• Title/Summary/Keyword: cyanobacterial

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Evaluation of Photosynthetic Squalene Production of Engineered Cyanobacteria Using the Chemical Inducer-Free Expression System (무-유도인자 단백질 발현 시스템을 이용한 재조합 시아노박테리아의 광합성 스쿠알렌 생산 평가)

  • Choi, Sun Young;Woo, Han Min
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.298-304
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    • 2021
  • Photosynthetic conversion through cyanobacteria and microalgae is an increasingly serious concern in the global warming crisis. Many value-added substances are produced through strain improvement, and much research and development is being conducted to determine its potential as an actual industrial strain. Economic barriers throughout processing production can be overcome to produce value-added chemicals by microalgal strains. In this study, we engineered cyanobacteria strains for the photosynthetic production of squalene and confirmed the continuous cultivation of CO2 and light conditions. The free-inducer system of gene expression was developed at the cyanobacterial strains. Then, the squalene production level and growth of the recombinant cyanobacteria were analyzed and discussed. For bio solar-cell factories, the ability to regulate genes based on the free-inducer gene expression system promotes metabolic engineering research and construction to produce value-added chemicals.

Discovery of novel Nodosilinea species (Cyanobacteria, Nodosilineales) isolated from terrestrial habitat in Ryukyus campus, Okinawa, Japan

  • Handung Nuryadi;Shimpei Sumimoto;Shoichiro Suda
    • ALGAE
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    • v.39 no.2
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    • pp.59-74
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    • 2024
  • Terrestrial cyanobacteria are extremely diverse. In urban areas, they can be found as black stains on the surface of building walls, stone monuments, or man-made structures. Many of the terrestrial cyanobacteria are still understudied. To expand knowledge of terrestrial cyanobacterial diversity, a polyphasic characterization was performed to identify 12 strains isolated from campus of University of the Ryukyus, Okinawa, Japan. Multigene phylogenetic analyses based on 16S rRNA gene and 16S-23S rRNA internal transcribed spacer (ITS) region showed that the isolated strains formed two independent subclades within Nodosilinea, and were distantly related to all described Nodosilinea species. The 16S-23S rRNA ITS secondary structures showed variations for D1-D1' and Box B domain, while V3 domain was almost identical among entire species of Nodosilinea, including the studied strains. In addition, a unique morphological character, i.e. forming nodule or spiral shape, was also observed in certain studied strains. According to polyphasic characterization, Nodosilinea coculeatus sp. nov. and Nodosilinea terrestrialis sp. nov., were proposed as two new species of terrestrial cyanobacteria from Okinawa.

Changes of the Environmental Factors in Upo Wetland (우포늪의 수환경요인 변화)

  • Lee, Jung-Joon;Lee, Jung-Ho
    • Korean Journal of Environmental Biology
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    • v.27 no.3
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    • pp.306-313
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    • 2009
  • In the Upo wetland, physico-chemical factors were observed during the period from March 2005 to December 2007 on a monthly basis. In the Upo wetland, water temperatures ranged $3.4{\sim}34.5^{\circ}C$. Conductivities were in the range of 133~806 ${\mu}S\;cm^{-1}$, which showed about 140 ${\mu}S\;cm^{-1}$ below in comparison with the precedent studies. The pH levels were between 6.7~9.1 with lower level in summertime. The dissolved oxygens were between 0.06~18.23 mg $L^{-1}$. COD ranged 4.9~20.8 mg $L^{-1}$, and showed a tendency to decrease every year. Nitrogen nutrients such as nitrate nitrogen ($NO_3-N$), ammonia nitrogen ($NH_3-N$) and total nitrogen (T-P) showed that they were generally decreased in comparison with those in the precedent studies. However the total nitrogen (T-N) is still considerably higher than the standard concentration level of eutrophication and algal blooming. Phosphate phosphorus ($PO_4-P$) and total phosphorus (T-P) were also shown as to be reduced considerably comparing with the values in the precedent studies. However, It was found out that total phosphorus (T-P) was dissolved over the criteria concentration of eutrophication. The average of TN/TP ratio was 18 in the Upo wetland, which proved that phosphorus was the limiting factor to the growth of phytoplankton in the Upo wetland. The chl-$\alpha$ was the highest in wintertime and the lowest in summertime, and especially in 2006 summer when the cyanobacterial bloom developed, it showed extremely high concentration.

A Novel Microcystin-degrading Bacterium, Microbacterium sp. MA21 (Microcystin을 분해하는 신균주 Microbacterium sp. MA21)

  • Ko, So-Ra;Lee, Young-Ki;Oh, Hee-Mock;Ahn, Chi-Yong
    • Korean Journal of Environmental Biology
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    • v.31 no.2
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    • pp.158-164
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    • 2013
  • A microcystin-degrading bacterium was isolated from Daechung reservoir, Korea. The isolated bacterium was identified as Microbacterium sp. by 16S rRNA gene sequence analysis, and designated as Microbacterium sp. MA21. This strain degraded cyanobacterial hepatotoxin, microcystin-LR, over 80% when incubated at $30^{\circ}C$ for 12 hr in R2A medium. Two unknown metabolites of microcystin were also identified during the degradation process. Although only Sphinogomonas and Actinobacteria have been known to degrade microcystin previously, this is the first report that Microbacterium sp. MA21 could degrade microcystin.

Molecular Classification of Commercial Spirulina Strains and Identification of Their Sulfolipid Biosynthesis Genes

  • Kwei, Chee Kuan;Lewis, David;King, Keith;Donohue, William;Neilan, Brett A.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.359-365
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    • 2011
  • Cyanobacterial strains of the genus Spirulina have recently been identified as an excellent source of sulfolipids, some of which possess anti-HIV properties. Thus, to investigate the distribution of sufolipid biosynthesis pathways in Spirulina, a genetic screening/phylogentic study was performed. Five different strains of Spirulina [Spirulina (Jiangmen), Spirulina sp., S. platensis, S. maxima, and Spirulina seawater] sourced from different locations were initially classified via 16S rDNA sequencing, and then screened for the presence of the sulfolipid biosynthesis genes sqdB and sqdX via a PCR. To assess the suitability of these strains for human consumption and safe therapeutic use, the strains were also screened for the presence of genes encoding nonribosomal peptide synthases (NRPSs) and polyketide synthases (PKSs), which are often associated with toxin pathways in cyanobacteria. The results of the 16S rDNA analysis and phylogenetic study indicated that Spirulina sp. is closely related to Halospirulina, whereas the other four Spirulina strains are closely related to Arthrospira. Homologs of sqdB and sqdX were identified in Spirulina (Jiangmen), Spirulina sp., S. platensis, and the Spirulina seawater. None of the Spirulina strains screened in this study tested positive for NRPS or PKS genes, suggesting that these strains do not produce NRP or PK toxins.

The Phylogenetic Relationship of Several Oscillatorian Cyanobacteria, Forming Blooms at Daecheong Reservoirs, Based on Partial 16S rRNA Gene Sequences

  • Lee, Wook-Jae;Bae, Kyung-Sook
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.504-507
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    • 2001
  • The partial 16S gene sequences of six filamentous cyanobacterial strains, Oscillatoria lmosa KCTC AG10168, Oscillatoria princeps KCTC AG10153, Oscillatoria sp. KCTC AG 10184, Phormidium tenue KCTC AG10158, Phormidium parchydematicum KCTC AG10164, and Lyngbya hieronymusii KCTC AG10199, which were isolated in the late summer at Daecheong Reservoirs, were determined and assigned their phylogenetic and taxonomic position among taxa of order Ocillatoriales whose partial 16S rRNA gene sequences aligned in this suty, were very heterogeneously clustered with other taxa. The two strains, Oscillatoria limosa KCTC AG10168 and O. princeps KCTC AG10153, formed a cluster with O. sancta PCC7515, which supported 64% of the bootstrap trees with high similarity (19-96.15%). Strain Oscillatoria sp. KCTC AG10184, that was known to produce a nasty substance, was closely related to the toxic Oscillatoria group. The study on morphological variation in various environments and toxin production will confirm the taxonomic status of these species. Phormidium tenus KCTC AG10158 and Phormidium parchydematicum KCTC AG10164 made a cluster with other oscillatorian species of Phormidium, Oscillatoria, and Leptolynbya, which supproted 100% of the bootstrap trees with a very high sequence smilarity (96.8-99.8%) in thsi study. The sequence analysis in this study also supported that taxa of oscillatoriales are not monophyletic. Some of the fractures, such as the presence or absence of sheath and cell shape, which were used to define them, would be inadequate and should be reconfirmed. We suggest that sequences of partial 16S rRNA gene fragments aligned in this study should be more useful than morphological features in the identification and reconfirmation of the taxonomic status of these oscillactorian cyanobacteria.

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Green Synthesis of Silver Nanoparticles Using Cell Extracts of Anabaena doliolum and Screening of Its Antibacterial and Antitumor Activity

  • Singh, Garvita;Babele, Piyoosh K.;Shahi, Shailesh K.;Sinha, Rajeshwar P.;Tyagi, Madhu B.;Kumar, Ashok
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1354-1367
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    • 2014
  • In the present work, we describe a simple, cheap, and unexplored method for "green" synthesis of silver nanoparticles using cell extracts of the cyanobacterium Anabaena doliolum. An attempt was also made to test the antimicrobial and antitumor activities of the synthesized nanoparticles. Analytical techniques, namely UV-vis spectroscopy, X-ray diffraction, Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM), and TEM-selected area electron diffraction, were used to elucidate the formation and characterization of silver-cyanobacterial nanoparticles (Ag-CNPs). Results showed that the original color of the cell extract changed from reddish blue to dark brown after addition of silver nitrate solution (1 mM) within 1 h, suggesting the synthesis of Ag-CNPs. That the formation Ag-CNPs indeed occurred was also evident from the spectroscopic analysis of the reaction mixture, wherein a prominent peak at 420 nm was noted. TEM images revealed well-dispersed, spherical Ag-CNPs with a particle size in the range of 10-50 nm. The X-ray diffraction spectrum suggested a crystalline nature of the Ag-CNPs. FTIR analysis indicated the utilization of a hydroxyl (-OH) group in the formation of Ag-CNPs. Ag-CNPs exhibited strong antibacterial activity against three multidrug-resistant bacteria. Additionally, Ag-CNPs strongly affected the survival of Dalton's lymphoma and human carcinoma colo205 cells at a very low concentration. The Ag-CNPs-induced loss of survival of both cell types may be due to the induction of reactive oxygen species generation and DNA fragmentation, resulting in apoptosis. Properties exhibited by the Ag-CNP suggest that it may be used as a potential antibacterial and antitumor agent.

Hydroxylation of Compactin (ML-236B) by CYP105D7 (SAV_7469) from Streptomyces avermitilis

  • Yao, Qiuping;Ma, Li;Liu, Ling;Ikeda, Haruo;Fushinobu, Shinya;Li, Shengying;Xu, Lian-Hua
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.956-964
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    • 2017
  • Compactin and pravastatin are competitive cholesterol biosynthesis inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase and belong to the statin drugs; however, the latter shows superior pharmacokinetic characteristics. Previously, we reported that the bacterial P450, CYP105D7, from Streptomyces avermitilis can catalyze the hydroxylation of 1-deoxypentalenic acid, diclofenac, and naringenin. Here, we demonstrate that CYP105D7 could also catalyze compactin hydroxylation in vitro. In the presence of both bacterial and cyanobacterial redox partner systems with an NADPH regeneration system, the reaction produced two hydroxylated products, including pravastatin (hydroxylated at the C6 position). The steady-state kinetic parameters were measured using the redox partners of putidaredoxin and its reductase. The $k_m$ and $k_{cat}$ values for compactin were $39.1{\pm}8.8{\mu}M$ and $1.12{\pm}0.09min^{-1}$, respectively. The $k_{cat}/K_m$ value for compactin ($0.029min^{-1}{\cdot}{\mu}M^{-1}$) was lower than that for diclofenac ($0.114min^{-1}{\cdot}{\mu}M^{-1}$). Spectroscopic analysis showed that CYP105D7 binds to compactin with a $K_d$ value of $17.5{\pm}3.6{\mu}M$. Molecular docking analysis was performed to build a possible binding model of compactin. Comparisons of different substrates with CYP105D7 were conclusively illustrated for the first time.

Application of Quantum-dot Nanocrystals for Cyanobacterial Toxin-Microcystin Detection (나노크리스탈 Quantum-dot을 적용한 남조류 독소 Microcystin 탐지 연구)

  • Lee, Jinwook;Yu, Hye-Weon;Kim, In S.
    • Journal of Korean Society on Water Environment
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    • v.23 no.5
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    • pp.705-711
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    • 2007
  • Green quantum-dot nanocrystal (QD525) with anti-microcystin monoclonal antibody was applied for detection of microcystin, a monocyclic peptide hepatotoxin, extracted from the culture of Microcystis aeruginosa. The presence of microcystin in the cell lysate was verified by HPLC analysis with UV absorbance at 238 nm. Microcystis cell extract exhibited fluorescence emission spectra, which peak was around 460 nm because of their complex organic substances. When a spherical QD525 antibody conjugates (10~20 nm in diameter) were bound to the microcystins in the Microcystis cell lysate, the fluorescence intensity of the primary peak at 525 nm diminished while the secondary emission peak at 460 nm slightly increased intensities. It is due to energy transfer from the primary (major) to the secondary (minor) peak, resulting from physical deformation of QD525 and different environmental factors. On the other hand, other cell extracts did not show any fluorescence emission change. This study is very available for detecting and monitoring the microcystin because it is one step assay without washing step and portable spectrophotometer makes on-site measurement possible. For health risk assessment of the microcystin, the reliable and rapid system to detect and quantify microcystin is seriously required.

Occurrence and Succession Pattern of Cyanobacteria in the Upper Region of the Nakdong River : Factors Influencing Aphanizomenon Bloom (낙동강 상류 수역에서 남조류 발생과 천이패턴 - Aphanizomenon 속을 중심으로 -)

  • Ryu, Hui-Seong;Park, Hae-Kyung;Lee, Hae-Jin;Shin, Ra-Young;Cheon, Se-Uk
    • Journal of Korean Society on Water Environment
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    • v.32 no.1
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    • pp.52-59
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    • 2016
  • This study investigated the occurrences and succession patterns of harmful cyanobacteria, as well as environmental factors, during a 3-year period (September 2012 to August 2015) in the upper region of the Nakdong River around Sangju weir. A total of 27 cyanobacterial taxa were observed in this study, and classified into 26 species and 1 variety belonging to 11 genera, 5 families, and 3 orders. Cell density ranged from 24 to 42,001 cells/ml, with a geometric mean of 33 cells/ml, during the survey period. The dominant orders differed depending on the survey year; order Oscillatoriales in 2013, Chroococcales in 2014 and Nostocales in 2015. An Aphanizomenon bloom occurred in June 2015 at which time the highest cell density of 36,873 cells/ml was detected in the upper region of the Nakdong River, where as the Aphanizomenon spp. cell density (190-1,704 cells/ml) had been low prior to that time. An Aphanizomenon bloom also occurred at around the same time downstream in the Young River, a major inflow branch of the Nakdong River. The Aphanizomenon cell density along the Nakdong River increased markedly after joining of the YoungRiver, indicating that the Aphanizomenon bloom in the YoungRiver caused a bloom in the Nakdong River. Meteorological and environmental parameters, such as very low precipitation, higher water temperature, pH, and TP concentration, and lower TN/TP ratio, in May and June of 2015 than in 2013 and 2014 exerted marked effects on the Aphanizomenon bloom in June 2015 in the Young River.