• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.268-272
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• 2003

Experiments were carried out to investigate the selection of industrial medium and optimization of fermentation process for the production of erythritol by Candida magnoliae SR101. In the batch fermentation, light steep water(LSW) was the best nitrogen source for the industrial production of erythritol. For the optimization of culture condition, the batch culture was performed. When the concentration of LSW was 65 mL/L in the defined medium containing 250 g/L of glucose, 44% of erythritol yield with 110 g/L of erythritol concentration and 0.66 g/L-hr of productivity, respectively were obtained. Two-stage fed-batch culture was performed to improve the volumetric productivity of erythritol. High density cell culture in the growth stage was performed by batch type with 100 g/L glucose and 500 mL/L LSW concentration, respectively. The cell yield was 0.72 g-cell/g-glucose. Productivity of erythritol was increased and concentration of organic acids such as gluconic acid and acetic acid were decreased when initial pH of 6.5 controlled by 28% ammonia water For increasing yield of erythritol, glucose concentration in the production stage was tested. 37% of total erythritol yield with 186 g/L of erythritol concentration and 1.66 g/L-hr of erythritol productivity were obtained when 820 g of glucose powder was directly added for making up 450 g/L of glucose at production stage.

• 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.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2005.06a
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• pp.230-232
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• 2005

The formation of insoluble aggregation of the recombinant kringle fragment of human apolipoprotein(a), rhLK8, in endoplasmic reticulum was identified as the rate-limiting step in the rhLK8 secretion in Saccharomyces cerevisiae. To analyze the protein secretion pathway, some of yeast genes closely related to protein secretion was rationally selected and their oligomer DNA were arrayed on the chip. The expression profiling of these genes during the induction of rhLK8 in fermentor fed-batch cultures revealed that several foldases including pdi1 gene were up-regulated in the early induction phase, whereas protein transport-related genes were up-regulated in the late induction phase. The coexpression of pdi1 gene increased rhLK8-folding capacity. Hence, the secretion efficiency of rhLK8 in the strain overexpressing pdi1 gene increased by 2-fold comparing in its parental strain. The oligomer DNA chip arrayed with minimum number of the genes selected in this study could be generally applicable to the monitoring system for the heterologous protein secretion and expression in Saccharomyces cerevisiae. With the optimization of fed-batch culture conditions and the alteration of genetic background of host, we obtained extracellular rhLK8 at higher yields than with Pichia pastoris systems, which was a 25-fold increased secretion level of rhLK8 compared to the secretion level at the initiation of this study.

• Korean Journal of Microbiology
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• v.37 no.2
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• pp.158-163
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• 2001

The effect of phosphate on the production of avermectin B1a was studied. Response surface methodology was applied to optimize the concentration of organic nitrogen sources. The portion of B1b in total avermectins was decreased from 5.8% to 3.0% by the addition of 1.5 g/ι inorganic phosphate to the production medium. Among organic nitrogen sources, soybean meal was the most effective on avermectin biosynthesis. Results showed that B1a productivity was increased by 44.8% in a laboratory scale fermenter cultivation of Streptomyces avermitilis YA99-40 through fed-batch process. A maximal B1a productivity was obtained by repeated 30 and 20 g/ι of glucose feeding at 136 and 206 hour, respectively. The B1a productivity was increased by 86.3% and the proportion of B1a in the total avermectins was improved from 38% to 45% with respect to the control process. These results would be very useful for enhancing productivity of B1a in an up-scaled processes.

• 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.

• KSBB Journal
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• v.17 no.1
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• pp.93-99
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• 2002

Two-stage fed-batch culture of Candide tropicalis that was designated primarily to cultivate the cell in the glucose medium (1st stage) and then produced the xylitol from xylose medium (2nd stage) was developed to improve a xylitol yield and productivity. In the growth stage, glucose was automatically supplied to the fermentor by pH-stat mode when the pH was up 5.7, When a feeding medium was added in order to reach the glucose and yeast extract concentrations up to 100 and 40 g/L, respectively, a high cell concentration and a relatively low ethanol concentration were obtained in 18.5 h culture. In the production stage, initial xylose concentration of 150 g/L was the most favorable for obtaining the final xylitol concentration and productivity. The addition of mineral salts was also enhanced a xylitol production. But the aeration rate was not significantly affected a xylitol production. When the addition of 16 g yeast extract and 232.5 g xylose powder at the production stage was used, xylitol yield and productivity were significantly increased. With these conditions, xylitol concentration, yield and productivity of 108.9 g/L, 74%) and 3.3 g/L·h, respectively, were obtained in a final volume of 1.58 L. The further addition of 16 g yeast extract and 232.5 g xylose powder increased the working volume partly (1.67 L) and resulted in a relatively high xylitol concentration, yield and productivity of 193 g/L, 70% and 3.6 g/L·h, respectively.

• KSBB Journal
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• v.21 no.2
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• pp.79-84
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• 2006

In order to minimize the reduction of lysine productivity by accumulation of lysine and byproducts in the end of fed-batch fermentations, a salt-tolerant mutant C14-49-3-15-7-3-20, which could grow at high concentrations of NaCl was isolated through mutagenesis from the Corynebacterium glutamicum mother strain I. In the evaluation of L-lysine productivity by fed-batch fermentations using a 5 L jar fermenter, the salt-tolerant mutant strain C14-49-3-15-7-3-20 produced 130.6 g/L of L-lysine with a 48.6% of yield. The mother strain I produced L-lysine concentration only 104.9 g/L with a yield 41.8%, implying the improvement of L-lysine productivity by introduction of salt-tolerance character.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.153-156
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• 2003

The MRS medium is widely used as an optimized medium for the growth of Lactobacillus spp. and also used for the growth of Leuconostoc spp. Leuconostoc mesenteroides shows quite different physicochemical properties compared to Lactobacilli spp. and it is one of the major strain of kimchi fermenting microorganisms with its usefulness in our traditional foods and availability in biotechnology in the future, specifically tailor-made medium is necessary for the growth of Leuconostoc mesenteroides. Sequential experimental designs (Plackett-Burman, fractional factorial, steepest ascent, central composite design and response surface methodology) were introduced to optimize and improve the Leuconostoc medium. Fifteen medium ingredients were investigated and fructose, sodium acetate and ammonium citrate were determined to give a critical and positive effect for cell-growth. The yield of biomass using the optimal medium was improved more than that of the MRS medium and the result of fed-batch culture showed the capability of the improvement in cell mass similar to the E.coli system.

• Microbiology and Biotechnology Letters
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• v.25 no.3
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• pp.311-316
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• 1997

A high xylitol producing yeast was isolated from the sludge of xylose manufacturing factory and then identified as Candida tropicalis DS-72 according to physiological properties. The strain was able to produce xylitol in a high concentration up to 72g/l from 100g/l xylose in 32 hours. Medium optimization for xylitol production by C. tropicalis DS-72 was performed. Effect of various nitrogen sources on xylitol production was investigated. Of nitrogenous compounds, yeast extract was the most suitable organic nitrogen nutrient for the enhancement of xylitol production. However, inorganic nitrogen resulted in a low cell concentration and did not produce xylitol. Effect of inorganic salts such as KH$_{2}$PO$_{4}$, and MgSO$_{4}$, 7H$_{2}$O on xylitol production was also studied. Optimal medium was selected as xylose 100g/l, yeast extract 10g/l, KH$_{2}$PO$_{4}$, 5 g/l and MgSO$_{4}$, 7H$_{2}$O 0.2 g/l. Xylitol of 88 g/l was produced from 100 g/l xylose in 30 hours using the optimal medium in a flask. In a fermentor, a fed-batch culture with 300g/l xylose was carried out. A final xylitol concentration of 240 g/l in the culture could be obtained in 43 hours of culture time by maintaining the high level of dissolved oxygen during growth phase and limiting the dissolved oxygen in the same culture during production phase. This result corresponded to a xylitol yield of 80% and a xylitol productivity of 5.58 g/1-h.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.71-77
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• 1998

The strain of Serratia marcescens QM B1466 produces selectively large amount of chitinolytic enzymes (about 1mg/L medium). Enzymatic hydrolysis of chitin to N-acetyl-${\beta}$-D-glucosamine (NAG) was performed with a system consisting of two hydrolases (chitinase and chitobiase) produced by optimization of a microbial host consuming chitin particles. For the development of Large-scale biological process for the production of NAG from chitinaceous waste, the selection and optimization of a microbial host, particle size of crab/shrimp chitin sources and initial induction time using chitin as a sole carbon source on chitinase/chitobiase production and NAG production were examined. Crab-shell chitin(1.5%) treated by dilute acid and , ball-milled with a normal diameter less than 250m gave the highest chitinase activity over a 7 days culture. Crude chitinase/ chitobiase solution obtained in a 10 L fed-batch fermentation showed a maximum activities of 23.6 U/mL and 5.1 U/mL, respectively with a feeding time of 3 hrs, near pH 8.5 at 30$^{\circ}C$.