• Journal of Microbiology and Biotechnology
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• v.31 no.4
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• pp.570-583
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• 2021

Pyrococcus furiosus α-amylase can hydrolyze α-1,4 linkages in starch and related carbohydrates under hyperthermophilic condition (~ 100℃), showing great potential in a wide range of industrial applications, while its relatively low productivity from heterologous hosts has limited the industrial applications. Bacillus subtilis, a gram-positive bacterium, has been widely used in industrial production for its non-pathogenic and powerful secretory characteristics. This study was conducted to increase production of P. furiosus α-amylase in B. subtilis through three strategies. Initial experiments showed that co-expression of P. furiosus molecular chaperone peptidyl-prolyl cis-trans isomerase through genomic integration mode, using a CRISPR/Cas9 system, increased soluble amylase production. Therefore, considering that native P. furiosus α-amylase is produced within a hyperthermophilic environment and is highly thermostable, heat treatment of intact culture at 90℃ for 15 min was performed, thereby greatly increasing soluble amylase production. After optimization of the culture conditions (nitrogen source, carbon source, metal ion, temperature and pH), experiments in a 3-L fermenter yielded a soluble activity of 3,806.7 U/ml, which was 3.3- and 28.2-fold those of a control without heat treatment (1,155.1 U/ml) and an empty expression vector control (135.1 U/ml), respectively. This represents the highest P. furiosus α-amylase production reported to date and should promote innovation in the starch liquefaction process and related industrial productions. Meanwhile, heat treatment, which may promote folding of aggregated P. furiosus α-amylase into a soluble, active form through the transfer of kinetic energy, may be of general benefit when producing proteins from thermophilic archaea.

• Journal of Microbiology
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• v.44 no.4
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• pp.439-446
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• 2006

In this study, the growth kinetics of Lactobacillus rhamnosus and lactic acid production in continuous culture were assessed at a range of dilution rates $(0.05 h^{-1}\;to\;0.40h^{-1})$ using a 2L stirred tank fermenter with a working volume of 600ml. Unstructured models, predicated on the Monod and Luedeking-Piret equations, were employed to simulate the growth of the bacterium, glucose consumption, and lactic acid production at different dilution rates in continuous cultures. The maximum specific growth rate of L. rhamnosus, ${\mu}_{max}$, was estimated at $0.40h^{-1}$I, and the Monod cell growth saturation constant, Ks, at approximately 0.25g/L. Maximum cell viability $(1.3{\times}10^{10}CFU/ml)$ was achieved in the dilution rate range of $D=0.28h^{-1}\;to\;0.35h^{-1}$. Both maximum viable cell yield and productivity were achieved at $D=0.35h^{-1}$. The continuous cultivation of L. rhamnosus at $D=0.35h^{-1}$ resulted in substantial improvements in cell productivity, of 267% (viable cell count) that achieved via batch cultivation.

• Korean Journal of Food Science and Technology
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• v.4 no.3
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• pp.200-205
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• 1972

Effects of several different petroleum fractions (LGO, HGO, VGO, Diesel oil, SP(E), HGO-wax, L/M-wax), stepwise addition of calculated amounts of HGO at defined intervals, recycling of spent media on cell growth of Candida tropicalis KIST 351 were studied using $2.5{\ell}$ fermenter by batch process. In addition, continuous cultivation of the yeast was also performed in the light of biomass production using $28{\ell}$ fermenter with LGO. 1) Cell concentration, yield on the basis of gas oil and n-paraffin with the petroleum fractions were in the range of $11{\sim}15g/{\ell}$, $10{\sim}12%$ and $77{\sim}82%$, respectively. 2) By stepwise addition of the gas oil, cell concentration and yield on the oil were increased up to 18.9 g/land 13%, respectively. 3) Spent medium slowed emulsifying ability of hydrocarbon and stimulating effect on the cell growth. Without additional supplementation of $Mg^{++}$ up to 20% of spent medium could be reused, while by adding of the $Mg^{++}$, 50% of medium could be recycled. 4) Optimum condition of continuous cultivation for biomass production was attained at the dilution rate of $D=0.1{\sim}0.125\;hr^{-1}$. Maximum yield coefficient on consumed n-paraffin was 0.94 at $D=0.1\;hr^{-1}$, however, 24% of supplied n-paraffin in the media was not utilized at this dilution rate.

• The Korean Journal of Mycology
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• v.30 no.2
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• pp.131-135
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• 2002

The optimum conditions for the production of Lentinus edodes mycelia and it's hypoglycemic effect was studied. Optimum pH and temperature for the production of mycelia in shaken flask culture were 5 and $30^{\circ}C$, respectively, for 24 days cultivation. Culture period for maximum production of mycelia (8.13 g/l) in 5-l jar fermenter cultivation was shortened as much as 6 days compared to shaken flask culture. The mycelial dose of 5 % proved almost equally effective in lowering the plasma glucose, total cholesterol, and triglyceride level as much as 23.0, 20.7, and 27.1%, respectively. The values of alanine transaminase and aspartate transaminase were also estimated and exhibited a substantial reduction (29.6 and 34.3%, respectively) in activity after the administration of dried mycelia.

• Korean Journal of Microbiology
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• v.50 no.1
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• pp.84-86
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• 2014

Mass production of biocontrol agent is an essential step for its commercial use. Media composition and culture conditions for production of Bacillus amyloliquefaciens SKU-78, a potential biocontrol agent against bacterial wilts, were optimized by a flask culture. Low cost media combining nitrogen and carbon sources were tested. Maximum cell growth (> $2{\times}10^9$ CFU/ml) was obtained in a medium of 5% soy flour combined with 3% corn starch after 24 h cultivation. The optimum initial pH, temperature and shaking speed was 5.5, $30^{\circ}C$ and 150-250 rpm, respectively. Fermentation of SKU-78 was scaled up in 30 L fermenter and the profiles of cell density, pH, dissolved oxygen and spore formation were recorded. After 8 h lag phase, exponential growth occurred and reached at maximum viable cell number ($1.2{\times}10^{11}$ CFU/ml) after 20 h. The SKU-78 strain grown in a low cost medium exhibited the high suppression of bacterial wilts. The results indicate that SKU-78 strain can be produced in a low cost medium and provide a basis for scaling up to industrial level.

• Korean Journal of Organic Agriculture
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• v.27 no.1
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• pp.65-76
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• 2019

Bacillus genus are found abundantly in various sites and their secondary metabolites were used as potential agents in agriculture, notably plant growth promoting and bio-control. The objective of this study was to develop the culture conditions of GH1-13 strain including higher cell growth, stable endospore-forming and enhancement of potential agents which are related with plant growth promoting and phytopathogen suppression. The optimal carbon and nitrogen sources were determined by glucose and soy bean flour, respectively, then resulted in $7.5{\times}10^9cells/mL$, $6.8{\times}10^9\;endospore\;cells/mL$ and sporulation yield of 90% after 30 h cultivation in 500 L submerged fermenter at $37^{\circ}C$, pH 7.0. Cells and cell-free supernatant of GH1-13 strains showed the potent antifungal activity against phytopathogenic fungi of Colletotrichum gloeosporioides. It was also confirmed that indole-3-acetic acid (IAA) production of GH1-13 strain was greatly increased by addition of 0.3% tryptophan.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.26-32
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• 2002

For commercial production of Korean fermented anchovy sauce through rapid fermentation, a bacterial strain which showed the high protease activity was isolated from a commercially fermented anchovy sauce. The isolate was Bacillus amyloliquefaciens, and named as B. amyloliquefaciens HTP-8. The incubation temperature, initial pH, and cultivation time for optimal production of protease by B. amyloliquefaciens HTP-8 were $30^{\circ}C$, 7.0, and 3 days, respectively. In jar fermenter, B. amyloliquefaciens HTP-8 showed higher protease activity when grown at pH 7.0. The protease was partially purified by 80% ammonium sulfate precipitation and CM-Sephadex C-50 ion exchange chromatography. The partially purified enzyme had specific activity of 103.3 units/mg, yield of 0.4%, and purification fold of 43.0. The optimal pH and temperature for the protease activity were 10.0 and $50^{\circ}C$, respectively. The protease was relatively stable at the pH range of 7.0~12.0 and at the temperatures below 4$0^{\circ}C$. The activity of the enzyme was inhibited by $Ag^{＋}$ /, $Ba^{2＋}$ and selectively inhibited by PMSF, suggesting that it is a serine protease.

• KSBB Journal
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• v.21 no.2
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• pp.96-102
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• 2006

Recently, it has been known that Hericium erinaceum is a one of the very useful functional materials with great attention in mushroom processing industry. In present study, a liquid culture which was not studied systematically until now, was conducted as a method of cultivation for H. erinaceum, and also examined the characteristics of the liquid culture and conditions of process optimization. A good basal medium was selected through the cultivation of 16 species mushroom media and the optimum condition for medium and cultivation were chosen by response surface method. From these results, the optimum condition of medium for mushroom was 3% glucose, 0.2% yeast extract/peptone(1:1) and 0.1% $KH_2PO_4/MgSO_4$(1:1) and also the optimal culture condition was obtained at inoculum of 13.42%, temperature of $22.3^{\circ}C$ and pH of 5.7. The mycelial dry weight of 9 g/I was obtained under these conditions and this amount was about 1.7 times higher than that which were cultivated in basal medium for 8 days.

• The Korean Journal of Mycology
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• v.29 no.2
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• pp.91-98
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• 2001

The mycelial morphology during submerged cultivation of Ganoderma ludium using by air-lift fermenter system were analyzed by image processing system and the characterization of mycelial morphology were investigated. In submerged culture using medium with different ammonium phosphate concentrations, the various morphological forms of G. lucidum mycelium were observed. The filamentous forms such as non-branched long filamentous mycelium, non-branched short mycelium, branched long filamentous mycelium, branched short mycelium, entangled mycelium and clump were observed, and also, and also, the pelleted forms such as smooth pellet, rough pellet and hollow rough pellet were observed. The mycelial morphology was changed from the filamentous to the pelleted forms by addition of ammonium phosphate. The fractal dimensions of pelleted and filamentous forms were 1.05 and 1.3, respectively, while the fractal dimension of mixtures of pelleted and filamentous forms was 1.16. Therefore, the fractal dimension was found to be more effective index for the detection of the mycelial morphology and morphological change during batch cultivation. The circularity was also found to be useful for evaluating the surface growth of pelleted mycelium.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.263-268
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• 2002

Bacillus polyfermenticus SCD, which is commonly called a 'Bisroot' strain, has been appropriately used for the treatment of long-term intestinal disorders, since the live strains, in the form of active endospores, can successfully reach the target intestine. Goal of this study was to develop an industrial medium for growth and sporulation of B. polyfermenticus SCD. From the results of effect of mixed carbon sources on growth and sporulation of B. polyfermenticus SCD, glucose 2% and starch 2% was particularly found to be the most effective for the maximum number of spore production, resulting in spore cells of $4.3{\times}10^9\;spores/mL$ with a sporulation yield of 91%. For the effect of nitrogen sources, the maximum spore cells of $5.7{\times}10^9\;spores/mL$ of B. polyfermenticus SCD with a sporulation yield of 97% was obtained when B. polyfermenticus SCD was cultivated in an optimum nitrogen source medium containing 5% soybean flour. A medium involving proper phosphate salt yielded the maximum number of a spore cells of $6.0{\times}10^9\;spores/mL$ with a sporulation yield of 95%. Finally, the efficacy of an industrial medium (KH5 medium) on growth and sporulation of B. polyfermenticus SCD was investigated in jar fermenter. The higher number of viable cells $(3.3{\times}10^{10}\;cells/mL)$ and spore cells $(3.0{\times}10^{10}\;spores/mL)$ were obtained in 5 L fermenter when compared with a 500 mL baffle flask cultivation. Thus, KH5 medium developed in this study shows promise as an industrial medium because of higher cells and sporulation yield.