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Enhanced Production of Carboxymethylcellulase by a Newly Isolated Marine Microorganism Bacillus atrophaeus LBH-18 Using Rice Bran, a Byproduct from the Rice Processing Industry (미강을 이용한 해양미생물 Bacillus atrophaeus LBH-18 유래의 carboxymethylcellulase 생산의 최적화)

  • Kim, Yi-Joon;Cao, Wa;Lee, Yu-Jeong;Lee, Sang-Un;Jeong, Jeong-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1295-1306
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    • 2012
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.

Isolation and Characteristics of a Phenol-degrading Bacterium, Rhodococcus pyridinovorans P21 (페놀분해세균 Rhodococcus pyridinovorans P21의 분리 및 페놀분해 특성)

  • Cho, Kwang-Sik;Lee, Sang-Mee;Shin, Myung-Jae;Park, Soo-Yun;Lee, Ye-Ram;Jang, Eun-Young;Son, Hong-Joo
    • Journal of Life Science
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    • v.24 no.9
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    • pp.988-994
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    • 2014
  • The effluents of chemical and petroleum industries often contain non-biodegradable aromatic compounds, with phenol being one of the major organic pollutants present among a wide variety of highly toxic organic chemicals. Phenol is toxic upon ingestion, contact, or inhalation, and it is lethal to fish even at concentrations as low as 0.005 ppm. Phenol biodegradation has been studied in detail using bacterial strains. However, these microorganisms suffer from substrate inhibition at high concentrations of phenol, whereby growth is inhibited. A phenol-degrading bacterium, P21, was isolated from oil-contaminated soil. The phenotypic characteristics and a phylogenetic analysis indicated the close relationship of strain P21 to Rhodococcus pyridinovorans. Phenol biodegradation by strain P21 was studied under shaking condition. The optimal conditions for phenol biodegradation by strain P21 were 0.09% $KNO_3$, 0.1% $K_2HPO_4$, 0.3% $NaH_2PO_4$, 0.015% $MgSO_4{\cdot}7H_2O$, 0.001% $FeSO_4{\cdot}7H_2O$, initial pH 9, and $20-30^{\circ}C$, respectively. When 1,000 ppm of phenol was added to the optimal medium, the strain P21 completely degraded it within two days. Rhodococcus pyridinovorans P21 could grow in up to 1,500 ppm of phenol as the sole carbon source in a batch culture, but it could not grow in a medium containing above 2,000 ppm. Moreover, strain P21 could utilize toxic compounds, such as toluene, xylene, and hexane, as a sole carbon source. However, no growth was detected on chloroform.

Adhesion Characteristics and the High Pressure Resistance of Biofilm Bacteria in Seawater Reverse Osmosis Desalination Process (역삼투 해수담수화 공정 내 바이오필름 형성 미생물의 부착 및 고압내성 특성)

  • Jung, Ji-Yeon;Lee, Jin-Wook;Kim, Sung-Youn;Kim, In-S.
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.1
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    • pp.51-57
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    • 2009
  • Biofouling in seawater reverse osmosis (SWRO) desalination process causes many problems such as flux decline, biodegradation of membrane, increased cleaning time, and increased energy consumption and operational cost. Therefore biofouling is considered as the most critical problem in system operation. To control biofouling in early stage, detection of the most problematic bacteria causing biofouling is required. In this study, six model bacteria were chosen; Bacillus sp., Flavobacterium sp., Mycobacterium sp., Pseudomonas aeruginosa, Pseudomonas fluorescens, and Rhodobacter sp. based on report in the literature and phylogenetic analysis of seawater intake and fouled RO membrane. The adhesion to RO membrane, the high pressure resistance, and the hydrophobicity of the six model bacteria were examined to find out their fouling potential. Rhodobacter sp. and Mycobacterium sp. were found to attach very well to RO membrane surface compared to others used in this study. The test of hydrophobicity revealed that the bacteria which have high hydrophobicity or similar contact angle with RO membrane ($63^{\circ}$ of contact angle) easily attached to RO membrane surface. P. aeruginosa which is highly hydrophilic ($23.07^{\circ}$ of contact angle) showed the least adhesion characteristic among six model bacteria. After applying a pressure of 800 psi to the sample, Rhodobacter sp. was found to show the highest reduction rate; with 59-73% of the cells removed from the membrane under pressure. P. fluorescens on the other hand analyzed as the most pressure resistant bacteria among six model bacteria. The difference between reduction rates using direct counting and plate counting indicates that the viability of each model bacteria was affected significantly from the high pressure. Most cells subjected to high pressure were unable to form colonies even thought they maintained their structural integrity.

The systematic implications of leaf micromorphological characteristics in the tribe Neillieae (Spiraeoideae, Rosaceae) (나도국수나무족(장미과) 잎 표피 미세형태학적 형질의 계통학적 유용성)

  • Song, Jun-Ho;Hong, Suk-Pyo
    • Korean Journal of Plant Taxonomy
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    • v.47 no.3
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    • pp.222-235
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    • 2017
  • A comparative study of the leaf epidermal micromorphology in the tribe Neillieae (Neillia: 4 species, 4 varieties; Physocarpus: 5 species; Stephanandra: 2 species) was carried out using scanning electron microscopy in order to evaluate the taxonomic and systematic implications of these characteristics. The leaves of the genera Neillia and Stephanandra were hypostomatic, whereas those of P. monogynus, P. opulifolius were amphistomatic. The range of the size of the stomata is $12.02-34.39{\times}10.76-27.13{\mu}m$; the smallest was found in N. thyrsiflora (average $13.98{\times}12.43{\mu}m$; $L{\times}W$), while the largest was measured in N. gracilis (average $26.82{\times}20.67{\mu}m$; $L{\times}W$). Paracytic stomata complexes are only found in N. affinis, and the anomocytic type was most commonly found. The papillate epidermal cell type was only observed on the abaxial surfaces of P. insularis. Platelet epicuticular waxes were found on the adaxial surfaces of N. affinis and S. tanakae. Four types (unicellular non-glandular, two- to five-armed, stellate, and glandular) of trichomes were found on the leaves. Stellates were observed in all species of Physocarpus except for P. insularis. Consequently, leaf epidermal micromorphological characteristics (e.g., the presence of papillate epidermal cells and stellate, and stomata complexes) may have high taxonomic and systematic value in Neillieae. Our results strongly support previous molecular phylogenetic and palynological hypotheses that Stephanandra and Neillia are a single genus and that Physocarpus insularis should be considered as a member of Spiraea.

Cloning of Low-molecular-weight Glutenin Subunit Genes and Identification of their Protein Products in Common Wheat (Triticum aestivum L.) (보통 밀에서 저분자글루테닌 유전자 클로닝 및 단백질 동정)

  • Lee, Jong-Yeol;Kim, Yeong-Tae;Kim, Bo-Mi;Lee, Jung-Hye;Lim, Sun-Hyung;Ha, Sun-Hwa;Ahn, Sang-Nag;Nam, Myung-Hee;Kim, Young-Mi
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.547-554
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    • 2010
  • Low-molecular-weight glutenin subunit (LMW-GS) in common wheat (Triticum aestivum L.) is important for quality processing of bread and noodles. The objectives of this study were to clarify the composition of LMW-GSs and to identify their corresponding proteins. Using LMW-GS specific primers we cloned and characterized 43 LMW-GS genes in the wheat cultivar 'Jokyoung'. Some of these genes contain polypeptides different in size due to the presence of various deletions or insertions within repetitive and glutamine-rich domains. The comparison of deduced amino acid sequence of the LMW-GS genes in Jokyoung with that of 12 groups LMW-GSs of wheat cultivar Norin 61 showed that the deduced amino acid sequences were nearly the same to LMW-GS groups of 1, 2, 3/4, 5, 7, 10 and 11. All LMW-GS genes contain eight cysteine residues, which are conserved among all of the typical LMW-GS sequences. The relative positions of cysteine residues are also conserved, except those of the first and seventh. Based on phylogenetic analysis, the 43 sequences with the same N-terminal and C-terminal amino acid sequences were clustered in the same group. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 7 spots of LMW-GSs of Jokyoung separated by two-dimensional gel electrophoresis (2DE). Of them, Glu-B3 (LMW-m and LMW-s) and Glu-D3 (LMW-m) were detected in two and three spots, respectively and the others were not clear. Collectively, we classified diverse LMW-GSs and identified their corresponding protein products. These results will be helpful in breeding programs for improvement of wheat flour quality.

Anti-oxidative and Anti-cancer Activities of Ethanol Extract of Litsea populifolia (인체 폐암 세포주 A549에서 Litsea populifolia 추출물의 항산화 및 항암활성 분석)

  • Jin, Soojung;Oh, You Na;Jeong, Hyun Young;Yun, Hee Jung;Park, Jung-ha;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.29 no.6
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    • pp.679-687
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    • 2019
  • Litsea populifolia, a plant species of the Lauraceae family, is widely distributed in the tropical and subtropical areas of Asia. The phylogenetic relationships and botanical characteristics of L. populifolia have been reported; however, its anti-oxidative and anti-cancer activities remain unclear. In this study, we evaluated the anti-oxidative and anti-cancer effects of ethanol extracts of L. populifolia (EELP) together with the molecular mechanism of its anti-cancer activity in human lung adenocarcinoma A549 cells. EELP showed significant anti-oxidative effects with a 50% inhibitory concentration at $11.71{\mu}g/ml$, which was measured by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. EELP exhibited cytotoxic activity and induced cell cycle arrest at the G1 phase in A549 cells in a dose-dependent manner, whereas EELP did not have the cytotoxic effect on the normal human lung cell line IMR90. Treatment with EELP also resulted in a decreased expression of G1/S transition-related molecules-including cyclin-dependent kinase (CDK) 2, CDK6, cyclin D1, and cyclin E-both for the transcription and translation levels. EELP-induced G1 arrest was associated with the phosphorylation of checkpoint kinase 2 (CHK2), p53, cell division cycle 25 homolog A (CDC25A), and the reduction of CDC25A expression in A549 cells. Collectively, these results suggest that EELP may exert an anti-cancer effect by cell cycle arrest at the G1 phase through both p53-dependent and p53-independent (ATM/CHK2/CDC25A/CDK2) pathways in A549 cells.

Isolation and characterization of cellulolytic yeast belonging to Moesziomyces sp. from the gut of Grasshopper (메뚜기의 내장에서 분리한 Moesziomyces 속에 속하는 셀룰로오스 분해 효모의 분리 및 특성)

  • Kim, Ju-Young;Jung, Hee-Young;Park, Jong-Seok;Cho, Sung-Jin;Lee, Hoon Bok;Sung, Gi-Ho;Subramani, Gayathri;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.234-241
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    • 2019
  • An intensive interaction between yeasts and insects has highlighted their relevance for attraction to food and for the insect's development and behavior. Yeast associated in the gut of insects secretes cellulase which aided in the food digestion (cellulose degradation). Three strains of cellulose-degrading yeast were isolated from the gut of adult grasshoppers collected in Gyeonggi Province, South Korea. The strains $ON22^T$, $G10^T$, and $G15^T$, showed positive cellulolytic activity in the carboxymethyl cellulose (CMC)-plate assay. The phylogenetic tree based on sequence analysis of D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer (ITS) regions revealed that the strains $ON22^T$ (100 and 98.4% sequence similarities in D1/D2 domains and ITS) and $G10^T$ (99.8 and 99.5% in D1/D2 domain and ITS region) were most closely related to the species Moesziomyces aphidis JCM $10318^T$; $G15^T$ (100% in D1/D2 domains and ITS) belongs to the species Moesziomyces antarcticus JCM $10317^T$, respectively. Morphology and biochemical test results are provided in the species description. Cellulase with its massive applicability has been used in various industrial processes such as biofuels like bioethanol productions. Therefore, this is the first report of the cellulolytic yeast strains $ON22^T$, $G10^T$, and $G15^T$ related to the genus Moesziomyces in the family Ustilaginaceae (Ustilaginales), in Korea.

Pseudomonas tolaasii bacteriophage-specific polyclonal antibody formation and its cross reactivity to various phages (Pseudomonas tolaasii 박테리오파지에 특이적인 다클론항체 형성 및 이를 이용한 파지 교차 반응성)

  • Yun, Yeong-Bae;Park, Soo-Jin;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.62 no.3
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    • pp.287-292
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    • 2019
  • Pseudomonas tolaasii causes brown blotch disease on the oyster mushroom (Pleurotus ostreatus). Various pathogenic strains of P. tolaasii were isolated and divided into three subtypes, $P1{\alpha}$, $P1{\beta}$, and $P1{\gamma}$. For phage therapy, bacteriophages against to these subtype strains were applied to mushroom cultivation and very successful to prevent from the disease. In this study, bacteriophages were isolated against the representative strains of subtype pathogens and their polyclonal antibodies were synthesized to investigate structural relationship among capsid proteins of phages. Phage preparations over $10^{10}pfu/mL$ were injected to rabbit thigh muscle and polyclonal antibodies were obtained after three times of boost injection. Titers of the antibodies obtained were over $2{\times}10^7Ab/mL$ for the phage ${\phi}6264$, $1{\times}10^6Ab/mL$ for the phage ${\phi}HK2$, and $1{\times}10^7Ab/mL$ for the phage ${\phi}HK19$ and phage ${\phi}HK23$. High specific activities were observed between antibodies and the corresponding bacteriophages. Some cross-reactivities between the antibodies and non-corresponding bacteriophages were also measured. Antibody $Ab{\phi}6264$ inactivated all phages of $P1{\alpha}$ subtype and only phage ${\phi}HK16$ among $P1{\beta}$ subtype phages. Antibody $Ab{\phi}HK23$ of $P1{\gamma}$ subtype neutralized all phages of $P1{\beta}$ subtype as well as the phage ${\phi}HK23$, showing the widest phage-inactivation range. When the structural-similarity studies of phages were investigated by using phage antibodies, closeness obtained by phylogenetic analysis of 16S rRNA genes of pathogenic strains were quite different from that of polyclonal antibody-specific structural similarity of phage capsid proteins. In conclusion, there is weak correlation between the host strain specificity of bacteriophage and its capsid structural similarity measured by phage antibodies.

First Report of Impatiens Necrotic Spot Virus on Plantago asiatica Cultivated in Open Fields (노지재배 질경이(Plantago asiatica)에서 봉선화괴저반점바이러스병 발생 국내 첫 보고)

  • Chung, Bong Nam;An, Tae Jin;Cho, In-Sook;Yoon, Ju-Yeon
    • Research in Plant Disease
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    • v.27 no.1
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    • pp.1-7
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    • 2021
  • In August 2020, necrotic ringspots on leaves were observed on 20 from 143 Plantago asiatica plants in open fields in Eumseong, Chungcheongbuk-do. Eight symptomatic Plantago asiatica plants were subjected to investigation on viral infection by reverse transcription and polymerase chain reaction. Impatiens necrotic spot virus, tomato spotted wilt virus and cucumber mosaic virus were detected from the symptomatic plants. Two impatiens necrotic spot virus (INSV) isolates ('INSV-plantain kr1' and '-plantain kr5') were sequenced and analyzed by comparing L genomic segment, nucleoprotein (N) gene and non-structural protein S (NSs) gene sequences. The nucleotide sequence of 'INSV-plantain kr1' isolate (MW114834) was most closely related to that of a 'Phalaenopsis' isolate (GQ336991) from China in the L genomic segment. 'INSV-plantain kr1' and '-plantain kr5' isolates shared the highest identities with those from 'Pepe' isolate (LC384872) and 'J' isolate (AB109100) in the NSs gene, respectively, and with that from 'YSMi-SH' isolate (FN400773) in the N gene. Phylogenetic analysis based on L genomic segment grouped the INSV-plantain kr1 isolate together with isolates from Korea (LC384870), China (GU112505, GQ336991), and Italy (DQ425094). This is the first report on INSV in P. asiatica from Korea.

Characteristics of Probiotics Isolated from Korean Traditional Foods and Antibacterial Activity of Synbiotics (한국전통발효식품에서 분리한 Probiotics의 특징 및 Synbiotics 항균활성 효과)

  • Moon, Chae-Yun;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.552-558
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    • 2021
  • Traditional foods are manufactured according to the characteristics of each region for the nations of the world. Korea has mainly farmed, and seasonings have developed around rice and vegetables. In fermented foods, lactic acid bacteria such as Lactobacillus sp. and Pediococcus sp. and Bacillus sp. were isolated and identified from fermented foods. In this study, lactic acid bacteria were isolated and identified from commercially available traditional Korean fermented foods, and candidate strains were selected through antibacterial activity tests on human and fish disease bacteria. Thereafter, the final strain was selected by examining the resistance to simulated gastric and intestinal fluids, and hemolysis. The three (M1, K1, C13) final selected latic acid bacteria were miced with prebiotics and the antibacterial activity of synbiotics was evaluated. As for the fist antibacterial activity result, C13 showed high antibacterial acitivity in human diseases and fish diseases. Then, M1, K1 and C13, which did not produce β-haemolysis and were resistant to simulated gastric and intestinal fluids, were subjected to the second antibacterial activity of synbiotics. When the three prebiotics (FOS, GOS, Inulin) and probiotics (M1, K1, C13) were mixed, the antibacterial activity was increased or inhibited. Based on the 16S rRNA gene sequencing results, K1 and M1 were analyzed as Bacillus tequiensis 99.72%, Bacillus subtilis 99.65%, Bacillus inaquosorum 99.72%, Bacillus cabrialesii 99.72%, Bacillus stercoris 99.58%, Bacillus spizizenii 99.58%, Bacillus halotolerans 99.58%, and Bacillus mojavensis 99.51%. And C13 was analyzed as Bacillus velezensis 99.71%, Bacillus nematocida 99.36%, Bacillus amyloliquefaciens 99.44%, Bacillus atrophaeus 99.22%, and Bacillus nakamurai 99.44%.