• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1384-1391
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• 2006

The signal transduction system is one of the most important devices involved in maintaining life, and many protein kinases are included in the cellular signal transduction system. Finding a protein kinase inhibitor is very valuable, as it can be used to study cell biology and applied to pharmaceuticals. For the efficient and rapid screening of protein kinase inhibitors, two assay systems were combined; the nonradioactive protein kinase assay system that uses an FITC-labeled IRS-2 peptide and the cell-based paper disc assay system that uses Streptomyces griseus as the indicator strain. Among 330 kinds of herb extracts tested, the extract of Dalbergia odorifera exhibited the strongest inhibitory activity in the two assay systems and was selected for further isolation. Based on solvent extraction and many steps of chromatography, seven compounds were finally separated to homogeneity and their structures determined by $^{1}H$ and $^{13}C$ NMR spectroscopies. Four were to be flavonoids and identified as butin ($C_{15}H_{12}O_5$, Mw=272.07), 3'-hydroxymelanetin ($C_{16}H_{12}O_6$, Mw=300.06), liquiritigenin ($C_{15}H_{12}O_4$, Mw=256.07), and 2'-hydroxyformononetin ($C_{16}H_{12}O_{5}$, Mw=284.07). 3'-Hydroxymelanetin inhibited the phosphorylation of the GSK3 protein by Akt to 37% at a concentration of $10{\mu}g/ml$ and showed the strongest cytotoxicity ($ED_{50}<50{\mu}g/ml$) against the human cancer cell line HCT116. Under the same conditions, liquiritigenin also inhibited the phosphorylation of GSK3 by Akt to 26%, and its cytotoxicity against the HCT116 cell line was lower than $100{\mu}g/ml$.

• Molecules and Cells
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• v.45 no.7
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• pp.502-511
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• 2022

Bacterial volatile compounds (BVCs) exert beneficial effects on plant protection both directly and indirectly. Although BVCs have been detected in vitro, their detection in situ remains challenging. The purpose of this study was to investigate the possibility of BVCs detection under in situ condition and estimate the potentials of in situ BVC to plants at below detection limit. We developed a method for detecting BVCs released by the soil bacteria Bacillus velezensis strain GB03 and Streptomyces griseus strain S4-7 in situ using solid-phase microextraction coupled with gas chromatography-mass spectrometry (SPME-GC-MS). Additionally, we evaluated the BVC detection limit in the rhizosphere and induction of systemic immune response in tomato plants grown in the greenhouse. Two signature BVCs, 2-nonanone and caryolan-1-ol, of GB03 and S4-7 respectively were successfully detected using the soil-vial system. However, these BVCs could not be detected in the rhizosphere pretreated with strains GB03 and S4-7. The detection limit of 2-nonanone in the tomato rhizosphere was 1 µM. Unexpectedly, drench application of 2-nonanone at 10 nM concentration, which is below its detection limit, protected tomato seedlings against Pseudomonas syringae pv. tomato. Our finding highlights that BVCs, including 2-nonanone, released by a soil bacterium are functional even when present at a concentration below the detection limit of SPME-GC-MS.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.3
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• pp.364-370
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• 2008

Chemical analysis and in vitro studies were conducted to investigate the nutritive value for ruminants of cell mass from lysine production (CMLP) which is a by-product of the lysine manufacturing process. Proximate analysis, protein fractionation, and in vitro protein degradation using protease from Streptomyces griseus and strained ruminal fluid were carried out to estimate ruminal protein degradability of CMLP with two reference feedstuffs-soybean meal (SBM) and fish meal (FM). Amino acid composition and pepsin-HCl degradability were also determined to evaluate postruminal availability. CMLP contained 67.8% crude protein with a major portion being soluble form (45.4% CP) which was composed of mainly ammonium nitrogen (81.8% soluble CP). The amount of nucleic acids was low (1.15% DM). The total amount of amino acids contained in CMLP was 40.60% DM, which was lower than SBM (47.69% DM) or FM (54.08% DM). CMLP was composed of mainly fraction A and fraction B2, while the protein fraction in SBM was mostly B2 and FM contained high proportions of B2 and B3 fractions. The proportion of B3 fraction, slowly degradable protein, in CP was the highest in fish meal (23.34%), followed by CMLP (7.68%) and SBM (1.46%). CMLP was degraded up to 51.40% at 18 h of incubation with Streptomyces protease, which was low compared to FM (55.23%) and SBM (83.01%). This may be due to the insoluble portion of CMLP protein being hardly degradable by the protease. The in vitro fermentation by strained ruminal fluid showed that the amount of soluble fraction was larger in CMLP (40.6%) than in SBM (17.8%). However, because the degradation rate constant of the potentially degradable fraction of CMLP (2.0%/h) was lower than that of SBM (5.8%/h), the effective ruminal protein degradability of CMLP (46.95%) was slightly lower than SBM (53.77%). Unavailable fraction in the rumen was higher in CMLP (34.0%) compared to SBM (8.8%). In vitro CP degradability of CMLP by pepsin was 80.37%, which was lower than SBM (94.42%) and FM (89.04%). The evaluation of protein degradability using different approaches indicated that soluble protein in CMLP may supply a large amount of ammonia in the rumen while insoluble protein can be by-passed from microbial attacks due to its low degradability. The results from this study suggest that CMLP can be used as a protein supplement to ruminants for supplying both non-protein nitrogen to rumen microbes and rumen undegradable protein to the host animal.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.4
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• pp.282-292
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• 1987

In order to exploit a new type of food source, enzamatically modified hydrolysates and the plasteins synthesized from the filefish (Nevoden modestus) protein hydrolysates by plastein reaction were investigated. The optimum conditions for enzymatic hydrolysis of filefish muscle and synthesis of plasteins using papain, pepsin, $\alpha-chymotrypsin$ and protease (from Streptomyces griceus) were determined. The optimum temperature and pH for the hydrolysis of filefish muscle by papain, pepsin, $\alpha-chymotrypsin$ and protease were $50^{\circ}C,\;40^{\circ}C,\;55^{\circ}C\;and\;50^{\circ}C$; and 6, 2, 7 and 8, respectively. Those for incubation time and enzyme concentration were 4hr, $0.5\%$ for papain and protease, 24hrs $1.0\%$ for pepsin and $\alpha-chymotrypsin$. The pepsin was found to be more reasonable substrate for plastein synthesis from the economic point of view. The enzyme-induced plastein reaction could be optimized, namely, pH 4 for pepsin, pH 7 for $\alpha-chymotrypsin$, pH 6 for papain and protease: substrate concentration $40\%$ for pepsin, $\alpha-chymotrypsin$ and protease, $50\%$ for papain; the time of incubation, 24hr; enzyme/substrate ratio, 1 : 100(W/V) ; incubation temperature, $50^{\circ}C$.

• Food Science of Animal Resources
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• v.30 no.3
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• pp.504-511
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• 2010

This study was undertaken to investigate the effects of feeding complex probiotics (Lactobacillus casei, Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus oryzae, Streptomyces griseus, $1.5{\times}10^{10}$ CFU/kg) and antibiotics (oxytetracycline (OTC), 110 ppm) on growth performance and meat quality characteristics of broiler chicks. In the experiment 1, 0.3% complex probiotics feeding level was chosen to be proper addition level due to better average daily gain (ADG), feed conversion (FC) and dressing percent (DP) results among 3 levels (0.1, 0.3 or 0.5%). In the experiment 2, 5 treatments (T1, no probiotics + no antibiotics; T2, probiotics 0.3% + no antibiotics; T3, probiotics 0.3% + antibiotics 50%; T4, probiotics 0.3% + antibiotics 100%; T5, no probiotics + antibiotics 100%) were investigated. In the growth performance of broilers, T5 (antibiotics 100% only) showed the highest (p<0.05) ADG and FC values while T1 (control) showed the worst growth performance. However, T3 (probiotics 0.3% + antibiotics 50%) showed higher ADG (p<0.05), FC (p<0.05) and DP (p>0.05) values compared to control. In the breast and leg meat quality, T3 showed similar pH, proximate composition, cooking loss and meat color values except shear force value compared to T5. Addition of 0.3% probiotics with 50% antibiotics (T3) tended to lower the blood cholesterol levels of broiler chicks and Escherichia coli or Salmonella counts in cecum microflora of broiler chicks compared to T5. In the residual antibiotics analysis, T3 contained 0.04 ppm of residual antibiotics in the breast meat while T4 or T5 contained 0.1 ppm of residual antibiotics and addition of 0.3% probiotics with 50% antibiotics in broiler diets could lower the residual antibiotics level to 40% in the meat. As a result, 0.3% probiotics addition with 50% antibiotics in the broiler diets could be recommended for the production of high quality broiler meat.

• Journal of Animal Science and Technology
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• v.44 no.5
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• pp.549-560
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• 2002

This study, including two in vitro experiments and an in vivo experiment were conducted to evaluate effects of Passtein$^{(R)}$ on crude protein degradability, ruminal fermentation characteristics and nutrient digestibility. In in vitro experiment protein degradability was examined using borate-phosphate buffer and neutral detergent, and using protease from Stroptomyces griseus at 39$^{\circ}C$ for 0, 2, 4, 8, 12, and 48 h. In addition, an in vivo experiment was conducted in a switch back design and ruminal fermentation and nutrient digestibility were determined. Four ruminal-fistulated Holstein cows weighing 300kg in mean body weight randomly allotted to 2 treatments (control and Passtein$^{(R)}$ supplementation). Although there was no significant difference on protein fraction between treatments, it appears that Passtein$^{(R)}$ supplementation decreased buffer soluble protein fraction compared to control. Protein degradability was not affected by Passtein$^{(R)}$ from 0 h to 4 h, but decreased at 12 h and 48 h compared to control. Degradation of immediately degradable fraction was higher in Passtein$^{(R)}$ treatment, but degradation of fermentable fraction was lower in Passtein$^{(R)}$ treatment compared to control. The pH and $NH_3$-N concentration tended to increase in Passtein$^{(R)}$ treatment, but VFA production, microbial counts and enzyme activity tended to decrease in Passtein$^{(R)}$ treatment compared to control. In addition, nutrient digestibility in the total tract tended to increase in Passtein$^{(R)}$ treatment compared to control.

• Korean Journal of Organic Agriculture
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• v.15 no.2
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• pp.195-205
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• 2007

This study was conducted to determine effects of probiotics on performance, nutrient digestibility and nitrogen retention in Korean black goats fed two diets differing in forage to concentrate (F:C) ratio (30:70 and 70:30), for the establishment of their more efficient feeding management system. The probiotics employed in this trial were mixtures of different microbial species, which consisted of Lactobacillus casei, Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus oryzae and Streptomyces griseus. Additional levels of probiotics to each F:C ratio (70:30 or 30:70) were 0 and 0.2%, respectively. Thus, twelve Korean black male goats were allotted to treatments in four groups of three goats per treatment and then they were housed in individual metabolism cages with a randomized complete block design for 21 days. Dry matter (DM) intakes were not affected by dietary F:C ratio and probiotics. Digestible DM amounts were significantly (p<0.05) decreased with increasing levels of dietary forage, but they were not affected by probiotics addition. Dry matter intakes per metabolic body weight and their ratio per body weight had a similar trend to DM intakes with no significant difference by F:C ratio and probiotics addition. The nutrient digestibility was significantly (p<0.05) increased with decreasing levels of forage in the diet, but it was not affected by probiotics addition. Urinary nitrogen loss was significantly (p<0.05) decreased with decreasing levels of dietary forage, but there was no significant difference between probiotics-supplemented and unsupplemented groups within the same F:C ratio. On the contrary, nitrogen retention was decreased with increasing levels of dietary forage, and probiotics supplementation to two diets differing in F:C ratio showed slightly increasing trends in the nitrogen retention. From the above results, probiotics supplementation to two diets differing in F:C ratio did not have the significant influence on feed intakes, nutrient digestibility and nitrogen retention. Consequently, these parameters of Korean black goats were dominated rather by F:C ratio than by dietary probiotics.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.10
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• pp.1377-1387
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• 2000

Two laboratory techniques: (1) an in vitro method with two procedures for measuring protein degradabilities and (2) an in vitro method with three procedures for measuring protein solubility, were investigated to determine which laboratory techniques could most accurately predict the quantity of rumen protein degradation kinetics of legume seeds after dry roasting under various conditions, in terms of (1) rumen protein disappearance ($D_j$, where j=0, 2, 4, 8, 12, 24 and 48 h incubation), (2) rumen protein effective degradability (EDCP), (3) the parameters describing rumen degradation characteristics (the soluble fraction: S, the potentially degradable fraction: D, undegradable fraction: U, lag time: T0 and the degradation rate: Kd) and (4) rumen bypass protein (BCP), which were determined by the method accepted internationally at present, in sacco nylon bag technique using the standardized Dutch method. Feeds evaluated were the raw and dry roasted whole faba (Vicia faba) beans (WFB) and whole lupin (Lupinus albus) seeds (WLS), each was dry roasted under various conditions (at 110, 130 or $150^{\circ}C$ for 15, 30 or 45 min). In vitro protein degradability ($D_1$_Auf and $D_{24}$_Auf) were determined using the modified Aufr re method by enzymatic hydrolysis for 1 h and 24 h using a protease extracted from Streptomyces griseus in a borate-phosphate buffer. In vitro protein solubility ($bf_1$_S, $bf_2$_S, $bf_3$_S) was measured in a borate-phosphate buffer with three different procedures. Results from laboratory techniques (in vitro) were correlated and linearly regressed with in sacco results. Of the three procedures of in vitro protein solubility evaluated, none of them could predict in sacco results with good precision. The highest Pearson correlation coefficient ($R^2$) was less than 0.50. Of two procedures of in vitro protein degradability studied, the $D_1$_Auf values were closely correlated with in sacco parameters: Kd, EDCP and %BCP with high R' values: 0.82, 0.85 and 0.85, respectively, and closely correlated with in sacco $D_j$ at 2, 4, 8 and 12 h rumen incubation with high $R^2$ values: 0.83, 0.91, 0.93 and 0.83, respectively. The $D_{24}$_Auf values could not predict in sacco results. The highest $R^2$ value was less then 0.40. These results indicated that in vitro protein solubility measured in borate-phosphate failed to identify differences in the rate and extent of protein degradation of legume seeds after dry roasting under various conditions and thus should not be used to predict rumen degradation, particularly for heat processed feedstuffs. But in vitro protein degradability using the modified Aufr re method by enzymatic hydrolysis for 1 h or possibly an intermediate time (>1 h and <24 h) is a promising laboratory procedure to detect effectiveness of dry roasting legume seeds on rumen protein degradation characteristics and could be used as a simple laboratory method to predict the rate and extent of protein degradation in the rumen in sacco with high accuracy. The equations to predict EDCP, Kd and BCP of dry roasted legume seeds (WLS and WFB) under various conditions are as follow: For both: EDCP (%)=-1.37+1.06*$D_1$_Auf ($R^2=0.85$, p<0.01). For both: Kd (%/h)=-21.81+0.49*$D_1$_Auf ($R^2=0.82$, p<0.01). For both: %BCP=103.37-1.07*$D_1$_Auf ($R^2=0.85$, p<0.01).

• Microbiology and Biotechnology Letters
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• v.1 no.1
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• pp.51-57
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• 1973

2,2'-Methylene bis (3,4,6-trichloroacetoxy benzene) had been synthesized from Hexachlorophene. The eleven new derivatives -(2,2'Methylene bis [3,4,6-trichloro o-(${\beta}$-anilinopropionoxy) benzene]: m. p 173∼4$^{\circ}C$, C$\_$31/H$\_$24/N$_2$O$_4$Cl$\_$6/, 2,2' Methylene bis [3,4,6-trichloro (${\beta}$-Cyclohexylaminopropionoxy) benzene]: M. P, 187∼8$^{\circ}C$, C$\_$31/H$\_$36/N$_2$O$_4$Cl$\_$6/, 2,2'-Methylene bis [3,4,6-trichloro (${\beta}$-phenyl hydrazinopropionoxy) benzene]; M. P. 151∼3$^{\circ}C$, C$\_$33/H$\_$28/N$_2$O$_4$Cl$\_$6/, 2,2'-Methylene bis [3,4,6-trichloro (${\beta}$-o-toluidinopropionoxy) benzene]: M. P. 172∼3$^{\circ}C$, C$\_$33/H$\_$28/N$_2$O$_4$Cl$\_$6/, 2,2'-Methylene bis [3,4,6-trichloro (${\beta}$-p-todudinopropionoxy) benzene]: 153∼4$^{\circ}C$, C$\_$33/H$\_$28/N$_2$O$_4$Cl$\_$6/, 2,2'-Methylene bis [3,4,6-trichloro (${\beta}$-o-chloro anilinopropionoxy) benzene]: 170∼2$^{\circ}C$, C$\_$31/H$\_$27/N$_2$O$_4$Cl$\_$8/, 2,2'-Methylene bis [3,4,6-trichloro (${\beta}$-p-sulfamilinopropionoxy) bengene]: M. P. 310-5$^{\circ}C$, C$\_$31/H$\_$24/N$_2$S$_2$O$\_$10/Cl$\_$8/, 2,2'-Methylene bis [3,4,6-trichloro (${\beta}$-piperidinopropionoxy) benzene]: M. P. 168∼2$^{\circ}C$, C$\_$29/H$\_$32/N$_2$O$_4$Cl$\_$6/, 2,2'-Methylene bis (3,4,6-trichloro (${\beta}$-morpholinopropionoxy) benzene]: M. P. 226∼8$^{\circ}C$, C$\_$27/H$\_$28/N$_2$O$\_$6/Cl$\_$6/, 2,2'-Methylene bis (3,4,6-trichloro (${\beta}$-2-amino pyridino propionoxy) benzene]; M. P. 145∼6$^{\circ}C$, C$\_$29/H$\_$22/N$_4$O$_4$Cl$\_$5/-were synthesized by Mannichs reaction as potential antimicrobial agents and their antimicrobial activity were tested against Bacillus subtilis, Pseudomonas ovalis, Escherichia coli, Staphylococcus aureus, aerogenes, Bacillus Acerobacter Polymyxa, Bacillus brevis, Streptomyces griseus, Candida troficalis, Rhodotorula glutinis, Candida utilis, Hansenula anamola, Saccharomyces cerevisiae in vitro. Among them, compounds of benzylamine and p-toludine showed an effective antimibrobial activity againt Bacillus subtilis and Pseudomonas ovalis.