Extracellular Substances Research Articles

The Role and Therapeutic Potential of Macropinocytosis in Cancer.

Macropinocytosis, a unique endocytosis pathway characterized by nonspecific internalization, has a vital role in the uptake of extracellular substances and antigen presentation. It is known to have dual effects on cancer cells, depending on cancer type and certain microenvironmental conditions. It helps cancer cells survive in nutrient-deficient environments, enhances resistance to anticancer drugs, and promotes invasion and metastasis. Conversely, overexpression of the RAS gene alongside drug treatment can lead to methuosis, a novel mode of cell death. The survival and proliferation of cancer cells is closely related to macropinocytosis in the tumor microenvironment (TME), but identifying how these cells interface with the TME is crucial for creating drugs that can limit cancer progression and metastasis. Substantial progress has been made in recent years on designing anticancer therapies that utilize the effects of macropinocytosis. Both the induction and inhibition of macropinocytosis are useful strategies for combating cancer cells. This article systematically reviews the general mechanisms of macropinocytosis, its specific functions in tumor cells, its occurrence in nontumor cells in the TME, and its application in tumor therapies. The aim is to elucidate the role and therapeutic potential of macropinocytosis in cancer treatment.

Bioaccessibility of Microplastic-Associated Antibiotics in Freshwater Organisms: Highlighting the Impacts of Biofilm Colonization via an In Vitro Protocol.

Microplastics in the environment can be colonized by microbes capable of forming biofilms, which may act as reactive coatings to affect the bioaccessibility of pollutants in organisms. This study investigated the dynamic evolution of biofilm colonization on microplastics and its impacts and mechanisms on the bioaccessibility of microplastic-associated sulfamethazine (SMT) via microcosm incubation in surface water and sediment. After 60 days of incubation, the microbial communities formed in microplastics were distinct and more diverse than those untethered in surroundings, and photoaging treatment decreased the affinity of biofilms on microplastics due to decreased hydrophobicity. Biofilm formation further enhanced the desorption and bioaccessibility of microplastic-sorbed SMT in organisms. In vitro experiments indicated that the critical effects were mainly related to the stronger interaction of gastrointestinal components (i.e., pepsin, bovine serum albumin (BSA), and NaT) with biofilm components (e.g., extracellular polymer substances) than with the pure surface of microplastics, which competed for binding sites in microplastics for SMT more significantly. Photoaging decreased the enhancing effects of biofilms due to their lower accumulation in aged microplastics. This study is the first attempt to reveal the role of biofilms in the bioaccessibility of microplastics with associated antibiotics and provide insights into the combined risk of microplastics in the environment.

Deciphering the underlying mechanism of MOF-808-based abiotic catalysis enhancing biodegradability of waste activated sludge: Insights from the effects on bioconversion of extracellular organic substances into methane

Extracellular organic substances (EOSs) usually control sludge biodegradability. Metal−organic framework-808 (MOF-808), a catalyst with a high ability of catalytic hydrolysis and proton transfer, has great potential to enhance biodegradability of EOSs. In this study, the underlying mechanism of MOF-808 enhancing the biodegradability of EOSs via abiotic catalysis was investigated. Experimental results showed that protein–like and humic acid–like substances were the main organic components in EOSs, and the MOF-808 enhanced the disintegration of protein-like substances rather than humic acid–like substances. Analyses of the changes in the functional group, the secondary structure of protein-like substances, and the electron transfer of catalytically degraded EOS samples revealed that the MOF-808 enhanced the hydrolysis of EOSs protein-like substances, loosened their secondary structure, and improved the electron transfer in EOSs. Further analyses of the MOF-808 before and after the catalysis reaction revealed that the coordination of Zr sites with protein-like substance-specific active sites (such as ZrN) in the MOF-808 substantially contributed to the high-efficiency catalysis. Biochemical methane potential assays confirmed that MOF-808–based abiotic catalysis promoted the generation efficiency of methane from the EOSs. These findings can elucidate the abiotic catalytic effect of MOF-808 on sludge biodegradability during the anaerobic digestion process.

Strategic thermosonication-mediated modulation of lactic acid bacteria acidification kinetics for enhanced (post)-fermentation performance

This study explored the feasibility of thermosonication (TS)-prestressed inoculum with different fermentation patterns for regulating microbial (post)-fermentation acidification kinetics. Through a Box-Behnken design, stimulative (20 min, 400 W, 33 kHz, 25 °C) and inhibitive (10 min, 600 W, 33 kHz, 20 °C) effects on the acidification capability of Lactobacillus plantarum A3 were achieved without observing greatly activated/inactivated strains growth, further confirmed by lactose fermentation performed by Streptococcus thermophilus and Lactobacillus bulgaricus. Lactic acid was the major contributing factor responsible for TS-induced acidification modifications corresponding to the potential fluctuations of CoA biosynthesis, fatty acid degradation and chain elongation pathways to TS prestress. Microscopy observations and quantitative extracellular substance assays showed palpable stress disturbance on microbes, but causing insignificant effects on product characteristics. This investigation demonstrated the potential of controlled sonication prestress strategies to achieve dual engineering effects on microbial metabolic behavior, for alleviating post-acidification problem or enhancing process efficiencies.

Research Progress on the Correlation Between Bacterial Biofilm Microenvironment and Charge Regulation

A bacterial biofilm (BF) is composed of microbial communities growing on the surface and extracellular substances secreted by itself, which is one of the main reasons for bacterial multidrug resistance. Analysis and treatment methods based on different principles have been widely used in BF research. Studies have proved that the communication behavior in bacterial BF is also controlled by electrochemical flow. Therefore, it is very effective to study the treatment strategy of new charge regulated anti-BF. Based on the electrical characteristics of bacteria and BF microenvironment, this article summarizes the progress of charge regulation in BF control and prospects the control methods based on the combination of charge regulation and other technologies.

Regulation of microglia phagocytosis and potential involvement of exercise.

Microglia are considered the main phagocytic cells in the central nervous system, remodeling neural circuits by pruning synapses during development. Microglial phagocytosis is also a crucial process in maintaining adult brain homeostasis and clearing potential toxic factors, which are recognized to be associated with neurodegenerative and neuroinflammatory disorders. For example, microglia can engulf amyloid-β plaques, myelin debris, apoptotic cells, and extracellular harmful substances by expressing a variety of specific receptors on the cell surface or by reprogramming intracellular glucose and lipid metabolism processes. Furthermore, physical exercise has been implicated to be one of the non-pharmaceutical treatments for various nervous system diseases, which is closely related to neuroplasticity and microglia functions including proliferation, activation, and phagocytosis. This review focuses on the central regulatory mechanisms related to microglia phagocytosis and the potential role of exercise training in this process.

Mechanism of dielectric barrier discharge plasma technology to improve the quantity of short-chain fatty acids in anaerobic fermentation of waste active sludge.

The mechanism of improving the anaerobic fermentation performance of waste active sludge by using dielectric barrier discharge (DBD) plasma pretreatment technology was investigated. The maximum accumulation of short-chain fatty acids (SCFAs) was observed on the 7th day of anaerobic fermentation when the DBD power was 76.50 W, which was 1726.70 mg COD/L, 1.50 times of the control group. The ratio of acetic acid in DBD group was 9.30% higher than that in the control. Further mechanism research indicated that DBD pretreatment can destroy the structure of extracellular polymer substances and release organic substances such as protein and polysaccharide. The dissolved organic matter analysis indicated that the DBD technique could increase the release of biodegradable organics (eg., tyrosine proteins, soluble microbial by-products), thus accelerate the biotransformation of organic substance. Bacterial community structure analysis showed that the increase in the abundance of Firmicutes and Bacteroidetes and the decrease in the abundance of Proteobacteria in DBD group were beneficial to the accumulation of SCFAs. Besides, further archaeal analysis indicated that the decrease of Methanosaeta sp. and Methanosarcina sp. abundance in the DBD group facilitate acetic acid accumulation. This study demonstrated that the DBD technique can be used as an effective and potential pretreatment method to improve sludge anaerobic fermentation performance.

Promotion of nitrogen removal in a zero-valent iron-mediated nitrogen removal system operated in co-substrate mode

In this study, the performance and mechanism of nitrogen removal were investigated in a zero-valent iron-mediated nitrogen removal system operated in co-substrate mode with sodium acetate as the organic carbon source. The results showed that the additional organic matter had the capacity to promote NH4+-N and total inorganic nitrogen (TIN) removal with efficiencies of 91.09% and 84.10%, and increases of 60.06% and 75.32% compared with the control group, respectively. The organic matter also stimulated the production of extracellular polymer substances that reduced the passivation and toxicity of iron to microorganisms. The ammonia oxidation activity was 2.5 times higher than that in the control group, and the anammonia oxidation activity and denitrification activity were substantially higher than in the control group with TIN removal efficiencies of 1.02 and 1.19 mgN/(gVSS·d), respectively. In addition, the organic matter increased the enrichment of the heterotrophic denitrification bacterium Diaphorobacter and facultative iron salt-based bacterium Dechloromonas.

Insight into biofilm-forming patterns: biofilm-forming conditions and dynamic changes in extracellular polymer substances.

The microbial biofilm adheres to the surface of the carrier, which protects the pollutant-degrading bacteria and resists harsh environments; thus, research on biofilm-forming patterns will help promote the application of biofilms in wastewater treatment. Herein, univariate analysis and response surface methodology (RSM) confirmed that glucose and mannose at 3-5g/L promoted biofilm formation. Notably, the microplate method demonstrated that compared to trivalent cations, divalent cations could more greatly enhance the activity (especially magnesium) of the biofilm matrix, and the period of biofilm formation in the three strains was divided into the following stages: initial attachment (0-10h), microcolony (10-24h), maturation (24-48h), and dispersion (36-72h). During maturation, large amounts of extracellular polysaccharides (EPs) and extracellular DNA (eDNA) were distributed in the extracellular and intracellular spaces, respectively, as observed by super-resolution structured illumination microscopy (SR-SIM). This study enhances the understanding of the characteristics and patterns of biofilm formation and can facilitate the application of biofilms in wastewater treatment.

Electrochemical impedance spectroscopy (EIS) reveals the role of microbial fuel cell-ceramic membrane bioreactor (MFC-CMBR): Electricity utilization and membrane fouling

Ceramic membrane has become a major concern due to creasing cost and competitive efficiency. Microbial fuel cell-ceramic membrane bioreactor (MFC-CMBR) is considered alternative technology for larger-scale industrial application because of its advantages of convenient detecting and control of membrane fouling. However, MFC-CMBR are highly susceptible to membrane fouling and harsh operating requirements in these wastewaters of different compositions. This research critically discusses that electrochemical response in different types of MFC-CMBRs and control of electricity utilization on ceramic membrane fouling. The experimental results indicated that the application of sludge acclimated in coupling system with higher external resistance could ensure that lower costs (electricity utilization and membrane cleaning) provided enough membrane fouling control. The improved performance of MFC-CMBR-1 could be attributed to its enhanced nitrification/denitrification activity and capacity of electrons migration between electrode and sludge mixture. The coupling system alleviated membrane fouling and impedance increasing by improving the characteristics of sludge (increased particle size, decreased adsorption adhesion free energy), EPS (decreased hydrophobicity, molecular weight distribution regulation). And filtration tests showed that roughness and contact angle for the MFC-CMBR tend for better development compared to CMBR, dependent on the changes in the chemical surface groups as a result of electric distribution ratio. In addition, correlation analysis and filtration experiments showed that the extracellular polymer substances (EPS) enhanced the charge transfer resistance (Rct), and the protein substance in EPS was the main fouling substance when external resistance was close to the internal resistance of MFC. In summary, the low internal resistance of ceramic membrane lead to obvious better fouling control and electricity utilization than organic membrane, and the paper provides insight into the MFC-CMBR systems for a wide range of detecting membrane fouling and applications of membrane fouling mitigation.

Delaying the decline of germination ability of pecan pollen by yeast Pichia fermentans 15B1

Microorganisms may be critical to the final outcome of pollination, but there are few studies on the effects of microorganisms on pollination process, especially the direct effects of microorganisms on pollen germination after pollination. Studies have shown that yeast can inhibit pollen germination regardless of in vitro germination and stigma germination, while bacteria like Acinetobacter pollinis can promote pollen germination. Pecan (Carya illinoinensis (Wangenh.) K. Koch) is a kind of wind-pollinated hybrid plant and the in vitro germination system of pecan pollen is well-established. In this paper, pecan pollen was used to reveal the special influence of yeast microorganisms on the pollen germination. It is well known that storage at room temperature will usually lead to a decrease in pollen viability and the study here showed that the germination ability of pecan pollen decreased by about 19.0% after two-day storage at room temperature. Although the culture of Pichia fermentans Lodder (strain 15B1) had no significant effect on the in vitro germination of pecan pollen without storage at room temperature, it could significantly improve the germination ability of pecan pollen after two-day storage at room temperature, with an average increase of 33.6%. It proved that the group with supernatant part of whole strain culture had the strongest promotion effect on pollen germination including pollen germination rate and pollen tube length among the whole strain culture, supernatant part and precipitation part, indicating that the pollen germination was mainly promoted by extracellular substances of strain rather than strain 15B1 itself. This promoting effect was more like a recovery of the lost germination ability of pecan pollen due to room temperature. Further experiments showed that the extracellular substances of strain 15B1 had good thermal stability. This study is of great significance to explore the biological role of microorganisms in pollination, and also provides an important idea for improving fruit pollination by microbial ways.

C058 and Other Functional Microorganisms Promote the Synthesis of Extracellular Polymer Substances in Mycelium Biofloc

The mycelium biofloc bioaugmented by Cordyceps strain C058 effectively purifies water, which may be related to the synthesis of extracellular polymer substances. To verify this conjecture, we analyzed the changes in extracellular polymer substances content in the mycelium biofloc under various hydraulic retention times (36 h, 18 h, and 11 h). The microstructure and microflora composition were analyzed using a scanning electron microscope and high-throughput sequencing. The ordinary biofloc without bioaugmentation was taken as a control. The results showed that under the above hydraulic retention time, the extracellular polymer substances contents of the mycelium biofloc were 51.20, 55.89, and 33.84 mg/g, respectively, higher than that of the ordinary biofloc (14.58, 15.72, and 18.19 mg/g). The protein content or the polysaccharide content also followed the same trend. Meanwhile, the sedimentation performance of the mycelium biofloc was better than that of the ordinary biofloc, attributed to the content of the extracellular polymer substances. It is worth noting that C058 is the main biofloc content, which promotes the synthesis of extracellular polymer substances in the mycelium biofloc. Other functional microorganisms in the mycelium biofloc were Janthinobacterium, Phormidium, Leptolyngbya, Hymenobacter, and Spirotrichea, which also promote the synthesis of extracellular polymer substances.

Removal Characteristics of Four Typical Antibiotics in Denitrification System

Wastewater treatment plants are an important gathering place for antibiotics, where denitrification plays a vital role in biological nitrogen removal. In order to explore the removal characteristics of antibiotics in a denitrifying sludge system, norfloxacin (NOR), oxytetracycline (OTC), sulfamethoxazole (SMX), and trimethoprim (TMP) were selected to investigate their transformation under different carbon source conditions in a denitrification system. The contribution of adsorption and biodegradation for antibiotic removal was also evaluated in this study. The results showed that a certain proportion of NOR, OTC, and TMP could be removed by denitrification, whereas NOR and OTC could act as the sole carbon source for denitrification. The removal of NOR and OTC showed a rapid adsorption and then slow biodegradation trend in the denitrification system. The contributions of adsorption were recorded as 83.5% and 58.9% for NOR and OTC removal, respectively. More than 40% were adsorbed by extracellular polymer substances (EPS), whereas the P450 enzyme played an important role in the OTC biodegradation process, with a contribution of 20%.

Order-of-magnitude enhancement in photocurrent generation of Synechocystis sp. PCC 6803 by outer membrane deprivation

Biophotovoltaics (BPV) generates electricity from reducing equivalent(s) produced by photosynthetic organisms by exploiting a phenomenon called extracellular electron transfer (EET), where reducing equivalent(s) is transferred to external electron acceptors. Although cyanobacteria have been extensively studied for BPV because of their high photosynthetic activity and ease of handling, their low EET activity poses a limitation. Here, we show an order-of-magnitude enhancement in photocurrent generation of the cyanobacterium Synechocystis sp. PCC 6803 by deprivation of the outer membrane, where electrons are suggested to stem from pathway(s) downstream of photosystem I. A marked enhancement of EET activity itself is verified by rapid reduction of exogenous electron acceptor, ferricyanide. The extracellular organic substances, including reducing equivalent(s), produced by this cyanobacterium serve as respiratory substrates for other heterotrophic bacteria. These findings demonstrate that the outer membrane is a barrier that limits EET. Therefore, depriving this membrane is an effective approach to exploit the cyanobacterial reducing equivalent(s).

A model describing the pressure drops across the membrane and cake layers for constant flux cross-flow microfiltration process

A combined model (pore-blocking/shrinking-cake filtration) was established based on the Darcy’s law, Hagen-Poiseuille equation and classic blocking law to study the fouling behavior of a 0.1 µm PAN membrane fouled with the extracellular polymer substance model solution (bovine serum albumin (BSA), sodium alginate (SA) and humic acid (HA)) under different operating conditions in the constant flux cross-flow filtration process. Results showed that the model predictions had good agreements with experimental data not only for single component solution (BSA, SA and HA), but also for the binary mixtures (BSA+SA, BSA+HA and SA+HA), ternary mixtures (BSA+SA+HA) and other membranes (0.1 µm PES and PVDF) (R2 ≥ 0.874). The dominant fouling mechanism is pore-blocking/shrinking at lower concentration, lower flux, or higher velocity in the whole stage. In contrast, the dominant fouling mechanism is pore-blocking/shrinking-cake formation in the first stage while it is cake formation in the second stage at a higher concentration, higher flux, or lower velocity. Meanwhile, the trans-membrane pressure (P) increased rapidly in the first stage for both situations while P kept constant for the first situation and P increased slowly for the second situation in the second stage. Furthermore, transition point and time from combined mechanism to individual mechanism were determined. This model can provide guides for industrial backwashing processes.

Contributions of MOF-808 to methane production from anaerobic digestion of waste activated sludge

The bioconversion of waste activated sludge (WAS) into methane is usually limited by the poor hydrolysis of sludge and/or poor syntrophic methanogenesis during anaerobic digestion (AD). In this study, the underlying mechanism of MOF-808 enhancing hydrolysis and syntrophic methanogenesis during AD process of WAS was investigated. Experimentally, with the effects of MOF-808 (150 mg MOF-808/g Volatile Solid (VS)), the methane production and the proportion of methane in biogas increased by approximately 26.7% and 15.6%, respectively, and the lag phase of methanogenesis decreased by 50.8%, which indicate that MOF-808 enhanced the generation efficiency of methane. The changes in activities of main hydrolytic enzymes with and without MOF-808 (150 mg MOF-808/g VS) during AD process revealed that MOF-808 improved the enzymatic hydrolysis of sludge, and the abiotic hydrolysis of sludge extracellular organic substances by MOF-808 shows that the maximum proportion and the initial increasing rate of low-molecular weight fractions increased by 60% and 583.7%, respectively, indicating that MOF-808 can greatly enhance the hydrolysis degree and rate of sludge via abiotic effect. These demonstrate that MOF-808 enhanced both biological and abiotic hydrolysis of sludge during AD. In addition, changes in the concentrations of acetate kinase and volatile fatty acids (VFAs) with and without MOF-808 (150 mg MOF-808/g VS) during AD process showed that MOF-808 accelerated the bioconversion of VFAs to methane, suggesting MOF-808 has a positive effect on syntrophic metabolism for methanogenesis. Moreover, further analyses of the microbial community structure of sludge samples with and without MOF-808 (150 mg MOF-808/g VS) showed that MOF-808 enriched hydrogen-producing bacteria and mixotrophic methanogens (i.e. Methanosarcina), and changed the methanogenic pathway via accelerating proton transfer between syntrophic anaerobes, especially improving the reduction of CO2 to methane, and resulting in highly efficient syntrophic methanogenesis. These findings, however, may provide an important reference for enhancing AD efficiency of WAS based on MOF-like materials.

The effect of TiO2 nanoparticles size on Cd (II) removal by the paddy crusts from waterbody

The influence of nanoparticles (NP) on the accumulation of heavy metals in the paddy crust (PC) was unclear. This experiment explored the potential effects and mechanism of TiO2 NP with different particle sizes on Cd removal by PC from waterbody. All the TiO2 with size of 25 nm (T25), 100 nm (T100) and 2000 nm (T2000) can promote the Cd uptake by the PC, and with the sequence: T100 > T25 > T1000. Especially, 0.1 mg/L T100 can enhanced ~45% Cd removal comparing with pristine PC, and the Cd uptake capacity reached as high as 92 mg/g. After treatment of 72 h, the PC+T100 can reduce Cd in the acidic coal drainage from 0.48 mg/L to 0.009 mg/L, with removal efficiency of 98%. The total concentration of surface adsorptive site number of PC+T100, PC+T25 and PC was 0.87 ± 0.03, 0.78 ± 0.07 and 0.67 ± 0.04, respectively. The TiO2 stimulation to PCs secreted the extracellular substance and then increased the content of amino/hydroxyl adsorptive sites by 30–70%. T100 can stimulate high EPS-producing algae and bacteria, whereas most T25 can only penetrate the cell walls of low EPS-producing microorganisms. T100 can stimulate not only the thick sheath layer of Oedogonium, the main component of PC, but also the thin layer of algae and bacterium. And induce it to produce a large amount of protein, phospholipids, etc. T25 can only stimulate the Microcystics, Chlorococcum, Scenedesmius and bacteria with thin cell membranes to produce low extracellular substance. However, T25 would easily enter the cell trough thin cell membranes, result in that the Cd attached to T25 to penetrate the intracell. Therefore, the non-EDTA-exchangeable Cd fraction in PC+T25 was 55.48%, higher than 36.82% of PC+T100, and 21.52% of PC. Our investigation demonstrated that the addition of TiO2 NP with appropriated size in PC was a potential efficient method to remove the heavy metal from waterbody.

Characterization of spatiotemporal electroactive anodic biofilm activity distribution using 1D simulations

Activity distribution limitation in electroactive biofilm remains an unclear phenomenon. Some observations using confocal microscopy have shown notable difference between activity close to the anode and activity at the liquid interface. A numerical model is developed in this work to describe biofilm growth and local biomass segregation in electroactive biofilm. Under our model hypothesis, metabolic activity distribution in the biofilm results from the competition between two limiting factors: acetate diffusion and electronic conduction in the biofilm. Influence of inactive biomass fraction (i.e. non-growing biomass fraction) properties (such as conductivity and density) is simulated to show variation in local biomass distribution. Introducing a dependence of effective diffusion to local density leads to a drastic biomass fraction segregation. Increasing density of inactive fraction reduces significantly acetate diffusion in biofilm, enhances biomass activity on the outer layer (liquid/biofilm interface) and maintains inner core largely inactive. High inactive fraction conductivity enhances biomass activity in the outer layer and enhances current production. Hence, investment in extracellular polymer substance (EPS), anchoring redox components, is benefit for biofilm electroactivity. However, under our model hypothesis it means that conductivity should be two order lower than biofilm conductivity reported in order to observe inner core active biomass segregation.

Contribution of Pseudomonas aeruginosa Exopolysaccharides Pel and Psl to Wound Infections.

Biofilms are the cause of most chronic bacterial infections. Living within the biofilm matrix, which is made of extracellular substances, including polysaccharides, proteins, eDNA, lipids and other molecules, provides microorganisms protection from antimicrobials and the host immune response. Exopolysaccharides are major structural components of bacterial biofilms and are thought to be vital to numerous aspects of biofilm formation and persistence, including adherence to surfaces, coherence with other biofilm-associated cells, mechanical stability, protection against desiccation, binding of enzymes, and nutrient acquisition and storage, as well as protection against antimicrobials, host immune cells and molecules, and environmental stressors. However, the contribution of specific exopolysaccharide types to the pathogenesis of biofilm infection is not well understood. In this study we examined whether the absence of the two main exopolysaccharides produced by the biofilm former Pseudomonas aeruginosa would affect wound infection in a mouse model. Using P. aeruginosa mutants that do not produce the exopolysaccharides Pel and/or Psl we observed that the severity of wound infections was not grossly affected; both the bacterial load in the wounds and the wound closure rates were unchanged. However, the size and spatial distribution of biofilm aggregates in the wound tissue were significantly different when Pel and Psl were not produced, and the ability of the mutants to survive antibiotic treatment was also impaired. Taken together, our data suggest that while the production of Pel and Psl do not appear to affect P. aeruginosa pathogenesis in mouse wound infections, they may have an important implication for bacterial persistence in vivo.

Characterization and algicidal activity of bacteria from the phycosphere of the harmful alga Karenia mikimotoi.

Harmful algal blooms (HABs) are detrimental to aquatic ecosystems; thus, economical and practical HAB control methods are needed. We analyzed a microbial community closely related to the alga Karenia mikimotoi, which has HABs that can be toxic to aquatic environments. We studied the relationship between algicidal bacteria and the microbial community of K. mikimotoi culture using culture-dependent and culture-independent methods. Bacterial strains Marinobacter sp. (O-7) and Pseudomonas sp. (D-2) were isolated from a K. mikimotoi seawater culture containing a mixed microbial community and determined to have algicidal activity. Both strains produced alga-lysing substances that were toxic to K. mikimotoi. The algicidal extracellular substances produced by D-2 were stable at temperatures ranging from - 80 to 120°C but sensitive to strong acidic/alkaline conditions. The substances produced by O-7 were inactivated at high temperatures and strong alkaline conditions. Extracellular substances produced by O-7 and D-2 caused K. mikimotoi and Prorocentrum donghaiense cell lysis, but no changes or inhibitory effects occurred in two other chlorophyta groups. O-7 and D-2 exhibited significantly greater algicidal activity during the logarithmic growth phase of K. mikimotoi growth compared to the stationary phase. Culture-independent analysis of the microbial community in association with K. mikimotoi was made using Illumina MiSeq sequencing. Phylogenetic analysis showed that Proteobacteria was the dominant bacterial population in the phycosphere of K. mikimotoi, containing Marinobacter sp. and Pseudomonas sp.. The Marinobacter was abundant and accounted for 5.3% of the total. Our results indicate that certain bacterial species from K. mikimotoi culture might be effective for removal of HABs.