Strain Burkholderia Sp Research Articles

Polysaccharides Produced by Plant Growth-Promoting Rhizobacteria Strain Burkholderia sp. BK01 Enhance Salt Stress Tolerance to Arabidopsis thaliana.

Salt stress is one of the most serious abiotic stresses leading to reduced agricultural productivity. Polysaccharides from seaweed have been used as biostimulants to promote crop growth and improve plant resistance to abiotic stress. In this study, PGPR strain Burkholderia sp. BK01 was isolated from the rhizosphere of wheat, and it was characterized for phosphorus (Pi) dissolution, indole-3-acetic acid (IAA) production, ammonia (NH3) and exopolysaccharides (EPS). In particular, strain BK01 can efficiently produce extracellular polysaccharide with a yield of 12.86 g/L, using sorbitol as carbon source. BK01 EPS was identified as an heteropolysaccharide with Mw 3.559 × 106 Da, composed of (D)-galactose (75.3%), (D)-glucose (5.5%), (L)-rhamnose (5.5%), (D)-galactouronic acid (4.9%) and (D)-glucuronic acid (8.8%). The present work aims to highlight the effect of the BK01 EPS on growth and biochemical changes in Arabidopsis thaliana under salt stress (100 mM). The purified BK01 EPS at a concentration of 100 mg/L efficiently promoted the growth of plants in pot assays, improved the chlorophyll content, enhanced the activities of SOD, POD and CAT, and decreased the content of MDA. This results suggested that the polysaccharides produced by PGPR strain Burkholderia sp. BK01 can be used as biostimulants to promote plant growth and improve plant resistance to salt stress.

Production of biohydrogen and green platform compound 2, 5-furandicarboxylic acid using rice straw hydrolysate

Lignocellulosic hydrolysate contains several inhibitors that hinder downstream biological processes. The immobilized strain Burkholderia sp. H-2 was used in this study to biodetoxify real rice straw hydrolysate and biotransform 5-hydroxymethylfurfural (5-HMF), an inhibitor, into 2, 5-furandicarboxylic acid (FDCA), a valuable platform compound. An electrodialysis with bipolar membranes (EDBM) system was applied to recover FDCA and perform secondary detoxification of the biodetoxified real rice straw hydrolysate. The rice straw hydrolysate after biodetoxification and EDBM treatment was used for anaerobic dark fermentative hydrogen production. The immobilized Burkholderia sp. H-2 could completely biotransform 1843 mg/L 5-HMF into 1532 mg/L FDCA within 24 h when the 2-fold diluted rice straw hydrolysate was used as the feedstock. The optimal EDBM system parameters to recover FDCA were a hydrolysate pH of 6, operated at a current density of 17.86 mA/cm2. The FDCA recovery efficiency was 44.6 %, when using the biodetoxified real rice straw hydrolysate as the EDBM feedstock. The rice straw hydrolysate after biodetoxification and EDBM treatment had better biohydrogen production performance than the undetoxified and biodetoxified rice straw hydrolysates.

Rapid Biodegradation of the Organophosphorus Insecticide Acephate by a Novel Strain Burkholderia sp. A11 and Its Impact on the Structure of the Indigenous Microbial Community.

The acephate-degrading microbes that are currently available are not optimal. In this study, Burkholderia sp. A11, an efficient degrader of acephate, presented an acephate-removal efficiency of 83.36% within 56 h (100 mg·L-1). The A11 strain has a broad substrate tolerance and presents a good removal effect in the concentration range 10-1600 mg·L-1. Six metabolites from the degradation of acephate were identified, among which the main products were methamidophos, acetamide, acetic acid, methanethiol, and dimethyl disulfide. The main degradation pathways involved include amide bond breaking and phosphate bond hydrolysis. Moreover, strain A11 successfully colonized and substantially accelerated acephate degradation in different soils, degrading over 90% of acephate (50-200 mg·kg-1) within 120 h. 16S rDNA sequencing results further confirmed that the strain A11 gradually occupied a dominant position in the soil microbial communities, causing slight changes in the diversity and composition of the indigenous soil microbial community structure.

A novel bacterial strain Burkholderia sp. F25 capable of degrading diffusible signal factor signal shows strong biocontrol potential.

Vast quantities of synthetic pesticides have been widely applied in various fields to kill plant pathogens, resulting in increased pathogen resistance and decreased effectiveness of such chemicals. In addition, the increased presence of pesticide residues affects living organisms and the environment largely on a global scale. To mitigate the impact of crop diseases more sustainably on plant health and productivity, there is a need for more safe and more eco-friendly strategies as compared to chemical prevention. Quorum sensing (QS) is an intercellular communication mechanism in a bacterial population, through which bacteria adjust their population density and behavior upon sensing the levels of signaling molecules in the environment. As an alternative, quorum quenching (QQ) is a promising new strategy for disease control, which interferes with QS by blocking intercellular communication between pathogenic bacteria to suppress the expression of disease-causing genes. Black rot caused by Xanthomonas campestris pv. campestris (Xcc) is associated with the diffusible signal factor (DSF). As detailed in this study, a new QQ strain F25, identified as Burkholderia sp., displayed a superior ability to completely degrade 2 mM of DSF within 72h. The main intermediate product in the biodegradation of DSF was identified as n-decanoic acid, based on gas chromatography-mass spectrometry (GC-MS). A metabolic pathway for DSF by strain F25 is proposed, based on the chemical structure of DSF and its intermediates, demonstrating the possible degradation of DSF via oxidation-reduction. The application of strain F25 and its crude enzyme as biocontrol agents significantly attenuated black rot caused by Xcc, and inhibited tissue maceration in the host plant Raphanus sativus L., without affecting the host plant. This suggests that agents produced from strain F25 and its crude enzyme have promising applications in controlling infectious diseases caused by DSF-dependent bacterial pathogens. These findings are expected to provide a new therapeutic strategy for controlling QS-mediated plant diseases.

Microplastic accelerate the phosphorus-related metabolism of bacteria to promote the decomposition of methylphosphonate to methane

Microplastic (MP) contaminants in marine water have become a global public health concern because of their persistence and potentially adverse effects on organisms. MP can affect the growth and metabolism of marine microorganisms and further impact the microbial environmental functions. The molecular impact mechanisms of MP on specific functional microbes with the capability of decomposing methylphosphonate (MPn) to release methane (CH4) in oxygenated water have rarely been reported upon. Herein, we investigated the effects of MP on microbes and concomitant methanogenesis via the microbial degradation of MPn. Furthermore, the specific perturbation was revealed at the molecular level combined with transcriptomics and metabolomics. The results showed that intracellular phosphorus utilization by MPn-degrading strain Burkholderia sp. HQL1813 was enhanced by accelerating the catabolism of MPn. Phosphorus transport-related genes (phnG-M, pstSCAB, phnCDE) were upregulated in the MP exposure groups. Amino acid metabolism, the phosphotransferase system and nucleotide metabolism were also perturbed after MP exposure. Notably, released CH4 increased by 24 %, 29 % and 14 % in the exposure group. In addition, the responses of the strain were dose-independent with increasing MP doses. These findings are beneficial for clarifying the effect of MP on specific functional microbes at the molecular level and their degradation of CH4 by MPn.

Cornstalk biochar promoted the denitrification performance and cellulose degradation rate of Burkholderia sp. CF6

A strain Burkholderia sp. CF6 with denitrification and cellulose degradation ability was isolated. When sodium carboxymethyl cellulose (CMC-Na) dosage was 5 g L−1, the nitrate removal efficiency reached 96.48% after 48 h and the nitrite accumulation was only 0.07 mg L−1. The exponential decay equation was used to match the kinetics of denitrification and the correlation coefficient (R2) was higher than 0.940. The denitrification and cellulose degradation capabilities were promoted by cornstalk biochar. Through the comparison of nitrogen balance of different biochars, biochar of 400 ℃ (BC400) had the highest gaseous nitrogen conversion efficiency (83.31%). BC400 can accelerate the nitrate removal rate (from 0.08 to 0.32 mg L−1 h−1) and shorten the peak time of cellulase activity (from 48 to 24 h). The fourier transform infrared spectroscopy (FTIR) manifested that the BC400 assisted biological denitrification was achieved through phenolic and quinone groups. From the results of three-dimensional excitation-emission matrix (3D-EEM), it can be concluded that soluble microbial by-product and aromatic protein have a significant correlation with the effect of BC400 on the biological process of strain CF6. This research will provide new insights for agricultural waste utilization to treat low carbon to nitrogen (C/N) ratio wastewater.

Biovalorization of lignin derived compounds with molasses as co-substrate for polyhydroxyalkanoate production

The bioconversion of 15 lignin-derived compounds (LDCs) into polyhydroxyalkanoate (PHA) was studied in the present study. PHA accumulating strain Burkholderia sp. ISTR5 was employed for the bioconversion of LDCs. Burkholderia sp. ISTR5 (R5) efficiently bioconverted LDCs into PHA. Due to low PHA yield on LDCs, the co-metabolism with agro-industrial waste molasses was studied, and it showed a considerable increase in the PHA production. With the supplementation of molasses, ∼ 20% increment in PHA production was observed on different LDCs. The highest increase in PHA percentage (w/w cell dry weight) after the addition of molasses was observed on 4-hydroxybenzoic acid (4-HBA) by 23%, followed by benzoic acid (22%) and gallic acid (21%). The Highest PHA accumulation was observed on p-coumaric acid (270 mg/L) followed by 4-HBA (260 mg/L) and gallic acid (256 mg/L). Produced PHA was characterized to be poly (hydroxybutyrate-co-hydroxyvalerate) (PHBV). The co-metabolism methodology considerably increased the PHA production and will be beneficial for setting up an agro-based efficient biorefinery. • Burkholderia sp. ISTR5 established for lignin derived compounds biovalorization. • Lignin derived compounds degradation resulted in the production of polyhydroxyalkanoate as major product. • Molasses as co-substrate played a crucial role for enhancement of PHA production. • Biovalorization of lignin derived compounds was presented with agro-industrial waste as co-substrate.

Bioconversion of Guaiacylglycerol-β-guaiacyl ether into vanillin by Burkholderia sp. ISTR5: A genomic and proteomic approach

Long-term research on biofuel production from lignocellulose did not progress much due to strong inter-unit linkages in lignin structure. Lignin contains β-O-4 linkage as a major bond (50–60%) between aromatic compounds to form a complex 3D-structure. The present study aims for the characterization of a novel strain Burkholderia sp. ISTR5 (R5) for β-O-4 linkage disintegration. Guaiacylglycerol-β-guaiacyl ether (GGE) was taken as a representative compound for β-O-4 linkage analysis. We showed that the strain R5 completely degraded GGE in 72 h. Further, gas chromatography-mass spectrometry (GC-MS) profile showed that vanillin is one of the highest produced chemicals during the degradation of GGE with small fractions of other metabolites in the reaction mixture. Nuclear Magnetic Resonance (NMR) analysis demonstrated the fragmentation of β-O-4 bonds and the advent of new chemical bonds. In addition to it, the whole genome analysis showed that R5 contains a superior enzyme chassis for the degradation of aromatic compounds. Enzymes such as glutathione-s-transferase, biphenyl dioxygenase, catalase/peroxidase, hydrolases, and dehydrogenases dominated the genome of R5. Proteomics analysis showed 2 glutathione lyase, 1 glutathione-s-transferase, 1 GMC oxidoreductase, 2 FAD binding oxidoreductase and several hydrolases, peroxidases, and transferases were upregulated on GGE. This study showed the capability of R5 to degrade and bioconvert the β-O-4 linkage lignin model compound GGE into vanillin.

Polyhydroxyalkanoate synthesis and characterization: A proteogenomic and process optimization study for biovalorization of industrial lignin

The strain Burkholderia sp. ISTR5 (R5) was studied for polyhydroxyalkanoate (PHA) production on Kraft lignin (KL) and lignosulfonate (LS) as substrates. During the initial screening, the maximum PHA mass fraction in biomass produced on KL and LS was 23% and 18%, respectively, at 96 h. PHA production on KL was further optimized using the Box-Behnken Design (BBD) model of Response Surface Methodology (RSM). After optimization, a 42.5% increase in PHA production and a 32.2% increase in the total cell biomass was observed. PHA was characterized by GC–MS, TEM, FTIR, NMR, and fluorescence microscopy. It was found to be a small chain length PHA with a copolymer of poly (hydroxybutyrate-co-hydroxyvalerate) (PHBV). The degradation of PHBV was also studied using this strain; it was observed that R5 completely degraded PHBV in 120 h. Genomic and proteomic analysis of R5 revealed numerous enzymes for the metabolism of lignin degradation and PHA production.

Complete Genome Sequence of a Chlorobenzene Degrader, Pandoraea pnomenusa MCB032.

Chlorobenzenes are ubiquitously distributed, highly persistent, and toxic environmental contaminants. Pandoraea pnomenusa MCB032 was isolated as a new dominant chlorobenzene-utilizing strain from a functionally stable bioreactor during the treatment of chlorobenzenes when strain Burkholderia sp. JS150 disappeared. In study, we report the complete genome sequence of strain MCB032 which consists of a circular chromosome and three plasmids, which are ~ 6Mb in length with 5450 open reading frames-12 encoding rRNAs and 77 encoding tRNAs. We further identified 17 putative genes encoding the enzymes involved in the methyl-accepting chemotaxis proteins in sensing chemical gradients during chemotaxis. The annotated complete genome sequence of this strain will provide genetic insights into the degradation of chlorinated aromatic compounds. The information will empower the elucidation of chlorobenzene affinity hierarchy and species succession in the bioreactor.

Characterization and functional gene analysis of a newly isolated indole-degrading bacterium Burkholderia sp. IDO3.

Indole is a common N-heterocyclic pollutant as well as a signaling molecule widespread in various environmental matrices. Several bacterial strains have been reported to be able to degrade indole, while the degradation capacity and functional enzymes are poorly documented. Herein, the degradation characteristics of a newly-isolated indole-degrading strain Burkholderia sp. IDO3 were carefully investigated. Strain IDO3 exhibited superior degradation ability which could completely remove 100 mg/L indole within 14 h in mineral salt medium. It maintained comparable degradation performance under conditions of pH 4.0-9.0, temperature 25-35 °C and rotary speed 0-250 r/min, and most of the tested heavy metals and organic pollutants did not significantly affect the degradation process. Two important intermediates, i.e. isatin and anthranilate, were identified in indole degradation process. The genomic clone library technique with indigo-based screening method was successfully applied to screen the functional genes. Heterologous expression assay proved that recombinant E. coli strain carrying indole oxygenase and reductase genes iifCD could transform indole to indigo. Bioinformatic analyses indicated that the newly obtained enzyme IifC_IDO3 formed a phylogenetically separate branch from other related aromatic oxygenases. This study provides new insights into our understanding of indole degradation by Burkholderia strains and offers efficient bacterial resource for indole bioremediation.

Catabolism of 2-Hydroxypyridine by Burkholderia sp. Strain MAK1: a 2-Hydroxypyridine 5-Monooxygenase Encoded by hpdABCDE Catalyzes the First Step of Biodegradation.

Microbial degradation of 2-hydroxypyridine usually results in the formation of a blue pigment (nicotine blue). In contrast, the Burkholderia sp. strain MAK1 bacterium utilizes 2-hydroxypyridine without the accumulation of nicotine blue. This scarcely investigated degradation pathway presumably employs 2-hydroxypyridine 5-monooxygenase, an elusive enzyme that has been hypothesized but has yet to be identified or characterized. The isolation of the mutant strain Burkholderia sp. MAK1 ΔP5 that is unable to utilize 2-hydroxypyridine has led to the identification of a gene cluster (designated hpd) which is responsible for the degradation of 2-hydroxypyridine. The activity of 2-hydroxypyridine 5-monooxygenase has been assigned to a soluble diiron monooxygenase (SDIMO) encoded by a five-gene cluster (hpdA, hpdB, hpdC, hpdD, and hpdE). A 4.5-kb DNA fragment containing all five genes has been successfully expressed in Burkholderia sp. MAK1 ΔP5 cells. We have proved that the recombinant HpdABCDE protein catalyzes the enzymatic turnover of 2-hydroxypyridine to 2,5-dihydroxypyridine. Moreover, we have confirmed that emerging 2,5-dihydroxypyridine is a substrate for HpdF, an enzyme similar to 2,5-dihydroxypyridine 5,6-dioxygenases that are involved in the catabolic pathways of nicotine and nicotinic acid. The proteins and genes identified in this study have allowed the identification of a novel degradation pathway of 2-hydroxypyridine. Our results provide a better understanding of the biodegradation of pyridine derivatives in nature. Also, the discovered 2-hydroxypyridine 5-monooxygenase may be an attractive catalyst for the regioselective synthesis of various N-heterocyclic compounds.IMPORTANCE The degradation pathway of 2-hydroxypyridine without the accumulation of a blue pigment is relatively unexplored, as, to our knowledge, no genetic data related to this process have ever been presented. In this paper, we describe genes and enzymes involved in this little-studied catabolic pathway. This work provides new insights into the metabolism of 2-hydroxypyridine in nature. A broad-range substrate specificity of 2-hydroxypyridine 5-monooxygenase, a key enzyme in the degradation, makes this biocatalyst attractive for the regioselective hydroxylation of pyridine derivatives.

Endogenous Stress Caused by Faulty Oxidation Reactions Fosters Evolution of 2,4-Dinitrotoluene-Degrading Bacteria

Environmental strain Burkholderia sp. DNT mineralizes the xenobiotic compound 2,4-dinitrotoluene (DNT) owing to the catabolic dnt genes borne by plasmid DNT, but the process fails to promote significant growth. To investigate this lack of physiological return of such an otherwise complete metabolic route, cells were exposed to DNT under various growth conditions and the endogenous formation of reactive oxygen species (ROS) monitored in single bacteria. These tests revealed the buildup of a strong oxidative stress in the population exposed to DNT. By either curing the DNT plasmid or by overproducing the second activity of the biodegradation route (DntB) we could trace a large share of ROS production to the first reaction of the route, which is executed by the multicomponent dioxygenase encoded by the dntA gene cluster. Naphthalene, the ancestral substrate of the dioxygenase from which DntA has evolved, also caused significant ROS formation. That both the old and the new substrate brought about a considerable cellular stress was indicative of a still-evolving DntA enzyme which is neither optimal any longer for naphthalene nor entirely advantageous yet for growth of the host strain on DNT. We could associate endogenous production of ROS with likely error-prone repair mechanisms of DNA damage, and the ensuing stress-induced mutagenesis in cells exposed to DNT. It is thus plausible that the evolutionary roadmap for biodegradation of xenobiotic compounds like DNT was largely elicited by mutagenic oxidative stress caused by faulty reactions of precursor enzymes with novel but structurally related substrates-to-be.

Филогенетический анализ генов деградации нафталина и фенантрена у штаммовBurkholderiasp.

The genetic systems responsible for naphthalene and phenanthrene catabolism have been analyzed in the five strains of Burkholderia sp. isolated from soil samples (West Siberia) contaminated by heavy residual fuel oil and in the strain Burkholderia sp. BS3702 from the laboratory collection isolated from soil samples of the coke gas works (Vidnoe, Moscow oblast). The results of this work demonstrate that naphthalene and phenanthrene degradation in the above strains is encoded by the sequences not homologous to the classical nah genes of pseudomonades. In the Burkholderia sp. BS3702 strain, the initial stages of phenanthrene degradation and the subsequent stages of salicylate degradation are controlled by the sequences of different evolutionary origins (phn and nag genes).

Immobilization of Burkholderia sp. lipase on a ferric silica nanocomposite for biodiesel production

In this work, lipase produced from an isolated strain Burkholderia sp. C20 was immobilized on magnetic nanoparticles to catalyze biodiesel synthesis. Core-shell nanoparticles were synthesized by coating Fe(3)O(4) core with silica shell. The nanoparticles treated with dimethyl octadecyl [3-(trimethoxysilyl) propyl] ammonium chloride were used as immobilization supporters. The Burkholderia lipase was then bound to the synthesized nanoparticles for immobilization. The protein binding efficiency on alkyl-functionalized Fe(3)O(4)-SiO(2) was estimated as 97%, while the efficiency was only 76% on non-modified Fe(3)O(4)-SiO(2). Maximum adsorption capacity of lipase on alkyl-functionalized Fe(3)O(4)-SiO(2) was estimated as 29.45 mg g(-1) based on Langmuir isotherm. The hydrolytic kinetics (using olive oil as substrate) of the lipase immobilized on alkyl-grafted Fe(3)O(4)-SiO(2) followed Michaelis-Menten model with a maximum reaction rate and a Michaelis constant of 6251 Ug(-1) and 3.65 mM, respectively. Physical and chemical properties of the nanoparticles and the immobilized lipase were characterized by Brunauer-Emmett-Teller (BET) analysis, scanning electron microscope (SEM), and Fourier transform infrared spectroscopy (FT-IR). Moreover, the immobilized lipase was used to catalyze the transesterification of olive oil with methanol to produce fatty acid methyl esters (FAMEs), attaining a FAMEs conversion of over 90% within 30 h in batch operation when 11 wt% immobilized lipase was employed. The immobilized lipase could be used for ten cycles without significant loss in its transesterification activity.

Studies revealing bioremediation potential of the strain Burkholderia sp. GB-01 for abamectin contaminated soils

Burkholderia sp. GB-01 strain was used to study different factors affecting its growth for inoculum production and then evaluated for abamectin degradation in soil for optimization under various conditions. The efficiency of abamectin degradation in soil by strain GB-01 was seen to be dependent on soil pH, temperature, initial abamectin concentration, and inoculum size along with inoculation frequency. Induction studies showed that abamectin depletion was faster when degrading cells were induced by pre-exposure to abamectin. Experiments performed with varying concentrations (2-160mg Kg(-1)) of abamectin-spiked soils showed that strain GB-01 could effectively degrade abamectin over the range of 2-40mg Kg(-1). The doses used were higher than the recommended dose for an agricultural application of abamectin, taking in account the over-use or spill situations. A cell density of approximately 10(8) viable cells g(-1) dry weight of soil was found to be suitable for bioremediation over a temperature range of 30-35°C and soil pH 7.5-8.5. This is the first report on bacterial degradation of abamectin in soil by a Burkholderia species, and our results indicated that this bacterium may be useful for efficient removal of abamectin from contaminated soils.

Burkholderia fungorum DBT1: a promising bacterial strain for bioremediation of PAHs-contaminated soils

An extensive taxonomic analysis of the bacterial strain Burkholderia sp. DBT1, previously isolated from an oil refinery wastewater drainage, is discussed here. This strain is capable of transforming dibenzothiophene through the 'destructive' oxidative pathway referred to as the Kodama pathway. Burkholderia DBT1 has also been proved to use fluorene, naphthalene and phenanthrene as carbon and energy sources, although growth on the first two compounds requires a preinduction step. This evidence suggests that the strain DBT1 exerts a versatile metabolism towards polycyclic aromatic hydrocarbons other than condensed thiophenes. Phylogenetic characterization using a polyphasic approach was carried out to clarify the actual taxonomic position of this strain, potentially exploitable in bioremediation. In particular, investigations were focused on the possible exclusion of Burkholderia sp. DBT1 from the Burkholderia cepacia complex. Analysis of the sequences of 16S, recA and gyrB genes along with the DNA-DNA hybridization procedure indicated that the strain DBT1 belongs to the species Burkholderia fungorum, suggesting the proposal of the taxonomic denomination B. fungorum DBT1.

Effect of Burkholderia sp. KCTC 11096BP on some physiochemical attributes of cucumber

Plant growth promoting rhizobacteria (PGPR) has been a focus of research for its potential as an eco-friendly alternative to chemical fertilizers in the agriculture industry. In current study, the effect of culture suspension (CS) of a novel gibberellins (GAs) producing bacterial strain Burkholderia sp. KCTC 11096BP, was observed on shoot length, shoot fresh and dry biomass, root fresh and dry biomass, chlorophyll contents, endogenous bioactive GAs (GA 1 and GA 4) and their immediate precursors, abscisic acid (ABA), soluble sugar contents and crude protein contents of cucumber ( Cucumis sativus L.). We found that growth attributes of cucumber were significantly promoted by the application of CS of Burkholderia sp. KCTC 11096BP. The quantity of GA 1 and GA 4 and their immediate precursors GA 20 and GA 9 respectively, were also significantly promoted as compared to their respective controls. Contrary to GAs, the quantity of endogenous free ABA in cucumber leaves was much lower in bacterial CS treated plants. Soluble sugar contents and crude protein contents of cucumber leaves were also significantly higher in bacterial CS treatments as compared to control. It was concluded that Burkholderia sp. KCTC 11096BP can be used as an eco-friendly bio-fertilizer in our farming systems.

Lipolytic activity of suspended and membrane immobilized lipase originating from indigenous Burkholderia sp. C20

In this work, a simple, inexpensive, and efficient method of preparing immobilized lipase is presented. The lipase originating from a newly isolated indigenous strain Burkholderia sp. C20 was immobilized onto cellulose nitrate (CN) membrane via filtration. The CN–immobilized lipase was able to retain 60% of its original activity after repeated uses for nine times. The thermal stability of the lipase was also slightly improved after immobilization. The optimal reaction conditions of CN–lipase were pH 9.0 and 55 °C, which are similar to those for the suspended lipase. Both suspended and immobilized lipase could hydrolyze the six oil substrates examined, while immobilized lipase displayed less specificity over the oil substrates. Kinetic analysis shows that the dependence of lipolytic activity of both suspended and immobilized lipase on oil substrate concentration can be described by Michaelis–Menten model with good agreement. The estimated kinetic constants for suspended lipase ( v max = 243.9 U/mg, K m = 0.024 mM) and immobilized lipase ( v max = 32.8 U/mg, K m = 5.61 mM) were quite different. Employment of immobilization seemed to result in a decrease in v max and an increase in K m, most likely due to the mass transfer resistance arising from formation of micelles during the lipase immobilization process.

Isolation of Fenitrothion-degrading Strain Burkholderia sp. FDS-1 and Cloning of mpd Gene

A short rod shaped, gram-negative bacterium strain Burkholderia sp. FDS-1 was isolated from the sludge of the wastewater treating system of an organophosphorus pesticides manufacturer. The isolate was capable of using fenitrothion as the sole carbon source for its growth. FDS-1 first hydrolyzed fenitrothion to 3-methyl-4-nitrophenol, which was further metabolized to nitrite and methylhydroquinone. The addition of other carbon source and omitting phosphorus source had little effect on the hydrolysis of fenitrothion. The gene encoding the organophosphorus hydrolytic enzyme was cloned and sequenced. The sequence was similar to mpd, a gene previously shown to encode a parathion-methyl-hydrolyzing enzyme in Plesiomonas sp. M6. The inoculation of strain FDS-1 (10(6) cells g(-1)) to soil treated with 100 mg fenitrothion emulsion kg(-1) resulted in a higher degradation rate than in noninoculated soils regardless of the soil sterilized or nonsterilized. These results highlight the potential of this bacterium to be used in the cleanup of contaminated pesticide waste in the environment.