1.Molecular mechanisms of microbial mercury resistance and their prospective applications in remediation of mercury-contaminated soils.
Di WANG ; Huan LUO ; Xiaojun SHI ; Zhenlun LI ; Ying MA
Chinese Journal of Biotechnology 2025;41(4):1323-1339
Mercury (Hg)-contaminated soil poses a significant threat to the environment and human health. Hg-resistant microorganisms have the ability to survive under the stress of inorganic and organic Hg and effectively reduce Hg levels and toxicity. Compared to physical and chemical remediation methods, microbial remediation technologies have garnered increasing attention in recent years due to their lower cost, remarkable efficacy, and minimal environmental impact. This paper systematically elucidates the molecular mechanisms of Hg resistance in microbes, with a focus on their potential applications in phytoremediation of Hg-contaminated soils through plant-microbe interactions. Furthermore, it highlights the critical role of microbes in enhancing the effectiveness of transgenic plants for Hg remediation, aiming to provide a theoretical foundation and scientific basis for the bioremediation of Hg-contaminated soils.
Mercury/toxicity*
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Biodegradation, Environmental
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Soil Pollutants/isolation & purification*
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Soil Microbiology
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Plants, Genetically Modified/metabolism*
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Bacteria/genetics*
2.Serratia marcescens TF-1 for biodegradation of chlorobenzene contaminants in soil and its application in in-situ remediation of chemical industrial sites.
Fang GOU ; Yunchun SHI ; Hao CHEN ; Wenting FU ; Liangjie LI ; Zhilin XING ; Jiangfeng GUO
Chinese Journal of Biotechnology 2025;41(6):2483-2497
Chlorobenzene contaminants (CBs) pose a threat to the eco-environment, and functional strains hold considerable potential for the remediation of CB-contaminated sites. To deeply explore the application potential of functional bacteria in the in-situ bioremediation of CBs, this study focused on the biodegradation characteristics and degradation kinetics of CB and 1, 2-dichlorobenzene (1, 2-DCB) in soil by the isolated strain Serratia marcescens TF-1. Additionally, an in-situ remediation trial was conducted with this strain at a chemical industrial site. Batch serum bottle experiments showed that the degradation rate of CB at the concentrations ranging from 20 to 200 mg/L by TF-1 was 0.22-0.66 mol/(gcell·h), following the Haldane model, with the optimal concentration at 23.12 mg/L. The results from simulated soil degradation experiments indicated that the combined use of TF-1 and sodium succinate (SS) significantly enhanced the degradation of CBs, with the maximum degradation rate of CB reaching 0.104 d-1 and a half-life of 6.66 d. For 1, 2-DCB, the maximum degradation rate constant was 0.068 7 d-1, with a half-life of 10.087 d. The in-situ remediation results at the chemically contaminated site demonstrated that the introduction of bacterial inoculant and SS significantly improved the removal of CBs, achieving the removal rates of 84.2%-100% after 10 d. CB, 1, 4-dichlorobenzene (1, 4-DCB), and benzo[a]pyrene were completely removed. Microbial diversity analysis revealed that the in-situ remediation facilitated the colonization of TF-1 and the enrichment of indigenous nitrogen-fixing Azoarcus, which may have played a key role in the degradation process. This study provides a theoretical basis and practical experience for the in situ bioremediation of CBs-contaminated sites.
Chlorobenzenes/isolation & purification*
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Biodegradation, Environmental
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Soil Pollutants/isolation & purification*
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Serratia marcescens/metabolism*
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Industrial Waste
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Soil Microbiology
3.Transgenic plant regeneration of tobacco (Nicotiana tabacum) haboring mammalian cyp2e1 gene.
Peihan LI ; Taihe XIANG ; Jun XIE ; Ting FENG ; Wenyi LU
Chinese Journal of Biotechnology 2012;28(10):1195-1204
CYP2E1 enzyme encoded by cyp2e1 gene plays an important role in metabolism of heterogeneous organics in mammalian liver cells. The transgenic plant with cyp2e1 can metabolize various low molecular weight organic pollutants. However, it is unclear the mechanism of expression control of cyp2e1 in transgenic plant. In this study, plasmid pSLD50-6 with cyp2e1 and pKH200 with gus as control were transformed into Agrobacterium tumefaciens GV3101 separately. Then, the cyp2e1 or gus genes were transferred into tobacco (Nicotiana tabacum) and the transgenic plants were regenerated via Agrobacterium tumefaciens method. Real-time quantitative PCR (qRT-PCR) was used to analyze the cyp2e1 gene expression. The expression of cyp2e1 in transgenic tobacco with cyp2e1 decreased obviously treated by ethyl alcohol and reduced slightly by benzene and toluene, while it enhanced by acetone, formaldehyde and oxygen deficit in different levels. In addition, the gene expression of NADPH-P450 oxidoreductase and cytochrome b5 enzyme in the transgenic tobacco with cyp2e1 were increased significantly treated by benzene, which showed that NADPH-P450 oxidoreductase and cytochrome b5 enzyme in transgenic tobacco have relation with CYP2E1 detoxication process. It suggested that the NADPH-P450 oxidoreductase and cytochrome b5 enzyme in transgenic plant formed the requirement in mammalian and participated in the electron transport chain of CYP2E1 enzyme catalytic process.
Agrobacterium tumefaciens
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genetics
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Animals
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Cytochrome P-450 CYP2E1
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biosynthesis
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genetics
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Gene Expression Regulation, Enzymologic
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genetics
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Gene Transfer Techniques
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Genetic Vectors
;
genetics
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Plants, Genetically Modified
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genetics
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Soil Pollutants
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isolation & purification
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Tobacco
;
enzymology
;
genetics
;
Transfection
4.Impact of anthracene addition on microbial community structure in soil microcosms from contaminated and uncontaminated sites.
QingFeng WANG ; ShuYing ZHANG ; Li ZOU ; ShuGuang XIE
Biomedical and Environmental Sciences 2011;24(5):543-549
OBJECTIVEThis paper aims to investigate the impact of anthracene addition on microbial community in agricultural soil irrigated with tap water or reclaimed wastewater.
METHODSThe changes of microbial community were characterized by terminal restriction fragment length polymorphism in combination with 16S rRNA gene clone library analysis.
RESULTSA significant change in microbial community composition was observed during the biodegradation of anthracene, with dominantly enriched members from the genus Methylophilus.
CONCLUSIONThis work might be useful for developing techniques for the isolation of novel putative PAH degrader.
Anthracenes ; analysis ; Bacteria ; classification ; genetics ; isolation & purification ; Biodegradation, Environmental ; DNA, Bacterial ; genetics ; isolation & purification ; Hazardous Waste ; Phylogeny ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S ; genetics ; Soil Microbiology ; Soil Pollutants ; analysis ; Waste Disposal, Fluid ; methods
5.Effect of cadmium, alone or in combination with CaCl2, on the growth, antioxidative enzyme activity and cadmium absorption of Solanum nigrum L. var pauciflorum hairy roots.
Heping SHI ; Eric Pokeung TSANG ; Yunling WANG ; Andrew Leewah CHAN
Chinese Journal of Biotechnology 2010;26(2):147-158
To study if Solanum nigrum hairy roots can be used for phytoremediation of Cd contamination, we investigated the effects of cadmium (Cd) alone, and in combination with different concentrations of CaCl2, on growth, activities of superoxide dismutase (SOD) and peroxidase (POD) and Cd absorption by hairy roots of S. nigrum L. var pauciflorum. The results showed that Cd concentrations of lower than 50 micromol/L enhanced the growth of hairy roots, while higher than 100 micromol/L inhibited growth and decreased the number of branched roots, also causing the root tips to become brown and shorter in length. In comparison with a control, the soluble protein content, the activities of SOD and POD in hairy roots cultures showed a trend of first increased and then gradually decreased, while the malondialdehyde (MDA) content significantly increased, when increasing the Cd concentrations. Cd concentration of 100 micromol/L or 300 micromol/L in combination with 10-30 mmol/L CaCl2 resulted in a decreased content of soluble protein and MDA in the hairy roots, but an enhanced SOD activity. The increased POD activities were observed when cultured in 100 micromol/L Cd and 10-30 mmol/L CaCl2 but decreased when cultured in 300 micromol/L Cd and 10-30 mmol/L CaCl2. Atomic Absorption Spectrometry determination showed that the Cd absorbed and adsorbed by the hairy roots increased along with the increase of Cd concentration. The exogenous addition of 10-30 mmol/L CaCl2 could reduce the toxicity of Cd. This was achieved on one hand by reducing the absorption of Cd, on the other hand by decreasing the lipid peroxidation through regulating the activities of antioxidant enzymes SOD and POD in the hairy roots.
Absorption
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Biodegradation, Environmental
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Cadmium
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isolation & purification
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metabolism
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Calcium Chloride
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metabolism
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Peroxidase
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metabolism
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Plant Roots
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growth & development
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physiology
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Soil Pollutants
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isolation & purification
;
metabolism
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Solanum nigrum
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enzymology
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growth & development
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physiology
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Superoxide Dismutase
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metabolism
6.Influence of vermicomposting on solid wastes decomposition kinetics in soils.
Journal of Zhejiang University. Science. B 2007;8(10):725-730
The effect of vermicomposting on kinetic behavior of the products is not well recognized. An incubation study was conducted to investigate C mineralization kinetics of cow manure, sugarcane filter cake and their vermicomposts. Two different soils were treated with the four solid wastes at a rate of 0.5 g solid waste C per kg soil with three replications. Soils were incubated for 56 d. The CO(2)-C respired was monitored periodically and a first-order kinetic model was used to calculate the kinetic parameters of C mineralization. Results indicated that the percentage of C mineralized during the incubation period ranged from 31.9% to 41.8% and 55.9% to 73.4% in the calcareous and acidic soils, respectively. The potentially mineralizable C (C(0)) of the treated soils was lower in the solid waste composts compared to their starting materials. Overall, it can be concluded that decomposable fraction of solid wastes has decreased due to vermicomposting.
Industrial Waste
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prevention & control
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Kinetics
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Refuse Disposal
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methods
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Soil
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analysis
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Soil Pollutants
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chemistry
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isolation & purification
7.Influences of copper speciation on toxicity to microorganisms in soils.
Shu-Juan SUN ; Jian XU ; Shu-Gui DAI ; Xue HAN
Biomedical and Environmental Sciences 2006;19(6):409-413
OBJECTIVETo investigate the relationship between copper speciation and microbial features (microbial communities and copper tolerance level) in order to determine the adverse effect of different forms of Cu on microorganisms.
METHODSTessier's sequential extraction procedure was used to qualify the different Cu forms (exchangeable, carbonate bound, Fe/Mn oxide bound, residue and organic matter bound), and the copper tolerance level (expressed as IC50, influence concentration) was measured by the plate-count method.
RESULTSBy simple correlation analysis, the IC50 was positively correlated with the concentration of exchangeable Cu (R2 = 0.8204), while weakly correlated with other forms of Cu.
CONCLUSIONThe bacterial community tolerance increases in the copper-contaminated soil while sensitive bacteria decrease in the copper-contaminated soils. The exchangeable Cu exerts high toxicity to microbial communities.
Bacteria ; drug effects ; isolation & purification ; China ; Copper ; analysis ; chemistry ; toxicity ; Environmental Monitoring ; Fungi ; drug effects ; isolation & purification ; Hydrogen-Ion Concentration ; Soil Microbiology ; Soil Pollutants ; chemistry ; toxicity
8.Mycorrhizoremediation--an enhanced form of phytoremediation.
Journal of Zhejiang University. Science. B 2006;7(7):503-514
Study of plant roots and the diversity of soil micro biota, such as bacteria, fungi and microfauna associated with them, is important for understanding the ecological complexities between diverse plants, microbes, soil and climates and their role in phytoremediation of contaminated soils. The arbuscular mycorrhizal fungi (AMF) are universal and ubiquitous rhizosphere microflora forming symbiosis with plant roots and acting as biofertilizers, bioprotactants, and biodegraders. In addition to AMF, soils also contain various antagonistic and beneficial bacteria such as root pathogens, plant growth promoting rhizobacteria including free-living and symbiotic N-fixers, and mycorrhiza helping bacteria. Their potential role in phytoremediation of heavy metal (HM) contaminated soils and water is becoming evident although there is need to completely understand the ecological complexities of the plant-microbe-soil interactions and their better exploitation as consortia in remediation strategies employed for contaminated soils. These multitrophic root microbial associations deserve multi-disciplinary investigations using molecular, biochemical, and physiological techniques. Ecosystem restoration of heavy metal contaminated soils practices need to incorporate microbial biotechnology research and development. This review highlights the ecological complexity and diversity of plant-microbe-soil combinations, particularly AM and provides an overview on the recent developments in this area. It also discusses the role AMF play in phytorestoration of HM contaminated soils, i.e. mycorrhizoremediation.
Biodegradation, Environmental
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Heavy Metal Poisoning, Nervous System
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metabolism
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Plant Roots
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metabolism
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microbiology
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Soil Microbiology
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Soil Pollutants
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isolation & purification
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pharmacokinetics
9.Screening of a triazophos-degrading strain and pathway of its degradation.
Li-Hong WANG ; Lin ZHANG ; Huan-Lin CHEN
Chinese Journal of Biotechnology 2005;21(6):954-959
A Triazophos-degrading strain, Klebsiella sp. E6, was identified by soil enrichment technology from the soil sampled from the vicinity of a factory manufacturing Triazophos (TAP). The nutrient requirement of the strain is simple. It can use TAP as the sole sources of carbon, nitrogen and phosphorus. Comparison of the degradation rates revealed that the strain degraded TAP most effectively when TAP was used as a sole nitrogen source. No inhibition effect occurred when TAP concentration was high as 1000 mg/L in the case of TAP was used as the sole nitrogen source. Analysis of the intermediates of TAP metabolism indicated that TAP is firstly hydrolyzed into 1-phenyl-3-hydroxy-1,2,4-triazole and O,O-diethyl phosphorothioic acid. 1-phenyl-3-hydroxy-1,2,4-triazole was further mineralized into inorganic compounds. A degradation pathway of TAP was proposed. The experiment results demonstrated that the strain has potential in biodegradation of TAP pollutions.
Biodegradation, Environmental
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Environmental Pollutants
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metabolism
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Klebsiella
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isolation & purification
;
metabolism
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Organothiophosphates
;
metabolism
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Pesticides
;
metabolism
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Soil Microbiology
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Triazoles
;
metabolism
10.Isolation of 2,4-dichlorophenol degrading bacterium strain and cloning and expression of its 2,4-dichlorophenol hydroxylase gene.
Wen-Hui ZHONG ; Ming SUN ; Guo-Qing HE ; Xiao-Shan FENG ; Zi-Niu YU
Chinese Journal of Biotechnology 2004;20(2):209-214
2,4-Dichlorophenol is toxic and biorefratory organic pollutant. A 2,4-dichlorophenol degrading bacterial strain GT241-1, identified as Pseudomonas sp., was isolated from soil samples which was collected from drainage area of several 2,4-dichlorophenol producing factories. Strain GT241-1 had strong 2,4-dichlorophenol degrading ability, it could decompose 91% 2, 4-dichlorophenol of 90 mg/L within 48 hours at 25 - 30 degrees C, and could utilize 2,4-dichlorophenol, 2,4-dichlorophenoxyacetic acid (2,4-D), benzoate and catechol as sole carbon and energy source. Southern blot showed that 2,4-dichlorophenol hydroxylase gene (dcpA) of strain GT241-1 locates on the about 10kb EcoR I/Xba I fragment. This fragment was recovered, linked to the vecter pUC19 and transformed into the E. coli DH5alpha. A aim transformant, Z539, was obtained by dot blotting from about 1200 transformants. PCR and the sequencing results shew that the whole dcpA gene is contained within the 10kb EcoR I /Xba I fragment of pZ539. This fragment was shortened to about 2.4kb by HindmIII. The shorted fragment was subcloned to vecter pRSET-B to get a transformant BS1-12. The subcloned fragment was sequenced. Sequencing results showed that the whole length of the subcloned fragment containing dcpA is 2389bp and the nucleotide span of coding region is from number 276 to number 2072 (1797 bp), with ATG and TAA as start and stop codon respectively. The sequence analysis of dcpA and the deduced amino acid encoded by dcpA showed that they are different from the relative sequences registered in the GenBank. The subcloned fragment carry the promoter of dcpA, this can deduce from the fact that the upflow length of dcpA coding region is 275bp, and further confirmed by the 2,4-dichlorophenol hydroxylase activity measurement results. The 2,4-dichlorophenol hydroxylase activity of transformant Z539 and BS1-12 were detected, the results showed these transformants have 2,4-dichlorophenol hydroxylase activity. By comparison, the activity of these transformants were lower than that of the strain GT241-1.
Amino Acid Sequence
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Bacterial Proteins
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genetics
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metabolism
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Biodegradation, Environmental
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Chlorophenols
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metabolism
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Cloning, Molecular
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Environmental Pollutants
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metabolism
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Mixed Function Oxygenases
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genetics
;
metabolism
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Molecular Sequence Data
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Pseudomonas
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enzymology
;
genetics
;
isolation & purification
;
Soil Microbiology

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