• Journal of the Korea Organic Resources Recycling Association
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• v.14 no.2
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• pp.91-100
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• 2006

Effects of the supplement of earthworm cast produced from the feeding of organic wastes and earthworm on the productivity and nutritional constituents of functional eggs were investigated. Compared with control experiments, the case supplemented with earthworm cast showed high ratios in egg production, selection and the reserved feed. According to the experiment with earthworm, both the number of jumbo eggs and the quantity of reserved feed were increased. Therefore, the nutritional effect of earthworm in the feed was positive. The optimum percentage of earthworm cast in the feed was 10%: the average laying increased to 96.8, which was a 5% increase; the ratio of the large eggs increased by 5% although the ratio of jumbo eggs and of extra large eggs decreased by 5% and 1.1%, respectively; the average reserved feed was 662.5g. Also, Beauveria bassiana was inoculated into the feed as valuable microorganisms to prevent the growth of pathogen and to obtain essential amino acid. With the inoculatio of B. bassiana KACC 40039, the average laying was 0.82/hen and with B. bassiana HYB, it was 0.77/hen. Those numbers were three to eight percentage over the control. As for the effect of inoculation of B. bassiana in the feed on the production of broken eggs, B. bassiana KACC 40039 produced no broken eggs. Analysis of nutritional contents of eggs showed the increase in protein content and decrease in lipid content when compared with the control. According to these results, increase in the income of farmers can be expected.

• Fisheries and Aquatic Sciences
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• v.22 no.10
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• pp.23.1-23.10
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• 2019

Background: Taurine is a conditional essential amino acid for fish. A study was conducted to investigate the compensating effect of supplemental taurine in diets for red seabream (Pagrus major) on impaired growth performance by fish meal (FM) replacement with soybean meal (SM) at low water temperature (14.15 ± 1.95 ℃). Methods: A FM-based diet was considered as a high FM diet and three other experimental diets were formulated to replace FM with SM by 20, 35, or 50% (HFM, SM20, SM35, or SM50, respectively) without taurine and other four diets were formulated by adding 1% taurine to the diets (HFM-T, SM20-T, SM35-T, or SM50-T, respectively). Triplicate groups of fish (108.9 ± 1.58 g/fish) were distributed into 24 polyvinyl circular tanks (215 L) with 20 fish per tank and fed one of the diets to satiation for 20 weeks. Results: Growth performance and feed utilization of red seabream were significantly improved by the dietary taurine supplementation. SM20-T and SM35-T diets increased fish growth that are comparable to HFM diet. Feed intake, feed conversion ratio, and protein efficiency ratio of fish fed SM20-T and SM35-T diets were not significantly different from those of HFM group. Dietary taurine supplementation in each FM replaced group numerically increased innate immunity of the fish. Lysozyme and superoxide dismutase activities were significantly decreased in fish fed SM35, SM50, and SM50-T diets compared to those of fish fed HFM diet while they were not significantly lower in SM20, SM20-T, SM35, and SM35-T groups. Glutathione peroxidase activity was significantly lower in fish group fed SM50 diet while SM50-T group did not significantly lower compared to that of HFM group. The relative expression level of hepatic IGF-1 mRNA was improved in fish fed taurine-supplemented diets compared to their respective SM diets. Conclusions: Growth performance and feed utilization of red seabream can be accelerated or restored by 1% taurine supplementation when they are fed high level of SM up to 35% in diets during low water temperature season.

• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.287-294
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• 2008

Three amino acid residues (His119, Glu164, and Glu338) in the active site of Thermus caldophilus GK24 ${\beta}$-glycosidase (Tca ${\beta}$-glycosidase), a family 1 glycosyl hydrolase, were mutated by site-directed mutagenesis. To verify the key catalytic residues, Glu164 and Glu338 were changed to Gly and Gln, respectively. The E164G mutation resulted in drastic reductions of both ${\beta}$-galactosidase and ${\beta}$-glucosidase activities, and the E338Q mutation caused complete loss of activity, confirming that the two residues are essential for the reaction process of glycosidic linkage hydrolysis. To investigate the role of His119 in substrate binding and enzyme activity, the residue was substituted with Gly. The H119G mutant showed 53-fold reduced activity on 5mM p-nitrophenyl ${\beta}$-D-galactopyranoside, when compared with the wild type; however, both the wild-type and mutant enzymes showed similar activity on 5mM p-nitrophenyl ${\beta}$-D-glucopyranoside at $75^{\circ}C$. Kinetic analysis with p-nitrophenyl ${\beta}$-D-galactopyranoside revealed that the $k_{cat}$ value of the H119G mutant was 76.3-fold lower than that of the wild type, but the $K_m$ of the mutant was 15.3-fold higher than that of the wild type owing to the much lower affinity of the mutant. Thus, the catalytic efficiency $(k_{cat}/K_m)$ of the mutant decreased to 0.08% to that of the wild type. The $k_{cat}$ value of the H119G mutant for p-nitrophenyl ${\beta}$-D-glucopyranoside was 5.l-fold higher than that of the wild type, but the catalytic efficiency of the mutant was 2.5% of that of the wild type. The H119G mutation gave rise to changes in optima pH (from 5.5-6.5 to 5.5) and temperature (from $90^{\circ}C\;to\;80-85^{\circ}C$). This difference of temperature optima originated in the decrease of H119G's thermostability. These results indicate that His119 is a crucial residue in ${\beta}$-galactosidase and ${\beta}$-glucosidase activities and also influences the enzyme's substrate binding affinity and thermostability.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.12 no.2
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• pp.103-111
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• 1979

For the effective utilization of the coastal fish resources in Korea, an investigation on the optimum processing conditions and the quality of a textured fish protein concentrate similar to the texture of animal meat has been carried out with the fish meat of Alaska pollack and mackerel. A noodle shaped product was prepared with the fish meat paste after the adjustment of pH and salt content. The product was soaked in $96\%$ ethyl alcohol to produce textured fish protein concentrate and then dried. The processing conditions were estimated with the rehydration capacity of the textured fish protein concentrate(FFC). The quality of the final product was evaluated with chemical composition, sensory test and texture measurement. The optimum pH and salt content of the fish meat for the processing of meaty textured FPC were 7.5 and $1.0\%$ respectively. The most effective soaking conditions were as follows:soaking time, 40 min. ; temperature of alcohol, 5 to $20^{\circ}C$;amount of alcohol, 4 times the weight of tile fish meat paste, number of soaking in alcohol, 4 times. The alcohol remaining in meaty textured FPC could be removed effectively by forced air drying. The yield and the contents of protein and lipid in the meaty textured FPC from Alaska pollack were $19.9\%\;84.3\%\;and\;0.5\%$ and those from mackerel were $29.8\%,\;78.1\%\;and\;3.6\%$ respectively. The content of essential amino acid in the meaty textured FPC from Alaska pollack and mackerel was not inferior to that of beef, textured soybean protein and FAO pattern. Beef meat can be substituted with the meaty textured FPC up to $50\%$ in processing meat balls withoutanysignificantlossinthetaste, ordor and texture.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.226-235
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• 2017

Carotenoids such as phytoene, lycopene, and ${\beta}-carotene$ are used as food colorants, animal feed supplements, and for human nutrition and cosmetic purposes. Previously, we reported the isolation of a novel marine bacterium, Kocuria gwangalliensis, which produces a pink-orange pigment. Phytoene desaturase (CrtI), encoded by the gene crtI, catalyzes lycopene formation from phytoene and is an essential enzyme in the early steps of carotenoid biosynthesis. CrtI is one of the key enzymes regulating carotenoid biosynthesis and has been implicated as a rate-limiting enzyme of the pathway in various carotenoid synthesizing organisms. Here, we report the cloning of the crtI gene responsible for lycopene biosynthesis from K. gwangalliensis. The gene consisted of 1,584 bases encoding 527 amino acid residues. The nucleotide sequence of the crtI gene was compared with that of other species, including Kocuria rhizophila and Myxococcus xanthus, and was found to be well conserved during evolution. An expression plasmid containing the crtI gene was constructed (pCcrt1), and Escherichia coli cells were transformed with this plasmid to produce a recombinant protein of approximately 57 kDa, corresponding to the molecular weight of phytoene desaturase. Lycopene biosynthesis was confirmed when the plasmid pCcrtI was co-transformed into E. coli containing the plasmid pRScrtEB carrying the crtE and crtB genes required for lycopene biosynthesis. The results from this study will provide valuable information on the primary structure of K. gwangalliensis CrtI at the molecular level.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.79-86
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• 2012

ErmSF is one of the Erm family proteins which catalyze S-adenosyl-$_L$-methionine dependent modification of a specific adenine residue (A2058, E. coli numbering) in bacterial 23S rRNA, thereby conferring resistance to clinically important macrolide, lincosamide and streptogramin B ($MLS_B$) antibiotics. $^{222}FXPXPXVXS^{230}$ (ErmSF numbering) sequence appears to be a consensus sequence among the Erm family. This sequence was supposed to be involved in direct interaction with the target adenine from the structural studies of Erm protein ErmC'. But in DNA methyltarnsferase M. Taq I, this interaction have been identified biochemically and from the complex structure with substrate. Arginine 223 and 227 in this sequence are not conserved among Erm proteins, but because of the basic nature of residues, it was expected to interact with RNA substrates. Two amino acid residues were replaced with Ala by site-directed mutagenesis. Two mutant proteins still maintained its activity in vivo and resistant to the antibiotic erythromycin. Compared to the wild-type ErmSF, R223A and R227A proteins retained about 50% and 88% of activity in vitro, respectively. Even though those arginine residues are not essential in the catalytic step, with their positive charge they may play an important role for RNA binding.

• Journal of Aquaculture
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• v.11 no.2
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• pp.231-240
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• 1998

Tetraselmis is widely used as a live food because of its easy handling, high nutrient, large size and wide tolerant range of temperature and salinity. In order to find the optimum Tetraselmis species for mass culture in Korea, five species of this microalgae were examined on size, optimum culture condition ${\textperthousand}$s, $^{\circ}C.$) and nutrient composition. The results obtained were as follows: Among five species of Tetraselmis, T. sp.(Haeundae) was the largest(major axis $17.6{\pm}1.87^{\mu}$m, mean cell volume 727${\mu}$m), and T. sp. (China) the smallest (major axis $14.6{\pm}1.46^{\mu}$m, mean cell volume 625m). Tetraselmis was very eurythermal and euryhaline species. But optimum temperature and salinity for growth were 24~$30^{\circ}C.$ and 27~30${\textperthousand}$, respectively. Among five species of Tetraselmis, T. sp. (China) seemed to be the most tolerant of high temperature over $30^{\circ}C.$, and T. tetrathele of low temperature below $6^{\circ}C.$. In culture density, T. suecica showed the highest growth rate among the among the five species. The cell density of this microalgae attained to $141{\times}10^4$cells/ml at $24^{\circ}C.$ and 30${\textperthousand}$ within 7 days. In chemical composition, crude protein amount was the highest in T. suecica (44.50%), and crude lipid amount it T. sp. (Haeundae, 7.13%). Total essential amino acid amount was the highest in T. sp. (Haeundae, 50.4%) and total polyunsaturated amount in T. sp. (China, 11.7%) The results on growth and chemical composition of five species of Tetraselmis indicated that T. suecica seemed to be the most suitable species for mass culture in Korea.

• IMMUNE NETWORK
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• v.3 no.2
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• pp.126-135
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• 2003

Background: One of the important factors in the prognosis of chronic hepatitis B patient is the degree of replication of hepatitis B virus (HBV). It has been known that HBV DNA polymerase plays the essential role in the replication of HBV. HBV DNA polymerase is composed of four domains, TP (Terminal protein), spacer, RT (Reverse transcriptase) and RNaseH. Among these domains, tyrosine, the $65^{th}$ residue of TP is an important residue in protein-priming reaction that initiates reverse transcription. If monoclonal antibody that recognizes around tyrosine residue were selected, it could be applied to further study of HBV replication. Methods: To produce TP-specific scFv (single-chain Fv) by phage display, mice were immunized using synthetic TP-peptide contains $57{\sim}80^{th}$ amino acid residues of TP domain. After isolation of mRNA of heavy-variable region ($V_H$) and light-chain variable region ($V_L$) from the spleen of the immunized mouse, DNA of $V_H$ and $V_L$ were obtained by RT-PCR and joined by a DNA linker encoding peptide (Gly4Ser)3 as a scFv DNA fragments. ScFv DNA fragments were cloned into a phagemid vector. scFv was expressed in E.coli TG1 as a fusion protein with E tag and phage gIII. To select the scFv that has specific affinity to TP-peptide from the phage-antibody library, we used two cycles of panning and colony lift assay. Results: The TP-peptide-specific scFv was isolated by selection process using TP-peptide as an antigen. Selected scFv had 30 kDa of protein size and its nucleotide sequences were analyzed. Indirect- and competitive-ELISA revealed that the selected scFv specifically recognized both TP-peptide and the HBV DNA polymerase. Conclusion: The scFv that recognizes the TP domain of the HBV DNA polymerase was isolated by phage display.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.59 no.3
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• pp.263-281
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• 2014

Transcription factors are essential for the regulation of gene expression in plant. They are binding to either enhancer or promoter region of DNA adjacent to the gene and are related to basal transcription regulation, differential enhancement of transcription, development, response to intercellular signals or environment, and cell cycle control. The mechanism in controlling gene expression of transcription can be understood through the assessment of the complete sequence for the maize genome. It is possible that the maize genome encodes 4,000 or more transcription factors because it has undergone whole duplication in the past. Previously, several transcription factors of maize have been characterized. In this review article, the transcription factors were selected using Pfam database, including many family members in comparison with other family and listed as follows: ABI3/VP1, AP2/EREBP, ARF, ARID, AS2, AUX/IAA, BES1, bHLH, bZIP, C2C2-CO-like, C2C2-Dof, C2C2-GATA, C2C2-YABBY, C2H2, E2F/DP, FHA, GARP-ARR-B, GeBP, GRAS, HMG, HSF, MADS, MYB, MYB-related, NAC, PHD, and WRKY family. For analyzing motifs, each amino acid sequence has been aligned with ClustalW and the conserved sequence was shown by sequence logo. This review article will contribute to further study of molecular biological analysis and breeding using the transcription factor of maize as a strategy for selecting target gene.

• Journal of Life Science
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• v.24 no.12
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• pp.1356-1363
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• 2014

Ghrelin is a 28 amino acid orexigenic peptide hormone that is secreted predominantly by tX/A cells in the stomach, and it plays a major role in energy homeostasis. Activated ghrelin has an n-octanoyl group covalently linked to the hydroxyl group of the Ser3 residue, which is critical for its binding to the G-protein coupled growth hormone secretagogue receptor-1a (GHS-R1a). According to recent reports, both ghrelin and its receptor, GHS-R1a, are expressed by a variety of immune cells, including T- and B-lymphocytes, monocytes, and dendritic cells, and ghrelin stimulation of leukocytes provides a potent immunomodulatory signal controlling systemic and age-associated inflammation and thymic involution. Here, we report that ghrelin protected murine thymocytes from dexamethasone (DEX)-induced cell death both in vivo and in vitro. Subsequently, we explored the molecular mechanisms of the antiapoptotic effect of ghrelin. According to our experiments, ghrelin inhibited the expression of proapoptotic proteins via the regulation of glucocorticoid receptor (GR) phosphorylation. As a result, ghrelin inhibited the proapoptotic activation of proteins, such as Caspase-3, PARP, and Bim. These data suggest that ghrelin, through GHS-R, inhibits the pathway to apoptosis by regulation of the proapoptotic protein activation signal pathway. They provide evidence that blocking apoptosis is an essential function of ghrelin during the development of thymocytes.