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1.
ScientificWorldJournal ; 2014: 834202, 2014.
Article in English | MEDLINE | ID: mdl-24977231

ABSTRACT

Luminescence-based assays for toxicants such as Microtox, ToxAlert, and Biotox have been used extensively worldwide. However, the use of these assays in near real time conditions is limited due to nonoptimal assay temperature for the tropical climate. An isolate that exhibits a high luminescence activity in a broad range of temperatures was successfully isolated from the mackerel, Rastrelliger kanagurta. This isolate was tentatively identified as Photobacterium sp. strain MIE, based on partial 16S rDNA molecular phylogeny. Optimum conditions that support high bioluminescence activity occurred between 24 and 30°C, with pH 5.5 to 7.5, 10 to 20 g/L of sodium chloride, 30 to 50 g/L of tryptone, and 4 g/L of glycerol as the carbon source. Assessment of near real time capability of this bacterial system, Xenoassay light to monitor heavy metals from a contaminated river running through the Juru River Basin shows near real time capability with assaying time of less than 30 minutes per samples. Samples returned to the lab were tested with a standard Microtox assay using Vibrio fishceri. Similar results were obtained to Xenoassay light that show temporal variation of copper concentration. Thus, this strain is suitable for near real time river monitoring of toxicants especially in the tropics.


Subject(s)
Biological Assay/instrumentation , Environmental Monitoring/instrumentation , Luminescent Measurements/instrumentation , Metals, Heavy/analysis , Photobacterium/drug effects , Rivers/chemistry , Water Pollutants, Chemical/analysis , Computer Systems , Equipment Design , Equipment Failure Analysis , Metals, Heavy/pharmacology , Rivers/microbiology , Water Pollutants, Chemical/pharmacology
2.
Biomed Res Int ; 2014: 503784, 2014.
Article in English | MEDLINE | ID: mdl-24696853

ABSTRACT

Arsenic is a toxic metalloid which is widely distributed in nature. It is normally present as arsenate under oxic conditions while arsenite is predominant under reducing condition. The major discharges of arsenic in the environment are mainly due to natural sources such as aquifers and anthropogenic sources. It is known that arsenite salts are more toxic than arsenate as it binds with vicinal thiols in pyruvate dehydrogenase while arsenate inhibits the oxidative phosphorylation process. The common mechanisms for arsenic detoxification are uptaken by phosphate transporters, aquaglyceroporins, and active extrusion system and reduced by arsenate reductases via dissimilatory reduction mechanism. Some species of autotrophic and heterotrophic microorganisms use arsenic oxyanions for their regeneration of energy. Certain species of microorganisms are able to use arsenate as their nutrient in respiratory process. Detoxification operons are a common form of arsenic resistance in microorganisms. Hence, the use of bioremediation could be an effective and economic way to reduce this pollutant from the environment.


Subject(s)
Arsenicals/isolation & purification , Environmental Restoration and Remediation/methods , Bacteria/metabolism , Biodegradation, Environmental , Environmental Pollution/analysis
3.
Biomed Res Int ; 2014: 853084, 2014.
Article in English | MEDLINE | ID: mdl-24724104

ABSTRACT

The first purification of the Mo-reducing enzyme from Serratia sp. strain DRY5 that is responsible for molybdenum reduction to molybdenum blue in the bacterium is reported. The monomeric enzyme has an apparent molecular weight of 105 kDalton. The isoelectric point of this enzyme was 7.55. The enzyme has an optimum pH of 6.0 and maximum activity between 25 and 35°C. The Mo-reducing enzyme was extremely sensitive to temperatures above 50°C (between 54 and 70°C). A plot of initial rates against substrate concentrations at 15 mM 12-MP registered a V max for NADH at 12.0 nmole Mo blue/min/mg protein. The apparent K m for NADH was 0.79 mM. At 5 mM NADH, the apparent V max and apparent K m values for 12-MP of 12.05 nmole/min/mg protein and 3.87 mM, respectively, were obtained. The catalytic efficiency (k cat/K m ) of the Mo-reducing enzyme was 5.47 M(-1) s(-1). The purification of this enzyme could probably help to solve the phenomenon of molybdenum reduction to molybdenum blue first reported in 1896 and would be useful for the understanding of the underlying mechanism in molybdenum bioremediation involving bioreduction.


Subject(s)
Bacterial Proteins/metabolism , Molybdenum/metabolism , Oxidoreductases/metabolism , Serratia/enzymology , Catalysis , Oxidation-Reduction
4.
J Environ Biol ; 35(2): 353-5, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24665761

ABSTRACT

Respiratory activity inhibition by toxic compounds in bacteria and yeast has been used to detect toxic compounds in the environment. Often the age of culture contributes towards the sensitivity of detection. In the present work, the effect of growth period on the sensitivity of an inhibitive assay for heavy metals using bacterial respiratory assay system based on the reduction of the water soluble tetrazolium dye MTT is reported. A silver-sensitive isolate was discovered to exhibit different sensitivities towards silver at different growth periods. An exponential decay model adequately described the inhibition due to silver. Analysis using ANOVA with post-hoc Tukey's test showed that the IC50 obtained by strain DRYS8 grown at the 12 hr- period in nutrient broth at 28 degrees C gave the lowest value compared to other growth periods. This study highlights the importance of taking into accounts growth conditions and age of culture in developing cellular-based bioassays.


Subject(s)
Rhizobium/drug effects , Silver/chemistry , Silver/pharmacology , Tetrazolium Salts/metabolism , Thiazoles/metabolism , Biological Assay , Oxygen Consumption/drug effects , Rhizobium/metabolism , Sensitivity and Specificity , Tetrazolium Salts/chemistry , Thiazoles/chemistry , Water Pollutants, Chemical/chemistry , Water Pollutants, Chemical/pharmacology
5.
J Environ Biol ; 35(2): 399-406, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24665769

ABSTRACT

A diesel-degrading bacterium was isolated from a diesel-contaminated site in Selangor, Malaysia. The isolate was tentatively identified as Acinetobacter sp. strain DRY12 based on partial 16S rDNA molecular phylogeny and Biolog GN microplate panels and Microlog database. Optimum growth occurred from 3 to 5% diesel and the strain was able to tolerate as high as 8% diesel. The optimal pH that supported growth of the bacterium was between pH 7.5 to 8.0. The isolate exhibited optimal growth in between 30 and 35 degrees C. The best nitrogen source was potassium nitrate (between 0.6 and 0.9% (w/v)) followed by ammonium chloride, sodium nitrite and ammonium sulphate in descending order. An almost complete removal of diesel components was seen from the reduction in hydrocarbon peaks observed using Solid Phase Microextraction Gas Chromatography analysis after 10 days of incubation. The best growth kinetic model to fit experimental data was the Haldane model of substrate inhibiting growth with a correlation coefficient value of 0.97. The maximum growth rate- micromax was 0.039 hr(-1) while the saturation constant or half velocity constant Ks and inhibition constant Ki, were 0.387% and 4.46%, respectively. MATH assays showed that 75% of the bacterium was found in the hexadecane phase indicating that the bacterium was hydrophobic. The characteristics of this bacterium make it useful for bioremediation works in the Tropics.


Subject(s)
Acinetobacter/metabolism , Gasoline , Acinetobacter/classification , Acinetobacter/genetics , Hydrogen-Ion Concentration , Phylogeny , Soil/chemistry , Soil Microbiology , Soil Pollutants/chemistry , Soil Pollutants/metabolism , Temperature , Time Factors
6.
Biomed Res Int ; 2013: 371058, 2013.
Article in English | MEDLINE | ID: mdl-24369531

ABSTRACT

Molybdenum is very toxic to agricultural animals. Mo-reducing bacterium can be used to immobilize soluble molybdenum to insoluble forms, reducing its toxicity in the process. In this work the isolation of a novel molybdate-reducing Gram positive bacterium tentatively identified as Bacillus sp. strain A.rzi from a metal-contaminated soil is reported. The cellular reduction of molybdate to molybdenum blue occurred optimally at 4 mM phosphate, using 1% (w/v) glucose, 50 mM molybdate, between 28 and 30 °C and at pH 7.3. The spectrum of the Mo-blue product showed a maximum peak at 865 nm and a shoulder at 700 nm. Inhibitors of bacterial electron transport system (ETS) such as rotenone, sodium azide, antimycin A, and potassium cyanide could not inhibit the molybdenum-reducing activity. At 0.1 mM, mercury, copper, cadmium, arsenic, lead, chromium, cobalt, and zinc showed strong inhibition on molybdate reduction by crude enzyme. The best model that fitted the experimental data well was Luong followed by Haldane and Monod. The calculated value for Luong's constants p max, K(s), S(m), and n was 5.88 µmole Mo-blue hr(-1), 70.36 mM, 108.22 mM, and 0.74, respectively. The characteristics of this bacterium make it an ideal tool for bioremediation of molybdenum pollution.


Subject(s)
Bacillus/metabolism , Biodegradation, Environmental , Molybdenum/metabolism , Agriculture , Animals , Bacillus/chemistry , Bacillus/isolation & purification , Environmental Pollutants , Kinetics , Molybdenum/chemistry , Molybdenum/toxicity , Phosphates/chemistry
7.
J Basic Microbiol ; 53(1): 9-19, 2013 Jan.
Article in English | MEDLINE | ID: mdl-22581645

ABSTRACT

In this work, we report on the isolation of a phenol-degrading Rhodococcus sp. with a high tolerance towards phenol. The isolate was identified as Rhodococcus sp. strain AQ5NOL 2, based on 16S rDNA analysis. The strain degraded phenol using the meta pathway, a trait shared by many phenol-degraders. In addition to phenol biodegradation, the strain was also capable of degrading diesel. Strain AQ5NOL 2 exhibited a broad optimum temperature for growth on phenol at between 20 °C and 35 °C. The best nitrogen sources were ammonium sulphate, glycine or phenylalanine, followed by proline, nitrate, leucine, and alanine (in decreasing efficiency). Strain AQ5NOL 2 showed a high tolerance and degradation capacity of phenol, for it was able to register growth in the presence of 2000 mg l(-1) phenol. The growth of this strain on phenol as sole carbon and energy source were modeled using Haldane kinetics with a maximal specific growth rate (µ(max)) of 0.1102 hr(-1), a half-saturation constant (K(s) ) of 99.03 mg l(-1) or 1.05 mmol l(-1), and a substrate inhibition constant (K(i)) of 354 mg l(-1) or 3.76 mmol l(-1). Aside from phenol, the strain could utilize diesel, 2,4-dinitrophenol and ρ-cresol as carbon sources for growth. Strain AQ5NOL 2 exhibited inhibition of phenol degradation by Zn(2+), Cu(2+), Cr(6+), Ag(+) and Hg(2+) at 1 mg l(-1).


Subject(s)
Phenols/metabolism , Rhodococcus/isolation & purification , Rhodococcus/metabolism , Analysis of Variance , Biodegradation, Environmental , Carbon/metabolism , DNA, Bacterial/analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Gasoline , Hydrogen-Ion Concentration , Kinetics , Metals, Heavy/metabolism , Nitrogen/metabolism , Phylogeny , RNA, Ribosomal, 16S/genetics , Rhodococcus/classification , Rhodococcus/genetics , Temperature
8.
Biomed Res Int ; 2013: 871941, 2013.
Article in English | MEDLINE | ID: mdl-24381945

ABSTRACT

A molybdenum-reducing bacterium from Antarctica has been isolated. The bacterium converts sodium molybdate or Mo6⁺ to molybdenum blue (Mo-blue). Electron donors such as glucose, sucrose, fructose, and lactose supported molybdate reduction. Ammonium sulphate was the best nitrogen source for molybdate reduction. Optimal conditions for molybdate reduction were between 30 and 50 mM molybdate, between 15 and 20°C, and initial pH between 6.5 and 7.5. The Mo-blue produced had a unique absorption spectrum with a peak maximum at 865 nm and a shoulder at 710 nm. Respiratory inhibitors such as antimycin A, sodium azide, potassium cyanide, and rotenone failed to inhibit the reducing activity. The Mo-reducing enzyme was partially purified using ion exchange and gel filtration chromatography. The partially purified enzyme showed optimal pH and temperature for activity at 6.0 and 20°C, respectively. Metal ions such as cadmium, chromium, copper, silver, lead, and mercury caused more than 95% inhibition of the molybdenum-reducing activity at 0.1 mM. The isolate was tentatively identified as Pseudomonas sp. strain DRY1 based on partial 16s rDNA molecular phylogenetic assessment and the Biolog microbial identification system. The characteristics of this strain would make it very useful in bioremediation works in the polar and temperate countries.


Subject(s)
Molybdenum/metabolism , Nitrogen/metabolism , Pseudomonas/isolation & purification , Pseudomonas/metabolism , Ammonium Sulfate/metabolism , Antarctic Regions , Chromatography , Hydrogen-Ion Concentration , Ions , Molybdenum/chemistry , Pseudomonas/genetics , RNA, Ribosomal, 16S/genetics , Temperature
9.
J Environ Biol ; 34(6): 1077-82, 2013 Nov.
Article in English | MEDLINE | ID: mdl-24555340

ABSTRACT

A bacterium capable of biodegrading surfactant sodium dodecyl sulphate (SDS) was isolated from Antarctic soil. The isolate was tentatively identified as Pseudomonas sp. strain DRY15 based on carbon utilization profiles using Biolog GN plates and partial 16S rDNA molecular phylogeny. Growth characteristic studies showed that the bacterium grew optimally at 10 degrees C, 7.25 pH, 1 g l(-1) SDS as a sole carbon source and 2 g l(-1) ammonium sulphate as nitrogen source. Growth was completely inhibited at 5 g l(-1) SDS. At a tolerable initial concentration of 2 g l(-1), approximately 90% of SDS was degraded after an incubation period of eight days. The best growth kinetic model to fit experimental data was the Haldane model of substrate inhibition with a correlation coefficient value of 0.97. The maximum growth rate was 0.372 hr(-1) while the saturation constant or half velocity constant (Ks) and inhibition constant (Ki), were 0.094% and 11.212 % SDS, respectively. Other detergent tested as carbon sources at 1 g l(-1) was Tergitol NP9, Tergitol 15S9, Witconol 2301 (methyl oleate), sodium dodecylbenzene sulfonate (SDBS), benzethonium chloride, and benzalkonium chloride showed Tergitol NP9, Tergitol 15S9, Witconol 2301 and the anionic SDBS supported growth with the highest growth exhibited by SDBS.


Subject(s)
Environmental Pollutants/metabolism , Pseudomonas/metabolism , Sodium Dodecyl Sulfate/metabolism , Soil Microbiology , Surface-Active Agents/metabolism , Antarctic Regions , Hydrogen-Ion Concentration , Nitrogen Compounds/metabolism , Pseudomonas/growth & development , Pseudomonas/isolation & purification , Temperature
10.
J Environ Biol ; 34(5): 967-70, 2013 Sep.
Article in English | MEDLINE | ID: mdl-24558814

ABSTRACT

Near-real-ime assay is anassay method that the whole process from sampling until results could be obtained in approximately Iess than one hour. The ElIman assay for acetyl cholinesterase (AChE) has near real-time potential due to its simplicity and fast assay time. The commercial acetylcholinesterase from Electrophorus electricus is well known for its uses in insecticides detection. A lesser known fact is AChE is also sensitive to heavy metals. A near real-time inhibitive assay for heavy metals using AChE from this source showed promising results. Several heavy metals such as copper, silver and mercury could be etected with IC50 values of1.212, 0.1185 and 0.097 mg I-1, respectively. The Limits of Detection (LOD) for copper, silver and mercury were 0.01, 0.015 and 0.01 mg I-1, respectively. TheLimits of quantitation (LOQ) or copper, silver and mercury were 0.196, 0.112 and 0.025 mg I-1, respectively. The LOQvalues for copper, silver and mercury were well below the maximum permissible limit for these metal ions as outlined by Malaysian Department of Environment. A polluted location demonstrated near real-time applicability of the assay with variation oftemporal levels of heavy metals detected. The results show that AChE from Electrophorus electricus has the potential to be used as a near real-time biomonitoring tool for heavy


Subject(s)
Acetylcholinesterase/metabolism , Electrophorus/metabolism , Environmental Monitoring/methods , Metals, Heavy/analysis , Water Pollutants, Chemical/analysis , Animals , Dithionitrobenzoic Acid/metabolism , Limit of Detection , Malaysia , Spectrophotometry, Atomic
11.
J Basic Microbiol ; 52(3): 296-305, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22052341

ABSTRACT

A novel molybdate-reducing bacterium, tentatively identified as Klebsiella sp. strain hkeem and based on partial 16s rDNA gene sequencing and phylogenetic analysis, has been isolated. Strain hkeem produced 3 times more molybdenum blue than Serratia sp. strain Dr.Y8; the most potent Mo-reducing bacterium isolated to date. Molybdate was optimally reduced to molybdenum blue using 4.5 mM phosphate, 80 mM molybdate and using 1% (w/v) fructose as a carbon source. Molybdate reduction was optimum at 30 °C and at pH 7.3. The molybdenum blue produced from cellular reduction exhibited absorption spectrum with a maximum peak at 865 nm and a shoulder at 700 nm. Inhibitors of electron transport system such as antimycin A, rotenone, sodium azide, and potassium cyanide did not inhibit the molybdenum-reducing enzyme. Mercury, silver, and copper at 1 ppm inhibited molybdenum blue formation in whole cells of strain hkeem.


Subject(s)
Klebsiella/metabolism , Molybdenum/metabolism , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Molecular Sequence Data , Oxidation-Reduction , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Serratia
12.
J Basic Microbiol ; 52(5): 573-81, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22144174

ABSTRACT

As well as for chemical and environmental reasons, acrylamide is widely used in many industrial applications. Due to its carcinogenicity and toxicity, its discharge into the environment causes adverse effects on humans and ecology alike. In this study, a novel acrylamide-degrading yeast has been isolated. The isolate was identified as Rhodotorula sp. strain MBH23 using ITS rRNA analysis. The results showed that the best carbon source for growth was glucose at 1.0% (w/v). The optimum acrylamide concentration, being a nitrogen source for cellular growth, was at 500 mg l(-1). The highest tolerable concentration of acrylamide was 1500 mg l(-1) whereas growth was completely inhibited at 2000 mg l(-1). At 500 mg l(-1), the strain MBH completely degraded acrylamide on day 5. Acrylic acid as a metabolite was detected in the media. Strain MBH23 grew well between pH 6.0 and 8.0 and between 27 and 30 °C. Amides such as 2-chloroacetamide, methacrylamide, nicotinamide, acrylamide, acetamide, and propionamide supported growth. Toxic heavy metals such as mercury, chromium, and cadmium inhibited growth on acrylamide.


Subject(s)
Acrylamide/metabolism , Rhodotorula/isolation & purification , Rhodotorula/physiology , Acrylates/metabolism , Biotransformation , Cluster Analysis , DNA, Fungal/chemistry , DNA, Fungal/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Ribosomal Spacer/genetics , Hydrogen-Ion Concentration , Metals, Heavy/toxicity , Molecular Sequence Data , Mycological Typing Techniques , Phylogeny , Rhodotorula/classification , Rhodotorula/genetics , Sequence Analysis, DNA , Temperature , Time Factors
13.
Folia Microbiol (Praha) ; 55(2): 137-43, 2010 Mar.
Article in English | MEDLINE | ID: mdl-20490756

ABSTRACT

A local molybdenum-reducing bacterium was isolated and tentatively identified as Acinetobacter calcoaceticus strain Dr.Y12 based on carbon utilization profiles using Biolog GN plates and 16S rDNA comparative analysis. Molybdate reduction was optimized under conditions of low dissolved oxygen (37 degrees C and pH 6.5). Of the electron donors tested, glucose, fructose, maltose and sucrose supported molybdate reduction after 1 d of incubation, glucose and fructose supporting the highest Mo-blue production. Optimum Mo-blue production was reached at 20 mmol/L molybdate and 5 mmol/L phosphate; increasing the phosphate concentrations inhibited the production. An increase in an overall absorption profiles, especially at peak maximum at 865 nm and the shoulder at 700 nm, was observed in direct correlation with the increased in Mo-blue amounts. Metal ions, such as chromium, cadmium, copper, mercury and lead (2 mmol/L final concentration) caused approximately 88, 53, 80, 100, and 20 % inhibition, respectively. Respiratory inhibitors, such as antimycin A, rotenone, sodium azide and cyanide showed in this bacterium no inhibition of the Mo-blue production, suggesting that the electron transport system is not a site of molybdate reduction.


Subject(s)
Acinetobacter calcoaceticus/metabolism , Molybdenum/metabolism , Acinetobacter calcoaceticus/genetics , Acinetobacter calcoaceticus/isolation & purification , Molecular Sequence Data , Oxidation-Reduction
14.
J Appl Microbiol ; 108(6): 2050-8, 2010 Jun.
Article in English | MEDLINE | ID: mdl-19968732

ABSTRACT

AIMS: To isolate and characterize a potent molybdenum-reducing bacterium. METHODS AND RESULTS: A minimal salt medium supplemented with 10 mmol l(-1) molybdate, glucose (1.0%, w/v) as a carbon source and ammonium sulfate (0.3%, w/v) as a nitrogen source was used in the screening process. A molybdenum-reducing bacterium was isolated and tentatively identified as Pseudomonas sp. strain DRY2 based on carbon utilization profiles using Biolog GN plates and partial 16S rDNA molecular phylogeny. Strain DRY2 produced 2.4, 3.2 and 6.2 times more molybdenum blue compared to Serratia marcescens strain DRY6, Enterobacter cloacae strain 48 and Eschericia coli K12, respectively. Molybdate reduction was optimum at 5 mmol l(-1) phosphate. The optimum molybdate concentration that supported molybdate reduction at 5 mmol l(-1) phosphate was between 15 and 25 mmol l(-1). Molybdate reduction was optimum at 40 degrees C and at pH 6.0. Phosphate concentrations higher than 5 mmol l(-1) strongly inhibited molybdate reduction. Inhibitors of electron transport system such as antimycin A, rotenone, sodium azide and cyanide did not inhibit the molybdenum-reducing enzyme activity. Chromium, copper, mercury and lead inhibited the molybdenum-reducing activity. CONCLUSIONS: A novel molybdenum-reducing bacterium with high molybdenum reduction capacity has been isolated. SIGNIFICANCE AND IMPACT OF THE STUDY: Molybdenum is an emerging global pollutant that is very toxic to ruminants. The characteristics of this bacterium suggest that it would be useful in the bioremediation of molybdenum pollutant.


Subject(s)
Molybdenum/metabolism , Pseudomonas/metabolism , Soil Microbiology , Soil Pollutants/metabolism , Biodegradation, Environmental , Carbon/metabolism , Culture Media , DNA, Bacterial/genetics , Nitrogen/metabolism , Oxidation-Reduction , Phosphates/metabolism , Phylogeny , Pseudomonas/genetics , Pseudomonas/isolation & purification , RNA, Ribosomal, 16S/genetics
15.
Article in English | WPRIM (Western Pacific) | ID: wpr-625754

ABSTRACT

Ganoderma lucidum is a fungus usually used in traditional Chinese medicine. The high value of G. lucidum is related to its polysaccharides content. Crude polysaccharides from G. lucidum (GLCP) were obtained using hot water extraction method. There is about 0.57 g of GLCP in 1 g crude of G. lucidum. The prebiotic potential of GLCP was tested against probiotic bacteria namely: Bifidobacterium longum BB536, Bifidobacterium pseudocatenulatum G4, Lactobacillus acidophilus and Lactobacillus casei Shirota. The prebiotic potentials were studied in 10 mL basal Trypticase Phytone Yeast (abbreviated as bTPY) medium (without glucose) supplemented with various concentrations of GLCP (abbreviated as bTPYglcp) (0.5%, 1.0%, 1.5% and 2.0%). bTPY medium supplemented with glucose (abbreviated as bTPYglu) and inulin (abbreviated bTPYinu) were used as comparison. Viable cell counts of the bacteria and the pH of the medium were determined during anaerobic incubation period of 0 h, 12 h, 24 h and 48 h at 37 °C. In the presence of carbohydrate source, cultures showed various degree of growth increment. With regards to the growth supporting property: bTPYglu, bTPYglu+glcp, bTPYglcp and bTPYinu were ranked first, second, third and fourth respectively. Interestingly, in bTPYglcp medium, bacterial growth increased with increasing GLCP concentrations when incubated until 24 h. B. longum BB536 was ranked first (10.53 log cfu/mL) in term of their growth in this medium. Growth of B.pseudocatenulatum G4 was ranked second with 10.40 log cfu/mL. This study shows that, GLCP could support the growth of the bacteria tested.

16.
J Basic Microbiol ; 49 Suppl 1: S43-54, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19455513

ABSTRACT

Extensive use of metals in various industrial applications has caused substantial environmental pollution. Molybdenum-reducing bacteria isolated from soils can be used to remove molybdenum from contaminated environments. In this work we have isolated a local bacterium with the capability to reduce soluble molybdate to the insoluble molybdenum blue. We studied several factors that would optimize molybdate reduction. Electron donor sources such as glucose, sucrose, lactose, maltose and fructose (in decreasing efficiency) supported molybdate reduction after 24 h of incubation with optimum glucose concentration for molybdate reduction at 1.5% (w/v). The optimum pH, phosphate and molybdate concentrations, and temperature for molybdate reduction were pH 6.5, 5.0, 25 to 50 mM and 37 degrees C, respectively. The Mo-blue produced by cellular reduction exhibited a unique absorption spectrum with a maximum peak at 865 nm and a shoulder at 700 nm. Metal ions such as chromium, cadmium, copper, silver and mercury caused approximately 73, 71, 81, 77 and 78% inhibition of the molybdenum-reducing activity, respectively. All of the respiratory inhibitors tested namely rotenone, azide, cyanide and antimycin A did not show any inhibition to the molybdenum-reducing activity suggesting components of the electron transport system are not responsible for the reducing activity. The isolate was tentatively identified as Enterobacter sp. strain Dr.Y13 based on carbon utilization profiles using Biolog GN plates and partial 16S rDNA molecular phylogeny.


Subject(s)
Enterobacter/metabolism , Molybdenum/metabolism , Soil Microbiology , Soil Pollutants/metabolism , Culture Media , DNA, Bacterial/genetics , Enterobacter/genetics , Enterobacter/isolation & purification , Hydrogen-Ion Concentration , Oxidation-Reduction , Phosphates/metabolism , Phylogeny , RNA, Ribosomal, 16S , Sequence Analysis, DNA , Temperature
17.
J Environ Biol ; 30(1): 1-6, 2009 Jan.
Article in English | MEDLINE | ID: mdl-20112855

ABSTRACT

A diesel-degrading bacterium from Antarctica has been isolated. The isolate was tentatively identified as Pseudomonas sp. strain DRYJ3 based on partial 16S rDNA molecular phylogeny and Biolog GN microplate panels and Microlog database. Growth on diesel was supported optimally by ammonium sulphate, nitrate and nitrite. The bacterium grew optimally in between 10 and 15 degrees C, pH 7.0 and 3.5% (v/v) diesel. The biodegradation of diesel oil by the strain increased in efficiency from the second to the sixth day of incubation from 1.4 to 18.8% before levelling off on the eighth day n-alkane oxidizing and aldehyde reductase activities were detected in the crude enzyme preparation suggesting the existence of terminal n-alkane oxidizing activity in this bacterium.


Subject(s)
Gasoline , Pseudomonas/metabolism , Antarctic Regions , Bacterial Proteins/metabolism , Biodegradation, Environmental , DNA, Ribosomal/chemistry , Hydrogen-Ion Concentration , Nitrogen/metabolism , Phylogeny , Pseudomonas/genetics , Pseudomonas/isolation & purification , Temperature
18.
J Environ Biol ; 30(1): 17-22, 2009 Jan.
Article in English | MEDLINE | ID: mdl-20112858

ABSTRACT

A new inhibitive heavy metals determination method using trypsin has been developed. The enzyme was assayed using the casein-Coomassie-dye-binding method. In the absence of inhibitors, casein was hydrolysed to completion and the Coomassie-dye was unable to stain the protein and the solution became brown. In the presence of metals, the hydrolysis of casein was inhibited and the solution remained blue. The bioassay was able to detect zinc and mercury with IC50 (concentration causing 50% inhibition) values of 5.78 and 16.38 mg l(-1) respectively. The limits of detection (LOD), for zinc and mercury were 0.06 mg l(-1) (0.05-0.07, 95% confidence interval) and 1.06 mg l(-1) (1.017-1.102, 95% confidence interval), respectively. The limits of quantitation (LOQ) for zinc and mercury were 0.61 mg l(-1) (0.51-0.74 at a 95% confidence interval) and 1.35 mg l(-1) (1.29-1.40 at a 95% confidence interval), respectively. The IC50 value for zinc was much higher than the IC50 values for papain and Rainbow trout, but was within the range of Daphnia magna and Microtox. The IC50 value for zinc was only lower than those for immobilized urease. Other toxic heavy metals, such as lead, silver arsenic, copper and cadmium, did not inhibit the enzyme at 20 mg l(-1). Using this assay we managed to detect elevated zinc concentrations in several environmental samples. Pesticides, such as carbaryl, flucythrinate, metolachlor glyphosate, diuron, diazinon, endosulfan sulphate, atrazine, coumaphos, imidacloprid, dicamba and paraquat, showed no effect on the activity of trypsin relative to control (One-way ANOVA, F(12,26)= 0.3527, p> 0.05). Of the 17 xenobiotics tested, only (sodium dodecyl sulphate) SDS gave positive interference with 150% activity higher than that of the control at 0.25% (v/v).


Subject(s)
Environmental Monitoring/methods , Trypsin/chemistry , Zinc/analysis , Inhibitory Concentration 50 , Metals, Heavy/chemistry , Pesticides/chemistry , Trypsin Inhibitors/analysis , Trypsin Inhibitors/chemistry , Xenobiotics/chemistry , Zinc/chemistry
19.
J Environ Biol ; 30(1): 39-44, 2009 Jan.
Article in English | MEDLINE | ID: mdl-20112861

ABSTRACT

In this work the development of an inhibitive assay for copper using the molybdenum-reducing enzyme assay is presented. The enzyme is assayed using 12-molybdophosphoric acid at pH 5.0 as an electron acceptor substrate and NADH as the electron donor substrate. The enzyme converts the yellowish solution into a deep blue solution. The assay is based on the ability of copper to inhibit the molybdenum-reducing enzyme from the molybdate-reducing Serratia sp. Strain DRY5. Other heavy metals tested did not inhibit the enzyme at 10 mg l(-1). The best model with high regression coefficient to measure copper inhibition is one-phase binding. The calculated IC50 (concentration causing 50% inhibition) is 0.099 mg l(-1) and the regression coefficient is 0.98. The comparative LC50, EC50 and IC50 data for copper in different toxicity tests show that the IC50 value for copper in this study is lower than those for immobilized urease, bromelain, Rainbow trout, R. meliloti, Baker's Yeast dehydrogenase activity Spirillum volutans, P. fluorescens, Aeromonas hydrophilia and synthetic activated sludge assays. However the IC50 value is higher than those for Ulva pertusa and papain assays, but within the reported range for Daphnia magna and Microtox assays.


Subject(s)
Copper/analysis , Environmental Monitoring/methods , Environmental Pollutants/analysis , Enzyme Assays/methods , Enzyme Inhibitors/analysis , Bacterial Proteins/antagonists & inhibitors , Copper/chemistry , Enzyme Inhibitors/chemistry , Molybdenum/chemistry , Molybdenum/metabolism
20.
J Environ Biol ; 30(1): 57-64, 2009 Jan.
Article in English | MEDLINE | ID: mdl-20112864

ABSTRACT

Several local acrylamide-degrading bacteria have been isolated. One of the isolate that exhibited the highest growth on acrylamide as a nitrogen source was then further characterized. The isolate was tentatively identified as Bacillus cereus strain DRY135 based on carbon utilization profiles using Biolog GP plates and partial 16S rDNA molecular phylogeny. The isolate grew optimally in between the temperatures of 25 and 30 degrees C and within the pH range of 6.8 to 7.0. Glucose, fructose, lactose, maltose, mannitol, citric acid and sucrose supported growth with glucose being the best carbon source. Different concentrations of acrylamide ranging from 100 to 4000 mg l(-1) incorporated into the growth media shows that the highest growth was obtained at acrylamide concentrations of between 500 to 1500 mg l(-1). At 1000 mg l(-1) of acrylamide, degradation was 90% completed after ten days of incubation with concomitant cell growth. The metabolite acrylic acid was detected in the media during degradation. Other amides such as methacrylamide, nicotinamide, acetamide, propionamide and urea supported growth with the highest growth supported by acetamide, propionamide and urea. Strain DRY135, however was not able to assimilate 2-chloroacetamide. The characteristics of this isolate suggest that it would be useful in the bioremediation of acrylamide.


Subject(s)
Acrylamide/metabolism , Bacillus cereus/metabolism , Environmental Pollutants/metabolism , Acrylamide/chemistry , Amides/chemistry , Amides/metabolism , Bacillus cereus/genetics , Bacillus cereus/growth & development , Bacillus cereus/isolation & purification , Biodegradation, Environmental , Carbon/metabolism , Cell Culture Techniques , Hydrogen-Ion Concentration , Phylogeny , Temperature
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