• Journal of Animal Science and Technology
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• v.52 no.2
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• pp.117-124
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• 2010

An in situ ruminal disappearance trial and an in vivo sheep metabolism trial were conducted to evaluate the nutritional value of spent mushroom substrate (SMS, originated from Pleurotus osteratus bed-type cultivation). The raw SMS was ensiled (ESMS) for 30 days with 5% (w/w, DM basis) molasses, 0.5% (v/w) yeast (Saccharomyces cerevisiae) and 0.5% (v/w) lactic acid bacteria (Lactobacillus plantarum). Two ruminally cannulated Holsteins (average BW 620 kg) were used to evaluate in situ disappearance. Six sheeps (average BW 48 kg) were fed, in $3{\times}3$ Latin square design, rice straw alone (Control), 25% (ESMS-25) and 50% (ESMS-50) of rice straw were replaced with ensiled SMS. For an in situ trial, ruminal DM and neutral detergent fiber (NDF) disappearance of SMS were increased after ensiling (P<0.05). For a sheep trial, protein and fiber (NDF, acid detergent fiber, crude fiber) digestibilities were decreased (P<0.05), crude ash digestibility was increased (P<0.001), and nitrogen retention was not affected (P>0.05) as rice straw was replaced with ensiled SMS. Ruminating time was decreased by an average of 28% by feeding ensiled SMS (P<0.05). Ensiled SMS (Bed-type cultivation) had 76% of energy value of rice straw. Consequently, ensiled SMS (Bed-type cultivation, 100% cotton waste) could be used as a roughage source appropriate for maintenance type rations for ruminants.

• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.839-846
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• 1997

Addition of three natural flavoring materials, hydrolyzed vegetable protein (HVP), hydrolyzed animal protein (HAP) and yeast extract (YE), into 0.2 M cystine-0.1 M lactose-0.1 M maltose solution (control) was studied for development of meat-like flavor by Maillard reaction. The HVP, HAP and YE were added individually at various concentrations and were mixed at selected concentration in order to compare their effects. The absorbance, color, sensory characteristics and volatile compounds of the solutions after the reaction at $100^{\circ}C$ for 8 hr were measured. The results showed that the absorbances of reaction solution at 420 nm and 278 nm were increased as reaction time and the concentration of the natural flavoring material increased. Also ‘L’ values of reaction solutions added with HVP, HAP or YE decreased while the ‘b’ value increased slightly. From the results of sensory evaluation 1.16% HVP, 0.94% HAP, 1.48% YE or 1.16% HVP + 0.94% HAP were selected as the appropriate substrates for the meat-like flavor development. The volatile compounds identified by GC/MS for the control and those added with 1.16% HVP or 1.16% HVP+0.94% HAP were 1 hydrocarbons, 9 aldehydes, 5 ketones, 1 ester, 5 alcohols, 2 aromatics(benzene), 2 furans, 1 sulfur compound.

• Journal of the Korean GEO-environmental Society
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• v.13 no.9
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• pp.53-59
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• 2012

This study investigated characteristics of decomposition of tetrabutoxysilane (TBOS) as a slow release substrate (SRS) and on effect of TBOS decompostion compounds (acetate and butylate) for anaerobic dechlorination of trichloroethylene (TCE). In the batch experiment, TCE, cis-dichloroethene (cis-DCE), 1-butanol and TBOS were analysed by GC/FID and acetate and butylate were measured by HPLC. 1M of TBOS transferred and accumulated 4M of 1-butanol by abiotically hydrolysis reaction. The hydrolysis rate was in a range of 0.186 ${\mu}M/day$. On other hand, 1-butanol fermented to butyrate and acetate with indigenous culture from natural sediments. This results showed that TBOS could be used a slow release substrate in the natural sites. The dechlorinated potential of TCE with acetate and butyrate was increased with a decreasing initial TCE concentrations. In addition, first order coefficients of dechlorination with acetate as electron donor was higher then that with butyrate. It is because that dechlorination of Geobacter lovleyi was affected by substrate affinity, biodegradability and microbial acclimation on various substrates. However, dechlorinated potential of Geobacter lovleyi was decreased with accumulation cis-DCE in the anaerobic decholoronation process. The overall results indicated that SRS with Geobacter lovleyi might be a promising material for enhancing dechlorination of TCE on natural site and cis-DCE should be treated by ZVI as reductive material or by coexisting other dechlorinated bacteria.

• BMB Reports
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• v.37 no.6
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• pp.720-725
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• 2004

A number of signaling molecules contain small pleckstrin homology (PH) domains capable of binding phosphoinositides or proteins. Phospholipase C (PLC)-${\gamma}1$ has two putative PH domains, an $NH_2$-terminal (PH1) and a split PH domain ($nPH_2$ and $cPH_2$). We previously reported that the split PH domain of PLC-${\gamma}1$ binds to phosphatidylinositol 4-phosphate (PI(4)P) and phosphatidylinositol 4,5-bisphosphate (PI(4,5)$P_2$) (Chang et al., 2002). To identify the amino acid residues responsible for binding with PI(4)P and PI(4,5)$P_2$, we used site-directed mutagenesis to replace each amino acid in the variable loop-1 (VL-1) region of the PLC-${\gamma}1$ $nPH_2$ domain with alanine (a neutral amino acid). The phosphoinositide-binding affinity of these mutant molecules was analyzed by Dot-blot assay followed by ECL detection. We found that two PLC-${\gamma}1$ nPH2 domain mutants, P500A and H503A, showed reduced affinities for phosphoinositide binding. Furthermore, these mutant PLC-${\gamma}1$ molecules showed reduced PI(4,5)$P_2$ hydrolysis. Using green fluorescent protein (GFP) fusion protein system, we showed that both $PH_1$ and $nPH_2$ domains are responsible for membrane-targeted translocation of PLC-${\gamma}1$ upon serum stimulation. Together, our data reveal that the amino acid residues $Pro^{500}$ and $His^{503}$ are critical for binding of PLC-${\gamma}1$ to one of its substrates, PI(4,5)$P_2$ in the membrane.

• Journal of Apiculture
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• v.32 no.1
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• pp.27-39
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• 2017

PCR-chip-based ultra-rapid multiplex PCRs for detection of six major infectious pathogens in honeybee were developed. The 6 kinds of major infectious pathogens in honeybee included Paenibacillus larvae causing American Foulbrood, Melissococcus plutonius causing European Foulbrood as bacteria, Ascosphaera apis (Chalkbrood), Aspergillus flavus (Stonebrood), Nosema apis and Nosema ceranae (Nosemosis) as fungi. The developed PCR-chip-based ultra-rapid multiplex PCR showed successful amplification for all six major pathogens in the presence of more than $10^3$ molecules. The time for confirming amplification (Threshold cycles; Ct-time) was about 7 minutes for two species, and about 9 minutes for four species. Total 40 cycles of PCR took 11 minutes 42 seconds and time for melting point analysis was 1 minute 15 seconds. Total time for whole PCR detection was estimated 12 minutes 57 seconds (40 cycles of PCR and melting point analysis). PCR-chip based ultra-rapid multiplex PCR using standard DNA substrates showed close to 100% accuracy and no false-amplification was found with honeybee genomic DNA. Ultra-rapid multiplex PCR is expected to be a fast and efficient pathogen detection method not only in the laboratory but also in the apiary field.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.327-333
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• 2005

Dihydroxy-acid dehydratase (DHAD, 2,3-dihydroxy-acid hydrolyase, EC 4.2.1.9) is one of the key enzymes involved in the biosynthetic pathway of the branched chain amino acid isoleucine and valine. Although the enzyme have been purified and characterized in various mesophiles including bacteria and eukarya, the biochemical properties of DHAD has bee not yet reported from hyperthermophilic archaea. In this study, we cloned, expressed, and purified a DHAD homologue from the thermoacidophilic archaeon Sulfolobus solfataricus P2, which grows optimally at $80\;^{\circ}C$ and pH 3, in E. coli. Characterization of the recombinant S. solfataricus DHAD (rSso_DHAD) revealed that it is the dimeric protein with a subunit molecular weight of 64,000 Da in native structure. rDHAD showed the highest activity toward 2,3-dihydroxyisovaleric acid among 17 aldonic acid substrates Interestingly, this enzyme also displayed 50 % activities toward some pentonic acids and hexonic acids when compared with the activity of this enzyme to the natural substrate. Moreover, rSso_DHAD indicated relatively higher activity toward D-gluconate than any other hexonic acids tested in substrates. $K_m$ and $V_{max}$ values of rSso_DHAD were calculated as $0.54\;{\pm}\;0.04\;mM$ toward 2,3dihydroxyisovalerate and $2.42\;{\pm}\;0.19\;mM$ toward D-gluconate, and as $21.6\;{\pm}\;0.4\;U/mg$ toward 2,3-dihydroxyisovalerate and $13.8\;{\pm}\;0.4\;U/mg$ toward D-gluconate, respectively. In the study for biochemical properties, the enzyme shows maximal activity between $70^{\circ}C$ and $80^{\circ}C$, and the pH range of pH 7.5 to 8.5. The half life time at $80^{\circ}C$ was 30 min. A divalent metal ion, $Mn^{2+}$, was only powerful activators, whereas other metal ions made the enzyme activity reduced. $Hg^{2+}$, organic mercury, and EDTA also strongly inhibited enzyme activities. Particularly, the rSso_DHAD activity was very stable under aerobic condition although the counterparts reported from mesophiles had been deactivated by oxygen.

• Journal of Bio-Environment Control
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• v.16 no.2
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• pp.101-107
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• 2007

This experiment was carried out to develop the substrate mixtures for the production of high quality plug seedlings of cucumber. Peatmoss based substrates, rice hull, carbonized rice hull, decomposed sawdust, perlite and granular rockwool were mixed by five different mixing ratioes (M1, M2, M3, M4 and M5). The cultivars used were cucumber (Cucumis sativus L. cv. Janghyung heukjinju) plants. The higher the content of peatmoss added, the higher the plant growth in terms of plant height, leaf area and total dry weight, which leading to the production of high quality plug seedlings. Seedlings growth of cucumber were greater in M5 mixtures [peatmoss:rice hull:decomposed sawdust=40:40:20(v/v)], M4 mixtures [peatmoss:rice hull:decomposed sawdust:granular rockwool=30:25:20:25(v/v)] and M2 mixtures [peatmoss:rice hull:decomposed sawdust:granular rockwool=20:20:15:25:20(v/v)] The concentrations of nutrient solution (EC) had a great influence on plant height, leaf area, total fresh and dry weight of cucumber seedlings growth. As the concentration of nutrient solution increased from 0.1 to $1.5dS{\cdot}m^{-1}$, the growth and seedling quality of cucumber in plant height, leaf area and dry weight were significantly improved.

• KSBB Journal
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• v.22 no.5
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• pp.362-365
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• 2007

Cyanobacteria have been identified as one of the most promising group producing novel biochemically active natural products. Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Large multienzyme complexes which are responsible for the non-ribosomal biosynthesis of peptides are modular for the addition of a single amino acid. An activation of amino acid substrates results in an amino adenylate occuring via an adenylation domain (A-domain). A-domains are responsible for the recognition of amino acids as substrates within NP synthesis. The A-domain contains ten conserved motifs, A1 to A10. In this study, ten conserved motifs from A1 to A10 were checked regarding their amino acid sequence of the NRPS-module of Microcystis aeruginosa PCC 7806. The part of the amino acid sequence chosen was that which contained as many conserved motives as possible, and then these amino sequence were compared between other cyanobacteria to design a new degenerate primer. A new degenerate primer (A3/A7 primer) was designed to detect any putative NP synthetase region in unkwon cyanobacteria by a reverse translation of the conserved amino acid sequence and a search for cyanobacterial DNA bank.

• Journal of Wetlands Research
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• v.13 no.2
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• pp.355-362
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• 2011

Bog bean (Menyanthes trifoliata L.) is an endangered species in Korea and a perennial macrophyte with long rhizome, inhabiting in oligotrophic fen or edges of montane lakes. To decide appropriate substrate type for restoration of this plant, we investigated the effect of substrates (e.g. water, Sphagnum mat, paddy soil) on growth of bog bean. There were two water conditions on paddy soils: saturated and flooded. We planted 10cm rhizome in mesocosms and measured coverage, leaf area, leaf number and rhizome biomass. Bog bean growed until August in water and Sphagnum mat and until October in paddy soil. Rhizome biomass at the end of November were 49, 77, 239, and 312g in water, Sphagnum mat, paddy soil with water saturated, and paddy soil with water flooded conditions, respectively. The results indicate that bog bean can grow better in paddy soil which have higher nutrient than water or Sphagnum mat which represents natural habitat condition of bog bean. This reveals that actual ecological niche of bog bean is different from fundamental ecological niche in substrate. For successful restoration of bog bean in nutrient rich area, it is necessary to know the competitiveness of bog bean in various substrate conditions.

• The Journal of Korean Academy of Prosthodontics
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• v.45 no.3
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• pp.382-388
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• 2007

Statement of problem. The role of calcium sulfate in stimulating the growth of gingival soft tissue has been reported in few studies. Such a unique property of calcium sulfate could serve as a trouble-solving broker in compensating for the lack of soft tissues in various oral surgeries. Purpose. The purpose of this study was to compare the proliferating activities of human gingival fibroblasts seeded on various bone graft barrier materials of calcium sulfate, collagen, and polytetrafluorethylene (PTFE). Material and methods. Two calcium sulfates ($CAPSET^{(R)}$. and $CalForma^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA), a resorbable natural collagen ($Bio-Gide^{(R)}$, Geistlich Pharma Ag., Wolhusen, Switzerland), and a non-resorbable PTFE ($TefGen-FD^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA) served as the human gingival fibroblasts' substrates and comprised the four experimental groups, whereas the untreated floors of culture plastics were used in the control group, in this study. Cells were trypsinized, seeded, and incubated for 48 h. The proliferating activities of fibroblasts were determined by XTT and SRB assay and absorbance (optical density, OD) was measured. One-way ANOVA was used to analyze the differences in the mean OD values between the groups of CAPSET, CalForma, Bio-Gide, TefGen, and the control (p<0.05). Results. From the XTT assay, the mean OD value of the control group, the highest, was significantly greater than that of any of the four experimental groups followed by CalForma, CAPSET, TefGen, and Bio-Gide. Further, the mean OD value of CalForma, was significantly greater compared to that of Bio-Gide. From the SRB assay, Calforma showed the highest mean OD value, which was significantly greater than that of any other groups, followed by the control, CAPSET, Bio-Gide, and TefGen. The mean OD values of both the control and CAPSET were significantly greater compared to that of TefGen (p<0.05). Conclusion. Assessment of the viability and proliferation of cultured fibroblasts seeded and incubated for 48 h on various barrier-material substrates using XTT and SRB assay showed that calcium sulfate $CalForma^{(R)}$ promotes the proliferating activity of human gingival fibroblasts.