CN101353634B - Klebsiella pneumoniae and use thereof - Google Patents

Klebsiella pneumoniae and use thereof Download PDF

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CN101353634B
CN101353634B CN 200810198451 CN200810198451A CN101353634B CN 101353634 B CN101353634 B CN 101353634B CN 200810198451 CN200810198451 CN 200810198451 CN 200810198451 A CN200810198451 A CN 200810198451A CN 101353634 B CN101353634 B CN 101353634B
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bacterium
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klebsiella pneumoniae
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CN101353634A (en
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周顺桂
王跃强
武春媛
庄莉
李芳柏
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Guangdong Institute of Eco Environmental Science and Technology
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Guangdong Institute of Eco Environment and Soil Sciences
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Abstract

The present invention discloses a klebsiella pneumoniae and application thereof; the invention obtains the klebsiella pneumoniae (MFC4) by separation and purification from the soil sample of an ancient forest of Zhao qing in Guangdong, which is preserved in China culture collection and management committee general microorganisms center, with the preservation number of CGMCC NO. 2454; the invention simultaneously provides a screening method of the strain and application thereof in the reduction of Fe(III) or humic acid; the strain can reduce iron oxides with high reduction and crystallinity, such as lepidocrocite and hematite, etc.

Description

One Klebsiella pneumoniae and application thereof
Technical field
The invention belongs to novel bacterial triage techniques and Environmental Biotechnology field, be specifically related to a Klebsiella pneumoniae (Klebsiella pneumoniae MFC4) and the application in Fe (III) reduction and humic acid reduction thereof.
Technical background
Fe (III) and humic acid are a large amount of materials that exist in the ecotopes such as settling, soil.Fe (III) mainly exists with the insoluble form of iron oxide, and the average content in the earth's crust is about 5.6%; Humic acid is to be dissolved in the part that alkali is insoluble to acid in the soil ulmin, is rich in quinonyl, heterogeneous natural organic mixture.Under the anaerobic condition, some microorganisms can pass through the biochemical reactions oxidation of organic compounds, and simultaneously with Fe (III) oxide compound or humic acid reduction, and the acquisition energy is grown.This quasi-microorganism is called as Fe (III) or humic acid reduction bacterium.
Microorganism is all significant in the biomass geochemistry working cycle of element to the reductive action of Fe (III) and humic acid, be embodied in: 1) Fe (III) or humic acid reduction bacterium is in the process of reduction Fe (III) or humic acid, can produce CO directly with organic matter in the environment or environment toxic substance oxidations such as toluene, vinylchlorid 2, participate in carbon cycle.2) reduction product Fe (II) or go back the ortho states humic acid and can the chemistry redox reaction take place with some oxidation state hazardous and noxious substances (Cr (VI), U (VI), nitroaromatic, azoic dyestuff and many halos pollutent) in the environment, thus the toxicity of these materials reduced.Therefore, microorganism has influence on the biomass geochemistry circulation of C in the environment, N, Fe, Mn and some trace metal elements to the reductive action of two kinds of materials, and can promote the detoxification of heavy metal and organic pollutant.Fe (III) or the latent effect of humic acid reduction bacterium in self purification of water body, the reparation of contaminated soil original position, sewage disposal have caused extensive concern.
Since the eighties in 19th century, from soil, settling, water body and mud, be separated to hundreds of Fe (III) reduction bacterium successively, comprise ground bacillus, Shiva Salmonella, methanogen, fermenting bacteria, thermophile bacteria etc.; The nineties in 20th century, multiple with humic acid or humic acid pattern thing anthraquinone-2,6-disulfonic acid (AQDS) is the also separated and evaluation of the bacterium of terminal electron acceptor, and wherein the part bacterial strain also has Fe (III) restoring function.But the bacterial strain that possesses Fe (III) reduction and humic acid restoring function when reporting is few in number, and does not find as yet that at present Klebsiella pneumonia can anaerobic reduction Fe (III) reduction and humic acid.
Summary of the invention
The object of the present invention is to provide a kind of new efficient bacterial strain: Klebsiella pneumonia (Klebsiella pneumoniae MFC4) with Fe (III)/humic acid restoring function.
Another object of the present invention is to provide the application of above-mentioned Klebsiella pneumonia in Fe (III) oxide compound and humic acid reduction.
The present invention is through a large amount of experimental studies, separation and purification obtains a Klebsiella pneumoniae from the ancient forest soil sample in Zhaoqing, Guangdong, have stronger Fe (III)/humic acid reducing power, the invention provides the application of described Klebsiella pneumonia aspect Fe (III) or humic acid reduction.
Klebsiella pneumonia provided by the invention (Klebsiella pneumoniae MFC4 or K.pneumoniae MFC4), be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on April 14th, 2008, preserving number is: CGMCC2454.
The enrichment of Klebsiella pneumonia of the present invention, separation and purification process may further comprise the steps:
(1) enrichment
Described enrichment is to pipette or take by weighing ancient forest soil sample to be inoculated in the liquid separation culture medium; Drum fills high-purity mixed gas (N in the nutrient solution 2/ CO 2=80/20) after, air-blowing finishes culture placed anaerobism workstation (N 2/ CO 2=80/20) leaves standstill cultivation.The colour-change situation that the productive rate of detection Fe (II) is observed nutrient solution simultaneously.When the output of Fe (II) constantly increased, the color of nutrient solution became black at last from original orange-yellow deepening gradually.When the output of Fe (II) surpass total iron content 50% the time, the inoculum size with 10% is forwarded in another fresh substratum, aforesaid operations three times repeatedly.
Contain in every liter of liquid separation culture medium: 1.0g glucose, 16g ironic citrate, 2.5gNaHCO 3, 0.25g NH 4Cl, 0.678g NaH 2PO 42H 2O, 0.1g KCl, vitamin solution 10.0mL and trace element solution 10.0mL.
Wherein, vitamin solution is to contain 2.0mg vitamin H, 2.0mg folic acid, 10.0mg vitamin B6,5.0mg VitB1,5.0mg riboflavin, 5.0mg nicotinic acid, 5.0mg calcium pantothenate, 0.1mg vitamin B12,5.0mg para-amino benzoic acid and 5.0mg Thioctic Acid in every premium on currency.
Described trace element solution is to contain 1.5g nitrilotriacetic acid(NTA), 3.0gMgSO in every premium on currency 47H 2O, 0.5g MnSO 4H 2O, 1.0g NaCl, 0.1g FeSO 47H 2O, 0.1gCoCl 26H 2O, 0.1g CaCl 2, 0.1g ZnSO 47H 2O, 0.01g CuSO 45H 2O, 0.01gAlK (SO 4) 212H 2O, 0.01gH 3BO 3With 0.01g Na 2MoO 42H 2O.
(2) purifying
Evenly coat after the reacted nutrient solution of the 4th generation enrichment suitably diluted (solid separation culture medium is to add the 18g agar powder in every liter of liquid separation culture medium) on the solid isolation medium, place anaerobism workstation (N 2/ CO 2=80/20) cultivate, picking list bacterium colony carries out single bacterium colony isolation and purification.
The present invention adopts following method to verify the Fe of described bacterial strain (III) restoring function.
Be seeded to the beef extract-peptone liquid nutrient medium from the inclined-plane of preserving bacterial strain of the present invention, in 30 ℃, shaking table activation thalline is 16～24 hours under 180 rev/mins of conditions, makes bacterial number reach exponential phase of growth, gets bacterial suspension.
1) according to separation and Culture based formulas and method, add different organism (formic acid, acetate, propionic acid, lactic acid, citric acid, ethanol, glycerol, glucose or sucrose) and ferriferous oxide (ferrihydrite, pyrrhosiderite, lepidocrocite or rhombohedral iron ore), inoculum size inoculation with 10% activates bacterium liquid in the isolation medium of liquid, anaerobism workstation (N 2/ CO 2=80/20) following 30 ℃ leave standstill cultivation;
2) at regular intervals (about 5～10 days) get the 4mL nutrient solution in 16mL0.5M HCl solution 180 rev/mins the concussion 1.5 hours; Precipitations such as thalline are filtered, adopt adjacent phenanthroline method to give the Fe (II) in the filtrate colour developing, adopt ultraviolet spectrophotometer to measure the content of Fe (II) at the 510nm place, the iron reducing activity of checking bacterial strain.
The bacterial strain with Fe (III) reduction and humic acid reducing power that provides a strain new is provided, with report bacterial strain and compare, described Klebsiella pneumonia bacterial strain has stronger Fe (III) reduction and humic acid reducing power, and the higher ferriferous oxide of the degree of crystallinity of reducing is lepidocrocite, rhombohedral iron ore etc. for example.
Description of drawings
Fig. 1 is 20000 times of scanning electron microscope diagram sheets of Klebsiella pneumonia (Klebsiella pneumoniae MFC4)
Embodiment
Enrichment, separation and the purifying of embodiment 1 Klebsiella pneumonia
Pipette 5mL or take by weighing the ancient forest soil sample of 5g and be inoculated in the liquid separation culture medium; Drum fills high-purity mixed gas (N in the nutrient solution 2/ CO 2=80/20), the air-blowing time is no less than 30 minutes; After air-blowing finishes culture placed anaerobism workstation (N 2/ CO 2=80/20) 30 ℃ leave standstill cultivation.The colour-change situation that the productive rate of detection Fe (II) is observed nutrient solution simultaneously.When the output of Fe (II) constantly increased, the color of nutrient solution became black at last from original orange-yellow deepening gradually.When the output of Fe (II) surpass total iron content 50% the time, the inoculum size with 10% is forwarded in another fresh substratum, aforesaid operations three times repeatedly.
Contain in every liter of liquid separation culture medium: 1.0g glucose, 16g ironic citrate, 2.5gNaHCO 3, 0.25g NH 4Cl, 0.678g NaH 2PO 42H 2O, 0.1g KCI, vitamin solution 10.0mL and trace element solution 10.0mL.
Wherein, vitamin solution is to contain 2.0mg vitamin H, 2.0mg folic acid, 10.0mg vitamin B6,5.0mg VitB1,5.0mg riboflavin, 5.0mg nicotinic acid, 5.0mg calcium pantothenate, 0.1mg vitamin B12,5.0mg para-amino benzoic acid and 5.0mg Thioctic Acid in every premium on currency;
Described trace element solution is to contain 1.5g nitrilotriacetic acid(NTA), 3.0gMgSO in every premium on currency 47H 2O, 0.5g MnSO 4H 2O, 1.0g NaCl, 0.1g FeSO 47H 2O, 0.1gCoCl 26H 2O, 0.1g CaCl 2, 0.1g ZnSO 47H 2O, 0.01g CuSO 45H 2O, 0.01gAlK (SO 4) 212H 2O, 0.01g H 3BO 3With 0.01g Na 2MoO 42H 2O.
Evenly coat on the solid isolation medium after the reacted nutrient solution of the 4th generation enrichment suitably diluted, place anaerobism workstation (N 2/ CO 2=80/20) cultivated 48 hours for 30 ℃, picking list bacterium colony carries out single bacterium colony isolation and purification.
The form of Klebsiella pneumonia of the present invention, physiology, biochemistry and 16S rDNA gene sequence characteristic:
1) thalline, colonial morphology characteristic: adopt conventional bacterium electron microscope observation, this bacterial strain is a Gram-negative bacteria, amphimicrobian, and rod-short, size is 1.0～1.7 * 0.6～0.8 μ m.Aerobic cultivation is after 24 hours on beef extract-peptone agar solid medium flat board, and colonial morphology is circular, neat in edge; Smooth surface, low convex surface; Opaque, white, colony diameter is 2～3mm.
2) physiological and biochemical property: this bacterium tool amphimicrobian energy for growth under anaerobic, can be an electron donor with glucose, sucrose, glycerine or citric acid, reductive water iron ore, pyrrhosiderite, lepidocrocite, rhombohedral iron ore and AQDS.Other physiological and biochemical properties after measured see Table 1.
Table 1 Klebsiella pneumonia (K.pneumoniae MFC4) physiological and biochemical property
Figure G2008101984515D00061
(annotate: "+" is expressed as positive reaction, and "-" is expressed as negative reaction)
3) 16S rDNA gene sequence characteristic: adopt ordinary method, use the SDS-Proteinase K, chloroform-primary isoamyl alcohol (volume ratio 24:1) extracting, the method for 0.6 volume isopropanol precipitating is extracted bacteria total DNA.Adopt the 16S rDNA of bacterial 16 S rDNA universal primer F27 and R1492 amplification bacterium, check order after pcr amplification product is reclaimed.Again the base sequence that obtains is carried out homologous sequence search (Blastsearch) in international nucleic acid sequence data storehouses such as GenBank, find out the type strain that homology is the highest in this bacterial strain and the database or be preserved in ATCC or the bacterial strain of international DSMZ such as DSM.According to comparison result, the 16S rDNA sequence tool highest homology of bacterial strain and Klebsiella pneumonia reaches 99.6%.
In conjunction with above-mentioned physio-biochemical characteristics, 16S rDNA sequence alignment result, bacterial strain of the present invention is named as Klebsiella pneumonia (Klebsiella pneumoniae MFC4).
Embodiment 2 Klebsiella pneumonia (Klebsiella pneumoniae MFC4) anaerobic reduction pyrrhosiderite
Culture medium A prescription: contain pyrrhosiderite 2.25g in every liter of deionized water, NaHCO 32.5g, NH 4Cl0.25g, NaH 2PO 42H 2O0.678g, KCl0.1g, glucose 1.0g, each 10.0mL of vitamin solution and trace element solution (vitamin solution and trace element solution composition are with embodiment 1 described liquid separation culture medium), in 121 ℃ of sterilizations 20 minutes, glucose was sterilized separately, and mix with other composition the sterilization back.
Preparation bacterial suspension: encircle the activatory bacterial strain in 200mL beef extract-peptone liquid nutrient medium with transfering loop picking one, in shaking table (30 ℃, 180 rev/mins) cultivated 16 hours, make bacterial number reach exponential phase of growth, 8000 rev/mins of centrifugal collection thalline, remove supernatant liquor, the thalline that precipitates is suspended in the 200mL culture medium A, make bacteria suspension.
Inoculated bacteria suspension makes its bacterium number reach 10 in nutrient solution A 8CFU/mL, drum fills high-purity mixed gas (N then 2/ CO 2=80/20), the drum time of filling is no less than 30 minutes, leaves standstill anaerobism in 30 ℃ and cultivates.Observe the colour-change situation of nutrient solution, got the 180 rev/mins of vibrations 1.5 hours in 16mL0.5mol/L HCl solution of the 4mL nutrient solution that mixes every 5～10 days, refilter, adopt adjacent phenanthroline method to give the Fe (II) in the filtrate colour developing, adopt ultraviolet spectrophotometer to measure the content of Fe (II) at the 510nm place, and be contrast with the nutrient solution that does not add bacterium, the iron reducing power of checking bacterial strain.Fe (II) content sees Table 2, and Fe (II) content unit is mg/L, represents the milligram number of contained Fe (II) in every liter of sample.As shown in Table 2, along with the prolongation of incubation time, the growing amount of Fe in the system (II) constantly increases, and apparently higher than not adding thalline system, illustrates that this bacterium has stronger reduction pyrrhosiderite function.
In the table 2 Klebsiella pneumonia MFC4 reduction pyrrhosiderite process
Fe (II) produces dynamically (mg/L of unit)
0 day 5 days 10 days 20 days 25 days
Do not add the bacterium contrast 25.47 23.88 20.52 32.86 38.70
Add bacterium 25.47 36.37 54.64 97.94 128.34
Embodiment 3 Klebsiella pneumonia (K.pneumoniae MFC4) anaerobic reduction ferrihydrite
Substratum B prescription: contain ferrihydrite 2.5g in every liter of deionized water, NaHCO 32.5g, NH 4Cl0.25g, NaH 2PO 42H 2O0.678g, KCl0.1g, glucose 1.0g, each 10.0mL of vitamin solution and trace element solution (vitamin solution and trace element solution composition are with embodiment 1 described liquid separation culture medium), in 121 ℃ of sterilizations 20 minutes, glucose was sterilized separately, and mix with other composition the sterilization back.
Preparation bacterial suspension: encircle the activatory bacterial strain in 200mL beef extract-peptone liquid nutrient medium with transfering loop picking one, in shaking table (30 ℃, 180 rev/mins) cultivated 16 hours, make bacterial number reach exponential phase of growth, 8000 rev/mins of centrifugal collection thalline, remove supernatant liquor, the thalline that precipitates is suspended among the 200mL substratum B, make bacteria suspension.
Inoculated bacteria suspension makes its bacterium number reach 10 in nutrient solution B 8CFU/mL, drum fills high-purity mixed gas (N then 2/ CO 2=80/20), the drum time of filling is no less than 30 minutes, leaves standstill anaerobism in 30 ℃ and cultivates.Observe the colour-change situation of nutrient solution, got the 180 rev/mins of vibrations 1.5 hours in 16mL0.5mol/L HCl solution of the 4mL nutrient solution that mixes every 5～10 days, refilter, adopt adjacent phenanthroline method to give the Fe (II) in the filtrate colour developing, adopt ultraviolet spectrophotometer to measure the content of Fe (II) at the 510nm place, and be contrast with the nutrient solution that does not add bacterium, the iron reducing power of checking bacterial strain.Fe (II) content sees Table 3, and Fe (II) content unit is mg/L, represents the milligram number of contained Fe (II) in every liter of sample.As shown in Table 3, the growing amount of Fe in the system (II) is apparently higher than the system that does not add bacterium, illustrate that this bacterium can the anaerobic reduction ferrihydrite, and reducing power is stronger.
In the table 3 Klebsiella pneumonia MFC4 reductive water iron ore process
Fe (II) produces dynamically (mg/L of unit)
0 day 5 days 10 days 20 days 25 days
Do not add the bacterium contrast 18.86 22.69 18.83 24.84 24.72
Add bacterium 18.86 36.43 77.64 129.16 164.11
Embodiment 4 Klebsiella pneumonia (K.pneumoniae MFC4) anaerobic reduction lepidocrocite
Culture medium C prescription: contain lepidocrocite 2.25g in every liter of deionized water, NaHCO 32.5g, NH 4Cl0.25g, NaH 2PO 42H 2O0.678g, KCl0.1g, glucose 1.0g, each 10.0mL of vitamin solution and trace element solution (vitamin solution and trace element solution composition are with embodiment 1 described liquid separation culture medium), in 121 ℃ of sterilizations 20 minutes, glucose was sterilized separately, and mix with other composition the sterilization back.
Preparation bacterial suspension: encircle the activatory bacterial strain in 200mL beef extract-peptone liquid nutrient medium with transfering loop picking one, in shaking table (30 ℃, 180 rev/mins) cultivated 16 hours, make bacterial number reach exponential phase of growth, 8000 rev/mins of centrifugal collection thalline, remove supernatant liquor, the thalline that precipitates is suspended in the 200mL culture medium C, make bacteria suspension.
Inoculated bacteria suspension makes its bacterium number reach 10 in nutrient solution C 8CFU/mL, drum fills high-purity mixed gas (N then 2/ CO 2=80/20), the drum time of filling is no less than 30 minutes, leaves standstill anaerobism in 30 ℃ and cultivates.Observe the colour-change situation of nutrient solution, got the 180 rev/mins of vibrations 1.5 hours in 16mL0.5mol/L HCl solution of the 4mL nutrient solution that mixes every 5～10 days, refilter, adopt adjacent phenanthroline method to give the Fe (II) in the filtrate colour developing, adopt ultraviolet spectrophotometer to measure the content of Fe (II) at the 510nm place, and be contrast with the nutrient solution that does not add bacterium, the iron reducing power of checking bacterial strain.Fe (II) content sees Table 4, and Fe (II) content unit is mg/L, represents the milligram number of contained Fe (II) in every liter of sample.As shown in Table 4, the growing amount of Fe in the system (II) is apparently higher than the system that does not add bacterium, illustrate that this bacterium can the anaerobic reduction lepidocrocite, and reducing power is stronger.
In the table 4 Klebsiella pneumonia MFC4 reduction lepidocrocite process
Fe (II) produces dynamically (mg/L of unit)
0 5 days 10 days 20 days 25 days
Do not add the bacterium contrast 18.89 18.28 26.51 39.93 18.34
Add bacterium 18.89 33.49 77.92 132.28 165.62
Embodiment 5 Klebsiella pneumonia (K.pneumoniae MFC4) anaerobic reduction rhombohedral iron ore
Substratum D prescription: contain rhombohedral iron ore 2.0g in every liter of deionized water, NaHCO 32.5g, NH 4Cl0.25g, NaH 2PO 42H 2O0.678g, KCl0.1g, glucose 1.0g, each 10.0mL of vitamin solution and trace element solution (vitamin solution and trace element solution composition are with embodiment 1 described liquid separation culture medium) was in 121 ℃ of sterilizations 20 minutes, glucose is sterilized separately, and mix with other composition the sterilization back.
Preparation bacterial suspension: encircle the activatory bacterial strain in 200mL beef extract-peptone liquid nutrient medium with transfering loop picking one, in shaking table (30 ℃, 180 rev/mins) cultivated 16 hours, make bacterial number reach exponential phase of growth, 8000 rev/mins of centrifugal collection thalline, remove supernatant liquor, the thalline that precipitates is suspended among the 200mL substratum D, make bacteria suspension.
Inoculated bacteria suspension makes its bacterium number reach 10 in nutrient solution D 8CFU/mL, drum fills high-purity mixed gas (N then 2/ CO 2=80/20), the drum time of filling is no less than 30 minutes, leaves standstill anaerobism in 30 ℃ and cultivates.Observe the colour-change situation of nutrient solution, got the 180 rev/mins of vibrations 1.5 hours in 16mL0.5mol/L HCl solution of the 4mL nutrient solution that mixes every 5～10 days, refilter, adopt adjacent phenanthroline method to give the Fe (II) in the filtrate colour developing, adopt ultraviolet spectrophotometer to measure the content of Fe (II) at the 510nm place, and be contrast with the nutrient solution that does not add bacterium, the iron reducing power of checking bacterial strain.Fe (II) content sees Table 5, and Fe (II) content unit is mg/L, represents the milligram number of contained Fe (II) in every liter of sample.As shown in Table 5, in the time of 25 days, the growing amount of Fe in the system (II) is not add nearly 5 times that thalline is, illustrates that this bacterium also has stronger reductive action to the good rhombohedral iron ore of crystalline structure.
Fe (II) produces dynamically (mg/L of unit) in the table 5 Klebsiella pneumonia MFC4 reducing hematite process
0 day 5 days 10 days 20 days 25 days
Do not add the bacterium contrast 1.95 2.31 2.34 2.37 9.03
Add bacterium 1.95 9.47 10.19 40.79 44.41
Embodiment 6 Klebsiella pneumonia (K.pneumoniae MFC4) are to the reduction of humic acid pattern thing AQDS
Substratum E prescription: contain AQDS2.0g in every liter of deionized water, NaHCO 32.5g, NH 4Cl0.25g, NaH 2PO 42H 2O0.678g, KCl0.1g, glucose 1.0g, each 10.0mL of vitamin solution and trace element solution (vitamin solution and trace element solution composition are with embodiment 1 described liquid separation culture medium) was in 121 ℃ of sterilizations 20 minutes, glucose is sterilized separately, and mix with other composition the sterilization back.
Preparation bacterial suspension: encircle the activatory bacterial strain in 200mL beef extract-peptone liquid nutrient medium with transfering loop picking one, in shaking table (30 ℃, 180 rev/mins) cultivated 16 hours, make bacterial number reach exponential phase of growth, 8000 rev/mins of centrifugal collection thalline, remove supernatant liquor, the thalline that precipitates is suspended among the 200mL substratum E, make bacteria suspension.
Inoculated bacteria suspension makes its bacterium number reach 10 in nutrient solution E 8CFU/mL, drum fills high-purity mixed gas (N then 2/ CO 2=80/20), the drum time of filling is no less than 30 minutes, leaves standstill anaerobism in 30 ℃ and cultivates.Observe the colour-change situation of nutrient solution, sampling in per 12 hours, in the anaerobism workstation, filter nutrient solution (precipitations such as thalline are filtered) with 0.22 μ m bacterial filter, the output of going back ortho states AQDS with ultraviolet spectrophotometer (wavelength 450nm place) mensuration, and with the system that do not add bacterium in contrast.Ortho states AQDS content (unit: mmol/L) see Table 6 also.As shown in Table 6, Klebsiella pneumonia can show that this bacterium was very strong to the reducing power of AQDS with 80% AQDS reduction in 48 hours.
Go back ortho states AQDS in the table 6 Klebsiella pneumonia MFC4 reduction AQDS process and produce dynamically (unit: mmol/L)
0 12 hours 24 hours 36 hours 48 hours
Do not add the bacterium contrast 0.03 0.02 0.03 0.05 0.05
Add bacterium 0.04 0.45 0.62 0.81 0.82

Claims (2)

1. a Klebsiella pneumoniae (Klebsiella pneumoniae) is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on April 14th, 2008, and preserving number is: CGMCC No.2454.
2. the application of the described Klebsiella pneumonia of claim 1 (Klebsiella pneumoniae) in reduction of iron (III) oxide compound and humic acid reduction.
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CN112501084B (en) * 2020-12-22 2022-04-08 山东农业大学 Rhizosphere probiotic Klebsiella ZH07 and application thereof
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