CN105861366B - Application of the polycyclic aromatic hydrocarbon-degrading bacteria FA1 in absorption heavy metal Pb ion - Google Patents
Application of the polycyclic aromatic hydrocarbon-degrading bacteria FA1 in absorption heavy metal Pb ion Download PDFInfo
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Abstract
The invention discloses application of the polycyclic aromatic hydrocarbon-degrading bacteria FA1 in absorption heavy metal Pb ion, the class of polycyclic aromatic hydrocarbon-degrading bacteria FA1 is named as Herbaspirillum chlorophenolicum, China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on May 4th, 2010, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, and deposit number is CGMCC No.3797.Discovery of the invention can provide bacterial strain and genetic material for the microorganism remediation of heavy metal and polycyclic aromatic hydrocarbon composite pollution environment.
Description
Technical field
Present invention relates particularly to application of the polycyclic aromatic hydrocarbon-degrading bacteria FA1 in absorption heavy metal Pb ion.
Background technique
Due to mining, exhaust gas discharge, solid waste accumulation and the factors such as sewage irrigation, a large amount of poisonous and harmful heavy metallics into
Enter into environment and continues to accumulate.Heavy metal contaminants are hardly degraded in the environment, and are enriched with food chain, have
Very strong bioconcentration, so that ecological environment and human health are faced and seriously threatened.As one of maximum heavy metal of toxicity,
Concentration over-standard phenomenon of the lead in China's soil and groundwater environment is continually seen in report.In recent years, using lead as representative
The environment and social concern that heavy metal pollution causes are more and more prominent, and therefore, the reparation of Heavy Metals In Environment is not worked as singly
One of the research hotspot of preceding field of environment protection, with more important realistic meaning.
Mainly there are peripheral doses, chemical remediation and biological prosthetic etc. for the restorative procedure of heavy metal at present.It is biological prosthetic
Method has the characteristics that efficient, low consumption, economy, environmental-friendly and easily operated management, compared with traditional physics, chemical restoration
It has a clear superiority, in particular for the heavy metal pollution situation of low concentration.The key of the technology be to filter out have compared with
The microorganism of strong absorption heavy metal ability.The microorganism that heavy metal can be adsorbed includes bacterium, fungi and algae etc., wherein thin
Bacterium due to its relatively stronger biochemical adaptability and more advantage.
It is worth noting that the heavy metal biological of existing report is repaired is directed to individual heavy metal pollution situation mostly, so
And the heavy metal pollution in environment often occurs with other organic pollutants simultaneously.Allusion quotation as heavy metal-organic matter combined pollution
Type represents, and heavy metal and polycyclic aromatic hydrocarbon composite pollution phenomenon are widely present in China's soil and groundwater.Two kinds of pollutants are total
The toxic effect of polycyclic aromatic hydrocarbon will cause to coerce to the microorganism that can adsorb heavy metal when depositing, and harms and arrives its suction to heavy metal
Attached efficiency.Therefore, for the heavy metal in actual environment-polycyclic aromatic hydrocarbon composite pollution status, it is necessary to filter out resistance to polycyclic virtue
Hydrocarbon toxicity is even capable of the heavy metal adsorption microorganism of degrading polycyclic aromatic hydrocarbons, studies its characterization of adsorption, is heavy metal and polycyclic virtue
The microorganism remediation of hydrocarbon combined pollution environment provides bacterial strain and genetic material.
Summary of the invention
Absorption property of the polycyclic aromatic hydrocarbon-degrading bacteria to heavy metal Pb ion for having different genera by investigating seminar, this
The polycyclic aromatic hydrocarbon-degrading bacteria FA1 filtered out is studied in invention discovery before has good tolerance and adsorptivity to heavy metal Pb.
The class of polycyclic aromatic hydrocarbon-degrading bacteria FA1 is named as Herbaspirillum chlorophenolicum, on May 4th, 2010
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address is Chaoyang District, Beijing City North Star west
The institute 3 of road 1, deposit number are CGMCC No.3797.The screening technique and bacterial strain performance of polycyclic aromatic hydrocarbon-degrading bacteria FA1 is shown in class
Article " the Degradation of fluoranthene by a newly isolated that topic group XU etc. is delivered for 2011
Strain of Herbaspirillum chlorophenolicum from activated sludge ",
Biodegradation, 22 (2), 335-345.
In view of its suction-operated for heavy metal Pb ion, present invention further proposes above-mentioned polycyclic aromatic hydrocarbon-degrading bacterias
FA1 is in the application administered in heavy metal-polycyclic aromatic hydrocarbon composite pollution.
Further, the invention also provides above-mentioned polycyclic aromatic hydrocarbon-degrading bacteria FA1 in the huge sum of money for preparing resistance to polycyclic aromatic hydrocarbon
Belong to the application in ion adsorbent, the adsorbent is prepared via a method which to obtain: polycyclic aromatic hydrocarbon-degrading bacteria FA1 is accessed into base
Enrichment culture is carried out in basal culture medium, then by the bacterium solution in logarithmic growth phase with 8000rpm refrigerated centrifuge 10min, with 100
The NaCl solution of~150mM/L is washed 3 times, and 4 DEG C of refrigerations are spare.
Preferably, the basal medium includes that beef extract 0.5%, peptone 1.0% and NaCl 0.5%, surplus are
Deionized water, pH7.0~7.2.
In above-mentioned application, when for guarantee adsorption effect, temperature of reaction system should to be adjusted when adsorbing heavy metal ion
It is 20~35 DEG C, pH is 2~6.
It is highly preferred that pH is 5~6 being 25~30 DEG C for when adsorbing heavy metal ion, adjusting temperature of reaction system.
It most preferably, is being 30 DEG C for when adsorbing heavy metal ion, adjusting temperature of reaction system, pH 5.5.
The utility model has the advantages that the present invention have unexpectedly discovered can simultaneously degrading polycyclic aromatic hydrocarbons and heavy metal ion polycyclic aromatic hydrocarbon
Degradation bacteria Herbaspirillum chlorophenolicum FA1, and by studying its characterization of adsorption, it is heavy metal and polycyclic
The microorganism remediation of aromatic hydrocarbons combined pollution environment provides bacterial strain and genetic material.
Detailed description of the invention
Fig. 1 is bacterial strain FA1 in various concentration Pb2+Growth curve in solution；
Fig. 2 is bacterial strain FA1 to Pb2+Adsorption dynamics adsorption kinetics；
Fig. 3 is bacterial strain FA1 to Pb2+Adsorption isotherm；
Fig. 4 is that temperature adsorbs Pb to bacterial strain FA12+The influence of efficiency；
Fig. 5 is that pH adsorbs Pb to bacterial strain FA12+The influence of efficiency.
Specific embodiment
The present invention may be better understood according to following embodiments.However, those skilled in the art will readily recognize that,
Content described in embodiment is merely to illustrate the present invention, described in detail without that should will not limit in claims
The present invention.
The preparation of 1 heavy metal absorbent of embodiment.
The polycyclic aromatic hydrocarbon-degrading bacteria Herbaspirillum that the present embodiment microorganism used therefor filters out before seminar
Chlorophenolicum FA1 (Xu et al.2011, Degradation of fluoranthene by a newly
Isolated strain of Herbaspirillum chlorophenolicum from activated sludge ",
Biodegradation, 22 (2), 335-345).Sequence accession number of the bacterial strain FA1 in the GenBank database of NCBI be
HM107777 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on May 4th, 2010, protects
Hide address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, number of registering on the books are
CGMCC No.3797。
Sorbent preparation method: before inoculation, bacterial strain FA1 being accessed in basal medium and carries out enrichment culture, then will place
Adsorbent is obtained with NaCl solution washing 3 times of 150mM/L with 8000rpm refrigerated centrifuge 10min in the bacterium solution of logarithmic growth phase,
4 DEG C of refrigerations are spare.
2 microorganism of embodiment is to heavy metal Pb2+Characterization of adsorption research.
All experimental groups of following characterization of adsorption researchs are equipped with 3 parallel and blank control groups.
(1) microorganism is to heavy metal Pb2+Tolerance
Take Pb (NO3)2, basal medium is added, so that Pb2+Concentration is respectively 0,10,20,50,100,200mg/L, and
Adjusting pH value of solution is 5.5, is inoculated with into bacterial strain FA1,30 DEG C in concussion and cultivate case, 125rpm constant temperature incubation, and timing sampling, with ultraviolet
The OD of spectrophotometric determination solution600Value is drawn using the time as the growth curve of abscissa.
Polycyclic aromatic hydrocarbon-degrading bacteria FA1 is in various concentration Pb2+Growth curve under existence condition is as shown in Figure 1.Be free of Pb2 +Control compare, as Pb in culture medium2+When concentration is 10mg/L, the growth of bacterial strain FA1 is barely affected.With Pb2+Concentration
Further increase, the speed of growth of bacterial strain gradually slows down, the corresponding OD of biomass600Value also becomes smaller therewith, illustrates Pb2+To bacterium
The growth and breeding of strain has certain inhibiting effect.It is to be noted, however, that the Pb of higher concentration2+Concentration (20-200mg/L)
Bacterial strain is not delayed to enter logarithmic growth phase.In Pb2+When at concentrations up to 200mg/L, the life of (15h) in logarithmic growth phase
Object amount is compared to without Pb2+For the control group of concentration, 28.3% is only only reduced, shows Herbaspirillum
Chlorophenolicum FA1 has good tolerance to heavy metal lead, has the value further studied.
(2) adsorption dynamics adsorption kinetics
Adsorbent prepared by embodiment 1 is accessed to the Pb (NO containing 30mg/L3)2The polytetrafluoroethylene (PTFE) centrifuge tube of solution
In, the access concentration of adsorbent is 60mg/L, sets the concentration of NaCl in background solution as 150mM/L, the initial pH of regulation system
It is 5.5.Adsorption reaction system is placed in isothermal vibration incubator, setting revolving speed is 125rpm, and temperature is 30 DEG C, every certain
Time sampling, filter membrane (0.22 μm), with Pb remaining in atomic absorption spectrophotometer (AAS) measurement solution2+Concentration.
The time of contact of cell and lead ion has a major impact polycyclic aromatic hydrocarbon-degrading bacteria FA1 absorption heavy metal lead.Using
Its adsorption dynamics adsorption kinetics of batch Research on experimental methods, as a result as shown in Figure 2.At absorption initial stage, suction of the bacterial strain to lead ion in solution
Attached 5h adsorbance is more than half for reaching total adsorbance when adsorption equilibrium, and adsorbance increase later slows down more rapidly, absorption
Rate decline.After 25h, adsorbance reaches 80% of total adsorbance when adsorption equilibrium, and bacterial strain is to Pb2+Be adsorbed on 80h after reach
Complete equipilibrium.
To Pb in bacterial strain FA1 adsorbent solution2+Dynamics data carry out analysis fitting, discovery pseudo-second order kinetic and
Elovich kinetic model compares that other models are preferable to the fitting effect of experimental result, the coefficient R of fitting2It is higher,
Respectively 0.986 and 0.9643.Meet pseudo-second order kinetic relation table detail bacterium FA1 to Pb in solution2+Absorption with chemistry inhale
Based on attached or ion exchange, and Elovich model well then illustrates Herbaspirillum to experimental result fitting
Chlorophenolicum FA1 is to Pb2+Biological adsorption be a heterogeneous diffusion process.
(3) adsorption isotherm experiment
The initial p b of series is set2+Concentration is 5,10,20,30,50,100,200mg/L, according to adsorption dynamics adsorption kinetics in 2
Experiment condition carries out the adsorption isotherm experiment at 30 DEG C, and the equilibrium concentration and adsorbance of lead is measured by sampling up to after balancing in absorption.
At 30 DEG C, bacterial strain FA1 (dosage 60mg/L) Pb in solution (pH=5.5)2+(initial concentration 30mg/L's)
Isothermal adsorption result is as shown in Figure 3.Corresponding equilibrium adsorption capacity data in foundation figure and the equilibrium concentration data in solution respectively,
Result is fitted with Langmuir model and Freundlich model, the results showed that, two models intend adsorption process
Conjunction is all relatively good, and wherein the fitting effect of Langmuir model is more preferable, corresponding coefficient R2Up to 0.9926, bacterial strain is implied
FA1 tends to mono layer adsorption to the absorption of heavy metal Pb.The maximal absorptive capacity obtained according to Langmuir models fitting is high
Up to 151.52mg/g, show polycyclic aromatic hydrocarbon-degrading bacteria Herbaspirillum chlorophenolicum FA1 used in this experiment
To Pb2+Adsorption capacity there is competitiveness very much.In view of bacterial strain FA1 has the property for capableing of effective degrading polycyclic aromatic hydrocarbons simultaneously
Can, bacterial strain Herbaspirillum chlorophenolicum FA1 used by this experiment has a good application prospect, right
Heavy metal-polycyclic aromatic hydrocarbon composite pollution environment is biological prosthetic significant.
(4) different factors adsorb Pb to bacterial strain FA12+Influence
1) temperature adsorbs Pb to bacterial strain FA12+Influence: in Pb2+Concentration is 30mg/L, throwing bacterium amount 60mg/L, pH=
Under conditions of 5.5, batch adsorption experiment is carried out at 20 DEG C, 25 DEG C, 30 DEG C, 35 DEG C respectively, absorption is measured by sampling up to after balancing
Pb2+Concentration, research temperature adsorb Pb to bacterial strain2+The influence of effect.
The adsorbent that this adsorption experiment uses is active microorganism, and the variation of temperature will directly influence the growth of microorganism
And metabolic process, therefore temperature adsorbs Pb to bacterial strain FA12+Efficiency influence significant (Fig. 4).Temperature is at 20-30 DEG C, with temperature
The raising of degree, microorganism obviously increase the removal rate of heavy metal therewith, corresponding to remove as temperature is further increased to 35 DEG C
Rate reduces instead.Accordingly, determine polycyclic aromatic hydrocarbon-degrading bacteria FA1 to Pb2+Optimal adsorption temperature be 30 DEG C.
2) pH adsorbs Pb to bacterial strain FA12+Influence: in Pb2+Concentration is 30mg/L, throwing bacterium amount 60mg/L, 30 DEG C of temperature
Under conditions of, the pH for adjusting solution respectively is 2.0,3.0,4.0,5.0,5.5, and Pb is measured by sampling up to after balancing in absorption2+Concentration,
It studies pH and Pb is adsorbed to bacterial strain2+The influence of effect.
PH adsorbs Pb to bacterial strain FA12+The influence of efficiency is as shown in Figure 5.In lower pH value, degradation bacteria FA1 is to Pb2+'s
Adsorption rate is very low, this is because in low ph solution there is a large amount of hydrogen ions and Pb2+Between there is cell surface suctions
Competition on attached site.With becoming larger for pH value of solution, the negative electrical charge of microbial cell surface increases, and is conducive to positively charged divalent
The absorption of lead ion reaches highest when pH is 5.5.In order to which pH variation is better described to microorganism adsorption Pb2+Influence, this
The setting for testing pH value is controlled 6.0 hereinafter, when because of pH=6.0, and precipitation equally will cause the Pb in solution2+Subtract
It is few.Accordingly, it is determined that polycyclic aromatic hydrocarbon-degrading bacteria FA1 adsorbs Pb2+Optimal pH be 5.5.
Wherein, in above-mentioned experiment, using Pb in atomic absorption spectrophotometer flame method measurement sample2+Concentration.
Biological adsorption amount calculation formula are as follows:
In formula: q is the adsorbance (mg/g) of bacterium, c0It is the initial concentration (mg/L) in solution, c is solution after absorption
Concentration (mg/L), v are the volumes of solution, and m is the biological adsorption agent quality (g) that solution is added.
Claims (3)
1. application of the polycyclic aromatic hydrocarbon-degrading bacteria FA1 in absorption heavy metal Pb ion, point of the polycyclic aromatic hydrocarbon-degrading bacteria FA1
Class is named asHerbaspirillum chlorophenolicum, Chinese microorganism strain is preserved on May 4th, 2010
Preservation administration committee common micro-organisms center, deposit number are CGMCC No. 3797.
2. application according to claim 1, which is characterized in that for adjusting reaction system when adsorbing heavy metal ion
Temperature is 20 ~ 35 DEG C, and pH is 2 ~ 6.
3. application according to claim 1, which is characterized in that for adjusting reaction system when adsorbing heavy metal ion
Temperature is 25 ~ 30 DEG C, and pH is 5 ~ 6.
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