• Journal of Life Science
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• v.18 no.7
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• pp.952-957
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• 2008

We evaluated the effects of various factors (e.g., serum, inhibitors of protein synthesis, and cytoskeleton and protein kinases) on early PMA-induced THP-1 cell adhesion using an adhesion assay with Sulforhodamine B (SRB) staining, which was used to assess the proliferation of the attached cells. THP-1 cell adhesion to a plastic substrate was detected 1 hr after exposure to Phorbol 12-Myristate 13-Acetate (PMA) and peaked after 18 hr. At concentrations > 25 nM PMA, the level of adhesion did not change. Based on our preliminary results, we used 25 nM PMA and 5 hr of culture as standard assay conditions. Early PMA-induced cell adhesion was not affected by the presence of serum or PD 98059 in the culture medium, but was affected by the addition of PKC inhibitors and cycloheximide. In the presence of actin inhibitor with PMA, the cell adhesion increased when comparing with PMA treatment only. Thus, early PMA-induced adhesion of THP-1 cells does not require serum in the culture medium, MAP-kinase activation, or actin polymerization, but does require de novo protein synthesis and PKC activation. Our SRB-based cell adhesion assay may be used to screen other PKC inhibitors.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.4
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• pp.343-351
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• 2001

This study was undertaken to investigate the mechanism of lipopolysaccharide (LPS) and nitric oxide (NO) as a regulator of vascular smooth muscle cell (VSMC) proliferation. VSMC was primarily cultured from rat aorta and confirmed by the immunocytochemistry with anti-smooth muscle myosin antibody. The number of viable VSMCs were counted, and lactate dehydrogenase (LDH) activity was measured to assess the degree of cell death. Concentrations of nitrite in the culture medium were measured as an indicator of NO production. LPS was introduced into the medium to induce the inducible nitric oxide synthase (iNOS) in VSMC, and Western blot for iNOS protein and RT-PCR for iNOS mRNA were performed to confirm the presence of iNOS. Inhibitors of iNOS and soluble guanylate cyclase (sGC), sodium nitroprusside (SNP) and L-arginine were employed to observe the action of LPS on the iNOS-NO-cGMP signalling pathway. LPS and SNP decreased number of VSMCs and increased the nitrite concentration in the culture medium, but there was no significant change in LDH activity. A cell permeable cGMP derivative, 8-Bromo-cGMP, decreased the number of VSMCs with no significant change in LDH activity. L-arginine, an NO substrate, alone tended to reduce cell count without affecting nitrite concentration or LDH level. Aminoguanidine, an iNOS specific inhibitor, inhibited LPS-induced reduction of cell numbers and reduced the nitrite concentration in the culture medium. LY 83583, a guanylate cyclase inhibitor, suppressed the inhibitory actions of LPS and SNP on VSMC proliferation. LPS increased amounts of iNOS protein and iNOS mRNA in a concentration-dependent manner. These results suggest that LPS inhibits the VSMC proliferation via production of NO by inducing iNOS gene expression. The cGMP which is produced by subsequent activation of guanylate cyclase would be a major mediator in the inhibitory action of iNOS-NO signalling on VSMC proliferation.

• Journal of Mushroom
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• v.13 no.1
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• pp.74-78
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• 2015

The 'Heuktari', a new mid-high temperature adaptable variety of oyster mushroom for the bottle culture, was bred by mating with monokaryons isolated from 'P11056' and 'MT07156'. The optimum temperature for the mycelial growth was $23{\sim}26^{\circ}C$ on PDA medium and that for the primordia formation and the growth of fruiting body of 'Heuktari' was $18{\sim}19^{\circ}C$ on sawdust substrate. In case of bottle cultivation, the period of mycelial growth was required about 30 days. In addition, the period of primordia formation and growth of fruiting body was 4 days and 5 days, respectively. In the characteristics of fruiting body, shape and color of pilei were round type and dark grayish brown, stipe color was white color and stipe shape was short and thick. The yield of fruiting bodies was 180 g/900 ml bottle which was 15% higher than that of Suhan-1ho. The gumminess and brittleness of stipe tissue were 110% and 140% stronger than those of Suhan-1ho, respectively.

• Journal of Life Science
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• v.22 no.6
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• pp.823-827
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• 2012

A bacterial strain producing a fibrinolytic enzyme, subtilisin CP-1, was isolated from Doen-Jang, a Korean traditional fermentation food. Based on the analysis of gene sequence of 16S rRNA and biochemical analysis, the strain was identified as Bacillus sp. and named as Bacillus sp. CP-1. To investigate the effect of the medium on the production of fibrinolytic enzyme from Bacillus sp. CP-1, two commercial bacterial culture media, tryptic soy broth (TSB) and Luria-Bertani (LB), were applied to the cultivation of Bacillus sp. CP-1. The strain secreted only one proteolytic enzyme (subtilisin CP-1) in the culture broth. The molecular weight of subtilisin CP-1 was estimated to be 28 kDa. Subtilisin CP-1 was optimally active at pH 9.0 and $45^{\circ}C$, and exhibited high specificity for Meo-Suc-Arg-Pro-Tyr-pNA (S-2586), a synthetic chromogenic substrate for chymotrypsin. The first eight amino acid residues of the N-terminal sequence of the enzyme are AQSVPYGI; this sequence is identical to that of subtilisin NAT and E.

• Microbiology and Biotechnology Letters
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• v.41 no.2
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• pp.145-152
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• 2013

The culture conditions to maximize the production of endoglucanase (EC 3.2.1.4) from the brown rot fungus Fomitopsis pinicola MKACC 54347 mycelia were investigated. Among the tested media for endoglucanase production, Mandel's mineral salts medium (MSM; 1% cellulose, 0.1% peptone, 0.14% $(NH_4)_2SO_4$, 0.03% urea, 0.2% $KH_2PO_4$, 0.03% $MgSO_4{\cdot}7H_2O$, 0.03% $CaCl_2$, and 0.1% trace metal solution (19.8 mM $FeSO_4$, 13.0 mM $MnSO_4$, 12.2 mM $ZnSO_4$, and 15.4 mM $CoCl_2$)) produced the highest activity of the enzyme. To optimize the medium composition for enzyme activity, the effects of various carbon, nitrogen, phosphorus, and inorganic sources were investigated in MSM. Maximal enzyme production was accomplished using a medium containing 2% carboxymethyl cellulose (CMC), 2% yeast extract, 0.2% $KH_2PO_4$, 0.03% $MnSO_4$, and 0.3% trace metal solution. Different physiological conditions, like incubation period and temperature, were also examined to assess their influence on enzyme production. Enzyme production from F. pinicola reached its highest level after cultivation for 8 days at $25^{\circ}C$. Nondenaturing polyacrylamide gel electrophoresis (PAGE), followed by the endoglucanase activity staining using CMC as the substrate, was performed to identify the endoglucanase under the culture conditions studied. Zymogram analysis of the culture supernatant revealed an endoglucanase band with a molecular mass of 52 kDa. The optimum pH and temperature for enzyme activity were $55^{\circ}C$ and pH 5.0, respectively.

• Applied Biological Chemistry
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• v.40 no.2
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• pp.89-94
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• 1997

In order to reduce energy input in direct ethanol production from raw starch by co-immobilized Aspergillus awamori(A) and Zymomonas mobilis(Z), A-Z 36 culture system which was changed to anaerobic after 36 h of aerobic fermentation without sterilization was investigated. This immobilized cell system can not be carried out under unsterile conditions because of growth of microbial contaminants from original medium. Among some food additives such as sorbic acid, benzoic acid, dehydroacetic acid, p-hydroxybenzoic acid, Vantocil IB and Neupectin-L, Vantocil IB and Neupectin-L were a potent antibacterial agent in A-Z 36 culture cell system and were not affected in hydrolysis of substrate as compared with the case of control. Ethanol yield(6.9 g/l) in system of addition of 0.1% Neupectin-L was slightly higher than that in control(6.4 g/l). When 2% starch was fed five times in fed-batch culture with 0.1% Neupectin-L, ethanol yield and productivity were 34 g/l and 2.0 g/l/day, respectively.

• Journal of Bio-Environment Control
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• v.10 no.2
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• pp.125-131
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• 2001

The objective of this study was to estimate the amount of nutrient and water taken up at different growth stages by cucumber (Cucumis sativus L. cv. Eunsung Backdadagi) grown in a closed substrate culture system. The amount of nutrient solution absorbed increased in proportion to days from planting at the first stage of growth and depended on the level of radiation after the mid stage of growth. After the mid growth stage, the amount of nutrient solution absorption was maintained at 80-100 mg.MJ$^{-1}$ . Total amount of absorbed inorganic ions except S increased since the nutrient solution absorption increased with the level of radiation, although the absorption rate of each inorganic ion declined. A highly significant correlation ($R^2$>0.9) was found between amount of inorganic ions absorbed and days after planting, LAI, total dry weight and leaf dry weight, but not with CGR. Correlation coefficient between days after planting and the amount of nutrient solution absorbed per unit radiation level was 0.92. Correlation coefficient between leaf area an the amount of nutrient solution absorbed per unit radiation level was 0.97. Regression of the amount of nutrient solution absorbed per unit radiation level and nutrient ions uptake showed a high significance ($R^2$>0.9).

• The Korean Journal of Mycology
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• v.22 no.3
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• pp.266-275
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• 1994

Brewer's yeast extract can be used as a good substrate for the culture of Lentinula edodes Mycelia(LEM). We found that it was better to filter the extract through three kinds of sieves and then to heat for hydrolysis and concentration at $90^{\circ}C$ prior to use. Also the maximum condition for the growth of LEM was investigated. We found that addition of inorganic salts such as calcium enhanced the growth of LEM. On the other hand, addition of carbon and nitrogen sources to the medium did not affect, and even inhibited under certain conditions, the growth of LEM. The maximum temperature for the growth of LEM was around $25^{\circ}C$. Also, it grows better when agitated by shaking at 100 rpm for airration. The appropriate concentration of the extract to use was 10%. Under these conditions, LEM could reach to the confluency after cultivation of 12 days. Our extract formula seems better than other available media for LEM growth, producing higher crude protein content and better taste.

• Journal of Mushroom
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• v.21 no.3
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• pp.140-144
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• 2023

The objective of this study was to achieve biological control of green mold disease in Pyogo mushrooms using antagonistic microorganisms. Bacillus subtilis BSM320 cells inhibited mycelial growth by 48-60% against three Trichodermaisolates including T. hazianumisolated from the substrates of Lentinula edodes, showing their antifungal activity.The bacteria were cultured to a high density of 4.2 × 10⁹±113.7 cfu/mlin aqueous extract of composted spent mushroom substrates of L. edodes containing 1% glucose and showed a higher growth rate than that observed when using the commercial medium, Luria-Bertani broth. The bacterial culture showed a 75% protective effect without damaging the mushroom fruiting bodies. These results suggest that B. subtilis BSM320culture is suitable for biological control of green mold disease during mushroom cultivation.

• Korean Journal of Agricultural Science
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• v.41 no.4
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• pp.321-326
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• 2014

The aim of this study was to investigate which hydroponic system is the optimum for growth and photosynthetic characteristics of Angelica gigas during experiment. Angelica gigas 'Manchu' were sowed and managed under a growth room chamber. The environmental conditions (temperature $22^{\circ}C/18^{\circ}C$ (day/night), relative humidity 50-70%, photosynthetic photon flux density (PPFD) $120{\pm}6{\mu}mol\;m^{-2}s^{-1}$) were maintained for 3 weeks. Forty eight seedlings with 4-5 leaves were transplanted in deep flow technique (DFT), substrate, and spray culture systems [culture bed: 800 (L) ${\times}$ 800 (W) ${\times}$ 400 mm(H)] under $150{\pm}5{\mu}mol\;m^{-2}s^{-1}$ PPFD provided with fluorescence lamps and cultivated for 11 weeks. At the end of the experiment, fresh and dry weights, leaf lenghth and width, SPAD, root fresh, and dry weights, and root volume of Anglica gigas were measured. Photosynthetic rate of Anglica gigas were measured with portable photosynthesis systems to investigate optimum PPFD, $CO_2$ concentration, and air temperature conditions. Fresh and dry weights of Anglica gigas grown in substrate were significantly greater than DFT-treated, but there were not significant with spray treatment. Leaf photosynthesis of Anglica gigas showed the tendency to sharply increase as PPFD was increased from 50 to $200{\mu}mol\;m^{-2}s^{-1}$. Though $CO_2$ saturation point was around $1000-1200{\mu}mol\;mol^{-1}$, increase in air temperature from 16 to $26^{\circ}C$ did not quite affect photosynthesis of Anglica gigas. In conclusion, Anglica gigas may be optimally cultivated with a spray culture system as air temperature, PPFD, and $CO_2$ concentration for environment are controlled at $20{\pm}3^{\circ}C$, $150{\mu}mol\;m^{-2}s^{-1}$, and around $1000{\mu}mol\;mol^{-1}$ for mass production.