• Journal of fish pathology
• /
• v.36 no.2
• /
• pp.263-275
• /
• 2023

Chum salmon (Oncorhynchus keta) is a species which returns to Korea for spawning and was produced as seed production at the Fisheries Resources Agency located in Uljin-gun, Gyeongsangbuk-do to preserve the species. However, farmed chum salmon showed symptoms of bacterial infection. Therefore, in this study, bacteria were isolated to identify the causative agent from chum salmon in October 2021. The isolated bacteria were identified based on the sequences of 16S rDNA, rpoD (RNA polymerase sigma factor σ⁷⁰), and vapA (A-layer) genes. Also, salinity-growth curve, biochemical characterization, antibiotic susceptibility test, and pathogenicity analysis were performed in four strains. As a result, four isolated strains were identified as Aeromonas salmonicida subsp. salmonicida. Additionally, the bacterial strains showed a decrease in growth as the salt concentration increased in the medium. All of the isolated strains exhibited γ-hemolysis, and the same biochemical properties. In the antimicrobial susceptibility test, all strains showed an inhibition zone of 40 to 44 mm for oxolinic acid, flumequine, and florfenicol. Pathogenic factors were assessed by RT-PCR at the mRNA level, and found that the four strains expresses the outer membrane ring of T3SS (ascV), inner membrane ring of T3SS (ascC), vapA, enterotoxin (act), and lipase (lip) genes which are well known to significantly contribute to the pathogenicity of A. salmonicida. The results of this study can be used as basic data to prevent A. salmonicida subsp. salmonicida occurring in sea-chum salmon in the future.

• Journal of Life Science
• /
• v.20 no.9
• /
• pp.1306-1313
• /
• 2010

The gene encoding esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est1R) was 2,465 bp in length, encoding a protein of 366 amino acid residues, and the molecular weight of the enzyme was 61,166 Da. Est1R of rumen cosmid library shared 5.9% amino acid identity with Est1R (P37967) of PNB carboxylesterase, 6.1% with Est1R (1EEAA) of acetylcholinesterase and 6.1% with Est1R (1H23A) of chain A. BlastP in NCBI database analysis of Est1R revealed that it was not homologous to previous known lipases and esterases. Est1R showed optimum activity at pH 7.0 and $40^{\circ}C$. On the other hand, the enzyme was found to be most active without organic solvent, followed by 95% activity with methanol, and the enzyme activity was highly affected by hexane (lost 51% activity). Therefore, the novel esterase gene est1R is likely obtainable from cow rumen metagenome and may be utilized for industrial purposes.

• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.12
• /
• pp.1957-1964
• /
• 2020

Objective: The aim of this study was to compare overfeeding performance, fatty acid composition, blood chemistry, enzymes and genes expression overfed Xupu and Landes geese. Methods: Sixty male Xupu geese (80 d) and Landes geese (80 d) were selected. After a period of one-week of pre-overfeeding, Xupu and Landes geese were overfed three meals of 550 and 350 g/d, respectively, of a high-carbohydrate diet in the first week of the overfeeding period. The next week, geese were given four meals of 1,200 and 850 g/d, respectively, over 8 to 14 d. Finally, geese were given five meals of 1,600 and 1,350 g/d, respectively, for the last two weeks. Results: After overfeeding for 28 d: Compared with Landes geese, Xupu geese liver weight and liver-to-body weight ratio decreased (p<0.05), while final weight, slaughter weight, total weight gain, abdominal fat weight, and feed-to-liver weight ratio increased (p<0.05). The levels of elaidic acid (C18:1t9), oleic acid (C18:1n-9), eicosenoic acid, and arachidonic acid in the liver of Xupu geese significantly increased (p<0.05), and the levels of myristic acid and stearic acid significantly decreased (p<0.05), while methyleicosanoate acid significantly increased (p<0.05). Xupu geese had higher plasma concentrations of triglyceride and very low density lipoprotein cholesterol (p<0.05), and decreased activities of alanine aminotransferase, aspartate aminotransferase, and lipase (LPS) (p<0.05). Landes geese had higher LPS activity (p<0.05), but lower cholinesterase activity (p<0.05) when compared with Xupu geese. The mRNA expression levels of fatty acid dehydrogenase (FADS) gene, elongase of long-chain fatty acid 1 (ELOVL1) gene, ELOVL5, and acyl-Co A: cholesterol acyltransferase 2 (ACAT2) gene were significantly upregulated (p<0.05) in Landes goose when compared with Xupu geese. Conclusion: This study demonstrates that the liver production performance of Landes geese was better than that of Xupu geese to some extent, which may be closely related to LPS activity, as well as the expression of FADS, ELOVL1, ELOVL5, and ACAT2.

• Asian-Australasian Journal of Animal Sciences
• /
• v.28 no.4
• /
• pp.544-550
• /
• 2015

This study was conducted to investigate effects of heat-oxidized soy protein isolate (HSPI) on growth performance, serum biochemical indices, apparent nutrient digestibility and digestive function of broiler chickens. A total of 320 1-day-old Arbor Acres chicks were randomly divided into 4 groups with 8 replicates of 10 birds, receiving diets containing soy protein isolate (SPI, control group) or the same amount of SPI heated in an oven at $100^{\circ}C$ for 1, 4, or 8 h, for 21 days, respectively. The results indicated that compared with the control group, body weight gain and feed intake of birds fed diet containing SPI heated for 8 h were significantly lower (p<0.05). Serum urea nitrogen concentration was higher in the broilers fed diet containing SPI heated for 4 or 8 h at d 21 (p<0.05). In contrast, serum glucose content was decreased by HSPI substitution at d 21 (p<0.05). The relative pancreas weight in HSPI groups was higher than that in the control group at d 21 (p<0.05). Meanwhile, the opposite effect was observed for relative weight of anterior intestine and ileum in broilers fed a diet containing SPI heated for 8 h (p<0.05). Birds fed diets containing SPI heated for 4 or 8 h had a decreased lipase activity in anterior intestinal content at d 14 and 21 (p<0.05), respectively. In addition, the same effect was also noted in broilers given diets containing SPI heated for 1 h at d 21 (p<0.05). Similarly, amylase, protease and trypsin activity in anterior intestinal content were lower in broilers fed diets containing SPI heated for 8 h at d 21 (p<0.05). The apparent digestibility of dry matter (DM) from d 8 to 10 and DM, crude protein (CP), and ether extract from d 15 to 17 were lower in broilers fed diets containing SPI heated for 8 h (p<0.05). Besides, birds given diets containing SPI heated for 4 h also exhibited lower CP apparent digestibility from d 15 to 17 (p<0.05). It was concluded that HSPI inclusion can exert a negative influence on the growth performance of broilers, which was likely to result from the simultaneously compromised digestive function.

• Asian-Australasian Journal of Animal Sciences
• /
• v.31 no.4
• /
• pp.595-606
• /
• 2018

Objective: The Fusarium mycotoxins of deoxynivalenol (DON) and zerolenone (ZEN) cause health hazards for both humans and farm animals. Therefore, the main intention of this study was to reveal DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the liver of piglets. Methods: In the present study, 15 six-week-old piglets were randomly assigned to the following three different dietary treatments for 4 weeks: control diet, diet containing 8 mg DON/kg feed, and diet containing 0.8 mg ZEN/kg feed. After 4 weeks, liver samples were collected and sequenced using RNA-Seq to investigate the effects of the mycotoxins on genes and gene networks associated with the immune systems of the piglets. Results: Our analysis identified a total of 249 differentially expressed genes (DEGs), which included 99 upregulated and 150 downregulated genes in both the DON and ZEN dietary treatment groups. After biological pathway analysis, the DEGs were determined to be significantly enriched in gene ontology terms associated with many biological pathways, including immune response and cellular and metabolic processes. Consistent with inflammatory stimulation due to the mycotoxin-contaminated diet, the following Kyoto encyclopedia of genes and genomes pathways, which were related to disease and immune responses, were found to be enriched in the DEGs: allograft rejection pathway, cell adhesion molecules, graft-versus-host disease, autoimmune thyroid disease (AITD), type I diabetes mellitus, human T-cell leukemia lymphoma virus infection, and viral carcinogenesis. Genome-wide expression analysis revealed that DON and ZEN treatments downregulated the expression of the majority of the DEGs that were associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9), proliferation (insulin-like growth factor 1, major facilitator superfamily domain containing 2A, insulin-like growth factor binding protein 2, lipase G, and salt inducible kinase 1), and other immune response networks (paired immunoglobulin-like type 2 receptor beta, Src-like-adaptor-1 [SLA1], SLA3, SLA5, SLA7, claudin 4, nicotinamide N-methyltransferase, thyrotropin-releasing hormone degrading enzyme, ubiquitin D, histone $H_2B$ type 1, and serum amyloid A). Conclusion: In summary, our results demonstrated that high concentrations DON and ZEN disrupt immune-related processes in the liver.

• Journal of Microbiology
• /
• v.43 no.3
• /
• pp.285-294
• /
• 2005

A bacterial strain M4-7 capable of degrading various polyesters, such as poly$(\varepsilon-caprolactone)$, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3-hydroxyoctanoate), and poly(3-hydroxy-5-phenylvalerate), was isolated from a marine environment and identified as Pseudomonas alcaligenes. The relative molecular mass of a purified extracellular medium-chain-length poly(3-hydroxyalkanoate) (MCL-PHA) depolymerase $(PhaZ_{palM4-7})$ from P. alcaligenes M4-7 was 28.0 kDa, as determined by SDS-PAGE. The $PhaZ_{palM4-7}$ was most active in 50 mM glycine-NaOH buffer (pH 9.0) at $35^{\circ}C$. It was insensitive to dithiothreitol, sodium azide, and iodoacetamide, but susceptible to p-hydroxymercuribenzoic acid, N-bromosuccinimide, acetic anhydride, EDTA, diisopropyl fluorophosphate, phenylmethylsulfonyl fluoride, Tween 80, and Triton X-100. In this study, the genes encoding MCL-PHA depolymerase were cloned, sequenced, and characterized from a soil bacterium, P. alcaligenes LB19 (Kim et al., 2002, Biomacro-molecules 3, 291-296) as well as P. alcaligenes M4-7. The structural gene $(phaZ_{palLB19})$ of MCL-PHA depolymerase of P. alcaligenes LB19 consisted of an 837 bp open reading frame (ORF) encoding a protein of 278 amino acids with a deduced $M_r$ of 30,188 Da. However, the MCL-PHA depolymerase gene $(phaZ_{palM4-7})$ of P. alcaligenes M4-7 was composed of an 834 bp ORF encoding a protein of 277 amino acids with a deduced Mr of 30,323 Da. Amino acid sequence analyses showed that, in the two different polypeptides, a substrate-binding domain and a catalytic domain are located in the N-terminus and in the C-terminus, respectively. The $PhaZ_{palLB19}$ and the $PhaZ_{palM4-7}$ commonly share the lipase box, GISSG, in their catalytic domains, and utilize $^{111}Asn$ and $^{110}Ser$ residues, respectively, as oxyanions that play an important role in transition-state stabilization of hydrolytic reactions.

• Korean Journal of Food Science and Technology
• /
• v.35 no.3
• /
• pp.503-509
• /
• 2003

For the development of the anti-obesity natural drug, the inhibitor of adipocyte differentiation was screened from Korean traditional medicinal plants. Phellodendri Cortex was selected as a candidate of adipocyte differentiation inhibitor. An inhibitory compound PC-4 was purified from the methanol (MeOH) extract of Phellodendri Cortex using silica gel and ODS RP-18 column chromatography and HPLC. PC-4 was obtained as yellow powder; UV ${\lambda}_{max}$ (MeOH): 230, 260, 340 and 430 nm. The chemical structure of PC-4 was determined as an isoquionoline alkaloid, berberine, on the basis of various NMR experiments including $^1H-\;and\;^{13}C-NMR$. The PC-4 inhibited the differentiation of preadipocyte NIH-3T3 L1 cells at a concentration of $1\;{\mu}g/mL$.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
• /
• 2000.06a
• /
• pp.26-86
• /
• 2000

마늘을 MAE(microwave-assisted extractiona)방법에 의하여 물, 30% 에탄올 및 50% 에탄올로 추출하고 그 특성을 비교하였다. 마늘과 각 용매의 비율은 1:2.5로 하여 마이크로웨이브 60W로 각각 5분 및 20분 동안 추출하였다. 마늘 올레오레진 수율은 50% 에탄올로 20분동안 추출한 추출물이 14.1%로 가장 높았으며 polyphenol함량도 물로 20분동안 추출한 경우가 579.0mg%로 가장 많은 함량을 나타내었다. 전자공여작용 효과 및 피르브산 함량도 물로 20분 동안 추출한 추출물이 각각 32.7% 및 마늘 1g당 13.4$\mu$molus로 가장 높게 나타났다. 각 용매에 대해 20분동안 추출한 추출물들을 37$^{\circ}C$에서 8일동안 저장하면서 피르브산 함량과 전자공여작용의 변화를 조사한 결과, 피르브산 함량은 물 추출물이 에탄올 추출물보다 높은 함량을 나타내었고 저장 시간이 증가할수록 그 함량은 감소하는 경향을 나타내었다. 전자공여작용도 물 추출물이 가장 높은 효과를 나타내었으며 추출시간에 비례하여 증가하다가 저장 6일째부터 감소하는 경향을 나타내었다. 각 용매에 대한 추출물의 색깔을 조사한 결과 물 추출물이 가장 밝고 옅은 갈색을 나타내었으며 30% 에탄올 추출물이 가장 어둡고 푸르스름한 노란색을 나타내었다. 또한 각 추출물들을 37$^{\circ}C$에서 10일간 저장하면서 갈색화 정도를 측정해 본 결과 물 추출물이 에탄올 추출물이 에탄올 추출물보다 갈색화가 많이 진행되었으며 0.1% cysteine을 첨가한 추출물과 에탄올 추출물의 갈색화는 비슷한 경향을 나타내었다. 마늘 추출물을 추출한 후에 0.1% cysteine을 첨가하여 저장한 것 보다 추출하기 전에 첨가하는 경우가 갈색화 억제 효과가 뚜렷하였다.문에 밀가루에 일부를 대용한 wheat flour dough를 사용하고 가정용 제빵기로 구워 최종 단계에까지의 제빵성 결과를 산출했다. amarans folur 5%의 대체에는 빵의 비용적이 비교적 증대했지만 그 이상 amarans flour을 대처하면 확연히 비용적은 감소했다. amarans flour 10% 대체에 hemicellulase 1250U 이상을 첨가하면 비용적은 눈에 띄게 증대했다. farinograph에 있어서 반죽의 안정성은 amarans flour 10% 대용에 현저히 감소했다. 반죽의 점탄성(아축응력, 탄성률, 점성계수)는 amarans flour 10%를 대용한 것이 무첨가한 것보다 많이 단단해졌음을 알 수 있었다. 혼합중의 반죽의 조사형 전자현미경 관찰로 amarans flour로 대체한 gluten이 단단해졌음을 알수 있었다. 유화제 stearly 칼슘, 혹은 hemicellulase를 amarans 10% 대체한 밀가루에 첨가하면 확연히 비용적을 증대시킬 수 있다는 사실을 알 수 있었다. quinoa는 명아주과 Chenopodium에 속하고 페루, 볼리비아 등의 고산지에서 재배 되어지는 것을 시료로 사용하였다. quinoa 분말은 중량의 5-20%을 quinoa를 대체하고 더욱이 분말중량에 대하여 0-200ppm의 lipase를 lipid(밀가루의 2-3배)에 대하여 품질개량제로서 이용했다. 그 결과 quinoa 대량 7.5%에서 비용적, gas cell이 가장 긍정적 결과를 산출했고 반죽의 조직구조가 강화되었다. 또 quinoa 대체에 의해 전분-지질 복합제의 흡열량이 증대된 것으로부터 전분-지질복합제의 형성 촉진이 시사되었다.이것으로 인하여 호화억제에 의한 노화 방지효과가 기대되었지만 실제로 빵의 노화는 현저히 진행되었다. 이것은 quinua 대체량 증가에 따른 반죽의 안정성이 저하되어 버린 것으로 생

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.34 no.7
• /
• pp.1052-1058
• /
• 2005

Structured lipid (SL) was synthesized by enzymatic interesterification with algae oil containing docosahexaenoic acid (DHA) and soybean oil in the stirred-batch type reactor. The reaction was performed for 15hr at $65^{\circ}C$ with 300 rpm catalyzed by sn-1,3 specific Lipozyme RM 1M lipase from Rhizomucor miehei ($11\%$ by weight of total substrates) in the absent organic solvent. SL contained $87.1\;area\%$ triacylglycerol (TAG), $12.1\;area\%$ diacylglycerol (DAG), $0.6\;area\%$ monoacylglycerol (MAG), and $0.2\;area\%$ free fatty acid (FFA). Major fatty acid profiles of SL were DHA $(15.7\;mol\%)$, linoleic $(31.1\;mol\%)$, palmitic $(20.2\;mol\%)$, oleic $(13.5\;mol\%)$ and eicosapentaenoic acid $(EPA,\;6.6 mol\%)$. SL contained the newly synthesized several peaks. Iodine and saponification of SL were 206.7 and 183.8. SL color showed darker and redder than soybean oil, and appeared the most yellowish color among SL, soybean, and algae oil.

• Journal of Life Science
• /
• v.32 no.7
• /
• pp.542-549
• /
• 2022

Polymethoxyflavones (PMFs) are flavonoids mainly found in citrus fruits and have been reported to exhibit a wide range of bioactivities, including anti-obesity, anti-cancer, and anti-inflammatory actions. To utilize PMFs as functional materials, it is necessary to develop a simple method of obtaining PMFs from citrus tissues containing a large amount of PMFs. It has been reported that Jinkyool (C. sunki Hort ex. Tanaka) peel contained a large amount of PMFs, but there are no studies on PMFs isolated from its leaves. In this study, we established a simple procedure for obtaining the PMF-rich fraction (PRF) from the leaves of Jinkyool and investigated the effects of PRF on lipid metabolism in 3T3-L1 cells. PRF inhibited lipogenesis during the differentiation of 3T3-L1 preadipocytes. It decreased the expression of peroxisome proliferator-activated receptor gamma (PPAR𝛾) and CCAAT/enhancer binding protein alpha (CEBP𝛼), FAS, and adipocyte fatty-acid-binding protein 2 (aP2). In mature 3T3-L1 adipocytes, PRF increases the phosphorylation of protein kinase A (PKA)/hormone-sensitive lipase (HSL), which are key factors involved in lipolysis. Moreover, it increases the phosphorylation of the AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) involved in fatty acid oxidation. These results suggest that PRF from Jinkyool leaves can be used as an anti-obesity agent with the action of inhibiting lipogenesis and promoting lipolysis and fatty acid oxidation in 3T3-L1 adipocytes.