• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.5
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• pp.612-624
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• 2022

A novel Gram-positive, motile, non-spore forming aerobic marine bacterium, designated 107-1^T was isolated from tidal mud collected in Gyehwa-do, South Korea. Cells of strain 107-1^T were short rod or coccoid, oxidase negative, catalase positive and grew at 10-40℃ (with optimum growth at 25-30℃). It utilized menaquinones MK-7 and 8 as its respiratory quinones and its major fatty acids were anteiso-C_15:0 (37.9%), iso-C_16:0 (14.9%), and iso-C_14:0 (10.8%). Phylogenetic analysis based on 16S rRNA gene sequences revealed a distinct clade containing strain 107-1^T and close species Planococcus ruber CW1^T(98.52% sequence similarity), P. faecalis KCTC 33580^T(97.67%), P. kocurii ATCC 43650T(97.48%), P. donghaensis DSM 22276T(97.47%), and P. halocryophilus DSM 24743T(97.37%). Strain 107-1^T contains one circular chromosome (3,513,248bp in length) with G+C content of 44.6 mol%. Estimated ranges for genome to genome distance, average nucleotide identity, and average amino acid identity comparing strain 107-1^T with close taxa were 20.3-34.8%, 77.9-86.9%, and 73.6-92.8%, respectively. Based on polyphasic analysis, strain 107-1^T represents a novel species belonging to the genus Planococcus.

• Journal of Life Science
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• v.32 no.10
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• pp.778-783
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• 2022

This study isolated a new agarase-producing bacterium and characterized its agarase. A new agar-degrading strain was isolated from the seashore of Namhae in Gyeongnam province, Korea, and was purely cultured using the Marine Agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-2 after 16S rRNA gene sequencing. The crude agarase was obtained from the culture medium of the Simiduia sp. SH-2 strain, and the agar-degrading activity was measured. The highest level of activity of the Simiduia sp. SH-2-derived agar-degrading enzyme was 625 U/l. Agar degradation activity was most significant at 40℃ and pH 7.0. Compared to the activity at 40℃, the relative activity was 31% at 20℃ and 71% at 30℃. Compared to the activity at pH 7.0, the relative activity was 94% and 89% at pH 6.0 and pH 8.0, respectively. Residual activity was greater than 96% after exposure to 20℃ and 30℃ for 2 hr and more than 49% after exposure to 40℃ for 2 hr. Simiduia sp. SH-2 was identified as a strain producing β-agarase that creates neoagarooligosaccharides, such as neoagarotetraose and neoagarohexaose. Therefore, the Simiduia sp. SH-2 strain and its β-agarase are expected to be useful functional material producers in the food, cosmetic, and pharmaceutical industries.

• Journal of Microbiology and Biotechnology
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• v.32 no.8
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• pp.1064-1071
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• 2022

Arabinogalactans have diverse biological properties and can be used as pharmaceutical agents. Most arabinogalactans are composed of β-(1→3)-galactan, so it is particularly important to identify β-1,3-galactanases that can selectively degrade them. In this study, a novel exo-β-1,3-galactanase, named PoGal3, was screened from Penicillium oxalicum sp. 68, and hetero-expressed in P. pastoris GS115 as a soluble protein. PoGal3 belongs to glycoside hydrolase family 43 (GH43) and has a 1,356-bp gene length that encodes 451 amino acids residues. To study the enzymatic properties and substrate selectivity of PoGal3, β-1,3-galactan (AG-P-I) from larch wood arabinogalactan (LWAG) was prepared and characterized by HPLC and NMR. Using AG-P-I as substrate, purified PoGal3 exhibited an optimal pH of 5.0 and temperature of 40℃. We also discovered that Zn²⁺ had the strongest promoting effect on enzyme activity, increasing it by 28.6%. Substrate specificity suggests that PoGal3 functions as an exo-β-1,3-galactanase, with its greatest catalytic activity observed on AG-P-I. Hydrolytic products of AG-P-I are mainly composed of galactose and β-1,6-galactobiose. In addition, PoGal3 can catalyze hydrolysis of LWAG to produce galacto-oligomers. PoGal3 is the first enzyme identified as an exo-β-1,3-galactanase that can be used in building glycan blocks of crucial glycoconjugates to assess their biological functions.

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.52-69
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• 2022

Ferulic acid (FA) is a phytochemical with various bioactive properties. It has recently been proposed that due to its phytogenic action it can be used as an alternative growth promoter additive to synthetic compounds. The objective of the present study was to evaluate the growth performance, carcass traits, fiber characterization and skeletal muscle gene expression on hair-lambs supplemented with two doses of FA. Thirty-two male lambs (n = 8 per treatment) were individually housed during a 32 d feeding trial to evaluate the effect of FA (300 and 600 mg d^-1) or zilpaterol hydrochloride (ZH; 6 mg d^-1) on growth performance, and then slaughtered to evaluate the effects on carcass traits, and muscle fibers morphometry from Longissimus thoracis (LT) and mRNA abundance of β₂-adrenergic receptor (β₂-AR), MHC-I, MHC-IIX and IGF-I genes. FA increased final weight and average daily gain with respect to non-supplemented animals (p < 0.05). The ZH supplementation increased LT muscle area, with respect to FA doses and control (p < 0.05). Cross-sectional area (CSA) of oxidative fibers was larger with FA doses and ZH (p < 0.05). Feeding ZH increased mRNA abundance for β₂-AR compared to FA and control (p < 0.05), and expression of MHC-I was affected by FA doses and ZH (p < 0.05). Overall, FA supplementation of male hair lambs enhanced productive variables due to skeletal muscle hypertrophy caused by MHC-I up-regulation. Results suggest that FA has the potential like a growth promoter in lambs.

• Animal Bioscience
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• v.35 no.2
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• pp.281-289
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• 2022

Objective: The aim of this study was to characterize the exopolysaccharides (EPS)-producing lactic acid bacteria from Taiwanese ropy fermented milk (TRFM) for developing a clean label low-fat fermented milk. Methods: Potential isolates from TRFM were selected based on the Gram staining test and observation of turbid suspension in the culture broth. Random amplified polymorphic DNA-polymerase chain reaction, 16S rRNA gene sequencing, and API CHL 50 test were used for strain identification. After evaluation of EPS concentration, target strains were introduced to low-fat milk fermentation for 24 h. Fermentation characters were checked: pH value, acidity, viable count, syneresis, and viscosity. Sensory evaluation of fermented products was carried out by 30 volunteers, while the storage test was performed for 21 days at 4℃. Results: Two EPS-producing strains (APL15 and APL16) were isolated from TRFM and identified as Lactococcus (Lc.) lactis subsp. cremoris. Their EPS concentrations in glucose and lactose media were higher than other published strains of Lc. lactis subsp. cremoris. Low-fat fermented milk separately prepared with APL15 and APL16 reached pH 4.3 and acidity 0.8% with a viable count of 9 log colony-forming units/mL. The physical properties of both products were superior to the control yogurt, showing significant improvements in syneresis and viscosity (p<0.05). Our low-fat products had appropriate sensory scores in appearance and texture according to sensory evaluation. Although decreasing viable cells of strains during the 21-day storage test, low-fat fermented milk made by APL15 exhibited stable physicochemical properties, including pH value, acidity, syneresis and sufficient viable cells throughout the storage period. Conclusion: This study demonstrated that Lc. lactis subsp. cremoris APL15 isolated from TRFM had good fermentation abilities to produce low-fat fermented milk. These data indicate that EPS-producing lactic acid bacteria have great potential to act as natural food stabilizers for low-fat fermented milk.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.84-84
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• 2003

DNA methylation is involved in epigenetic processes such as X-chromosome inactivation, imprinting and silencing of transposons. DNA methylation is a highly plastic and critical component of mammalian development The DNA methyltransferases (Dnmts) are responsible for the generation of genomic methylation patterns, which lead to transcriptional silencing. The maintenance DNA methyltransferase enzyme, Dnmt 1, and the de novo methyltransferase, Dnmt3a and Dnmt3b, are indispensable for development because mice homozygous for the targeted disruption of any of these genes are not viable. The occurrence of DNA methylation is not random, and it can result in gene silencing The mechanisms underlying these processes are poorly understood. It is well established that DNA methylation and histone deacetylation operate along a common mechanistic pathway to repress transcription through the action of methyl-binding domain proteins (MBDs), which are components of, or recruit, histone deacetylase (HDAC) complexes to methylated DNA. As a basis for future studies on the role of the DNA-methyl-transferase in porcine development, we have isolated and characterized a partial cDNA coding for the porcine Dnmt1. Total RNA of testis, lung and ovary was isolated with TRlzol according to the manufacture's specifications. 5 ug of total RNA was reverse transcribed with Super Script II in the presence of porcine Dnmt 1 specific primers. Standard PCRs were performed in a total volume of 50 ul with cDNA as template. Two DNA fragmenets in different position were produced about 700bp, 1500bp and were cloned into pCR II-TOPO according to the manufacture's specification. Assembly of all sequences resulted in a cDNA from 158bp of 5'to 4861bp of 3'compare with the known human maintenance methyltransferase. Now, we are cloning the unknown Dnmt 1 region by 5'-RACE method and expression of Dnmt 1 in tissues from adult porcine animals.

• Korean Journal of Environmental Biology
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• v.40 no.3
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• pp.255-266
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• 2022

In crustaceans, molting is regulated by interactions between ecdysteroid and juvenile hormone (JH) signaling pathway-related genes. Unlike the ecdysteroid signaling pathway, little information on the role of JH signaling pathway-related genes in molting is available in zooplanktonic crustaceans. In this study, three genes (juvenile hormone acid O-methyltransferase (JHAMT), methoprene-tolerant (Met), and juvenile hormone epoxide hydrolase (JHEH)) which are involved in the synthesis, receptor-binding, and degradation of JH were identified using sequence and phylogenetic analysis in the brackish water flea, Diaphanosoma celebensis. Transcriptional changes in these genes during the molting cycle in D. celebensis were analyzed. Sequence and phylogenetic analysis revealed that these putative proteins may be functionally conserved along with those of insects and other crustaceans. In addition, the expression of the three genes was correlated with the molting cycle of D. celebensis, indicating that these genes may be involved in the synthesis and degradation of JH, resulting in normal molting. This study will provide information for a better understanding of the role of JH signaling pathway-related genes during the molting process in Cladocera.

• Korean Journal of Environmental Biology
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• v.39 no.4
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• pp.445-451
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• 2021

In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

• Journal of Life Science
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• v.33 no.6
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• pp.506-511
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• 2023

The purposes of this study were to isolate and characterize lactic acid bacteria with antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) from fermented food and to confirm the antibacterial activities of fermented rice products using the isolated lactic acid bacterium. Three bacteria, namely, Weissella sp. ISF-1, ISF-2, and ISF-3, were selected from fermented squid based on the 16S rRNA gene sequence. All three strains grew well in an MRS medium containing 5% (w/v) NaCl and showed antibacterial activity against Bacillus cereus, Staphylococcus aureus, and MRSA. Their growth was excellent at 0% ~ 5% (w/v) NaCl and relatively good up to 7% (w/v) NaCl. The initial pH of 8 was optimal for their growth, and good growth was also observed at pH 6, 7, and 9. The lyophilisates of the fermented rice using Weissella sp. ISF-1 showed antibacterial activities against B. cereus, S. aureus, and MRSA. We inferred that isolated lactic acid bacteria could be useful in the development of probiotics and biopreservatives for foods and in the treatment of MRSA and may increase the value of rice products.

• Korean Journal of Breeding Science
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• v.51 no.4
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• pp.318-325
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• 2019

Salt stress is a significant factor limiting growth and productivity in crops. However, little is known about the response and resistance mechanism to salt stress in maize. The objective of this research was to develop an enhanced salt-tolerant silage maize by mutagenesis with gamma radiation. To generate gamma radiation-induced salt-tolerant silage maize, we irradiated a KS140 inbred line with 100 Gy gamma rays. Salt tolerance was determined by evaluating plant growth, morphological changes, and gene expression under NaCl stress. We screened 10 salt-tolerant maize inbred lines from 2,248 M₂ mutant populations and selected a line showing better growth under salt stress conditions. The selected 140RS516 mutant exhibited improved seed germination and plant growth when compared with the wild-type under salt stress conditions. Enhanced salt tolerance of the 140RS516 mutant was attributed to higher stomatal conductance and proline content. Using whole-genome re-sequencing analysis, a total of 328 single nucleotide polymorphisms and insertions or deletions were identified in the 140RS516 mutant. We found that the expression of the genes involved in salt stress tolerance, ABP9, CIPK21, and CIPK31, was increased by salt stress in the 140RS516 mutant. Our results suggest that the 140RS516 mutant induced by gamma rays could be a good material for developing cultivars with salt tolerance in maize.