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한국미생물생명공학회(구 한국산업미생물학회)> Journal of Microbiology and Biotechnology> Crystal Structure of Acyl-CoA Oxidase 3 from Yarrowia lipolytica with Specificity for Short-Chain Acyl-CoA

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Crystal Structure of Acyl-CoA Oxidase 3 from Yarrowia lipolytica with Specificity for Short-Chain Acyl-CoA

Sangwoo Kim , Kyung-jin Kim
  • : 한국미생물생명공학회
  • : Journal of Microbiology and Biotechnology 28권4호
  • : 연속간행물
  • : 2018년 04월
  • : 597-605(9pages)
Journal of Microbiology and Biotechnology

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Acyl-CoA oxidases (ACOXs) play important roles in lipid metabolism, including peroxisomal fatty acid β-oxidation by the conversion of acyl-CoAs to 2-trans-enoyl-CoAs. The yeast Yarrowia lipolytica can utilize fatty acids as a carbon source and thus has extensive biotechnological applications. The crystal structure of ACOX3 from Y. lipolytica (YlACOX3) was determined at a resolution of 2.5 A. It contained two molecules per asymmetric unit, and the monomeric structure was folded into four domains; Nα, Nβ, Cα1, and Cα2 domains. The cofactor flavin adenine dinucleotide was bound in the dimer interface. The substrate-binding pocket was located near the cofactor, and formed at the interface between the Nα, Nβ, and Cα1 domains. Comparisons with other ACOX structures provided structural insights into how YlACOX has a substrate preference for short-chain acyl-CoA. In addition, the structure of YlACOX3 was compared with those of medium- and long-chain ACOXs, and the structural basis for their differences in substrate specificity was discussed.

UCI(KEPA)

I410-ECN-0102-2018-400-004252156

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  • : 자연과학분야  > 생물
  • : KCI등재
  • : SCI,SCOPUS
  • : 월간
  • : 1017-7825
  • : 1738-8872
  • : 학술지
  • : 연속간행물
  • : 1991-2021
  • : 5714


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1Two Enteropathogenic Escherichia coli Strains Representing Novel Serotypes and Investigation of Their Roles in Adhesion

저자 : Jing Wang , Hongbo Jiao , Xinfeng Zhang , Yuanqing Zhang , Na Sun , Ying Yang , Yi Wei , Bin Hu , Xi Guo

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1191-1199 (9 pages)

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Enteropathogenic Escherichia coli (EPEC), which belongs to the attaching and effacing diarrheagenic E. coli strains, is a major causative agent of life-threatening diarrhea in infants in developing countries. Most EPEC isolates correspond to certain O serotypes; however, many strains are nontypeable. Two EPEC strains, EPEC001 and EPEC080, which could not be serotyped during routine detection, were isolated. In this study, we conducted an in-depth characterization of their putative O-antigen gene clusters (O-AGCs) and also performed constructed mutagenesis of the O-AGCs for functional analysis of O-antigen (OAg) synthesis. Sequence analysis revealed that the occurrence of O-AGCs in EPEC001 and E. coli O132 may be mediated by recombination between them, and EPEC080 and E. coli O2/O50 might acquire each O-AGC from uncommon ancestors. We also indicated that OAgknockout bacteria were highly adhesive in vitro, except for the EPEC001 wzy derivative, whose adherent capability was less than that of its wild-type strain, providing direct evidence that OAg plays a key role in EPEC pathogenesis. Together, we identified two EPEC O serotypes in silico and experimentally, and we also studied the adherent capabilities of their OAgs, which highlighted the fundamental and pathogenic role of OAg in EPEC.

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2Aloe-Emodin-Mediated Photodynamic Therapy Attenuates Sepsis-Associated Toxins in Selected Gram-Positive Bacteria In Vitro

저자 : Woodvine Otieno , Chengcheng Liu , Yanhong Ji

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1200-1209 (10 pages)

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Sepsis is an acute inflammatory response that leads to life-threatening complications if not quickly and adequately treated. Cytolysin, hemolysin, and pneumolysin are toxins produced by grampositive bacteria and are responsible for resistance to antimicrobial drugs, cause virulence and lead to sepsis. This work assessed the effects of aloe-emodin (AE) and photodynamic therapy (PDT) on sepsis-associated gram-positive bacterial toxins. Standard and antibiotic-resistant Enterococcus faecalis, Staphylococcus aureus, and Streptococcus pneumonia bacterial strains were cultured in the dark with varying AE concentrations and later irradiated with 72 J/cm-2 light. Colony and biofilm formation was determined. CCK-8, Griess reagent reaction, and ELISA assays were done on bacteriainfected RAW264.7 cells to determine the cell viability, NO, and IL-1β and IL-6 pro-inflammatory cytokines responses, respectively. Hemolysis and western blot assays were done to determine the effect of treatment on hemolysis activity and sepsis-associated toxins expressions. AE-mediated PDT reduced bacterial survival in a dose-dependent manner with 32 μg/ml of AE almost eliminating their survival. Cell proliferation, NO, IL-1β, and IL-6 cytokines production were also significantly downregulated. Further, the hemolytic activities and expressions of cytolysin, hemolysin, and pneumolysin were significantly reduced following AE-mediated PDT. In conclusion, combined use of AE and light (435 ± 10 nm) inactivates MRSA, S. aureus (ATCC 29213), S. pneumoniae (ATCC 49619), MDR-S. pneumoniae, E. faecalis (ATCC 29212), and VRE (ATCC 51299) in an AE-dose dependent manner. AE and light are also effective in reducing biofilm formations, suppressing proinflammatory cytokines, hemolytic activities, and inhibiting the expressions of toxins that cause sepsis.

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3Ramlibacter terrae sp. nov. and Ramlibacter montanisoli sp. nov., Isolated from Soil

저자 : Shehzad Abid Khan , Hyung Min Kim , Ju Hye Baek , Hye Su Jung , Che Ok Jeon

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1210-1217 (8 pages)

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Two gram-negative, catalase-positive, strictly aerobic, and white colony-forming bacteria, strains H242T and B156T, were isolated from soil in South Korea. Cells of strain H242T were oxidase-positive and non-motile short rods, while those of strain B156T were oxidase-negative and long non-motile rods. Ubiquinone-8 was identified as the sole isoprenoid quinone in both strains. C16:0, cyclo-C17:0, and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol were identified in both strains as the major cellular fatty acids and polar lipids, respectively. The DNA G+C contents of strains H242T and B156T were 69.4 mol% and 69.3 mol%, respectively. Phylogenetic analyses based on 16S rRNA and 92 concatenated core gene sequences revealed that strains H242T and B156T formed distinct phylogenic lineages from other Ramlibacter type strains. The DNA-DNA hybridization (DDH) value between strains H242T and B156T was 24.6%. Strains H242T and B156T were most closely related to Ramlibacter ginsenosidimutans BXN5-27T and Ramlibacter monticola G-3-2T with 98.4% and 98.6% 16S rRNA gene sequence similarities, respectively. Digital DDH values between strain H242T and R. ginsenosidimutans and between strain B156T and R. monticola were 23.5% and 26.1%, respectively. Phenotypic, chemotaxonomic, and molecular analyses indicated that strains H242T and B156T represent two novel species of the genus Ramlibacter, for which the names Ramlibacter terrae sp. nov. and Ramlibacter montanisoli sp. nov., respectively, are proposed. The type strains of R. terrae and R. montanisoli are H242T (=KACC 21667 T =JCM 33922T) and B156T (=KACC 21665 T =JCM 33920T), respectively.

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4Isolation and Characterization of Cold- Adapted PGPB and Their Effect on Plant Growth Promotion

저자 : Mingyuan Li , Jilian Wang , Tuo Yao , Zhenlong Wang , Huirong Zhang , Changning Li

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1218-1230 (13 pages)

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Cold-adapted plant growth-promoting bacteria (PGPB) with multiple functions are an important resource for microbial fertilizers with low-temperature application. In this study, culturable coldadapted PGPB strains with nitrogen fixation and phosphorus solubilization abilities were isolated. They were screened from root and rhizosphere of four dominant grass species in nondegraded alpine grasslands of the Qilian Mountains, China. Their other growth-promoting characteristics, including secretion of indole-3-acetic acid (IAA), production of siderophores and ACC deaminase, and antifungal activity, were further studied by qualitative and quantitative methods. In addition, whether the PGPB strains could still exert plant growth-promoting activity at 4°C was verified. The results showed that 67 isolates could maintain one or more growth-promoting traits at 4°C, and these isolates were defined as cold-adapted PGPB. They were divided into 8 genera by 16S rRNA gene sequencing and phylogenetic analysis, of which Pseudomonas (64.2%) and Serratia (13.4%) were the common dominant genera, and a few specific genera varied among the plant species. A test-tube culture showed that inoculation of Elymus nutans seedlings with cold-adapted PGPB possessing different functional characteristics had a significant growth-promoting effect under controlled low-temperature conditions, including the development of the roots and aboveground parts. Pearson correlation analysis revealed that different growth-promoting characteristics made different contributions to the development of the roots and aboveground parts. These cold-adapted PGPB can be used as excellent strain resources suitable for the near-natural restoration of degraded alpine grasslands or agriculture stock production in cold areas.

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5Profiles of Bacillus spp. Isolated from the Rhizosphere of Suaeda glauca and Their Potential to Promote Plant Growth and Suppress Fungal Phytopathogens

저자 : Ping Lu , Ke Jiang , Ya-qiao Hao , Wan-ying Chu , Yu-dong Xu , Jia-yao Yang , Jia-le Chen , Guo-hong Zeng , Zhou-hang Gu , Hong-xin Zhao

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1231-1240 (10 pages)

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Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.

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6Suppression of Fusarium Wilt Caused by Fusarium oxysporum f. sp. lactucae and Growth Promotion on Lettuce Using Bacterial Isolates

저자 : Dil Raj Yadav , Mahesh Adhikari , Sang Woo Kim , Hyun Seung Kim , Youn Su Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1241-1255 (15 pages)

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This study was carried out to explore a non-chemical strategy for enhancing productivity by employing some antagonistic rhizobacteria. One hundred eighteen bacterial isolates were obtained from the rhizospheric zone of various crop fields of Gangwon-do, Korea, and screened for antifungal activity against Fusarium wilt (Fusarium oxysporum f. sp. lactucae) in lettuce crop under in vitro and in vivo conditions. In broth-based dual culture assay, fourteen bacterial isolates showed significant inhibition of mycelial growth of F. oxysporium f. sp. lactucae. All of the antagonistic isolates were further characterized for the antagonistic traits under in vitro conditions. The isolates were identified on the basis of biochemical characteristics and confirmed at their species level by 16S rRNA gene sequencing analysis. Arthrobacter sulfonivorans, Bacillus siamensis, Bacillus amyloliquefaciens, Pseudomonas proteolytica, four Paenibacillus peoriae strains, and Bacillus subtilis were identified from the biochemical characterization and 16S rRNA gene sequencing analysis. The isolates EN21 and EN23 showed significant decrease in disease severity on lettuce compared to infected control and other bacterial treatments under greenhouse conditions. Two bacterial isolates, EN4 and EN21, were evaluated to assess their disease reduction and growth promotion in lettuce in field conditions. The consortium of EN4 and EN21 showed significant enhancement of growth on lettuce by suppressing disease caused by F. oxysporum f. sp. lactucae respectively. This study clearly indicates that the promising isolates, EN4 (P. proteolytica) and EN21 (Bacillus siamensis), can be commercialized and used as biofertilizer and/or biopesticide for sustainable crop production.

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7Freeze-Dried Powder of Rubus coreanus Miquel Ameliorates Isoproterenol-Induced Oxidative Stress and Tissue Damage in Rats

저자 : Jin Tae Kim , Shuai Qiu , Yimeng Zhou , Ji Hyun Moon , Seung Beom Lee , Ho Jin Park , Hong Jin Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1256-1261 (6 pages)

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Rubus coreanus Miquel (bokbunja), Korean black raspberry, is known to possess various phytochemicals that exert antioxidative, anti-inflammatory, and anti-cancer effects. However, most studies on Rubus coreanus Miquel have been performed with the solvent extracts and/or a single component to demonstrate the efficacy, while studies evaluating the effect of the whole fructus of Rubus coreanus Miquel are limited. In this study, therefore, we employed the isoproterenol (IPN)- induced myocardial infarction model and investigated the effect of freeze-dried powder of Rubus coreanus Miquel (RCP) on oxidative stress and prevention of organ damage. Oral administration of RCP reduced the level of toxicity markers, alanine transaminase (ALT), aspartate transaminase (AST), and lactate dehydrogenase (LDH) without affecting body weight and diet intake. The oxidative stress marker glutathione (GSH) increased about 45% and malonaldehyde (MDA) decreased about 27% compared to the IPN group with RCP-H (3%) administration. By histological analysis, IPN induced significant myocardial damage in the heart and vascular injury in the liver, and RCP administration ameliorated the damages in a dose-dependent manner. Taken together, RCP activated the antioxidant system leading to prevention of damage to organs by IPN in rats, making it possible to expect beneficial efficacies by consuming the whole fructus of Rubus coreanus Miquel.

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8Bioconversion of Untreated Corn Hull into L-Malic Acid by Trifunctional Xylanolytic Enzyme from Paenibacillus curdlanolyticus B-6 and Acetobacter tropicalis H-1

저자 : Thi Bich Huong Duong , Prattana Ketbot , Paripok Phitsuwan , Rattiya Waeonukul , Chakrit Tachaapaikoon , Akihiko Kosugi , Khanok Ratanakhanokchai , Patthra Pason

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1262-1271 (10 pages)

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L-Malic acid (L-MA) is widely used in food and non-food products. However, few microorganisms have been able to efficiently produce L-MA from xylose derived from lignocellulosic biomass (LB). The objective of this work is to convert LB into L-MA with the concept of a bioeconomy and environmentally friendly process. The unique trifunctional xylanolytic enzyme, PcAxy43A from Paenibacillus curdlanolyticus B-6, effectively hydrolyzed xylan in untreated LB, especially corn hull to xylose, in one step. Furthermore, the newly isolated, Acetobacter tropicalis strain H1 was able to convert high concentrations of xylose derived from corn hull into L-MA as the main product, which can be easily purified. The strain H1 successfully produced a high L-MA titer of 77.09 g/l, with a yield of 0.77 g/g and a productivity of 0.64 g/l/h from the xylose derived from corn hull. The process presented in this research is an efficient, low-cost and environmentally friendly biological process for the green production of L-MA from LB.

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9Aqueous Extract of Lysimachia christinae Hance Prevents Cholesterol Gallstone in Mice by Affecting the Intestinal Microflora

저자 : Shijia Liu , Quji Luorong , Kaizhi Hu , Weiguo Cao , Wei Tao , Handeng Liu , Dan Zhang

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1272-1280 (9 pages)

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With changes in human dietary patterns, the proportion of high-fat and high-cholesterol foods in the daily diet has increased. As a result, the incidence rate of cholelithiasis is increasing rapidly. Many studies have reported on the crucial role that the intestinal microflora plays in the progression of gallstones. Although the whole herb of Lysimachia christinae, a traditional Chinese medicine, has long been extensively used as a remedy for cholelithiasis in China, its effects on the intestinal microflora remain unknown. Hence, in this study, we investigated the ability of the aqueous extract of L. christinae (LAE) to prevent cholesterol gallstones (CGSs) in model animals by affecting the intestinal microflora. The effects of LAE on body weight, serum lipid profile, visceral organ indexes, and histomorphology were studied in male C57BL/6J mice, which were induced by a lithogenic diet. After the 8-week study, CGSs formation was greatly reduced after LAE treatment. LAE also reduced body weight gain and hyperlipidemia and restored the histomorphological changes. Moreover, the intestinal microflora exhibited significant variation. In the model group fed the lithogenic diet, the abundances of the genera unclassified Porphyromonadaceae, Lactobacillus and Alloprevotella decreased, but in contrast, Akkermansia dramatically increased compared with the control check group, which was fed a normal diet; the administration of LAE reversed these changes. These results imply that L. christinae can be considered an efficient therapy for eliminating CGSs induced by a high-fat and high-cholesterol diet, which may be achieved by influencing the intestinal microflora.

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10The Effect of Lactobacillus gasseri BNR17 on Postmenopausal Symptoms in Ovariectomized Rats

저자 : Sol Lee , Dong Hoon Jung , Miri Park , Seung-woo Yeon , Sang-hyuk Jung , Sung-il Yun , Han-oh Park , Wonbeak Yoo

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 31권 9호 발행 연도 : 2021 페이지 : pp. 1281-1287 (7 pages)

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Clinical and preclinical studies have reported that Lactobacillus gasseri BNR17, a probiotic bacterial strain isolated from human breast milk, reduces body weight and white adipose tissue volume. In order to further explore the actions of L. gasseri BNR17, we investigated the anti-menopausal effects of L. gasseri BNR17 in an ovariectomized (OVX) rat model. The serum alanine aminotransferase levels of the rats in the OVX-BNR17 group were lower than those of the rats in the OVX-vehicle only (OVX-Veh) group. Upon administration of L. gasseri BNR17 after ovariectomy, calcitonin and Serotonin 2A levels increased significantly, whereas serum osteocalcin levels showed a decreasing tendency. Compared to the rats in the OVX-Veh group, those in the OVX-BNR17 group showed lower urine deoxypyridinoline levels, lower pain sensitivity, and improved vaginal cornification. Furthermore, L. gasseri BNR17 administration increased bone mineral density in the rats with OVX-induced femoral bone loss. These results suggest that L. gasseri BNR17 administration could alleviate menopausal symptoms, indicating that this bacterium could be a good functional probiotic for managing the health of older women.

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1Improving the Microbial Safety of Fresh-Cut Endive with a Combined Treatment of Cinnamon Leaf Oil Emulsion Containing Cationic Surfactants and Ultrasound

저자 : Jun-beom Park , Ji-hoon Kang , Kyung Bin Song

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 503-509 (7 pages)

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Endive is widely consumed in a fresh-cut form owing to its rich nutritional content. However, fresh-cut vegetables are susceptible to contamination by pathogenic bacteria. This study investigated the antibacterial activities of the combined treatment of cinnamon leaf oil emulsion containing cetylpyridinium chloride or benzalkonium chloride (CLC and CLB, respectively) as a cationic surfactant and ultrasound (US) against Listeria monocytogenes and Escherichia coli O157:H7 on endive. The combined treatment of CLC or CLB with US reduced the population of L. monocytogenes by 1.58 and 1.47 log colony forming units (CFU)/g, respectively, and that of E. coli O157:H7 by 1.60 and 1.46 log CFU/g, respectively, as compared with water washing treatment. The reduction levels of both pathogens were higher than those observed with 0.2 mg/ml sodium hypochlorite. In addition, the combined treatment showed no effect on the quality of the fresh-cut endive (FCE). In particular, the degree of browning in FCE was less for the treatment group than for the control and water washing treatment groups. Thus, cationic surfactant-based cinnamon leaf oil emulsions combined with US may be an effective washing treatment for the microbial safety of FCE.

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2A New Method of Producing a Natural Antibacterial Peptide by Encapsulated Probiotics Internalized with Inulin Nanoparticles as Prebiotics

저자 : Lian-hua Cui , Chang-guo Yan , Hui-shan Li , Whee-soo Kim , Liang Hong , Sang-kee Kang , Yun-jaie Choi , Chong-su Cho

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 510-519 (10 pages)

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Synbiotics are a combination of probiotics and prebiotics, which lead to synergistic benefits in host welfare. Probiotics have been used as an alternative to antibiotics. Among the probiotics, Pediococcus acidilactici (PA) has shown excellent antimicrobial activity against Salmonella Gallinarum (SG) as a major poultry pathogen and has improved the production performances of animals. Inulin is widely used as a prebiotic for the improvement of animal health and growth. The main aim of this study was to investigate the antimicrobial activity of inulin nanoparticle (IN)-internalized PA encapsulated into alginate/chitosan/alginate (ACA) microcapsules (MCs) for future in vivo application. The prepared phthalyl INs (PINs) were characterized by DLS and FE-SEM. The contents of phthal groups in the PINs were estimated by 1H-NMR measurement as 25.1 mol.-%. The sizes of the PINs measured by DLS were approximately 203 nm. Internalization into PA was confirmed by confocal microscopy and flow cytometry. The antimicrobial activity of PIN-internalized probiotics encapsulated into ACA MCs was measured by coculture antimicrobial assays on SG. PIN-internalized probiotics had a higher antimicrobial ability than that of ACA MCs loaded with PA/inulin or PA. Interestingly, when PINs were treated with PA and encapsulated into ACA MCs, as a natural antimicrobial peptide, pediocin was produced much more in the culture medium compared with other groups with inulin-loaded ACA MCs and PA encapsulated into ACA MCs.

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3Conessine Treatment Reduces Dexamethasone-Induced Muscle Atrophy by Regulating MuRF1 and Atrogin-1 Expression

저자 : Hyunju Kim , Minsu Jang , Rackhyun Park , Daum Jo , Inho Choi , Joonho Choe , Won Keun Oh , Junsoo Park

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 520-526 (7 pages)

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Conessine, a steroidal alkaloid, is a potent histamine H3 antagonist with antimalarial activity. We recently reported that conessine treatment interferes with H2O2-induced cell death by regulating autophagy. However, the cellular signaling pathways involved in conessine treatment are not fully understood. Here, we report that conessine reduces muscle atrophy by interfering with the expression of atrophy-related ubiquitin ligases MuRF-1 and atrogin-1. Promoter reporter assay revealed that conessine treatment inhibits FoxO3a-dependent transcription, NF-κB-dependent transcription, and p53-dependent transcription. We also showed by quantitative RT-PCR and western blot assays that conessine treatment reduced dexamethasone-induced expression of MuRF1 and atrogin-1. Finally, we demonstrated that conessine treatment reduced dexamethasone-induced muscle atrophy using differentiated C2C12 cells. These results collectively suggest that conessine is potentially useful in the treatment of muscle atrophy.

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4Protection of Radiation-Induced DNA Damage by Functional Cosmeceutical Poly-Gamma-Glutamate

저자 : Yu-jin Oh , Mi-sun Kwak , Moon-hee Sung

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 527-533 (7 pages)

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This study compared the radioprotective effects of high-molecular-weight poly-gammaglutamate (γ-PGA, average molecular mass 3,000 kDa) and a reduced form of glutathione (GSH, a known radioprotector) on calf thymus DNA damage. The radiation-induced DNA damage was measured on the basis of the decreased fluorescence intensity after binding the DNA with ethidium bromide. All the experiments used 60Co gamma radiation at 1,252 Gy, representing 50% DNA damage. When increasing the concentration of γ-PGA from 0.33 to 1.65 μM, the DNA protection from radiation-induced damage also increased, with a maximum of 87% protection. Meanwhile, the maximal DNA protection when increasing the concentration of GSH was only 70%. Therefore, γ-PGA exhibited significant radioprotective effects against gamma irradiation.

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5Properties of Kimchi Fermented with GABA-Producing Lactic Acid Bacteria as a Starter

저자 : Kang Wook Lee , Jae Min Shim , Zhuang Yao , Jeong A Kim , Jeong Hwan Kim

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 534-541 (8 pages)

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Kimchi (a traditional Korean fermented vegetable) was prepared with a starter, Lactobacillus zymae GU240 producing γ-aminobutyric acid (GABA), and one precursor of GABA (glutamic acid, glutamic acid monosodium salt (MSG), or kelp extract). L. zymae GU240, an isolate from kimchi, can grow at 7% NaCl and low temperature. Five different kimchi samples were fermented for 20 weeks at -1°C. Kimchi with starter alone could not produce GABA. The GABA content was highest in kimchi with co-inoculation of the starter and MSG (1% (w/w)). Kimchi co-inoculated with the starter and kelp extract powder (3% (w/w)) had the second highest GABA content. Addition of glutamic acid powder (1% (w/w)) caused a reduction in the pH level of kimchi and growth inhibition of lactic acid bacteria and yeasts. Kimchi samples with MSG or kelp extract showed improvement of sensory evaluation scores. The results demonstrate the possibility to produce kimchi with improved functionality and taste by using L. zymae GU240 as a starter along with a suitable precursor such as MSG or kelp extract.

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6Non-Benzoquinone Geldanamycin Analog, WK-88-1, Induces Apoptosis in Human Breast Cancer Cell Lines

저자 : Yu-ru Zhao , Hong-mei Li , Meilin Zhu , Jing Li , Tao Ma , Qiang Huo , Young-soo Hong , Cheng-zhu Wu

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 542-550 (9 pages)

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Heat shock protein 90 (Hsp90) is treated as a molecular therapeutic target for the prevention and treatment of cancer. Geldanamycin (GA) was the first identified natural Hsp90 inhibitor, but hepatotoxicity has limited its clinical application. Nevertheless, a new GA analog (WK-88- 1) with the non-benzoquinone skeleton, obtained from genetically engineered Streptomyces hygroscopicus, was found to have anticancer activity against two human breast cancer cell lines. WK-88-1 produced concentration-dependent inhibition of cell proliferation, cell cycle arrest, and apoptosis in estrogen receptor (ER)-positive MCF-7 and ER-negative MDA-MB-231 cell lines. Detailed analysis showed that WK-88-1 downregulated some key cell cycle molecules (CDK1 and cyclin B1) and lead to G2/M cell cycle arrest. Further studies also showed that WK- 88-1 could induce human breast cancer cell apoptosis by downregulating Hsp90 client proteins (Akt, p-Akt, IKK, c-Raf, and Bcl-2), decreasing the ATP level, increasing reactive oxygen species production, and lowering the mitochondrial membrane potential. Meanwhile, we discovered that WK-88-1 significantly decreased the levels of Her-2 and ER-α in MCF-7 cells but not in MDA-MB-231 cells. In addition, WK-88-1 significantly increased caspase-3, -8, and -9 activities and the cleavage of PARP in a concentration-dependent manner (with the exception of caspase-3 and PARP in MCF-7 cells). Taken together, our preliminary results suggest that WK-88-1 has the potential to play a role in breast cancer therapy.

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7Analysis of Microbiota in Bellflower Root, Platycodon grandiflorum, Obtained from South Korea

저자 : Daeho Kim , Sanghyun Hong , Hongjun Na , Jihwan Chun , Robin B. Guevarra , You-tae Kim , Sangryeol Ryu , Hyeun Bum Kim , Ju-hoon Lee

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 551-560 (10 pages)

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Bellflower root (Platycodon grandiflorum), which belongs to the Campanulaceae family, is a perennial grass that grows naturally in Korea, northeastern China, and Japan. Bellflower is widely consumed as both food and medicine owing to its high nutritional value and potential therapeutic effects. Since foodborne disease outbreaks often come from vegetables, understanding the public health risk of microorganisms on fresh vegetables is pivotal to predict and prevent foodborne disease outbreaks. We investigated the microbial communities on the bellflower root (n = 10). 16S rRNA gene amplicon sequencing targeting the V6-V9 regions of 16S rRNA genes was conducted via the 454-Titanium platform. The sequence quality was checked and phylogenetic assessments were performed using the RDP classifier implemented in QIIME with a bootstrap cutoff of 80%. Principal coordinate analysis was performed using the weighted Fast UniFrac distance. The average number of sequence reads generated per sample was 67,192 sequences. At the phylum level, bacterial communities from the bellflower root were composed primarily of Proteobacteria, Firmicutes, and Actinobacteria in March and September samples. Genera Serratia, Pseudomonas, and Pantoea comprised more than 54% of the total bellflower root bacteria. Principal coordinate analysis plots demonstrated that the microbial community of bellflower root in March samples was different from those in September samples. Potential pathogenic genera, such as Pantoea, were detected in bellflower root samples. Even though further studies will be required to determine if these species are associated with foodborne illness, our results indicate that the 16S rRNA gene-based sequencing approach can be used to detect pathogenic bacteria on fresh vegetables.

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8Hydroxylation of Resveratrol with DoxA In Vitro: An Enzyme with the Potential for the Bioconversion of a Bioactive Stilbene

저자 : Hemraj Rimal , Sang-cheol Yu , Joo-ho Lee , Tokutaro Yamaguchi , Tae-jin Oh

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 561-565 (5 pages)

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The late-stage doxorubicin biosynthesis pathway acting enzyme (DoxA) from Streptomyces peucetius CYP129A2 exhibited substrate promiscuity towards the stilbene group of compounds such as resveratrol. DoxA along with two accessory enzymes ferrdoxin reductase and ferredoxin from spinach hydroxylated resveratrol at the 3'-position in vitro to produce piceatannol. The product was identified by HPLC-PDA and high-resolution HR-qTOF-ESI/ MS analyses in positive mode. The ESI/MS fragments resembled the hydroxylated product of resveratrol.

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9Synthesis of Aesculetin and Aesculin Glycosides Using Engineered Escherichia coli Expressing Neisseria polysaccharea Amylosucrase

저자 : Soyoon Park , Keumok Moon , Cheon-seok Park , Dong-hyun Jung , Jaeho Cha

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 566-570 (5 pages)

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Because glycosylation of aesculetin and its 6-glucoside, aesculin, enhances their biological activities and physicochemical properties, whole-cell biotransformation and enzymatic synthesis methodologies using Neisseria polysaccharea amylosucrase were compared to determine the optimal production method for glycoside derivatives. High-performance liquid chromatography analysis of reaction products revealed two glycosylated products (AGG1 and AGG2) when aesculin was used as an acceptor, and three products (AG1, AG2, and AG3) when using aesculetin. The whole-cell biotransformation production yields of the major transfer products for each acceptor (AGG1 and AG1) were 85% and 25%, respectively, compared with 68% and 14% for enzymatic synthesis. These results indicate that whole-cell biotransformation is more efficient than enzymatic synthesis for the production of glycoside derivatives.

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10Crystal Structure and Biochemical Characterization of Xylose Isomerase from Piromyces sp. E2

저자 : Hyeoncheol Francis Son , Sun-mi Lee , Kyung-jin Kim

발행기관 : 한국미생물생명공학회 간행물 : Journal of Microbiology and Biotechnology 28권 4호 발행 연도 : 2018 페이지 : pp. 571-578 (8 pages)

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Biofuel production using lignocellulosic biomass is gaining attention because it can be substituted for fossil fuels without competing with edible resources. However, because Saccharomyces cerevisiae does not have a D-xylose metabolic pathway, oxidoreductase or isomerase pathways must be introduced to utilize D-xylose from lignocellulosic biomass in S. cerevisiae. To elucidate the biochemical properties of xylose isomerase (XI) from Piromyces sp. E2 (PsXI), we determine its crystal structure in complex with substrate mimic glycerol. An amino- acid sequence comparison with other reported XIs and relative activity measurements using five kinds of divalent metal ions confirmed that PsXI belongs to class II XIs. Moreover kinetic analysis of PsXI was also performed using Mn2+, the preferred divalent metal ion for PsXI. In addition, the substrate-binding mode of PsXI could be predicted with the substrate mimic glycerol bound to the active site. These studies may provide structural information to enhance D-xylose utilization for biofuel production.

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