• Journal of the Korea Organic Resources Recycling Association
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• v.18 no.3
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• pp.31-37
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• 2010

Chitinases (EC 3.2.1.14) hydrolyze the ${\beta}$-1,4-linkages in chitin, the second most abundant polymer of N-acetyl-${\beta}$-D-glucosamine which is a structural component of protective biological matrices such as fungal cell walls and insect exoskeletons. The glycosyl hydrolases 18 family including chitinases is an ancient gene family widely expressed in archea, prokaryotes and eukaryotes. Since earthworms live in the soil with a lot of microbial activities and fungi are supposed to be a major component of the diet of earthworm, it has been reported that there would be appropriate immune system to protect themselves from microorganisms attacks. In this study, the novel chitinase, EaChi, from the midgut of earthworm, Eisenia andrei, were identified and characterized. To obtain full-length cDNA sequence of chitinase, RT-PCR and RACE-PCR analyses were carried out by using the previously identified EST sequence amongst cDNA library established from the midgut of E. andrei. EaChi, a partial chitinase gene, was composed of 927 nucleotides encoding 309 amino acids. By the multiple sequence alignments of amino acids with other different species, it was revealed that EaCHI is a member of glycosyl hydrolases 18 family, which has two highly conserved domains, substrate binding and catalytic domain.

• Journal of the Korean Chemical Society
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• v.37 no.2
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• pp.237-243
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• 1993

The primary and secondary structure of the 5S rRNA isolated from Xanthomonas celebensis were determined by enzymatic and chemical degradation methods. It consists of 119 nucleotides and contains no modified nucleosides. As with the 5S rRNAs of X. maltophilia and X. citri, it contains an additional uridine residue on the 5'-terminus. Its secondary structure was almost identical to the models previously proposed by us for the 5S rRNA of two Xanthomonas species. Its secondary structure consists of five helices, five loops and two bulges. The tertiary interactions in the 5S rRNA molecule were analyzed by Fe(II)-EDTA treatment and hybridization method using deoxyhexamer. From the fact that some adenine residues in loop M, region $I_1-C$, loop $H_1$, and loop $H_2$ become susceptible to diethylpyrocarbonate when the 5S rRNA was hybridized with deoxyhexamer complementary to the sequence $U_{35}CCCAU_{40}$ and that some nucleotide residues in loop M, loop $H_1$ and region $D-I_2$ become resistant Fe(II)-EDTA cleavage in the presence of $Mg^{2+}$, it is presumed that loops $H_1$ and $H_2$ interact with loop M in some way. In the tertiary interaction, the regions $I_1-C$ and $D-I_2$ seem to act as hinges in folding the stems $B-I_1-C$ and $D-I_2-E.$ It was found that loop $H_1$ changes into a smaller loop of three bases by forming noncanonical A : C base-pairs ih acidic environment.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.99-104
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• 2009

The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.

• Journal of Life Science
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• v.26 no.3
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• pp.275-281
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• 2016

Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

• Journal of Animal Science and Technology
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• v.44 no.2
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• pp.181-190
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• 2002

This study was performed to determine sequences of the mt DNA D-loop region, including $tRNA^{Pro}$ and $tRNA^{Pre}$ and to analysis sequence variation polymorphism in Korean cattle. The resulting sequencies were compared with previously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify an 1142bp between nucleotides 15061 and 404 within the D-loop region of mt DNA using specific primers. Korean cattle showed 24 polymorphic sites by nucleotide substitutions and insertions of single base pairs. About 50% of polymorphic sites were found in positions 16042 to 16122 with the most variable region. Among these polymorphic sites, variations at 16055, 16230 and 16260 bp were detected as new sequence variants in Korean cattle. These specific polymorphic sites have not been reported in the Japanese black cattle and European cattle. Therefore, mt DNA variants in the D-loop region may be used as genetic markers for specifying Korean cattle. The frequencies of positions 169, 16302, 16093, 16042, 16119 with a high level of sequence polymorphism were 0.81, 0.56, 0.56, 0.50 and 0.43, respectively. In comparison of genetic distances, Korean cattle showed the more closely to European cattle as Bos taurus than Bos indicus such as African and India breeds. In conclusion, these mt DNA sequence polymorphisms in the D-loop region for Korean cattle may be useful for the analysis of cytoplasmic genetic variation and associations with economic important traits and genetic analysis of maternal lineage.

• Korean Journal of Food Science and Technology
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• v.9 no.4
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• pp.255-263
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• 1977

Fermented squid, Loligo kobiensis, is widely used and occupies an important position in foods of this country. But no study on its taste compounds has been reported. This study was attempted to establish the basic data for evaluating taste compounds of fermented squid. The changes of such compounds during fermentation as free amino acids, nucleotides and their related compounds, TMAO, TMA and betaine were analysed. The sample was prepared with 20% salt content and fermented at a controlled temperature of $15{\pm}3^{\circ}C$. ADP, AMP and inosine tended to degrade rapidly while hypoxanthine increased more than four times as compared with raw sample at 91 day fermentation. In the free amino acid composition of fresh squid, abundant amino acids were proline, taurine, alanine, arginine, serine, glutamic acid, lysine, glycine, leucine and valine in order. Such amino acids like phenylalanine, methionine, tyrosine, isoleucine, and histidine were poor. In squid extract, proline and taurine were dominant holding 40.2% and 32.0% of total free amino acids respectively. The total free amino acid nitrogen in fresh squid was 33.6% of its extract nitrogen. The changes of free amino acid composition in the extract of squid during fermentation was not observed. In the extract of fermented product, abundant amino acids were proline, leucine, lysine, serine, arginine, alanine, valine, isoleucine, phenylalanine, methionine and glycine in order. Glutamic acid and histidine were poor and taurine and tyrosine were trace in content. The increase of total free amino acids during 63 day fermentation reached approximately wore than 1.8 times as compared with that of raw sample and than decreased slowly. The amount of betaine increased more than 1.2 times as compared with that of raw sample during 91 day fermentation. TMA increased while TMAO decreased during fermentation. The amount of TMAO nitrogen in 91 days fermented squid was 402.4mg% on moisture and salt free base. Betaine and TMAO known as sweet compounds were abundant in fermented squid. It is supposed that these compounds could also play a role as important taste compounds of fermented squid. It is concluded that the major taste compounds of fermented squid were amino acids like proline, leucine, serine, lysine, arginine, alanine and betaine. Other compounds such as valine, isoleucine and TMAO and hypoxanthine could also not be excluded as taste supporters in fermented squid.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.2
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• pp.109-117
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• 1986

For the purpose of obtaining basic data which can be applied to processing of retort pouched shellfishes, retort pouched seasoned ark shell, Anadara broughtonii, was prepared. The frozen ark shell was thawed and seasoned with a mixed seasoning powder prepared with $10.0\%$ of sorbitol, $2.0\%$ of table salt and $0.5\%$ of monosodium glutamate at $5^{\circ}C$ for 10 hours, and then dried at $45^{\circ}C$ for 4 hours. The dried seasoned ark shell was coated with $1.0\%$ sodium alginate solution, dried with cola air blast for 2 hours and then vacuum-packed in the laminated plastic film bag (polyester/casted polypropylene= $12{\mu}m/70{\mu}m,\;15{\times}16cm$), and finally sterilized up to Fo=6.0 in hot water circulating retort at $121^{\circ}C$ for 10 minutes. The major fatty acids of raw ark shell and retort pouched seasoned ark shell products were 16:0, 20:5, 22:6, 18:0 and 18:3, and predominant free amino acids of those were lysine, arginine, glycine, alanine, glutamic acid and leucine. In nucleotides and its related compounds of raw ark shell and retort pouched seasoned ark shell products, the most abundant one was AMP, and total extract-N of those was chiefly consisted of free amino acids, betaine and nucleotide and its related compounds. During the processing procedure such as drying and sterilization, unsaturated fatty acids slightly decreased while saturated fatty acids increased, and total extract-N content decreased about a half. From the results of chemical and microbial experiments during storage, it was concluded that the products could be preserved in a good condition for 100 days at room temperature, and their duality could be improved by the coating treatment of sodium alginate solution.

• Korean Journal of Poultry Science
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• v.36 no.3
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• pp.207-213
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• 2009

Adenylosuccinate lyase (ADSL) deficiency is a disease of purine metabolism which affects patients both biochemicall and behaviorally. An obstacle of this purine nucleotide cycle(PNC) can be caused brain functional disorder and growth disorder. So ADSL deficiency, which is associated with sever mental retardation, autistic features and energy metabolism. This study was performed to identify SNP on ADSL gene in chicken. The nucleotides were observed as T to C ($7724^{th}$ nucleotide), C to T ($7732^{nd}$ nucleotide), G to T ($10108^{th}$ nucleotide), A to T ($10356^{th}$ nucleotide), G to A($10375^{th}$ nucleotide), A to C ($10402^{nd}$ nucleotide), A to T ($12716^{th}$ nucleotide), T to A ($12717^{th}$ nucleotide), C to T ($15491^{st}$ nucleotide), C to T ($15542^{nd}$ nucleotide) and C to T ($15550^{th}$ nucleotide). The nucleotide substitutions at $15542^{nd}$ and $15550^{th}$ (GeneBank accession no. AY665559) were found as missense mutation (alanine$\rightarrow$valine, proline$\rightarrow$serine, respectively). This study will be useful for farther researches for identifying association between these SNPs and energy metabolism in chicken. The C15550T SNP showed three genotypes, CC, CT, TT by digestion with the genotype TT had significantly faster the first lay day (150.0) than CT (162.0, P<0.05) and genotype TT (150.0, P<0.05) had significantly higher the egg production rate than CT (172.4, P<0.05). According to result of this study, a C15550T was found to have a significantly effect first lay day and mean egg production. It will be possible to use SNP marker on selecting chicken to improve important economic traits, which is the first lay day and mean egg production.

• Development and Reproduction
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• v.12 no.1
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• pp.19-30
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• 2008

Proper development of fertilized oocyte to blastocyst is a key step in mammalian development to implantation. During development of preimplantation embryos, the mammalian embryo needs supply the energy substrate for keep viability. Usually mammalian oocyte get substrate especially energy substrate from oviduct and uterus, because it does not store much substrate into cytoplasm during oogenesis. Carbohydrates are known as a main energy substrate for preimplantation stage embryos. Glucose, lactate and pyruvate are essential component in preimplantation embryo culture media and there are stage specific preferences to them. Glucose transporter and $H^+$-monocarboxylate cotransporter are a main mediator for carbohydrate transport and those expression levels are primarily under the control of intrinsic or extrinsic factors like insulin and glucose. Other organic substances, amino acids, lipids and nucleotides are used as energy substance and cellular regulation factor. Though since 1960s, successful development of fertilized embryo to blastocyst has been accomplished with chemically defined medium for example BWW and give rise to normal offspring in mammals, the role of metabolites and the regulation of intermediary metabolism are still poorly understood. Glucose may permit expression of metabolic enzymes and transporters in compacting morula, capable of generating the energy required for blastocyst formation. In addition, it has been suggested that the cytokines can modulate the metabolic rate of carbohydrate in embryos and regulate the preimplantation embryonic development through control the metabolic rate. Recently we showed that lactate can be used as a mediator for preimplantation embryonic development. Those observations indicate that metabolites of carbohydrate are required by the early embryo, not only as an energy source, but also as a key substrate for other regulatory and biosynthetic pathways. In addition metabolites of carbohydrate may involve in cellular activity during development of preimplantation embryos. It is suggested that through these regulation and with other regulation mechanisms, embryo and uterus can prepare the embryo implantation and further development, properly.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.5
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• pp.459-468
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• 1986

This study was attempted to process low-sodium salt fermented small shrimp as substitutes for traditional high-sodium salt fermented one which has widely been favored and consumed in Korea. Low-salt fermented small shrimp was prepared with $4\%$ sodium chloride and $4\%$ potassium chloride, and various additives such as $0.5\%$ lactic acid, $6\%$ sorbitol and $4\%$ ethylalcohol extract of red pepper as preservatives and flavor enhancers. And the changes of taste compounds, volatile compounds and fatty acid composition in low-salt fermented small shrimp were analyzed and compared with those of conventional $20\%$ sodium salt fermented one during the fermentation of 120 days at $25{\pm}3^{\circ}C$. The most favorable taste for fermented small shrimp were reached at 60 days of fermentation. Judging from sensory evaluation, little difference of taste was detected between the low-salt fermented small shrimp and high-sodium salt fermented one. The principal taste compounds in fermented small shrimp were free amino acids, and betaine and nucleotides and their related compounds played an assistant role. The major amino acids in fermented small shrimp were glutamic acid, leucine, proline, glycine, lysine and aspartic acid. The major fatty acids in fermented small shrimp samples were 16:0, 20:5, 22:6, 16:1 and 18:1, and unsaturated fatty acids decreased slightly while saturated fatty acids increased during fermentation. At 60 days of fermentation 8 kinds of volatile fatty acids (acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid, isocarproic acid, carproic acid), 6 kinds of carbonyl compounds (ethanal, propanal, 2-methylpropanal, 3-methylbutanal, pentanal, 2-methylpentanal), and 3 kinds of volatile amines (methylamine, trimethylamine, isopropylamine) were identified.