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1.
J Hazard Mater ; 471: 134352, 2024 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-38677120

RESUMEN

Microcystis typically forms colonies under natural conditions, which contributes to occurrence and prevalence of algal blooms. The colonies consist of Microcystis and associated bacteria (AB), embedded in extracellular polymeric substances (EPS). Previous studies indicate that AB can induce Microcystis to form colonies, however the efficiency is generally low and results in a uniform morphotype. In this study, by using filtrated natural water, several AB strains induced unicellular M. aeruginosa to form colonies resembling several Microcystis morphotypes. The mechanisms were investigated with Methylobacterium sp. Z5. Ca2+ was necessary for Z5 to induce Microcystis to form colonies, while dissolved organic matters (DOM) facilitated AB to agglomerate Microcystis to form large colonies. EPS of living Z5, mainly the aromatic protein components, played a key role in colony induction. Z5 initially aggregated Microcystis via the bridging effects of Ca2+ and DOM, followed by the induction of EPS synthesis and secretion in Microcystis. In this process, the colony forming mode shifted from cell adhesion to a combination of cell adhesion and cell division. Intriguingly, Z5 drove the genomic rearrangement of Microcystis by upregulating some transposase genes. This study unveiled a novel mechanism about Microcystis colony formation and identified a new driver of Microcystis genomic evolution.


Asunto(s)
Calcio , Matriz Extracelular de Sustancias Poliméricas , Microcystis , Microcystis/metabolismo , Calcio/metabolismo , Matriz Extracelular de Sustancias Poliméricas/metabolismo , Methylobacterium/metabolismo , Methylobacterium/genética
2.
J Appl Microbiol ; 135(1)2024 Jan 02.
Artículo en Inglés | MEDLINE | ID: mdl-38061837

RESUMEN

AIMS: This study explores the phosphate (Pi)-solubilizing characteristics and mechanisms of a novel phosphate-solubilizing bacterium, Agrobacterium deltaense C1 (C1 hereafter). METHODS AND RESULTS: The growth-promoting effects of C1 were investigated by gnotobiotic experiments, and the Pi-solubilizing mechanism was revealed by extracellular metabolomics, liquid chromatography analysis, and reverse transcription quantitative polymerase chain reaction. Results showed that C1 significantly increased Arabidopsis biomass and total phosphorus (P) content under P deficiency. Under Ca3(PO4)2 condition, the presence of C1 resulted in a significant and negative correlation between available P content and medium pH changes, implying that Pi dissolution occurs through acid release. Metabolomics revealed C1's ability to release 99 organic acids, with gluconic acid (GA), citric acid, and α-ketoglutaric acid contributing 64.86%, 9.58%, and 0.94%, respectively, to Pi solubilization. These acids were significantly induced by P deficiency. Moreover, C1's Pi solubilization may remain significant even in the presence of available P, as evidenced by substantial pH reduction and high gcd gene expression. Additionally, C1 produced over 10 plant growth-promoting substances. CONCLUSIONS: C1 dissolves Pi primarily by releasing GA, which enhances plant growth under P deficiency. Notably, its Pi solubilization effect is not significantly limited by available Pi.


Asunto(s)
Fosfatos , Microbiología del Suelo , Fosfatos/metabolismo , Fósforo/metabolismo , Agrobacterium/genética , Agrobacterium/metabolismo , Bacterias/genética
3.
Chemosphere ; 310: 136767, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-36241112

RESUMEN

Cyanobacterial blooms negatively affect aquatic ecosystems and human health. Algicidal bacteria can efficiently kill bloom-causing cyanobacteria. Bacillus altitudinis G3 isolated from Dianchi Lake shows high algicidal activity against Microcystis aeruginosa. In this study, we investigated its algicidal characteristics including attack mode, photosynthesis responses, and source and the contribution of reactive oxygen species (ROS). The results showed that G3 efficiently and specifically killed M. aeruginosa mainly by releasing both thermolabile and thermostable algicidal substances, which exhibited the highest algicidal activity (99.8%, 72 h) in bacterial mid-logarithmic growth phase. The algicidal ratio under full-light conditions (99.5%, 60 h) was significantly higher than under dark conditions (<20%, P < 0.001). G3 filtrate caused photosystem dysfunction by decreasing photosynthetic efficiency, as indicated by significantly decreased Fv/Fm and PIABS (P < 0.001) values. It also inhibited photosynthetic electron transfer as indicated by significantly decreased rETR (P < 0.001), especially QA- downstream, as revealed by significantly decreased φEo and ψo, and increased Mo (P < 0.001). These results indicated that the algicidal activity of G3 filtrate is light-dependent, and the cyanobacterial photosystem is an important target. Cyanobacterial ROS and malondialdehyde contents greatly increased by 37.1% and 208% at 36 h, respectively. ROS levels decreased by 49.2% (9 h) when diuron (3-(3-4-dichlorophenyl)-1,1-dimethylurea) partially blocked photosynthetic electron transport from QA to QB. Therefore, excessive ROS were produced from disrupted photosynthesis, especially the inhibited electron transport area in QA- downstream, and caused severe lipid peroxidation with significantly increased MDA content and oxidative stress in cyanobacteria. The ROS scavenger N-acetyl-l-cysteine significantly decreased both cyanobacterial ROS levels (34%) and algicidal ratio (52%, P < 0.05) at 39 h. Thus, excessive ROS production due to G3 filtrate administration significantly contributed to its algicidal effect. G3 could be an excellent algicide to control M. aeruginosa blooms in waters under suitable light conditions.


Asunto(s)
Bacillus , Microcystis , Humanos , Especies Reactivas de Oxígeno/farmacología , Ecosistema , Floraciones de Algas Nocivas
4.
Sci Total Environ ; 806(Pt 4): 150719, 2022 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-34606873

RESUMEN

Cyanobacterial blooms significantly decrease water quality and can damage ecosystems and, as such, require efficient control methods. Algicidal bacteria and their associated substances are promising tools for controlling cyanobacterial blooms; however, their specific algicidal mechanisms remain unclear. Therefore, the current study sought to investigate the algicidal mechanism of tryptoline (1,2,3,4-tetrahydro-9 h-pyrido[3,4-b]indole) against Microcystis aeruginosa, with a specific focus on the contribution made by reactive oxygen species (ROS), the underlying mechanisms of ROS increase, as well as the photosystem response. Results show that the algicidal ratio of tryptoline significantly and positively correlates with algal ROS. Moreover, 93.79% of the algicidal ratio variation is attributed to ROS in the tryptoline group, while only 47.75% can be attributed to ROS in the tryptoline + N-acetyl-L-cysteine (NAC) group, where ROS are partially scavenged by NAC. In the presence of tryptoline, algicidal effect and ROS levels were significantly enhanced in the presence of light as compared to those in the dark (P < 0.001). Hence, the increase in ROS production attributed to tryptoline is primarily affected by the presence of light and photosynthesis. Additionally, tryptoline significantly reduces Fv/Fm, PIABS, ETo/RC, and the expression of psaB and psbA genes related to photosynthesis, while increasing Vj and DIo/RC (P < 0.05). These results suggest that tryptoline hinders algal photosynthesis by significantly decreasing photosynthetic efficiency and carbon assimilation, inhibiting photochemical electron transfer, and increasing closed reaction centers and energy loss. Moreover, following partial blockade of the photosynthetic electron transfer from QA to QB by diuron (3-(3-4-dichlorophenyl)-1,1-dimethylurea), the ROS of algae exposed to tryptoline is significantly decreased. Thus, tryptoline inhibits electron transfer downstream of QA, which increase the number of escaping electron and thereby increase ROS generation. Collectively, this study describes the algicidal mechanism of tryptoline against M. aeruginosa and highlights the critical factors associated with induction of algicidal activity.


Asunto(s)
Microcystis , Carbolinas , Ecosistema , Floraciones de Algas Nocivas , Fotosíntesis , Especies Reactivas de Oxígeno
5.
Sci Total Environ ; 769: 144489, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33465632

RESUMEN

Cyanobacterial harmful algal blooms (CyanoHABs) bring economic loss, damage aquatic ecosystems, and produce cyanobacterial toxins that threaten human health. Algicidal bacteria as pathogens can expediate the decline of CyanoHABs. In this study, a Streptomyces globisporus strain (designated G9), isolated from soil near a eutrophic pond, showed high algicidal activity against Microcystis aeruginosa. Experimental results show that G9 preyed on Microcystis through cell-to-cell contact: (1) the hyphae of G9 killed cyanobacterial cells by twining around them, while cells beyond the reach of G9 hyphae were in normal shapes; (2) No algicides were detectable in the supernatant of G9 cultures or G9-Microcystis cocultures. The algicidal ratio of G9 to M. aeruginosa reached 96.7% after 6 days. G9 selectively killed the tested cyanobacterial strains, while it had only minor impacts on the growth of tested Chlorophyceae. Differential gene expression studies show that G9 affected the expression of key genes of M. aeruginosa involved in photosynthesis, microcystin synthesis and cellular emergency responses. Further, the microcystin-LR content decreased gradually with G9 treatment. As the first reported Streptomyces sp. with algicidal (predation) activity requiring cell-to-cell contact with target prey, G9 is a good candidate for the exploration of additional cyanobacteria-bacteria interactions and the development of novel strategies to control CyanoHABs.


Asunto(s)
Microcystis , Streptomyces , Ecosistema , Floraciones de Algas Nocivas , Humanos , Microcistinas
6.
World J Microbiol Biotechnol ; 36(12): 188, 2020 Nov 26.
Artículo en Inglés | MEDLINE | ID: mdl-33241509

RESUMEN

Cyanobacterial blooms are a worldwide problem, especially in freshwaters. As one of the most abundant co-existing organisms of algae, bacteria play critical roles in cyanobacteria growth, particularly the cyanobactericidal bacteria which can efficiently kill cyanobacteria. Recent years, cyanobactericidal bacteria are highly recognized as a method that could potentially block cyanobacterial blooms. Many studies have been conducted to assess their effects on the termination of cyanobacteria blooms and explore their cyanobactericidal mechanisms, e.g., attacking by cell to cell or releasing specific compounds, the physiological, metabolic, and transcriptional disturbance on cyanobacteria. In this review, the present state of research on cyanobactericidal bacteria for the bloom-causing cyanobacteria species is summarized. The challenges in applying cyanobactericidal bacteria in the control of natural cyanobacterial blooms are discussed.


Asunto(s)
Agentes de Control Biológico , Cianobacterias/efectos de los fármacos , Cianobacterias/fisiología , Eutrofización , Agua Dulce/microbiología , Interacciones Microbianas
7.
Chemosphere ; 249: 126097, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32078851

RESUMEN

Benz(a)anthracene (BaA) is a polycyclic aromatic hydrocarbons (PAHs), that belongs to a group of carcinogenic and mutagenic persistent organic pollutants found in a variety of ecological habitats. In this study, the efficient biodegradation of BaA by a green alga Chlamydomonas reinhardtii (C. reinhardtii) CC-503 was investigated. The results showed that the growth of C. reinhardtii was hardly affected with an initial concentration of 10 mg/L, but was inhibited significantly under higher concentrations of BaA (>30 mg/L) (p < 0.05). We demonstrated that the relatively high concentration of 10 mg/L BaA was degraded completely in 11 days, which indicated that C. reinhardtii had an efficient degradation system. During the degradation, the intermediate metabolites were determined to be isomeric phenanthrene or anthracene, 2,6-diisopropylnaphthalene, 1,3-diisopropylnaphthalene, 1,7-diisopropylnaphthalene, and cyclohexanol. The enzymes involved in the degradation included the homogentisate 1,2-dioxygenase (HGD), the carboxymethylenebutenolidase, the ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and the ubiquinol oxidase. The respective genes encoding these proteins were significantly up-regulated ranging from 3.17 fold to 13.03 fold and the activity of enzymes, such as HGD and Rubisco, was significantly induced up to 4.53 and 1.46 fold (p < 0.05), during the BaA metabolism. This efficient degradation ability suggests that the green alga C. reinhardtii CC-503 may be a sustainable candidate for PAHs remediation.


Asunto(s)
Antracenos/metabolismo , Biodegradación Ambiental , Chlamydomonas reinhardtii/metabolismo , Contaminantes Ambientales/metabolismo , Benzo(a)Antracenos/metabolismo , Carcinógenos/metabolismo , Dioxigenasas/metabolismo , Fenantrenos , Hidrocarburos Policíclicos Aromáticos/metabolismo
8.
J Basic Microbiol ; 59(11): 1112-1124, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31502316

RESUMEN

Microcystis aeruginosa blooms are a worldwide serious environmental problem and bloom control with bacteria is promising. In this study, a Bacillus licheniformis strain Sp34 with potent algicidal and inhibitory effects on the microcystins synthesis against fast-growing M. aeruginosa was isolated from Dianchi Lake. Sp34 killed the bloom-causing algal strain M. aeruginosa DCM4 of Dianchi Lake with an initial Chlorophyll-a concentration of 2.0 mg/L at a cell density of no less than 1.35 × 105 CFU/ml. It can also efficiently kill some other harmful algal species, such as M. wesenbergii and Phormidium sp. The algicidal activity of Sp34 relied on the release of algicidal substances, which had good heat (-20°C to 121°C) and acid-base (pH 3-11) resistance. In addition, the high algicidal activity depended on the good growth of algae indicated by the significantly positive correlations between algal growth and algicidal ratio (p < .001). The algicidal effect of Sp34 involved causing oxidative stress, lipid peroxidation, and morphological injury of algal cells, along with DNA damage and dysfunction of DNA-repair function, weakening the photosynthesis system, and inhibiting microcystin synthesis. In general, Sp34 can kill fast-growing M. aeruginosa and inhibit algal microcystin synthesis efficiently, so, it is a promising biocontrol agent to mitigate cyanobacterial blooms.


Asunto(s)
Bacillus licheniformis/metabolismo , Agentes de Control Biológico/metabolismo , Agentes de Control Biológico/farmacología , Microcystis/efectos de los fármacos , Antibiosis , Bacillus licheniformis/clasificación , Bacillus licheniformis/genética , Bacillus licheniformis/crecimiento & desarrollo , Clorofila/análogos & derivados , Clorofila/biosíntesis , Clorofila/genética , Eutrofización/efectos de los fármacos , Lagos/microbiología , Microcistinas/biosíntesis , Microcistinas/genética , Microcystis/genética , Microcystis/crecimiento & desarrollo , Microcystis/metabolismo , Estrés Oxidativo/efectos de los fármacos , Transcripción Genética/efectos de los fármacos
9.
Sci Total Environ ; 650(Pt 1): 34-43, 2019 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-30195130

RESUMEN

Microcystis aeruginosa and hepatotoxic microcystins produced by it have posed a severe threat to aquatic ecological security and human health. In this study a Streptomyces amritsarensis HG-16, showing high algicidal activity against M. aeruginosa and strong inhibitory effect on microcystin synthesis, was obtained by screening some anti-Fusarium sp. microbial strains isolated before in our laboratory. HG-16 bound cyanobacterial cells by mycelia to form flocs and killed M. aeruginosa by secreting active substances, which were proteinase K resistant and stable in the temperature range of 35-75 °C and pH range of 3-11. HG-16 removed M. aeruginosa of 105 and 106 cell mL-1 cell densities in similar rate and was active against all the tested harmful unicellular and filamentous cyanobacteria. Results of differential gene expression analysis indicated that HG-16 affected the photosynthesis system and microcystin synthesis of M. aeruginosa. Accordingly, the algicidal activity of HG-16 was light-dependent, and microcystin synthesis of M. aeruginosa decreased by 91.2% with HG-16 treatment. Thus, it is promising to utilize HG-16 to mitigate harmful cyanobacterial blooms, inhibit microcystin synthesis and control plant disease caused by Fusarium.spp. through irrigating farmland with eutrophic water applied HG-16.


Asunto(s)
Herbicidas/farmacología , Microcistinas/biosíntesis , Microcystis/efectos de los fármacos , Streptomyces/química
10.
J Basic Microbiol ; 57(11): 941-949, 2017 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-28833312

RESUMEN

A Comamonas testosterone bacterial strain, named as DB-7, capable of utilizing dimethyl phthalate (DMP) as sole carbon source and energy for growth was isolated from soil with plastic film mulching by an enrichment culture technique. This bacterium was identified as C. testosterone by 16S rRNA sequence analysis and phospholipid fatty acid profile. DB-7 could degrade more than 99% of 450 mg L-1 DMP within 14 hours, and degraded DMP of different concentrations rapidly. The optimal degradation temperature and pH were 30-35 °C and pH 9.0, respectively. The degradation rate of DMP was positively related to inoculum volume of the bacterium. The result of HPLC and LC/MS analysis of metabolic products indicated that the major degrading intermediates were mono-methyl phthalate (MMP) and phthalic acid (PA) during the degradation of DMP by DB-7. Partial sequences of three genes involved in PA metabolism were detected in DB-7, and the expression of phthalate 4, 5-dioxygenase was drastically induced in the presence of DMP and PA. DB-7 is promising to be applied to DMP bioremediation because of its high degrading efficiency.


Asunto(s)
Biodegradación Ambiental , Comamonas testosteroni/aislamiento & purificación , Comamonas testosteroni/metabolismo , Ácidos Ftálicos/metabolismo , Microbiología del Suelo , Comamonas , Comamonas testosteroni/genética , Comamonas testosteroni/crecimiento & desarrollo , Ácidos Grasos , Genes Bacterianos/genética , Concentración de Iones de Hidrógeno , Metabolismo/genética , Oxigenasas/genética , Oxigenasas/metabolismo , ARN Ribosómico 16S/genética , Análisis de Secuencia , Temperatura , Factores de Tiempo
11.
Front Microbiol ; 8: 1202, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28713340

RESUMEN

Particle-associated bacteria (PAB) and free-living bacteria (FLB) from aquatic environments during phytoplankton blooms differ in their physical distance from algae. Both the interactions within PAB and FLB community fractions and their relationship with the surrounding environmental properties are largely unknown. Here, by using high-throughput sequencing and network-based analyses, we compared the community and network characteristics of PAB and FLB from a plateau lake during a Microcystis aeruginosa bloom. Results showed that PAB and FLB differed significantly in diversity, structure and microbial connecting network. PAB communities were characterized by highly similar bacterial community structure in different sites, tighter network connections, important topological roles for the bloom-causing M. aeruginosa and Alphaproteobacteria, especially for the potentially nitrogen-fixing (Pleomorphomonas) and algicidal bacteria (Brevundimonas sp.). FLB communities were sensitive to the detected environmental factors and were characterized by significantly higher bacterial diversity, less connectivity, larger network size and marginal role of M. aeruginosa. In both networks, covariation among bacterial taxa was extensive (>88% positive connections), and bacteria potentially affiliated with biogeochemical cycling of nitrogen (i.e., denitrification, nitrogen-fixation and nitrite-oxidization) were important in occupying module hubs, such as Meganema, Pleomorphomonas, and Nitrospira. These findings highlight the importance of considering microbial network interactions for the understanding of blooms.

12.
Sci Total Environ ; 583: 214-221, 2017 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-28104332

RESUMEN

Cyanobacterial harmful algal blooms (CyanoHABs) cause severe environmental problems, economic losses and threaten human health seriously. In the present study, a Bacillus sp. strain, designated as AF-1, with strong antagonistic activity against plant pathogenic fungus Fusarium graminearum was isolated from purple soil. Bacillus sp. AF-1 selectively killed Microcystis aeruginosa at low cell density (1.6×103cfu/mL), and showed the strongest bactericidal activity against M. aeruginosa NIES-843 (Ae=93%, t=6d). The algicidal substances originated from strain AF-1 were stable in the temperature range of 35-100°C, and pH range of 3-11. Cell-free filtrate of AF-1 culture caused excessive accumulation of intracellular reactive oxygen species (ROS), cell death and the efflux of intracellular components of M. aeruginosa NIES-843 cells. The expression of genes recA, psbA1, psbD1, rbcL and mcyB, involved in DNA repair, photosynthesis and microcystin synthesis of NIES 843, were significantly influenced by the cell-free filtrate of AF-1 culture. Bacillus sp. AF-1 has the potential to be developed as a bifunctional biocontrol agent to control CyanoHABs and F. graminearum caused plant disease.


Asunto(s)
Bacillus/fisiología , Fusarium/fisiología , Floraciones de Algas Nocivas/fisiología , Microcystis/fisiología , Contaminación del Agua/prevención & control
13.
Environ Sci Pollut Res Int ; 24(5): 5001-5011, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-28000068

RESUMEN

Previous research found that the water-level fluctuating zone (WLFZ) of the Three Gorges Reservoir (TGR) was an Hg-sensitive area. However, little research has been conducted on the distribution of Hg-methylating microorganisms in this area. The goal of this research was to provide an initial description of the distribution of the dsrB (for sulfate-reducing bacteria) and hgcA (one gene confirmed for Hg methylation) genes. Different types of soil were selected to analyze the abundance of the dsrB and hgcA in different periods, in inundated soil (SI, ≤155 m, which becomes sediment during the wet period, SS) and in non-inundated soil (≥175 m, SN) from Shibao, a typical WLFZ of the TGR. A significant positive correlation was observed between dsrB and hgcA abundance and MeHg concentrations, suggesting that microorganisms with these genes contribute to Hg methylation. Principal component analysis (PCA) indicated that dsrB diversity was highest in SI, followed by SS; SS had the highest diversity of hcgA. Six phylogenetic trees were constructed and showed that more strains were present in SI than in SS. HgcA sequences in SS were confined to three evolutionarily distant clades, δ-Proteobacteria, a methanogen group, and a Clostridia group, which was relatively rare among most clades.


Asunto(s)
Genes Bacterianos , Mercurio/metabolismo , Microbiología del Suelo , Metilación , Filogenia , Suelo , Contaminantes del Suelo , Agua/análisis , Contaminantes Químicos del Agua
14.
Wei Sheng Wu Xue Bao ; 54(3): 261-8, 2014 Mar 04.
Artículo en Chino | MEDLINE | ID: mdl-24984517

RESUMEN

Beta-Barrel outer membrane proteins are the major components of the outer membrane of Gram-negative bacteria, which are in contact with the extracellular environment directly. Beta-barrel outer membrane proteins play key roles in nutrients absorption and keeping membrane integrity. They are also involved in the pathogenicity and multiple-antibiotic resistance of pathogenic bacteria. Therefore, beta-barrel outer membrane proteins are very important for the survive of bacteria cells, and full understanding of the biosynthesis, folding and membrane insertion of these proteins is of great significance for fighting against pathogenic bacteria and utilization of beneficial bacteria. In this article, the research progress on the biosynthesis in cytoplasm, the translocation across the inner membrane, transportation to periplasm and the folding and membrane insertion of beta-barrel outer membrane proteins are reviewed, and the progress on the study of OMPs assembly machinery is emphasized.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/metabolismo , Membrana Celular/metabolismo , Bacterias Gramnegativas/metabolismo , Proteínas de la Membrana Bacteriana Externa/química , Proteínas de la Membrana Bacteriana Externa/genética , Membrana Celular/química , Membrana Celular/genética , Bacterias Gramnegativas/química , Bacterias Gramnegativas/genética , Pliegue de Proteína , Transporte de Proteínas
15.
Wei Sheng Wu Xue Bao ; 54(11): 1241-7, 2014 Nov 04.
Artículo en Chino | MEDLINE | ID: mdl-25752130

RESUMEN

A cell of gram-negative bacteria is surrounded by two layers of membrane, the inner membrane and the outer membrane. Proteins are the major composition of outer membrane. Many outer membrane proteins carry a trans-membrane ß-barrel structure that formed by multiple anti-parallel ß-strands connected with hydrogen bonds. These proteins can act as porins, transporters, enzymes, receptors, virulence factors and structural proteins. Therefore, their correct folding and membrane integration are important for the survival of gram-negative bacteria. Most ß-barrel outer membrane proteins could be easily expressed recombinantly and refolded in vitro under certain conditions. The in vitro folding processes could be monitored and investigated through many ways, which makes outer membrane proteins become a model system to study the effects of abiotic and biological factors on the folding of membrane proteins. In this article, the research progress on the in vitro refolding of outer membrane proteins are reviewed from the aspects of refolding methods, the factors that affect folding processes and experimental methods. Finally, the research prospects in this field are discussed.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/química , Proteínas de la Membrana Bacteriana Externa/metabolismo , Bacterias Gramnegativas/química , Bacterias Gramnegativas/metabolismo , Modelos Moleculares , Replegamiento Proteico , Estructura Secundaria de Proteína
16.
Biochem Biophys Res Commun ; 425(2): 363-7, 2012 Aug 24.
Artículo en Inglés | MEDLINE | ID: mdl-22842563

RESUMEN

Polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA), are found in various cold-adapted microorganisms. We previously demonstrated that EPA-containing phospholipids (EPA-PLs) synthesized by the psychrotrophic bacterium Shewanella livingstonensis Ac10 support cell division, membrane biogenesis, and the production of membrane proteins at low temperatures. In this article, we demonstrate the effects of EPA-PLs on the folding and conformational transition of Omp74, a major outer membrane cold-inducible protein in this bacterium. Omp74 from an EPA-less mutant migrated differently from that of the parent strain on SDS-polyacrylamide gel, suggesting that EPA-PLs affect the conformation of Omp74 in vivo. To examine the effects of EPA-PLs on Omp74 protein folding, in vitro refolding of recombinant Omp74 was carried out with liposomes composed of 1,2-dipalmitoleoyl-sn-glycero-3-phosphoglycerol and 1,2-dipalmitoleoyl-sn-glycero-3-phosphoethanolamine (1:1 molar ratio) with or without EPA-PLs as guest lipids. SDS-PAGE analysis of liposome-reconstituted Omp74 revealed more rapid folding in the presence of EPA-PLs. CD spectroscopy of Omp74 folding kinetics at 4 °C showed that EPA-PLs accelerated ß-sheet formation. These results suggest that EPA-PLs act as chemical chaperones, accelerating membrane insertion and secondary structure formation of Omp74 at low temperatures.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/química , Proteínas de la Membrana Bacteriana Externa/metabolismo , Ácido Eicosapentaenoico/metabolismo , Shewanella/metabolismo , Ácido Eicosapentaenoico/biosíntesis , Liposomas/química , Liposomas/metabolismo , Pliegue de Proteína , Estructura Secundaria de Proteína
17.
Sheng Wu Gong Cheng Xue Bao ; 23(5): 789-93, 2007 Sep.
Artículo en Chino | MEDLINE | ID: mdl-18051853

RESUMEN

Atrazine could be used as the sole carbon, nitrogen and energy sources for growth by strain Arthrobacter sp. AG1, and the atrazine-degrading genes of AG1 were found to be the combination of trzN, atzB and atzC. The atrazine chloride hydrolysase gene trzN was cloned by PCR amplification,whose sequence shared 99% identity with that of Norcardioides sp. C190. Two large plasmids were found in AG1,and trzN and atzB were confirmed to be localized on the larger plasmid pAG1 by the method of southern hybridization. Subculture of AG1 in liquid LB for three generations, 34% of the subsequent cells were found to lose degrading activity, however, neither plasmid was lost. PCR amplification results showed that the mutants had only lost the trzN gene instead of atzB and atzC. It was deduced that mutation might be due to the trzN gene deletion from the plasmid. This study provided new evidence that atrazine metabolic genotypes were resulted from horizontal gene transfer between different bacteria under environmental selective pressure.


Asunto(s)
Arthrobacter/genética , Atrazina/metabolismo , Genes Bacterianos/genética , Biodegradación Ambiental , Herbicidas/metabolismo
18.
Wei Sheng Wu Xue Bao ; 47(3): 544-7, 2007 Jun.
Artículo en Chino | MEDLINE | ID: mdl-17672324

RESUMEN

Atrazine (AT), a kind of herbicide for the pre and post-emergence control of annual and broad leaved weeds and perennial grasses, had been widely used in the world. However, the extensive use of atrazine had led to widespread environmental pollution. A bacterium strain SA1, which could degrade AT completely, was isolated from an atrazine-degrading consortium by long-time repeated alternative cultivation and plate striking. Combining cultural and physiobiochemical characteristics with 16S rDNA sequence analysis, SA1 was identified as Pseudomonas sp.. SAl could use atrazine as the sole carbon, nitrogen and energy sources for growth, and the main product of AT biodegradation was cyanuric acid. AT degrading activity of SA1 was not affected by the addition of nitrogen resources. However, cyanuric acid could be degraded quickly to an undetectable level when glucose was added. The optimal temperature and pH value for SAl growth was 37 degrees C and pH7, respectively. Atrazine could be degraded efficiently by the resting cells of SAl under the conditions of 10 degrees C - 40 degrees C or pH value 4-11, and SA1 had a wide range of temperature and pH value for AT degradation when compared with ADP. atzABCD and conserved sequence of tnpA gene of IS1071 could be amplified from SA1, and these genes could be lost during subculture.


Asunto(s)
Atrazina/metabolismo , Herbicidas/metabolismo , Pseudomonas/aislamiento & purificación , Pseudomonas/metabolismo , Microbiología del Suelo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , ADN Bacteriano/genética , ADN Ribosómico/genética , Datos de Secuencia Molecular , Filogenia , Pseudomonas/clasificación , Pseudomonas/genética , ARN Ribosómico 16S/genética
19.
Ying Yong Sheng Tai Xue Bao ; 16(8): 1518-22, 2005 Aug.
Artículo en Chino | MEDLINE | ID: mdl-16262070

RESUMEN

The study showed that the application of atrazine stimulated soil microorganisms obviously. In comparing with control(without atrazine), the respiration intensity of soil applied with 50 mg atrazine kg(-1) soil increased greatly, the concentrations of soil NH4(+)-N and NO3(-)-N changed significantly, and the individuals of soil microbes, especially bacteria and fungi, also increased greatly. The application of strain BTAH1 could degrade 98% of applied atrazine within one week, and led to the decrease of soil respiration intensity. Under BTAH1 application, the individuals of actinomyces and fungi decreased, while those of bacteria did not, and the concentrations of soil NH4(+)-N and NO3(-)-N came back to the level of the control. ARDRA analysis on the 16s rDNA library of soil bacteria suggested that the application of atrazine could decrease the biodiversity of soil microorganisms, while applying BTAH1 could recover the biodiversity.


Asunto(s)
Atrazina/metabolismo , Bacterias/efectos de los fármacos , Bacterias/metabolismo , Microbiología del Suelo , Atrazina/farmacología , Biodegradación Ambiental , Biodiversidad , Herbicidas/metabolismo , Herbicidas/farmacología
20.
Yi Chuan ; 27(5): 845-51, 2005 Sep.
Artículo en Chino | MEDLINE | ID: mdl-16257921

RESUMEN

Horizontal gene transfer, unlike vertical gene transfer, is a means of genetic communication in bacteria. In the special polluted environment, horizontal transfer of polluted-degrading gene has significant functions. Study on horizontal transfer of degrading gene in polluted environment may deepen our understanding of the mechanism of bacterial adaptation to the organic-polluted environment. In the practical application in bioremediation, horizontal transfer of degrading gene can be regulated to promote degrading ability of microorganisms. In this article, we will review the advances in the study on mechanisms of genetic interactions among bacteria, the effect of degrading gene transfer in contaminated environment on microorganisms'adaptation to contaminated environment and the degradation of the pollutants.


Asunto(s)
Bacterias/metabolismo , Proteínas Bacterianas/metabolismo , Contaminantes Ambientales/metabolismo , Transferencia de Gen Horizontal , Adaptación Fisiológica/genética , Adaptación Fisiológica/fisiología , Bacterias/genética , Bacterias/crecimiento & desarrollo , Proteínas Bacterianas/genética , Restauración y Remediación Ambiental/métodos
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