• Journal of Applied Biological Chemistry
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• v.65 no.4
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• pp.439-446
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• 2022

Soybean is a crop with high-quality of protein and oil, and it is one of the most widely used genetically modified (GM) crops in the world today. In South Korea, Kwangan is the most utilized variety as a parental line for GM soybean development. In this study, untargeted LC-MS metabolomic approaches were used to compare metabolite profiles of Kwangan and three other commercial varieties cultivated in Gunwi and Jeonju in 2020 year. Metabolomic studies revealed that the 4 soybean varieties were distinct based on the partial least squares-discriminant analysis (PLS-DA) score plots; 18 metabolites contributed to variety distinction, including phenylalanine, isoflavones, and fatty acids. All varieties were clearly differentiated by location on the PLS-DA score plot, indicating that the growing environment is also attributable to metabolite variability. In particular, isoflavones and linolenic acid levels in Kwangan were significantly lower and higher, respectively compared to those of the three varieties. It was discussed that it might need to include more diverse conventional varieties as comparators in regard to metabolic characteristics of Kwangan for the assessment of substantial equivalence biogenetically engineered soybeans in a Kwangan-variety background.

• The Korean Journal of Physiology and Pharmacology
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• v.27 no.4
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• pp.417-426
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• 2023

The TRPM4 gene encodes a Ca²⁺-activated monovalent cation channel called transient receptor potential melastatin 4 (TRPM4) that is expressed in various tissues. Dysregulation or abnormal expression of TRPM4 has been linked to a range of diseases. We introduced the hemagglutinin (HA) tag into the extracellular S6 loop of TRPM4, resulting in an HA-tagged version called TRPM4-HA. This TRPM4-HA was developed to investigate the purification, localization, and function of TRPM4 in different physiological and pathological conditions. TRPM4-HA was successfully expressed in the intact cell membrane and exhibited similar electrophysiological properties, such as the current-voltage relationship, rapid desensitization, and current size, compared to the wild-type TRPM4. The presence of the TRPM4 inhibitor 9-phenanthrol did not affect these properties. Furthermore, a wound-healing assay showed that TRPM4-HA induced cell proliferation and migration, similar to the native TRPM4. Co-expression of protein tyrosine phosphatase, non-receptor type 6 (PTPN6 or SHP1) with TRPM4-HA led to the translocation of TRPM4-HA to the cytosol. To investigate the interaction between PTPN6 and tyrosine residues of TRPM4 in enhancing channel activity, we generated four mutants in which tyrosine (Y) residues were substituted with phenylalanine (F) at the N-terminus of TRPM4. The YF mutants displayed properties and functions similar to TRPM4-HA, except for the Y256F mutant, which showed resistance to 9-phenanthrol, suggesting that Y256 may be involved in the binding site for 9-phenanthrol. Overall, the creation of HA-tagged TRPM4 provides researchers with a valuable tool to study the role of TRPM4 in different conditions and its potential interactions with other proteins, such as PTPN6.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.3
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• pp.195-211
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• 1986

The object of this study is to obtain fundamental data on cultured fishes produced in Korea to improve their food components. For this purpose, the food components of cultured fresh water fishes such as eel, Anguilla japonica, snakehead, Channa argus, and common carp, Cyprinus carpio, were investigated and compared with those of the wild ones. The results obtained are summarized as follows: 1. Common characteristics in the proximate composition were that wild fish was higher in crude protein content and lower in crude lipid content than those of cultured one. 2. Among the 9 kinds of minerals analyzed in all the samples, sodium, potassium, calcium and magnesium contents were absolutely predominant being more than $99.52\%$. These four elements in feedstuff also occupied $99.68{\sim}99.92%$ of total minerals. 3. The neutral lipids of wild and cultured eel, snakehead and common carp occupied $55.7{\sim}95.8%$ of lipid fractions, while the content of the phospholipids in snakehead was particularly higher than those of others. 4. The neutral lipids of wild and cultured eel, snakehead and common carp mainly consisted of triglycerides ($85{\sim}95%$), and a little quantity of diglycerides, monoglycerides, free sterol ester and hydrocarbon were also identified in the neutral lipid. 5. The phospolipids of eel and common carp were mainly occupied by phosphatidyl choline ($71.3{\sim}83.9%$), followed by phosphatidyl ethanolamine ($12.1{\sim}23.5%$) and phosphatidyl serine ($7.5{\sim}13.8%$). The phospholipids of snakhead consisted of phosphatidyl choline ($50.7{\sim}64.5%$), phosphatidyl ethanolamine ($28.0{\sim}35.5%$) and phosphatidyl serine ($7.5{\sim}13.8%$). Generally, phosphatidyl choline content was higher in wild fish than in cultured one, while phosphatidyl ethanolamine and phosphatidyl serine contents were higher in cultured one. 6. The major fatty acids in total lipid of wild eel, snakehead and common carp were $C_{16:0}\;and\;C_{20:5}$, while those in cultured ones were $C_{18:1},\;C_{18:2}\;and\;C_{22:6}$. The fatty acid composition of neutral lipids showed similar tendency to that of total lipid, and the main fatty acids in phospholipids of cultured fishes were $C_{18:1}\;and\;C_{18:2}$. In glycolipids, $C_{20:5}\;and\;C_{22:6}$ were higher in wild fishes, while $C_{18:2}$ were higher in cultured ones. 7. Total amino acids contents of wild and cultured eel were nearly the same, being $16.65\%$ ana $15.99\%$ respectively. The major amino acids of wild and cultured fish were glutamic acid, leucine, aspartic acid and lysine in order. In snakehead, the contents of aspartic acid and proline in cultured fish were higher than those in wild one, while the contents of glutamic acid, alanine, glycine were higher in the wild one. Total amino acid content of cultured common carp was $21.7\%$ compared with $17.08\%$ in wild one. The contents of glutamic acid, aspartic acid, glycine, proline and alanine occupied higher quantities in cultured common carp compared with those in wild one while the other amino acids revealed no significant difference. 8. Aspartic acid in free amino acids of cultured eel held $1.0\%$ of total free amino acids, while that in wild eel held $2.9\%$. Histidine, arginine and tyrosine content of cultured fish were two times higher than those of wild one. But free amino acid composition of samples seemed to be no marked differences according to cultured places. The contents of arginine, aspartic acid, glutamic acid, methionine and phenylalanine of snakehead ware higher in wild one than in cultured one, while the contents of lysine, histidine, glycine, and alanine ware higher in cultured one. In free amino acids content of wild common carp, histidine, glycine and lysine occupied $76.9\%$ of total free amino acids. Lysine, histidine, aspartic acid, alanine, valine and leucine were higher in wild one compared with those of cultured one, while glycine and tyrosine contents were higher in cultured fish.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.5
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• pp.615-623
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• 2001

A study of mammary function in relation to glucose metabolism of first lactation Bali cows on grass-legume diets was carried out using 12 primiparous cows (initial BW $263.79{\pm}21.66kg$) for 16 weeks starting immediately post calving. The animals were randomly allocated into 4 dietary treatment groups R1, R2, R3 and R4, receiving from the last 2 months of pregnancy onwards, rations based on a mixture of locally available grass and legume feed ad libitum. On a DM basis R1 contained 70% elephant grass (PP, Penicetum purpureum) plus 30% Gliricidia sepia leaves (GS), R2 was 30% PP plus 25% GS supplemented with 55% Hibiscus tilliacius leaves (HT, defaunating effect), R3 and R4 were 22.5% PP+41.25% GS+11.25% HT+25% concentrate, with R4 supplemented with zinc-diacetate. TDN, CP and zinc contents of the diets were 58.2%, 12.05% and 18.3 mg/kg respectively for R1, 65.05%, 16.9% and 25.6 mg/kg respectively for R2, 66.03%, 16.71% and 29.02 mg/kg respectively for R3 and 66.03%, 16.71% and 60.47 mg/kg respectively for R4. Milk production and body weights were monitored, an energy and protein balance trial conducted, overall glucose kinetics parameters assessed, mammary blood flow (MBF) and metabolite arteriovenous differences (${\Delta}AVs$) measured to get uptake data and mammary performance relationships. Parameters of glucose kinetics at peak lactation or during dry condition were not affected by ration quality. Glucose pool size, space of distribution and flux increased by 61.77, 62.26 and 82.08%, respectively, during lactation compared to the dry period. Mean glucose flux of lactating Bali cows was $5.52mg/min.kgBW^{0.807}$ which resembles the range of values of temperate dairy cows. Calculation showed that glucose requirements for maintenance, milk lactose and fat-glycerol synthesis, and the formation of NADPH reached 461.69 g for a yield of 1 kg/d or equal to 320.62 mg/min, which was less than the average glucose flux of lactating Bali cows of 481.35 mg/min. Mammary blood flow (MBF) values ranged from 56 to 83 l/h for the different treatments and the ratio MBF per kg milk produced improved from av. 1540 l/kg for R1 to av. 967 l/kg for R4 treated cows. Mammary glucose uptake ranged from 6.27 to 12.03 g/h or 120 to 140 g/kg milk. Glucose uptake was mass-wise 2 to 4 times the amount secreted as lactose, which indicated values less than the calculated mammary glucose needs and that little lactose was synthesized. The excess glucose taken-up was used for other metabolic processes. Linear relationships between metabolite ${\Delta}AVs$ and arterial blood plasma concentration [A] showed that in Bali cows triglycerides (TG), phenylalanine (Phe) and tyrosine (Tyr) have high coefficients of determination, i.e. 0.77, 0.81 and 0.69, respectively. For glucose, the relationship is quadratic with an $R^2$ value of 0.49. It was concluded that lactose synthesis was inadequate, which led to a speculation that milk yield could be improved by increased lactose synthesis.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.1
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• pp.99-106
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• 2009

For Exp. 1, 120 ((Yorkshire${\times}$Landrace)${\times}$Duroc) weaned pigs (7.96${\pm}$0.01 kg average initial BW, 21 days weaning) were used in a 28 d-growth assay to determine the effects of replacing soy protein concentrate (SPC) with fermented soy protein (FSP) in a starter diet (d 0 to 7) on the growth performance, apparent fecal amino acid digestibility and subsequent performance in weaned pigs. Dietary treatments included: i) FSP0 (basal diet; whey-skim milk powder-SPC based diet); ii) FSP5 (replacing SPC with 5% FSP); iii) FSP10 (replacing SPC with 10% FSP). Pigs were fed the phase I diet for 7 days, and then each group was fed a common commercial diet for 21 days to determine the effect of previous diet on subsequent performance. Average daily gain (ADG) from d 5 to 7 (linear effect, p = 0.01) and d 7 to 14 (linear effect, p<0.001) were increased as FSP level increased. The pigs fed with FSP was heavier than the pigs fed with SPC at d 5 to 7 and d 7 to 14 after weaning (p<0.05). In the entire period (d 0 to 28), there were no significant differences in weight gain and final weight between SPC and FSP diets (p>0.05). Average daily feed intake (ADFI) was higher in pigs fed with the 5% FSP diet than those fed with the other diets at d 0 to 2 post-weaning (quadratic effect, p = 0.05). Also, for the entire period of phase I (d 0 to 7), pigs consumed more 5% FSP diet compared to other treatments (quadratic effect, p = 0.03). Gain/feed (G/F) was not affected by dietary SPC or FSP in phase I and subsequent periods, but G/F from d 5 to 7 after weaning was improved linearly (p = 0.04) as dietary FSP level increased. Pigs fed with 10% FSP also improved G/F compared with those fed only SPC (p<0.05). At d 7, there were linear increments in fecal dry matter (DM) (p<0.1) and nitrogen (N) (p<0.01) digestibilities as the dietary FSP level increased. The digestibilities of fecal essential and total amino acids were increased as the FSP level increased (linear effect, p<0.1). For Exp. 2, three ((Yorkshire${\times}$Landrace)${\times}$Duroc) weaned barrows (average initial BW of 7.32 kg) were surgically fitted with a simple T-cannula approximately 15 cm prior to the ileo-cecal junction. The experimental designs were 3${\times}$3 latin squares with pigs and periods as blocking criteria. Dietary treatments and composition were the same as in Exp. 1. Apparent ileal N digestibility was increased as FSP level was increased (linear effect, p<0.05). The dietary treatments (SPC and FSP) did not affect apparent ileal DM digestibility (p>0.05). Among essential amino acids, apparent digestibility of ileal arginine (Arg), lysine (Lys), methionine (Met) and phenylalanine (Phe) were improved as the FSP level increased (linear effect, p<0.1). Also, apparent ileal total essential, non-essential and total amino acid digestibilities were increased linearly (p<0.1). In conclusion, replacing SPC with fermented soy protein appeared beneficial in growth performance, N and amino acid digestibility during the early 7 days after weaning, and an equivalent effect showed on growth performance in subsequent period of 7 to 28 days after weaning.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.27-28
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• 2014

Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

• Journal of the Korean Society of Food Culture
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• v.11 no.2
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• pp.229-233
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• 1996

This study was carried out to analyze the changes in nitrogen compounds, free-amino acids and volatile organic acids of traditional Korean Soy Sauce with two different fermentation jars and varying Meju concentration. Total nitrogen content in the glass jar was higher than that of the clay jar. However, the glass jar contained more nitrogen in ammonia type nitrogen and less in amino type nitrogen than clay jar, resulting in inferior quality. Total free amino acids content was highest on the 150th day. Among free amino acids, the concentration of glutamic acids, aspartic acid, serine, alanine and lysine, which give sweet and savory taste, were higher than that of the others. Phenylalanine, valine, leucine and isoleucine, which give bitter taste, were also present in significant quantities. Among identified volitile organic acids, acetic acid was present in the hightest concentration, and it's concentration was higher in the jar than in the glass jar. Meju concentration 1:4 showed slow increse while 1.3:4 showed similar trends in the glass jar 1:4 and clay jar 1.3:4, and it's concentration decreased after the ripening period in all samples. In addition valerie acid and capric acid were also present in small quantities.

• Korean Journal of Food Science and Technology
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• v.29 no.2
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• pp.211-217
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• 1997

The physical and chemical stability of anthocyanins from a Korean pigmented rice variety was investigated at various conditions of pH, temperature, metal ion, sugar, organic acid and light. The anthocyanin pigments were relatively stable with half-lives of 36 days (pH 2.0) and 17 days (pH 3.0), while they were decomposed in a day at neutral and basic pH of 7.0 and 9.0 at $25^{\circ}C$. The anthocyanins also showed high thermal stability at pH 3.0; the half-lives were 7.4 hr, 23.6 hr and 96.3 hr at $95^{\circ}C,\;75^{\circ}C\;and\;50^{\circ}$, respectively. Addition of di- and tri-valent metal ions at pH 3.0 resulted in the increase of color intensity and stability throughout 21 days of storage periods at $25^{\circ}C$. Most sugars added accelerated the degradation of anthocyanin pigments, so that fructose showed the greatest degradation effect on the pigments. Addition of citric acid at pH 3.0 increased stability of anthocyanins, while tartaric acid decreased stability. The anthocyanins were very sensitive on light irradiation with a degradation half-life of 14 hr under 20,000 lux-light dosage at pH 3.0.

• Journal of fish pathology
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• v.6 no.2
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• pp.93-101
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• 1993

The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rna1-1 mutant gene from rna1-1 mutant yeast strain(R49 ; trpl, ura3-52, rna1-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3~4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rna1-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNA I gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating $pUC{\Delta}$/RNA I. After constructing $pYEP{\Delta}RNA$ I (consists of -103nt deleting RNA I gene, URA3 gene, $2{\mu}m$ rep. origin), pYEPrna1-1(consists of Xba I fragment of pUCrna1-1. URA3 gene, $2{\mu}m$ rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rna1-1) by electroporation, respectively. Yeast transformant carrying $pYEP{\Delta}RNA$ I did not complement the thermal sensitivity of rna1-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.1
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• pp.40-50
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• 1990

The purpose of this paper is to develop a colorant from krill, Euphausia superba, process wastes for use in food products. Carotenoproteins were extracted from preboiled krill processing offal(PKPO) and raw frozen krill processing offal(RKPO) with the aid of proteolytic enzymes. The long-term stability of the astaxanthin associated with the carotenoprotein by the addition of pretense inhibitor and antioxidant to the product were also investigated. Total astaxanthin contents of PKPO and RKPO were $35.1mg\%,\;22.1mg\%$ and those in carotenoproteins were $98.6mg\%,\;61.9mg\%$, respectively. The chitin contents of PKPO and RKPO were $6.9\%,\;4.5\%$, however, those of carotenoproteins were not determined. When $0.5\%$ trypsin was added to the extraction medium containing 0.5M $Na_3EDTA$ at $4^{\circ}C,\;74\%$ of astaxanthin and $83\%$ of the protein of PKPO were recovered as carotenoprotein in 24hrs. The amino acid profile in carotenoprotein was mainly composed of glutamic acid, methionine, aspartic acid and isoleurine. Their contents amounted to about 40% of the total amino acids, followed by alanine, phenylalanine, Iysine, leucine, threonine and tyrosine in that order, with a small amount of cysteine and tryptophan. The levels of essential amino acids were high as much as $38.3\%\~43.6\%$ of the total amino acids. The maximum observance of the carotenoid fraction from krill processing offal and from carotenoprotein was 469nm in petroleum ether. The separated components of carotenoprotein by TLC had Rfs $0.20\~0.23\;0.56\~0.60$ and $0.88\~0.91$. The carotenoids were comprised of astaxanthin, astaxanthin monoester and asthaxanthin diester in $25\~30\%\;,35\~40\%$and $40\~45\%$, respectively. The loss of carotenoids in the carotenoprotein can be prevented by the addition of pro-tease inhibitor(trasylol) and antioxidant(BHT) below $4^{\circ}C$.