• Microbiology and Biotechnology Letters
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• v.19 no.5
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• pp.491-496
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• 1991

To maximize the production of aminoglycoside-3'-phosphotransferase of E. coli ATCC 21990 carrying R factor which encodes aminoglycoside-3'-phosphotransferase (APH(3')) phosphorylating the 3'-hydroxyl group of aminoglycoside, mutants M1 and M2, media composition and several factors affecting the enzyme production during fermentation were studied. Although the specific activity of APH(3') produced by a mutant M1 was increased as much as four times than that of E. coii ATCC 21990, the growth rate was decreased. The increase of the enzyme production was obtained by increased biomass during fermentation. A mutant M2 was obtained to increase the cell growth rate. Mutant M2 cells were cultivated with optimal media and pure oxygen gas in a fed-batch mode of fermentor operation. The specific activity of APH(3') was decreased, but total enzyme activity of APH(3') was increased as much as two point five times than that of mutant MI.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.356-363
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• 2016

Human papillomavirus (HPV), a non-enveloped, double-stranded DNA tumor virus, is a primary etiological agent of cervical cancer development. As a potential tool for prophylactic vaccination, the development of virus-like particles (VLPs) containing the HPV16 L1 capsid protein is highly desired. In this study, we developed a high-level expression system of the HPV16 L1 in Escherichia coli for the purpose of VLP development. The native gene of HPV16 L1 has many rare codons that cause the early termination of translation and result in the production of truncated forms. First, we optimized the codon of the HPV16 L1 gene to the preferable codons of E. coli, and we succeeded in producing the full-size HPV16 L1 protein without early termination. Next, to find the best host for the production of HPV16 L1, we examined a total of eight E. coli strains, and E. coli BL21(DE3) with the highest yield among the strains was selected. With the selected host-vector system, we did a fed-batch cultivation in a lab-scale bioreactor. Two different feeding solutions (complex and defined feeding solutions) were examined and, when the complex feeding solution was used, a 6-fold higher production yield (4.6 g/l) was obtained compared with that with the defined feeding solution.

• Korean Journal of Food Science and Technology
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• v.46 no.4
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• pp.418-424
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• 2014

High-acidity vinegar was manufactured using pear concentrate by fed-batch fermentation without additional nutrients, and the physicochemical properties and volatile components were investigated at different fermentation stages (Stages 1-4) and at various initial alcohol concentrations (IAC; 6-9%). The levels of reducing sugar, free amino acids, total phenolic content, total flavonoid content, and radical scavenging ability increased slightly during Stage 4 (high-acidity vinegar), which was affected by alcohol feeding. The contents of approximately 20 types of volatile compounds differed between the moderate- and high-acidity vinegar samples, as determined by solid-phase microextraction/gas chromatography-mass spectroscopy. The level of acetic acid in high-acidity vinegar increased according to the initial alcoholic content applied. The high-acidity vinegar produced by fed-batch culture at an IAC of 6-7% showed improved physicochemical and volatile properties as compared to the moderate-acidity vinegar.

• Korean Journal of Food Science and Technology
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• v.34 no.4
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• pp.610-616
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• 2002

In order to improve a complicated Cheongju manufacturing process, saccharification process with gelatinized rice flour was employed during a Cheongju fermentation. High sugar content without unsaccharified residue appeared to impede the yeast growth and fermentation. To solve this problem, addition of zeolite to the saccharifying solution containing 20% (w/v) sugar and fed-batch system were employed. These adjustments resulted in a increase of yeast viability and 40% time-saving alterations of fermentation. The Cheongju, having 18% (v/v) of ethanol content and fresh and rich flavor, could be made in 12 days. Yeast cells recovered from the fermentation precipitates could be reused up to four times without any adverse effect on cell viability, alcohol production, and flavor of the product. The complicated conventional brewing process of Cheongju can thus be simplified effectively.

• Journal of Microbiology and Biotechnology
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• v.32 no.9
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• pp.1178-1185
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• 2022

Steroids are a class of compounds with cyclopentane polyhydrophenanthrene as the parent nucleus, and they usually have unique biological and pharmacological activities. Most of the biosynthesis of steroids is completed by a series of enzymatic reactions starting from cholesterol. Synthetic biology can be used to synthesize cholesterol in engineered microorganisms, but the production of cholesterol is too low to further produce other high-value steroids from cholesterol as the raw material and precursor. In this work, combinational strategies were established to increase the production of cholesterol in engineered Saccharomyces cerevisiae RH6829. The basic medium for high cholesterol production was selected by screening 8 kinds of culture media. Single-factor optimization of the carbon and nitrogen sources of the culture medium, and the addition of calcium ions, zinc ions and citric acid, further increased the cholesterol production to 192.53 mg/l. In the 5-L bioreactor, through the establishment of strategies for glucose and citric acid feeding and dissolved oxygen regulation, the cholesterol production was further increased to 339.87 mg/l, which was 734% higher than that in the original medium. This is the highest titer of cholesterol produced by microorganisms currently reported. The fermentation program has also been conducted in a 50-L bioreactor to prove its stability and feasibility.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.378-381
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• 2001

In the batch culture experiments, the addition of $MnSO_4$, was examined in flask culture, and then the optimal amounts of $MnSO_4$ was investigated in 2.5- L jar-fermenter. As a results, 0.005% $MnSO_4$ was found to enhance the ARA yield of 1.14- fold. Also the addition of $KH_2PO_4_4$ was investigated in 2.5-L jar-fermenter and the ARA yield was enhanced 1.20-fold. Fed batch culture study shown a relatively high productivity of cell mass (62.1 g/L) and ARA content (12.0 g/L) when 14% ammonia solution were alternatively used to control the pH and nitrogen source in the cultivation period.

• Microbiology and Biotechnology Letters
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• v.23 no.1
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• pp.6-11
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• 1995

In order to screen microorganism producing lactic acid with high productivity from nature, we used a medium containing 100 g/l glucose and selected several microorganisms producing more than 80 g/l L-lactic acid. We investigated their physiological characteristics and compared them. The best microorganism was identified as Lactobacillus casei subsp. rhamnosus. The optimum pH for growth and production of lactic acid was 6.0 and this strain showed the highest growth rate at around 30$\circ$C , but the optimum temperature for lactic acid production was 45$\circ$C . The growth was inhibited proportionally from 50 g/l to 300 g/l of glucose and the maximal cell mass increased according to increasing the concentration of corn steep liquor (CSL) protein up to 30 g/l. In batch fermentation for lactic acid production, we produced 128 g/l L-lactic acid with 20 g/l CSL protein and 150 g/l glucose in 35 hours. In pH-stat fed-batch fermentation, we were able to produce 183 g/l L-lactic acid.

• Microbiology and Biotechnology Letters
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• v.25 no.3
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• pp.235-239
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• 1997

Chlorococcum littorale has been grown in high $CO_2$ concentrations to utilize $CO_2$ gas in the polluted air. The effect of incident light intensity on the specific growth rate is expressed by a photoinhibition model, showing half- saturation constant, $K_0\;as\;8\;(W/m^2)$ and inhibition constant, Ki as 35 $(W/m^2)$. The maximum specific growth rate was also estimated as 0.095 (1/day) under this condition. This strain maintained the optimum growth rate in 20% of $CO_2$ gas but 50% of input $CO_2$ gas is the maximum concentration considering the economical efficiency. The maximum Specific $CO_2$ consumption rate, $qCO_2$ was measured as 17.48 (mg $CO_2/g$ dry wt./day) in batch cultivation, 11.2 (mg $CO_2/g$ dry wt./day) in fed-batch cultivation and 10.87 (mg $CO_2/g$ dry wt./day) at 0.065 (1/day) of dilution rate in continuous cultivation. The chemical composition of the biomass obtained from this process showed 32.5% of protein, 27.5% of lipid, 16.5% of carbohydrate and ash 11.7%.

• Journal of Microbiology and Biotechnology
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• v.28 no.9
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• pp.1473-1481
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• 2018

A cellulase hyperproducing mutant strain, JNDY-13, was obtained using the ARTP mutation system and with Trichoderma reesei RUT-C30 as the parent strain. Whole-genome sequencing of JNDY-13 confirmed that 105 of the 653 SNPs were point mutations, 336 mutations were deletions and 165 were insertions. Moreover, 99 mutations were insertions and duplications. Among all the mutations, the one that occurred in the galactokinase gene might be related to the production of cellulases in T. reesei JNDY-13. Moreover, the up-regulation of cellulase and hemicellulase genes in JNDY-13 might contribute to higher cellulases production. Under optimal conditions, the highest cellulase activity by batch fermentation reached 4.35 U/ml, and the highest activity of fed-batch fermentation achieved was 5.40 U/ml.

• Journal of Microbiology and Biotechnology
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• v.3 no.3
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• pp.168-173
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• 1993

The expression kinetics of human lipocortin I (LCI), a potential anti-inflammatory agent, was studied in the shake-flask and fermenter cultures of Saccharomyces cerevisiae carrying a galactose-inducible expression system. The cell growth, expression level of LCI, and the plasmid stability were investigted under various galactose induction conditions. The expression of LCI was repressed by the presence of a very small amount of dextrose in the culture medium, but it was induced by galactose after dextrose became completely depleted. The optimal ratio of dextrose to galactose for lipocortin I production was found to be 1.0 (10 g/l dextrose and 10 g/l galactose). With optimal D/G ratio of 1.0 and the addition of galactose prior to dextrose depletion, LCI of about 100~130 mg/l was produced. LCI at a concentration of 174 mg/l was porduced in the fed-batch culture, which was nearly a twice as much of that produced in the batch culture. The plasmid stability was very high in all culture cases, and thus was considered to be not an important parameter in the expression of LCI.