• Korean Journal of Food Science and Technology
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• v.24 no.5
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• pp.477-481
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• 1992

As a series of fundamental research projects to produce doenjang (Korean fermented soy paste) of better quality, two kinds of doenjang were manufactured from a traditional meju (Korean soy bean koji) and the mixed with Aspergillus oryzae and Bacillus natto, and histological changes in the cell structures of soy bean of the two were reported doenjang samples were observed and compared during the entire period of fermentation processes. Cell walls of the soy bean were ruptured by pressure and heat during the pressure cooking process and some of them were observed to have the ghost-like shapes. Remarkable differences in the plasmolysis of the cytoplasms were observed between the seed coat and the inner part of soy bean. Small vacuoles resulting from the fusion of the glycoprotein globules by protease and from the hydrolysis of the starch granules by amylase were also observed. Penetration of microorganisms was transferred from the seed coat to the inside of soy bean as the fermentation proceeded. Slimy substances were observed on the seed coat and the parenchyma cells of soy bean fermented with the mixed with Aspergillus oryzae and Bacillus natto. Cell walls of soy bean became difficult to stain and they showed unusual, polygonal shapes as the fermentation proceeded. Samples fermented with the mixed with Aspergillus oryzae and Bacillus natto showed more remarkable tendencies than traditional meju.

• Microbiology and Biotechnology Letters
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• v.13 no.3
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• pp.179-184
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• 1985

Sexually compatible heterothallic haploids and diploids therefrom of Saccharomycopsis lipolytica were compared with respect to production. Diploids constructed through mating were confirmed by random spore analysis, whereas those constructed through protoplast fusion were confirmed by haploidization. ATCC 44601 and IFO 1209 produced larger amount of citrate and isocitrate than IFO 1550 and IFO 1551. A mode of citrate production by diploids was intermediate of the parental haploid strains. The specific activities of citrate synthase and isocitrate lyase in IFO 1550 and IFO 1551 were higher than those in ATCC 44601 and IFO 1209, indicating little correlation between citrate production and specific activities of these enzymes.

• Korean Journal of Agricultural Science
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• v.41 no.3
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• pp.237-243
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• 2014

Human Glucagon like peptide-1 (GLP-1) is an incretin hormone that promotes secretion of insulin. In order to eliminate the formation of the soluble aggregate, Ala19 in GLP-1 was substituted with Thr, resulting in a GLP-1 mutant GLP-1A19T. The gene synthesis of GLP-1A19T and the fusion of 6-lysine tagged ubiquitin gene were accomplished by using the overlap extension polymerase chain reaction. The ubiquitin fused GLP-1A19T (K6UbGLP-1A19T) is expressed as form of inclusion body with little formation of the soluble aggregation in recombinant E. coli. In order to produce K6UbGLP-1A19T in large amounts, fed-batch fermentation was carried out in a pH-stat feeding strategy. Maximum dry cell weight of 87.7 g/L and 20.4% of specific K6UbGLP-1A19T content were obtained. Solid-phase refolding using a cation exchanger was carried out to renature K6UbGLP-1A19T. The refolded K6UbGLP-1A19T aggregated little and was released GLP-1A19T by on-column cleavage with ubiquitin-specific protease-1. The molecular mass of GLP-1A19T showed an accurate agreement with its theoretical molecular mass.

• Journal of Microbiology and Biotechnology
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• v.23 no.7
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• pp.923-931
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• 2013

Rapamycin, known as an inhibitor of Target of Rapamycin (TOR), is an immunosuppressant drug used to prevent rejection in organ transplantation. Despite the close association of the TOR signaling cascade with various scopes of metabolism, it has not yet been thoroughly investigated at the metabolome level. In our current study, we applied mass spectrometric analysis for profiling primary metabolism in order to capture the responsive dynamics of the Chlamydomonas metabolome to the inhibition of TOR by rapamycin. Accordingly, we identified the impact of the rapamycin treatment at the level of metabolomic phenotypes that were clearly distinguished by multivariate statistical analysis. Pathway analysis pinpointed that inactivation of the TCA cycle was accompanied by the inhibition of cellular growth. Relative to the constant suppression of the TCA cycle, most amino acids were significantly increased in a time-dependent manner by longer exposure to rapamycin treatment, after an initial down-regulation at the early stage of exposure. Finally, we explored the isolation of the responsive metabolic factors into the rapamycin treatment and the culture duration, respectively.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.611-617
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• 1990

This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.50-56
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• 2012

The Wnt/${\beta}$-catenin signaling pathway regulates diverse developmental processes and adult tissue homeostasis. Inappropriate regulation of this pathway has been associated with human diseases, such as cancers, osteoporosis, and Alzheimer's disease. Using a cell-based chemical screening with natural compounds, we discovered silybin, a plant flavonoid isolated from the Silybum marianum, which activated the Wnt/${\beta}$-catenin signaling pathway in a synergy with Wnt3a-conditioned medium (Wnt3a-CM). In the presence of Wnt3a-CM, silybin up-regulated ${\beta}$-catenin response transcription (CRT) in HEK293-FL reporter cells and 3T3-L1 preadipocytes through stabilization of intracellular ${\beta}$-catenin protein. Silybin and Wnt3a-CM synergistically reduced expression of important adipocyte marker genes including peroxisome-proliferator-activated $receptor{\gamma}$ ($PPAR{\gamma}$) and CAATT enhancer-binding protein ${\alpha}$ (C/$EBP{\alpha}$) in 3T3-L1 preadipocytes, accompanied by the activation of Wnt/${\beta}$-catenin signaling pathway. Taken together, our findings indicate that silybin is a small-molecule synergist of the Wnt/${\beta}$-catenin signaling pathway and can be used as a controllable reagent for investigating biological processes that involve the Wnt/${\beta}$-catenin signaling pathway.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.550-557
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• 2010

Escherichia coli (E. coli) heat-labile enterotoxin B subunit (LTB) was regarded as one of the most powerful mucosal immunoadjuvants eliciting strong immunoresponse to coadministered antigens. In the research, the high-level secretory expression of functional LTB was achieved in P. pastoris through high-density fermentation in a 5-1 fermentor. Meanwhile, the protein was expressed in E. coli by the way of inclusion body, although the gene was cloned from E. coli. Some positive yeast and E. coli transformants were obtained respectively by a series of screenings and identifications. Fusion proteins LTB-6$\times$His could be secreted into the supernatant of the medium after the recombinant P. pastoris was induced by 0.5% (v/v) methanol at $30^{\circ}C$, whereas E. coli transformants expressed target protein in inclusion body after being induced by 1 mM IPTG at $37^{\circ}C$. The expression level increased dramatically to 250-300 mg/l supernatant of fermentation in the former and 80-100 mg/l in the latter. The LTB-6$\times$His were purified to 95% purity by affinity chromatography and characterized by SDS-PAGE and Western blot. Adjuvant activity of target protein was analyzed by binding ability with GMI gangliosides. The MW of LTB-6$\times$His expressed in P. pastoris was greater than that in E. coli, which was equal to the expected 11 kDa, possibly resulted from glycosylation by P. pastoris that would enhance the immunogenicity of co-administered antigens. These data demonstrated that P. pastoris producing heterologous LTB has significant advantages in higher expression level and in adjuvant activity compared with the homologous E. coli system.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.5
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• pp.779-784
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• 1995

The effective production of 5'-GMP(5'-Guanylic acid) by immobilized 5'-GMP producing fusant RC102(intergeneric protoplast fusion between Brevibacterium ammoniagenes ATCC21263 and Corynebacterium glutamicum ATCC21171) was investigated. The Fusant RC102 was immobilized by entrapping in -carrageenan, agar, polyacrylamide or Ca-alginate. 3% k-carrageenan was selected as the most suitable matrix. In the production of 5'-GMP using the immobilized whole cells of fusant RC102, the optimum conditions were $32^{\circ}C$, pH 8.0, $30\mu\textrm{g}/L\;of\;Mn^{2+},\;1{\times}10^{-6}%\;of\;Zn^{2+}$. In order to use fermentation medium containing CSL(Corn Steep Liquor) plentiful in $Mn^{2+}$, the optimum conditions of penicillin G, D-cycloserine and POESA(polyoxyethylene stearylamine) for production of 5'-GMP were 0.8unit/ml, 0.8unit/ml, 0.8unit/ml and 5mg/ml, respectively. Cationic surfactant, POESA was effective and superior to the antibiotics, penicillin G or D-cyloserine in 5'-GMP productivity. The condinuous fermentation using immobilized fusant RC102 showed that 5'-GMP productivity was stable for more than 15 days.

• Microbiology and Biotechnology Letters
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• v.28 no.2
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• pp.80-86
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• 2000

To develop thermotolerant ethanol producing yeast strains, the protoplasts of Saccharomyces carlsbergensis having good fermentability at $30^{\circ}C$ and Kluyveromyces marxianus able to grow at $42^{\circ}C$ were fused. Under the optimal conditions for protoplast formation, the frequency of protoplast formation of S. carlsbergensis was 92 - 94% and that of K. marxianus was 98%. Fusion frequency between S. carlsbergensis and K. marxianus was $1.4\times10^{-6}-4.8$\times10^{-7}$. Among the 27 fusants obtained, 6 fusants were able to grow at $42^{\circ}C$. While the parental strains produced 3.2-3.4%(w/v) ethanol after 3 days from the fermentation medium containing glucose, fusants SK41-4 and SK53-22 produced 5.2%(w/v) ethanol in the same condition. The thermotolerance of SK53-22 was not high, but that of SK41-4 was quite high.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.8
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• pp.1031-1037
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• 2007

Changes in red ginseng fusion cheonggukjang properties under various hydrolytic conditions were investigated for its possible application to different types of food products. Among the four types of protease that were analyzed, protease (KMF -G) produced the highest hydrolysis rate, calcium binding capacity, and total phenolic compound content. In addition, the highest fibrinolytic activity and ACE inhibitory activity were also exhibited at 87.10 units and 67.17%, respectively. Among a number of different protease concentrations, a 0.02% concentration of protease (KMF-G) was found to be appropriate for the purposes of the study. The best results for red ginseng cheonggukjang hydrolysis were observed at the 60 and 90 min intervals. However, there was not a significant difference between the results at the two time points. The unpleasant odor and bitter taste associated with red ginseng fusion cheonggukjang improved with hydrolytic activity exceeding 60 min. Thus, the optimal hydrolysis time was determined to be 60 min. The total ginsenoside content of red ginseng cheonggukjang was 9.197 mg/g and the hydrolysate content was 11.707 mg/g. Based on the results, it was determined that the addition of 0.02% protease (KMF -G) and treatment for 60 min are the optimal hydrolytic conditions for red ginseng cheonggukjang to improve its biochemical characteristics, including fibronolytic activity and ACE inhibitory activity.