CN101397544A - Manganese oxidizing bacteria bacillus strain WH4 and use thereof - Google Patents

Abstract

The invention relates to a manganese oxidizing bacterium, in particular to a manganese oxidizing bacterium-bacillus sp.WH4 with the preservation number of CGMCC No.2089 and applications thereof. The invention provides strain resources for biosynthetic manganese oxides used in the treatment of toxic metallic elements and organic pollunts.

Description

Manganese oxidizing bacteria Bacillus strain WH4 and application thereof

Technical field

The present invention relates to the environmental microorganism field.Particularly, the present invention relates to the biosynthetic bacterial classification material of a kind of metal oxide, be Bacillus strain WH4 (Bacillus sp.WH4), by China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, preservation date is on June 14th, 2007, and preserving number is CGMCC No.2089.The invention still further relates to its application.

Background technology

Manganese (Mn) content in the earth's crust abundant (occupying the 10th) is the common heavy metal element that abundance is only second to iron.Mn oxide (oxide compound, hydroxy compound and the oxyhydroxide that comprise manganese) (Tebo etal., 2004) be mineral forms with height reactive behavior, as one of natural strong oxidizer, Mn oxide participates in the redox reaction of the organic and mineral compound of occurring in nature widely; Mn oxide has very big specific surface area and very strong adsorptive power simultaneously, can adsorb various heavy and trace element, determining these poisonous and distribution and the biological effectiveness thereof of important element in the surface knowledge system of land, be described as " street cleaner of ocean " always.

For a long time, Mn oxide is used as important mining mineral resource utilization always.Nature has been found the oxide/hydroxide mineral of more than 30 kind of manganese at present, is used to many fields, comprises water treatment and sewage disposal; The reparation of metal and organic substance in soil and the settling; The removal of metal and recovery.The sorbent material (Casale et al.2002) that is widely used as the filtration medium of tap water and removes Mn (II), Fe (II), hydrogen sulfide and arsenic specially as manganese green sand (glaukonine that promptly contains the different valence state Mn oxide); For another improving fixing (Hettiarachchi et al., 2000) plumbous in soil and the residual mining area sample significantly by adding phosphorus and Mn oxide cryptomelane.As important Mineral resources, Mn oxide also is widely used in the industrial production.Is crucial composition during steel industry and high-strength alloy are made as manganese as Oxygen Scavenger and sweetening agent.Manganese ore consumption on the our times in the Iron And Steel Industry has accounted for the 80-90% of manganese ore ultimate production; The content average out to 0.6% (weight percent) of manganese in the iron and steel then reaches 10% even also higher (Lankford et al., 1984) in high-intensity steel.Natural Mn oxide also is used as the negative material of zinc-carbon dry cell.Recent years, the alkaline cell that is material produce with Mn oxide synthetic, electrolytic has also occupied Battery Market rapidly.In addition, Mn oxide also is used to the production of particular aluminum alloy, coating and catalyzer etc.

The manganese oxide mineral is distributed widely in continent and big ocean floor, and the reserves of big ocean floor are tens to several thousand times of the corresponding reserves in land, and the exploitation difficulty is bigger.To 1996, China land found out that there are 213 places in the manganese ore district, possessed manganese ore reserves 5.66 hundred million t, occupied the 6th in the world.But generally speaking, the manganese resource skewness of China, the mineral products scale is less; In the manganese ore district, 213 place, large-scale have only 7 places, and all the other are medium and small mineral deposit; Ore quality is relatively poor, and based on lean ore, rich manganese ore reserves only account for 6.4%; Ore component complexity, mineral deposit belong to deposition or sedimentary metamorphic type more, and exploitation utilizes difficulty big.

The formation of Mn oxide comprises abiotic oxidation and bio-oxidation.The abiotic oxidation of Mn (II) comprises chemical oxidation, surface catalysis and disproportionation reaction, influenced by Eh and pH.Generally believe that Mn (II) is oxidized to Mn (VI) and is made up of two steps, generate Mn (III) earlier, as Mn ₃O ₄, γ-MnOOH, then through disproportionation or protonated generation Mn (VI) (Tebo et al., 1997).Though it is thermodynamic (al) spontaneous processes that chemical oxidation Mn (II) ion forms Mn (III)/Mn (IV) mineral, the kinetics speed of oxidising process is slower.The oxidation of catalysis Mn (II) and the generation of Mn oxide strongly of microorganism, particularly bacterium, (soon to 100,000 times) (Tebo etal., 1991 that make the generating rate specific surface chemical catalysis of Mn (IV) faster; 1997; 2004).Therefore the essential factor that Mn oxide formed during bio-oxidation was considered to control environment.From ocean and lake environment, separated at present and obtained some manganese oxidizing bacteria bacterial strains, and to three quasi-mode bacterial strains wherein, promptly the plate-like silk is sent out bacterium (Leptothrix discophora) SS-1 and SP-6, pseudomonas putida (Pseudomonas putida) MnB1 and GB-1, genus bacillus (Bacillussp.) SG-1 and has been carried out extensive studies.Result of study shows, has similar feature by the Mn oxide that forms naturally in microorganism catalysis synthetic Mn oxide (being called bio-oxidation manganese) and the environment, many different mineral structures (comprising lamellated or tunnel-shaped structure) are arranged, and its major ingredient is the lobate mineral such as the vernadite (δ-MnO of the low crystal form of 4 valencys ₂), acid birnessite or like manganite etc.Compare with the manganese oxide mineral of artificial chemosynthesis, a little less than the crystallization of bio-oxidation manganese, particle diameter is little, Mn valence state height, and octahedral voids is many in the structure, thereby has surfactivity (Toner et al., 2005 such as stronger absorption, oxidation; 2006).U (VI) enters the biological manganese oxide compound of growing, and form the biological manganese ore similar to todorokite, and similar transformation (Teboet al., 2004) can not take place the stratiform manganese ore of synthetic.Pb ²⁺Send out adsorptive capacity on the Mn oxide that bacterium SS-1 generates much larger than synthetic manganese mineral (Nelson, et al., 1999a at the plate-like silk; 1999b).The Zn of the Mn oxide absorption that pseudomonas putida MnB1 generates ²⁺Be positioned at the upper and lower in hole, maximal absorptive capacity reaches 4.1mol/kg, and is very identical with total hole number wherein; Biofilm loads etc. just adsorb Zn after the saturated absorption of Mn oxide ²⁺, adsorptive capacity can reach 38% (Toner et al., 2006) of total adsorptive capacity.More research shows that also biosynthetic Mn oxide is to Co ²⁺, Ni ²⁺, As ³⁺Deng metallic element stronger adsorptive power being arranged also, is new adsorbent, ion-exchanger and catalystic material (Nelson etal., the 1999a with very big exploitation potential; 2002; Tani et al., 2004; Villalobos et al., 2005), be the hot fields of present international research.Microbe species with manganese oxidation capacity is abundant, be distributed widely in the physical environment, but only limit to from the isolating manganese oxidizing microorganisms of marine facies for the bio-oxidation synthetic research of manganese in the world at present, will provide new microorganism resource for the biosynthesizing Mn oxide the research of the manganese oxidizing microorganisms in soil iron-manganese concretion source.

Summary of the invention

Technical problem to be solved by this invention provides the preparation method of synthetic bacterial strain of a kind of Mn oxide efficiently and biosynthesizing Mn oxide, and the synthetic microorganism resource and the method for providing of biological manganese oxide is provided, the present invention also comprises the application of above-mentioned bacterial strains.

In one aspect of the invention, the invention provides genus bacillus (Bacillus sp.) WH4 bacterial strain, its preserving number is CGMCC No.2089.

In one aspect of the invention, the invention provides the application that described bacterial strain is used for the biosynthesizing Mn oxide.

In one embodiment of the invention, described Mn oxide comprises the oxide compound of manganese, hydroxy compound and oxyhydroxide.

In another embodiment of the invention, described Mn oxide is used for the improvement of toxic metal element or organic pollutant.

In another embodiment of the invention, wherein said Mn oxide is used to prepare the sorbent material of metallic element.

In another embodiment of the invention, wherein said Mn oxide is used to prepare the ion-exchanger of metallic element.

In another embodiment of the invention, wherein said Mn oxide is used to prepare the catalyzer of metallic element.

The technical scheme that the present invention realizes comprises following several steps:

A. bacterium separation and Culture: preparation oligotrophic substratum, the iron-manganese concretion sample is carried out grinding to form powdery after the surface sterilization, with sterilized water do to coat behind 10 times of serial dilutions cultivate on the substratum 2 week back picking list bacterium colonies carry out separation and purification.

B. manganese oxidation bacterial strain screening: the bacterial strain of purifying is carried out color reaction with p-diaminodiphenyl, and the bacterial strain of choosing reacting positive carries out the Mn oxide synthetic test, the higher bacterial strain of screening manganese oxidation capacity.

C. scanning electron microscopic observation bound energy spectrum analysis (SEM-EDX) is confirmed the oxidation capacity of bacterial strain to manganese.

D. strain identification: morphological specificity is carried out in conjunction with 16S rDNA sequential analysis.

The effect of invention

Carry out separation and purification and p-diaminodiphenyl mensuration through single bacterium colony, obtain 3 apparent positive strains altogether, these 3 bacterial strains are being contained different concns MnCl a large amount of enrichment culture ₂Liquid nutrient medium in cultivate to do further screening, the result as shown in Figure 1, the MnCl 1 to 40mM ₂In the concentration range, 3 bacterial strains (WH4, WH21 and WH12) can both detect Mn ⁴⁺Generation, but bacterial strain WH4 synthetic MnO ₂Concentration be significantly higher than two other bacterial strain, it is at the MnCl of 20mM ₂The MnO that generates under the concentration ₂Concentration be up to 2000 μ M, be 100 μ M Mn/ days to the oxidation capacity of manganese, apparently higher than the oxidation capacity of the manganese oxidizing bacteria that obtains from water of bibliographical information: at the initial MnCl of 10mM ₂Be up to 25-35 μ M Mn/ days (Fernandes et al., 2005) under the concentration.MnCl at 40mM ₂Under the concentration, the MnO that bacterial strain WH4 generates ₂Concentration decreases during than 20mM, is 1719 μ M; Work as MnCl ₂When concentration reached 60mM, WH4 obviously was reduced to 45 μ M to the oxidation capacity of manganese.In physical environment, Mn ²⁺Concentration generally be no more than 1-5 μ M, Mn in environment ²⁺Concentration during greater than 10 μ M, Mn ²⁺Promptly to the Production by Bacteria toxigenicity and bacteria growing inhibiting (Chapnick et al., 1982).The manganese oxidizing bacteria oxidation Mn that in the past reported from the marine environment acquisition ²⁺Optimum concentration range (Nealson, 2006 between 10 μ M-10mM; Fernandes et al., 2005).As seen, bacterial strain WH4 not only has stronger manganese oxidation capacity, and is also very strong to the tolerance of manganese, can be used as Mn oxide synthetic good material.

Previous studies shows, the manganese oxidizing bacteria is by different modes: enter substratum, form glycocalix or participate in the catalytic oxidation process (Tebo et al., 2004) of manganese by being distributed in enzyme on the perispore outside cytolemma as secretion manganese oxidation factor.The result of scanning electron microscope bound energy spectrum analysis can detect higher manganese oxide content on the thalline of bacterial strain WH4 shown in Fig. 2 and table 1, reach 28.48%.As seen bacterial strain WH4 can form the aggregate of Mn oxide at perispore, and it may be by being distributed in the oxidising process of the protein factor participation manganese on the perispore.

The strain identification result: bacterial strain WH4 bacterium colony oyster white is containing MnCl ₂Substratum on cultivated for 2 weeks after because the manganese oxide that forms is rich long-pending around thalline, bacterium colony changes into brown to brown gradually, and prolongs colony colour with incubation time and deepen.Gram-positive microorganism, cell is shaft-like, and big or small 0.4-0.6 μ m * 1-2 μ m produces gemma, the gemma column.The homology analysis of sequence shows among the 16S rDNA sequence of this bacterial strain and the GenBank, among the GenBank with it the bacterial classification that reaches more than 90% of sequence homology mainly be genus bacillus (Bacillus), and genus bacillus (Bacillus sp.41KF2b, Bacillus sp.ROO40B) and the sequence similarity of bacillus pumilus (Bacillus pumilus) and bacterial strain WH4 reach 100% (table 2), so determine that this bacterium is a genus bacillus, tentative genus bacillus WH4 (Bacillus sp.WH4) by name.

Description of drawings

The bacterial strain that Fig. 1 display separation arrives is at different MnCl ₂Under the concentration to the oxidation capacity of manganese.

Fig. 2 shows that genus bacillus WH4 is containing 1mM MnCl ₂Solid medium on cultivate the sem photograph (5000 *) after 20 days and the energy spectrum analysis figure of gauge point correspondence.Wherein, arrow is depicted as gauge point.

Fig. 3 shows the 16S rDNA sequence (1460bp) of genus bacillus WH4.

The following examples are further illustrated aspect of the present invention.Yet they never limit instruction of the present invention or the disclosure that provides as this paper.

Embodiment

Embodiment 1. materials are prepared: A. oligotrophic substratum (improved L2 substratum):

Peptone (peptone): 2g Yeast extract (yeast extract): 0.5g

NaCl：8.766g KCl：0.373g

MgSO ₄：6.162g CaCl ₂：0.555g

Trace element: 1ml distilled water: 1L

PH 7.5 agar: 1.5% (m/v)

Trace element:

CuSO ₄.5H ₂O：10mM ZnSO ₄.7H ₂O：44mM

CoCl ₂.6H ₂O：20mM Na ₂MoO ₄.2H ₂O：13mM

Substratum preparation is finished the back and is regulated pH to 7.5 with NaOH, and 115 ℃ of moist heat sterilization 30min treat the HEPES (pH 7.5) and the MnCl of adding filtration sterilization (0.25 μ m) when substratum is cooled to 60 ℃ of left and right sides ₂Solution makes HEPES and MnCl ₂Final concentration be respectively 20mM and 1mM.Preparation is dull and stereotyped behind the mixing.With above-mentioned composition do not add agar prepare the corresponding liquid substratum.

B. iron-manganese concretion sample

Sample is taken from the claypan Udalf sublayer in Wuhan, Hubei, and soil parent material is following another name for Sichuan Province loess (Q3).With aseptic nipper iron-manganese concretion sample (diameter 5-9mm, subsphaeroidal) is chosen from soil and to be put into valve bag, transport the laboratory on ice back, deposit for 4 ℃.

Embodiment 2. bacterium separation and Culture:

The iron-manganese concretion sample is removed the tuberculosis surface soil with aseptic washed several times with water, with aseptic filter paper inhale remove unnecessary moisture content after, get the 10g sample and carry out surface sterilization 1min with 0.1% NaClO, wash the NaClO of several with aseptic deionized water again with flush away remnants, with mortar sample is ground to form powdery under the aseptic condition, add the 10ml sterilized water, get 1ml behind the mixing and coat above-mentioned culture plate, 25 ℃ of 2 weeks of cultivation to getting the 0.2ml suspension after the 9ml sterilized water is made 10 times of serial dilutions.The culture dish picking of choosing colony number and be 30-100 has single bacterium colony of different colony characteristicses, the line separation and Culture, and culture condition is the same.

Embodiment 3. has the bacterial isolates screening of manganese oxidation activity

A. p-diaminodiphenyl qualitative detection:

P-diaminodiphenyl (the going back ortho states) solution of new preparation is light blue, when around Mn is arranged ⁴⁺When existing, Mn ⁴⁺To go back and show mazarine after the oxidation of ortho states p-diaminodiphenyl, and cultivate the p-diaminodiphenyl solution that drips the new preparation of 1% (m/v) on the bacterium lawn in 2 weeks to flat board, it is positive that lawn becomes mazarine person thereupon, and expression has Mn ⁴⁺Generate.After testing, obtain 3 apparent positive strains (WH4, WH21 and WH12) altogether.

B.LBB (Leukoberbelin blue) development process quantitative screening:

Microorganism synthetic bio-oxidation manganese is mainly hydrated state or hydroxylated MnO ₂, so MnO ₂Content can characterize what of synthetic bio-oxidation manganese.Also the Leukoberbelin blue (LBB) of ortho states is having 3 valencys and the Mn (Mn more than 3 valencys ³+, Mn ⁴⁺Or Mn ⁷⁺) oxidized apparent blueness when existing, under 620nm, have maximum light absorption value, with KMnO ₄For standard substance is made typical curve, by the KMnO of 40 μ M ₄The MnO that is equivalent to 100 μ M ₂Can convert MnO ₂Content (Krumbein﹠amp; Altmann, 1973).Scrape with transfering loop and to get among the A single bacterium colony of 3 positive strains and be inoculated into respectively to cultivate in the aforesaid liquid substratum and made seed liquor in 36 hours, seed liquor is inoculated in the ratio of 2% (v/v) respectively contains 0,1,5,10,20,40,60mM MnCl ₂The aforesaid liquid substratum, in 25 ℃ cultivate 20 days after sampling respectively, with MnO in the LBB determination of color nutrient solution ₂Concentration, not inoculate the MnCl that contains respective concentration of bacterium liquid ₂The aforesaid liquid substratum be blank, each is handled and to establish 4 repetitions.The result is referring to Fig. 1: the MnCl 1 to 40mM ₂In the concentration range, 3 bacterial strains (WH4, WH21 and WH12) can both detect Mn ⁴⁺Generation, but MnO in the bacterial strain WH4 synthetic bio-oxidation manganese ₂Concentration is significantly higher than two other bacterial strain, and it is at the MnCl of 20mM ₂The MnO that generates under the concentration ₂Concentration be up to 2000 μ M, be 100 μ M Mn/ days to the oxidation capacity of manganese, apparently higher than the oxidation capacity of the manganese oxidizing bacteria that obtains from water of bibliographical information: at the initial MnCl of 10mM ₂Be up to 25-35 μ M Mn/ days (Fernandes et al., 2005) under the concentration.

The scanning electron microscopic observation (SEM) of embodiment 4. bacterial strain WH4 and energy spectrum analysis (EDX)

Scrape to be taken at and contain 1mM MnCl ₂Oligotrophic substratum solid plate on the lawn cultivated 20 days prepare sample by biological sample Electronic Speculum preparation requirement (Ying Guohua, 1993), carry out scanning electron microscopic observation and energy spectrum analysis (S-3000N, HITACHI Co., Japan).The result of scanning electron microscope bound energy spectrum analysis can detect higher manganese oxide content on the thalline of bacterial strain WH4 shown in Fig. 2 and table 1, reach 28.48%.As seen bacterial strain WH4 can form the aggregate of Mn oxide at perispore, and it may be by being distributed in the oxidising process of the protein factor participation manganese on the perispore.

Table 1 EDAX results

Annotate: ^aK refers to the energy that atom K layer excites, and all the other together.

Embodiment 5. bacterial genomes DNA extraction and 16S rDNA sequential analysis

With SDS alkaline lysis-phenol: chloroform: (25:24:1, v/v/v) method of extracting-ethanol sedimentation (Maloy, 1990) is extracted bacterial genomes DNA to primary isoamyl alcohol.(27F:AGAGTTTGATCMTGGCTCAG and 1492R:TACGGYTACCTTGTTACGACTT) carries out pcr amplification with bacterial 16 S rDNA universal primer, obtain the PCR product of the about 1500bp of clip size, send Sinogenomax Co., Ltd.'s order-checking behind the PCR product purification.

The sequence that records is carried out carrying out nucleotide homology relatively in GenBank with NCBI (National Center for Biotechnology Information) blast program after sequence assembly and the artificial check and correction with the DNAStar software package.The result shows that the bacterial classification that reaches more than 90% with bacterial strain WH4 homology among the GenBank mainly is genus bacillus (Bacillus), genus bacillus (Bacillus sp.41KF2b, Bacillus sp.ROO40B) and the sequence similarity of bacillus pumilus (Bacillus pumilus) and bacterial strain WH4 reach 100% (table 2), so determine that this bacterium is a genus bacillus, tentative genus bacillus WH4 (Bacillus sp.WH4) by name.

The BLAST analytical results of the 16S rDNA of table 2 bacterial strain WH4

Genus bacillus WH4 (Bacillus sp.WH4) is deposited in (No. 13, one in Haidian District, Beijing City north, China Committee for Culture Collection of Microorganisms common micro-organisms center on June 14th, 2007, Institute of Microorganism, Academia Sinica), preserving number is CGMCC No.2089.

Reference

1.Casale，R.J，LeChevallier，M.W.，Pontius，F.W.2002.ManganeseControl?and?Related?Issues.Denver，C.AWWA?Res.Found.Am.WaterWorks?Assoc.187pp.

2.Chapnick?S.D.，Moore；W.S.，Nealson，K.H.1982.Microbially?mediatedmanganese?oxidation?in?afreshwater?lake.Limnology?and?Oceanography27，1004-1014.

3.Fernandes，S.O.，Krishnan，K.P.，Khedekar，V.D.，Loka?Bharathi，P.A..2005.Manganese?oxidation?by?bacterial?isolates?from?the?Indian?ridgesystem.Biometals?18：483-492.

4.Hettiarachchi，G.M.，Pierzynski，G.M.，Ransom，M.D.2000.In?situstabilization?of?soil?lead?using?phosphorus?and?manganese?oxide.Environmental?Science?and?Technology?34：4614-4619.

5.Krumbein，W.E.，Altmann，H.J.1973.A?new?method?for?the?detectionand?enumeration?of?manganese?oxidizing?and?reducing?microorganisms.Helgoland?Marine?Research?25：347-356.

6.Lankford，W.T.Jr，Samways，N.L.，Craven，R.F.，McGannon，H.E.1984.The?Making，Shaping，and?Treating?of?Steel(Assoc.of?Iron?and?SteelEngineers，Pittsburgh，PA).

7.Maloy，S.R..Experimental?Techniques?in?Bacterial?Genetics，Boston：Jones?and?Bartlett?Publishers，1990.

8.Nealson.K.H.2006.The?Manganese-oxidizing?bacteria.In?TheProkaryotes.Springer?New?York.

9.Nelson，Y.M.，Lion，L.W.，Ghiorse，W.C.，Shuler，M.L.1999a.Production?of?biogenic?Mn?oxides?by?Leptothrix?discophora?SS-1?in?achemically?defined?growth?medium?and?evaluation?of?their?Pb?adsorptioncharacteristics.Applied?and?Environmental?Microbiology?65，175-180.

10.Nelson，Y.M.，Lion，L.W.，Shuler，M.L.，Ghiorse，W.C.1999b.Leadbinding?to?metal?oxide?and?organic?phases?of?natural?aquatic?biofilms.Limnol?Oceanogr?4：1715-1729.

11.Nelson，Y.M.；Lion，L.W.；Shuler，M.L.；Ghiorse，W.C.2002.Effect?ofoxide?formation?mechanisms?on?lead?adsorption?by?biogenic?manganese(hydr)oxides，iron(hydr)oxides，and?their?mixtures.EnvironmentalScience?and?Technology?36，421-425.

12.Tani，Y.，Ohashi，M.，Miyata，N.，Seyama，H.，Iwahori，K.，Soma，M.2004.Sorption?of?Co(II)，Ni(II)，and?Zn(II)on?biogenic?manganese?oxidesproduced?by?a?Mn-oxidizing?fungus，strain?KR21-2.Journal?ofEnvironmental?Science?and?Health?A.39，2641-2660.

13.Tebo?B?M.1991.Manganese(II)oxidation?in?the?suboxic?zone?of?theBlack?Sea.Deep?Sea?Research?38(Suppl)：883-905.

14.Tebo，B.M.，Bargar，J.R.，Clement，B.G.，Dick，G.J.，Murray，K.J.，Parker，D.，Verity，R.，Webb，S.M.2004.Biogenic?manganese?oxides：Properties?and?mechanisms?of?formation.Annual?Review?Earth?PlanetScience?32：287-328.

15.Tebo，B.M.，Ghiorse，W.C.，Waasbergen，van?L.G.，Siering，P.L.，Caspi，R.1997.Bacterially-mediated?mineral?formation：Insights?intomanganese(II)oxidation?from?molecular?genetic?and?biochemical?studies.In?Geomicrobiology：Interactions?between?Microbes?and?Minerals.Banfield，J.F.，and?Nealson，K.H.(eds).Mineralogical?Society?ofAmerica，Washington，D.C.，pp.225-266.

16.Toner，B.，Fakra，S.，Villalobos，M.，Warwick，T.，Sposito，G.2005.Spatially?resolved?characterization?of?biogenic?manganese?oxideproduction?within?a?bacterial?biofilm.Applied?and?EnvironmentalMicrobiology?71：1300-1310.

17.Toner，B.，Manceau，A.，Webb，S.M.，Sposito，G.2006.Zinc?sorption?tobiogenic?hexagonal-birnessite?particles?within?a?hydrated?bacterial?biofilm.GEOCHIMICA?ET?COSMOCHIMICA?ACTA?70：27-43.

18.Villalobos，M.，Tebo，B.M.2005.Introduction：Advances?in?thegeomicrobiology?and?biogeochemistry?of?manganese?and?iron?oxidation.Geomicrobiology?Journal?22：77-78.

19. Ying Guohua, the chief editor. Medical Biology electron microscopy and cell ultrastructure, the modern press in Hong Kong, 1993, pp59-63.

Sequence table

＜110〉Ecological Environment Research Center, Chinese Academy of Sciences

＜120〉manganese oxidizing bacteria Bacillus strain WH4 and application thereof

<130>IB076991

<160>1

<170>PatentIn?version?3.1

<210>1

<211>1459

<212>DNA

<213>Bacillus?sp.WH4

<400>1

Claims (7)

1. genus bacillus (Bacillus sp.) WH4 bacterial strain, its preserving number is CGMCC No.2089.

2. the application of the bacterial strain of claim 1 in the biosynthesizing Mn oxide.

The application of 3 claims 2, wherein said Mn oxide comprises the oxide compound of manganese, hydroxy compound and oxyhydroxide.

The application of 4 claims 2, wherein said Mn oxide is used for the improvement of toxic metal element or organic pollutant.

The application of 5 claims 4, wherein said Mn oxide is used to prepare the sorbent material of metallic element.

6. the application of claim 4, wherein said Mn oxide is used to prepare the ion-exchanger of metallic element.

7. the application of claim 4, wherein said Mn oxide is used to prepare the catalyzer of metallic element.

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