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Frontiers of Environmental Science & Engineering    2020, Vol. 14 Issue (1) : 9-     https://doi.org/10.1007/s11783-019-1188-2
REVIEW ARTICLE
Transmembrane transport of polycyclic aromatic hydrocarbons by bacteria and functional regulation of membrane proteins
Hongqi Wang1(), Ruhan Jiang1, Dekang Kong1, Zili Liu1, Xiaoxiong Wu1, Jie Xu1, Yi Li2,3()
1. College of Water Sciences, Beijing Normal University, Beijing 100875, China
2. Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection (Ministry of Education), Guangxi Normal University, Guilin 541004, China
3. College of Environment and Resource, Guangxi Normal University, Guilin 541004, China
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Abstract

• Explaintheadsorption, uptake and transmembrane transport of PAHs by bacteria.

• Analyze functional regulation of membrane proteins inthe transmembrane transport.

• Proteomics technology such as iTRAQ labeling was used to access expressed proteins.

• Single cell analysis technology wereused to study the morphological structure.

In recent years, increasing research has been conducted on transmembrane transport processes and the mechanisms behind the microbial breakdown of polycyclic aromatic hydrocarbons (PAHs), including the role of membrane proteins in transmembrane transport and the mode of transmission. This article explains the adsorption, uptake and transmembrane transport of PAHs by bacteria, the regulation of membrane protein function during the transmembrane transport. There are three different regulation mechanisms for uptake, depending on the state and size of the oil droplets relative to the size of the microbial cells, which are (i) direct adhesion, (ii) emulsification and pseudosolubilization, and (iii) interfacial uptake. Furthermore, two main transmembrane transport modes are introduced, which are (i) active transport and (ii) passive uptake and active efflux mechanism. Meanwhile, introduce the proteomics and single cell analysis technology used to address these areas of research, such as Isobaric tags for relative and absolute quantitation (iTRAQ) technology and Nano Secondary ion mass spectrometry (Nano-SIMS). Additionally, analyze the changes in morphology and structure and the characteristics of microbial cell membranes in the process of transmembrane transport. Finally, recognize the microscopic mechanism of PAHs biodegradation in terms of cell and membrane proteins are of great theoretical and practical significance for understanding the factors that influence the efficient degradation of PAHs contaminants in soil and for remediating the PAHs contamination in this area with biotechnology.

Keywords Polycyclic aromatic hydrocarbons      Transmembrane transport      Adsorption and uptake of hydrocarbons      Proteomics      Functional regulation of membrane protein      Single cell analysis technology     
发布日期: 2019-10-31
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作者相关文章
Hongqi Wang
Ruhan Jiang
Dekang Kong
Zili Liu
Xiaoxiong Wu
Jie Xu
Yi Li
引用本文:   
Hongqi Wang,Ruhan Jiang,Dekang Kong, et al. Transmembrane transport of polycyclic aromatic hydrocarbons by bacteria and functional regulation of membrane proteins[J]. Front. Environ. Sci. Eng., 2020, 14(1): 9.
网址:  
https://journal.hep.com.cn/fese/EN/10.1007/s11783-019-1188-2     OR     https://journal.hep.com.cn/fese/EN/Y2020/V14/I1/9
Fig.1  Scheme of mycelia fluoranthene biosorption, transport and release (Schamfuss et al., 2013).
Adsorption type Microbial species Hydrocarbons species Parameters and results References
Direct adhesion Pseudomonas sp. M1 n-Hexadecane The degradation was not significantly affected by 4 mM EDTA. Goswami and Singh (1991)
Acinetobacter sp. CR n-Heneicosane Higher impeller speed resulted in both lower microbial growth and lower n-alkane degradation rate of the bacterium, although it increased the specific surface area of the oil, which was measured by a previously developed device. This result was due to the decreased number of cells adhering to the oil surface, i.e., intense agitation inhibited the adhesion of cells to the oil surface. Hori et al. (2002)
Rhodococcus equi OU2 Hexadecane The surfactant is unnecessary for hexadecane to be degraded by OU2 Bouchez-Naitali and Vandecasteele (2008)
Rhodococcus baikonurensis EN3 Diesel oil Tergitol 15 S, APG, LAE 9 and Tween 80 were effective only to a small extent in the enhancement of the biodegradation of diesel oil, and SDS inhibited rather than stimulated the degradation of diesel oil. Lee et al. (2006)
Emulsification and pseudosolubilization Pseudomonas sp. N1 n-Hexadecane The degradation was strongly inhibited by the 4 mM EDTA. Goswami and Singh (1991)
Pseudomonas aeruginosa GL1 Hexadecane The surfactant is necessary for hexadecane to be degraded by GL1. Bouchez-Naitali and Vandecasteele (2008)
Pseudomonas aeruginosa AT10 Total petroleum hydrocarbons, the group of isoprenoids from the aliphatic fraction and the alkylated PAHs from the aromatic fraction The rhamnolipid MAT10 reduced the surface tension (cST) up to 26.8 mN/m and the interfacial tension (cIT) against hexadecane to 1 mN/m. The CMC of MAT10 was 150 mg/L. Geddes et al. (2008)
Mixed consortia of Pseudomonas aeruginosa strain PG201, Rhodococcus sp. H131A, and a Pseudomonas strain which produced the rhamnolipid Dyna270 Hexadecane, dodecane, benzene, toluene, iso-octane, pristane (2,6,10,14-tetramethyl pentadecane), naphthalene, and phenanthrene The rhamnolipid biosurfactants enhanced the rate of linear alkane biodegradation more than the biodegradation rate of the monoaromatics. Inakollu et al. (2004)
Rhodococcus baikonurensis EN3 Diesel oil These hydrocarbon solubilization studies for pure hydrocarbons and mixed hydrocarbon systems have indicated that the biosurfactant shows hydrocarbon specificity, with hydrocarbon substrate packed within the micelle core and is influenced by the size and shape of the hydrocarbon substrate in the mixed waste systems. Lee et al. (2006)
Rhodococcus sp. Pyrene The experimental kinetic data were fitted well with a mathematical model showing PAHs uptake both from the interface and from the aqueous medium by a population consisting of adsorbed cells in dynamic equilibrium with cells in the aqueous medium; interfacial uptake was predominant in these experiments. Bouchez et al. (1997)
Mycobacterium PYR-1 PAHs Results showed that the agitation rate affected cell growth and PAH degradation rates, while the substrate concentration did not; these are two characteristics of systems that exhibit an interfacial uptake mechanism. Moreover, detailed examination using fluorescence microscopy revealed that, in addition to associating with the aqueous-organic interface, this bacterium exists exclusively on the organic side of the interface. MacLeod and Daugulis (2005)
Candida lipolytica Hexadecane Interfacial tension between oil and water and Sauter mean drop size decreased as cultivation proceeded. Nakahara et al. (1977)
Candida lipolytica NRRL Y-6795
Candida intermedia IF0 0761
Candida tropicalis ATCC 20336
Saccharomyces cerevisiae HANSEN IF0 0305
n-Decane C. intermedia and C. tropicalis, which can utilize hydrocarbon, adhere well to hydrocarbon, but S. cerevisiae, which cannot utilize hydrocarbon, did not adhere. Miura et al. (1977)
Tab.1  Three different regulation mechanisms in the adsorption and uptake of hydrocarbons by bacteria
Fig.2  Schematic overview of the interactions between bacteria, soil, pollutants, and surfactants (Volkering et al., 1997).
Fig.3  Uptake of PAHs by Mycobacterium PYR-1 exanimated using fluorescence microscopy (MacLeod and Daugulis, 2005).
Fig.4  Effect of different inhibitors on the trans-membrane transport of 14C-fluoranthene by Rhodococcus sp. BAP-1 was added as an inhibitors at 0 min (a) and 5 min (b) (Li et al., 2014).
Type of transport Microbial species Hydrocarbon species Features and conclusions References
Active transport Pseudomonas putida Toluene The results show that the efflux system in P. putida S12 is specific for organic solvents and does not export antibiotics or other known substrates of multidrug-resistant pumps. Isken and De Bont (2000)
Pseudomonas putida KT2442 Toluene Solute molecules are transported across the cell membrane by consuming energy of an inverse concentration gradient with the participation of pump proteins. Fukumori et al. (1998)
Pseudomonas putida DOT-T1E 1,2,4-[C-14] Trichlorobenzene, toluene, xylenes, benzene The concentration of organic molecules at the cell membrane increased with the addition of energy inhibitors. Ramos et al. (1998)
Pseudomonas sp. DG17 Octadecane Addition of the energy inhibitor NaN3 inhibited the uptake of 14C-octadecane by Pseudomonas sp. DG17. Hua and Wang (2014)
Rhodococcus sp. BAP-1 Fluoranthene Demonstrated that the mechanism for fluoranthene travel across the cell membrane of Rhodococcus sp. BAP-1 requires energy Li et al. (2014)
Passive uptake and active efflux mechanism Mixed microbial consortium
Yersinia enterocolitica, Pseudomonas aeruginosa, Serratia liquefaciens, Pseudomonas fluorescens
Phenol This study was carried out to understand the effect of varying sublethal concentrations of phenol on isolated individual bacterial cultures. Sharma et al. (2002)
Assisted diffusion of ingestible material and an active efflux mechanism Pseudomonas fluorescens LP6a Phenanthrene The mechanism of transport of PAHs by Pseudomonas fluorescens LP6a, a PAM-degrading bacterium, was studied by inhibiting membrane transport and measuring the resulting change in cellular uptake.
The data were consistent with the presence of two conflicting transport mechanisms: uptake by passive diffusion and an energy-driven efflux system to transport PAHs out of the cell.
Bugg et al. (2000)
Tab.2  Passive and active transmembrane transport of hydrocarbons by bacteria
Fig.5  Structural features of TodX, TbuX, and FadL [TodX (left), TbuX (center), FadL (right)] (Ramos-González et al., 2002).
Fig.6  Structural features of OmpW (Hong et al., 2006).
Fig.7  Functional analysis of differentially expressed proteins in every cluster (Wang et al., 2019).
Fig.8  The number of upregulated and downregulated proteins in different KEGG-identified pathways (Wang et al., 2019).
Specific technology Microbial species Substances that interact with cells Features and conclusions References
iTRAQ Streptococcus suis Erythromycin Seventy-nine differentially expressed proteins were identified in sub-MIC erythromycin inhibiting planktonic cell.
Several cell surface proteins, as well as those involved in quorum-sensing, were found to be implicated in biofilm formation.
Zhao et al. (2015)
Rice suspension cells Pathogenic bacteria of bacterial blight (Xanthomonas oryzae pv. oryzae). Proteomics analysis of rice PM can be applied to identify potential PM components involved in the rice defense response to microbes. Chen et al. (2007)
Resistance to fusarium Pathogenic bacteria of head blight (FHB) The first report of the application of proteomic techniques in studying the interaction between a series of barley genotypes representing various levels of resistance to FHB and the F. graminearum pathogen. The induced plant defense responses following fungal infection were diverse among resistant, intermediate and susceptible barley genotypes. Geddes et al. (2008)
Madam Vinous’ sweet orange plants Pathogenic bacteria of citrus Huanglongbing (HLB) iTRAQ technology has been used to in identified 20 proteins and 10 differentially expressed proteins. These proteins may be good candidates for biomarkers to identify HLB-diseased plants prior to the expression of typical symptoms. However, similar changes in protein expression may be caused by other abiotic stresses, such as wounding and pathogen infections other than those induced by CLas. Fan et al. (2011)
Abomasal mucosa Nematode, Hemonchus contorts The study identified and quantified more than 4400 unique proteins, of which 158 proteins showed>1.5-fold difference between resistant and susceptible sheep. Trefoil factor 2, a member of RAS oncogene family (RAP1A) and ring finger protein 126 were among the proteins found to be highly abundant in the abomasal surface of resistant sheep, whereas adenosine deaminase and the gastrokine-3-like precursor were found at higher levels in susceptible sheep. Nagaraj et al. (2012)
E. chaffeensis Tick-borne rickettsial pathogen, Ehrlichia chaffeensis The study documents the impact of mutations on the global expression of pathogen protein and the influence of protein abundance on attenuation of mutations and protection of vertebrate host against infection. Kondethimmanahalli et al. (2019)
Rubrivivax benzoatilyticus JA2 Glucose The present study deciphered the molecular/metabolic events associated with glucose-grown cells of strain JA2 and unraveled how a carbon source modulates metabolic phenotypes. Proteomic profiling revealed extensive metabolic remodeling in the glucose-grown cells wherein signal transduction, selective transcription, DNA repair, protein transport and quality control processes were upregulated to cope with the changing milieu. Proteins involved in DNA replication, translation, electron transport, photosynthetic machinery were downregulated, possibly to conserve energy. Gupta et al. (2019)
Clostridium difficile Metronidazole This study provided an in-depth proteomic analysis of a stable, metronidazole-resistant C. difficile isolate. The results suggested that a multifactorial response may be associated with high level metronidazole resistance in C. difficile, including the possible roles in altered iron metabolism and/or DNA repair. Chong et al. (2014)
1-DE/LC-MS/MS Novosphingobium pentaromativorans US6-1 PAHs 1-DE/LC-MS/MS analysis of N. pentaromativorans US6-1 cultured in the presence of different PAHs and monocyclic aromatic hydrocarbons (MAHs) identified approximately 1,000 and 1,400 proteins, respectively. Yun et al. (2014)
2-DE Cyanobacterium Anabaena PD-1 Polychlorobiphenyl Twenty-five upregulated proteins were identified using 2-DE coupled with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). These proteins were involved in (i) PCB degradation, (ii) transport processes, (iii) energetic metabolism, (iv) electron transport, (v) general stress response, (vi) carbon metabolism, and (vii) nitrogen reductase. Zhang et al. (2014)
Tab.3  Proteomics technology
GO Protein sources Interference factors Protein functions References
Cellular components Synechocystis sp. PCC 6803 Butanol Cytoplasm, cytoplasmic part, protein transport, establishment of protein localization and protein localization. Tian et al., (2013)
Fillets treated by microwave thawing (MT) Magnetic nanoparticles plus microwave exposure Membrane, membrane par, cell, cell part, organelle, organelle part. Cao et al. (2019)
Vibrio parahemolyticus ATCC 17802 Food preservative and low temperature Cell outer membrane. Zhong et al. (2018)
Cotton Roots and Leaves Salt stress Cell, cell part, intracellular part, intracellular organelle, organelle-related components. Chen et al. (2016)
Aspergillus flavus Water activity Cell, cell part, extracellular region, extracellular region part. Zhang et al. (2015)
Rhodococcus sp. BAP-1 Fluoranthene Cell, macromolecular complex, membrane. Wang et al. (2019)
Molecular functions Synechocystis sp. PCC 6803 Butanol Ribonucleoprotein complex, macromolecular complex, structural molecule activity, RNA binding, nucleic acid binding, gene expression and macromolecule metabolic processes. Tian et al. 2013)
Fillets treated by microwave thawing (MT) Magnetic nanoparticles plus microwave Binding, catalytic activity, transporter activity. Cao et al. 2019)
Vibrio parahemolyticus ATCC 17802 Food preservative and low temperature Siderophore transmembrane transporter activity, receptor activity, iron ion binding, receptor activity. Zhong et al. (2018)
Rice (Oryza sativa) Embryogenesis Metabolic enzymes, binding (protein binding and nucleotide binding), transporters. Zi et al. (2013)
Aspergillus flavus Water activity Catalytic activity, binding, molecular transducer activity, nucleic acid binding transcription factor activity, nutrient reservoir activity, receptor activity, structural molecule activity. Zhang et al. (2015)
Rhodococcus sp. BAP-1 Fluoranthene Catalytic activity, binding followed by transporter activity. Wang et al. (2019)
Biological processes Synechocystis sp. PCC 6803 Butanol Inorganic cation transmembrane transporter activity, hydrogen ion transmembrane transporter activity, monovalent inorganic cation transmembrane transporter activity, intracellular protein transport. Tian et al. (2013)
Fillets treated by microwave thawing (MT) Magnetic nanoparticles plus microwave Biological regulation, cellular processes, regulation biological processes, response to stimuli and signaling, metabolic processes, developmental processes. Cao et al. (2019)
Vibrio parahemolyticus ATCC 17802 Food preservative and low temperature Choline transport, iron ion transport and homeostasis. Zhong et al. (2018)
Rice (Oryza sativa) Embryogenesis Proteins that participate in metabolism and respond to stimuli. Zi et al. (2013)
The resurrection plant Xerophyta viscosa Dehydration stress Cellular processes, binding. Abdalla and Rafudeen (2012)
Cotton Roots and Leaves Salt stress Metabolic processes, organic substance metabolic processes, primary metabolic processes, cellular metabolic processes. Chen et al. (2016)
Aspergillus flavus Water activity Metabolic processes, cellular processes, single-organism processes. Zhang et al. (2015)
Rhodococcus sp. BAP-1 Fluoranthene Metabolic processes, cellular processes, single-organism processes. Wang et al. (2019)
Tab.4  Functional classification of the combination of differentially expressed proteins based on GO
Fig.9  High resolution Nano-SIMS images of cultured BDG-3 (Su et al., 2016).
Specific technology Microbial species Features and conclusions References
FISH ? FISH not only enables the detection of culturable microorganisms but also yet-to-be cultured (so-called unculturable) organisms and can therefore help in understanding complex microbial communities. In this review, methodological aspects, as well as problems and pitfalls of FISH, are discussed in an examination of past, present and future applications. Moter and Göbel (2000)
Marine bacteria FISH with horseradish peroxidase (HRP)-labeled oligonucleotide probes and tyramide signal amplification, also known as catalyzed reporter deposition (CARD), is currently not generally applicable to heterotrophic bacteria in marine samples. The enhanced fluorescence intensities and signal-to-background ratios make CARD-FISH superior to FISH with directly labeled oligonucleotides for the staining of bacteria with low rRNA content in the marine environment.” (Pernthaler et al. (2002)
Bacterial populations in human feces FISH with group-specific 16S rRNA-targeted oligonucleotide probes. The combination of the two Bacteroides-specific probes detected a mean of 5.4 × 1010 cells·g?1 (dry weight) in feces; the Clostridium coccoides-Eubacterium rectale group-specific probe detected a mean of 7.2 × 1010 cells·g?1 (dry weight) in feces. The Clostridium histolyicum, Clostridium lituseburense, and Streptococcus-Lactococcus group-specific probes detected only numbers ranging from 1 × 107 to 7 × 108 cells·g?1 (dry weight) in feces. Franks et al. (1998)
Nano-SIMS E. coli E. coli exposed to food-grade TiO2 showed some internalization of TiO2 (7% of cells), as observed with high-resolution nano secondary ion mass spectrometry (nano-SIMS) chemical imaging. Radziwill-Bienkowska et al. (2018)
Spleen cells Nano-SIMS makes it possible to both capture images of and quantify molecules labeled with stable or radioactive isotopes within subcellular compartments. Lechene et al.(2006)
Individual bacteria within eukaryotic host cells In this study, with multi-isotope imaging mass spectrometry we directly captured images and measured nitrogen fixation by individual bacteria within eukaryotic host cells and demonstrated that fixed nitrogen is used for host metabolism. Lechene et al. (2007)
E. coli E. coli in water were treated with OD radicals, and D atom incorporation into cells was visualized using time-of-flight SIMS and nano-SIMS. The results show that D atoms from NTPJ reach the cytoplasm of E. coli in H2O, indicating the usefulness of this OD-tracking method for the study of radical interactions within living cells. Lee et al. (2014)
Paracoccus sp. strain HPD-2 Nano-SIMS results provided a direct evidence for the contribution of hematite to the formation of iron ions inside of HPD-2 cell, which resulted in the prominent generation of ROS (including EPFRs) that has been demonstrated to be able to affect cells. Gan et al. (2018)
Tab.5  Single cell analysis technology—FISH and Nano-SIMS
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