that the more that wheat bran was contained in the medium, the more mycelia was produced for A. oryzae and A. niger. The highest enzyme activities were obtained with a 10~15% adding rate of wheat bran for both strains. Changes in the enzyme activities of the liquid starters during various incubation times (0, 24, 48, 72 and 96 hrs), indicated that the highest enzyme activities were seen between 48 and 72 hrs of culture. In addition, a comparative study was carried out on the production of enzymes using wheat as a substrate in nuruk, with liquid starter made from fermented agents according to the same concentrations used with the wheat bran. The pH, acidity, amino acidity, reducing sugar content and enzyme activity (${\alpha}$-amylase, glucoamylase, acidic protease) of wheat nuruk made with liquid starter were compared with those of wheat nuruk made with solid starter. The results suggest that the liquid starter is superior in both cases.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.198-204
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• 2005

Seeds of neem were collected from different parts of India and analyzed for their azadirachtin content by High Performance Liquid Chromatography (HPLC). In order to assess the effects of genotypic and geographical variation on azadirachtin content in cell cultures, callus development was attempted in the seeds containing high and low concentration of azadirachtin. The concentration of azadirachtin in callus cultures was significantly affected by the explant source. Seed kernels with higher azadirachtin content produced higher azadirachtin content in callus cultures and lower azadirachtin content was seen in callus cultures produced from seed kernels with low azadirachtin content. The protocol for development of elite stock culture of Azadirachta indica was established with the objective of selecting a high azadirachtin-producing cell line. The highest azadirachtin-producing cell line was selected and the effects of different media and illumination conditions on growth and azadirachtin production were studied in shake flask suspension culture. Detailed batch growth kinetics was also established. These studies provided elite starter culture and associated protocols for cultivation of A. indica plant cell culture in the bioreactor.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.75-80
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• 2008

In vitro culture of shoot tip of garlic (Allium sativium L. cv. Seosan) was carried out to find medium condition of the induction of multiple shoots and bulbing for muliproduction of virus-free seed bulbs. For this work, tank culture was introduced. In shoot tip culture on MS solid medium the induction of multiple shoots and bulbing were better by adding 3% sucrose than 8%. Supplementation with 2mg/L 2ip and 0.2 mg/L IAA in this medium was effective. Three point three shoots including 2.7 bulbs were formed from a shoot tip after cultivation for 30 days on this medium. Bulbing of garlic in liquid culture with plastic water tank of 20L supplied air at the side of the lower part was better by adding 3% sucrose than 8% by subculture for 45 days with shoots obtained from shoot tip culture for 30 days on soid MS medium. Shoot growth was vigorous at 3% sucrose however bulb growth was more effective on the medium of 8% sucrose. Because of the effectiveness on solid medium added 3% sucrose, 2 mg/L 2ip and 0.2 mg/L IAA for initial production of multi-shoot in stem tip culture and the effectiveness in liquid culture with water tank for growth of bulbs, the method of two-step culture could be introduced for the multiple production of seed bulb of high quality. It was more desirable by supply of 0.2 mg/L BA and 0.02 mg/L NAA at tank culture time. But growth of the bulbs became poor by increasing concentration of NAA of the medium.

• Journal of Mushroom
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• v.16 no.3
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• pp.162-170
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• 2018

To improve the productivity of Pleurotus ostreatus, different conditions of liquid spawn culture were tested. The optimum culture conditions were potato dextrose broth, incubation temperature of $22^{\circ}C$, and pH 6. Fifteen different media ('A' to 'O') containing 0.3% soybean meal (SM) were prepared by varying sugar and glucose contents. The cultures were propagated in SM media for 14 days, at $22^{\circ}C$, and pH 6. Their absorbances were higher after 14 days of incubation in the media containing both sugar and glucose. In particular, the absorbance of media containing 5 to 20% of glucose alone ('C', 'D', 'E', and 'F') tended to increase in the incubation period. Dry cell weight was lower in media containing less than 20% sugar or glucose alone than in media containing 30% sugar ('A') or 30% glucose ('B') alone. In sawdust media, in 900 mL-bottle, the optimum inoculation volume of liquid spawn was 15 to 20 mL. The texture of the mushroom cultivated with the liquid spawn was superior to that cultivated in the solid spawn.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.45-45
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• 2018

We had already reported the successful germination for green pods of purple lady's slipper orchid (Cypripedium macranthos Sw.). The green pod methods is to take immature seeds in green capsules, sterilize the capsule, and take out the sterile seeds. This method, however, needs very critical time of harvest. The critical time of seed harvest changes depending upon the species, condition of the specimen, and climatic influence, and the right time lies between 5 and 12 weeks after fertilization. In this study, the mature seeds were collected after 120-130 days with hand-polination of lady's slipper orchids. Mature seeds are usually dormant and it has to be overcome, either with hormone or storing the seeds near freezing for two or three months to break dormancy. The seeds were first surface sterilized with 70% ethanol and then transferred 1% NaOCl for 10-15 minutes, followed by rinses 3 times with sterilized distilled water. The cypripedium seeds consists of an embryo within a seed coat known as a testa. The testa is water repellent and the seed has a large air space between the embryo and testa so the seed tends to float on water. We had resolved the problems with vacuum pump to soak water into the testa before sterilization. The seeds were placed on liquid or agar solidified germination media. Cultures were incubated at $24{\pm}1^{\circ}C$ in dark. The seeds were germinated in 6-8 weeks in liquid suspension culture (germination percentage over 18%); however, the seeds on agar solidified media took more than 5 months to germinate and the germination percentage less than 5%. The most effective media for liquid culture was 1/4 strength Murashige and Skoog (MS) medium with 50 ml/l coconut water ($4brix^{\circ}$) at pH 5.8.

• Journal of Plant Biology
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• v.32 no.4
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• pp.313-322
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• 1989

A system was established for induction of multi-shoots and plant regeneration from mesophyll protoplasts of alfalfa, Medicago sativa L. cv. Vernal. Different hormonal effects were tested at each step of protoplast culture, i.e. cell division in modified Kao's liquid medium (K566-7). calli formation on SH semi solid medium, and multi-shoot regeneration from calli on SHa and SHb solid media. Frequency of multi-shoots and plant regeneration was affected by various combinations of phytohormones in final step. The evaluation of multi-shoots induction systems via protoplast culture was discused.