CN103013847B

CN103013847B - Ammonia-producing mineral leaching bacterium as well as culture method and application of ammonia-producing mineral leaching bacterium

Info

Publication number: CN103013847B
Application number: CN2012102386182A
Authority: CN
Inventors: 王洪江; 吴爱祥; 熊有为; 尹升华
Original assignee: University of Science and Technology Beijing USTB
Current assignee: University of Science and Technology Beijing USTB
Priority date: 2012-07-10
Filing date: 2012-07-10
Publication date: 2013-12-11
Anticipated expiration: 2032-07-10
Also published as: CN103013847A

Abstract

The invention discloses an ammonia-producing mineral leaching bacterium JAT-1 as well as a culture method and application of the ammonia-producing mineral leaching bacterium JAT-1. The ammonia-producing mineral leaching bacterium JAT-1 is gram-negative and rod-shaped, belongs to Providencia.sp; the collection name of the ammonia-producing mineral leaching bacterium is alkaline ammonia-producing copper leaching bacterium, and the collection number is 6214. The Providencia JAT-1 is separated from wet soil, takes carbamide as a decomposition matrix and a nitrogen source and sodium citrate or glucose as a carbon source, is at the optimal growth pH value of 8.0-9.5 and is aerobiotic or facultative anaerobic. According to the invention, high-alkalinity gangue copper ores are leached by adopting the JAT-1, and therefore the copper leaching efficiency is greatly enhanced. The application of the ammonia-producing mineral leaching bacterium JAT-1 disclosed by the invention provides an economic, energy-saving and environment-friendly effective way for the leaching of high-alkalinity copper ores which are difficult to treat and plays important roles in extending microbial leaching fields and enriching leaching microbe kinds.

Description

Ammonia ore-leaching bacteria and cultural method and application are produced in one strain

Technical field

The invention belongs to the microbial leaching field, relate to product ammonia ore-leaching bacteria and cultural method and application that a strain can change urea into ammoniacal liquor.

Background technology

At present, for copper mine, leach, acid system leaches and acidophilic bacteria occupies dominant position.But refractory copper ore and copper mine tailing for height containing Basic gangue mineral, under the employing acid system, the technological line of Chemical Leaching or Bioleaching is obviously infeasible.On the one hand, for keep leaching microbacteria (as thiobacillus ferrooxidans, Thiobacillus thiooxidans, Leptospirillums ferrooxidansdeng) normal growth breeding and good oxidation activity, need to adopt in advance sour agent to carry out drip washing to ore, the Copper Ores of high alkalinity gangue or the increase greatly of the copper mine tailing unit of making acid consumption, unreasonable economically; On the other hand, acid system can produce the micro-soluble materials such as a large amount of calcium sulfate and sal epsom, because stopping up the leaching passage, reduces leaching rate.

With the acid system Leaching Systems, compare, alkaline Leaching have metal selective strong, the erosion resistance of equipment is required low, little to the pollution of environment, be a main developing direction, for example ammoniacal liquor leaches.The volatility of ammoniacal liquor is large, causes processing cost to increase, and environmental pollution is serious, can't carry out large-scale dump leaching; Adopt pressure ammonia leaching feasible technically, but energy consumption is high, facility investment is large, and economic benefit is poor.What is more important, also be not applicable to the special-purpose Pseudomonas that ammonia soaks at present, hampers applying of ammonia leaching-out technique.Produce ammonium bacteria if can seek a strain, make this bacterium be applied to ammonia process and soak copper, the defect that can avoid normal pressure ammonia to soak, and improve ammonia and soak efficiency, enriched the ore immersion bacterial culture storehouse, promote ammonia to soak the progress of technology, significant.

Summary of the invention

In order to address the above problem, the object of the invention be to provide a strain can ureaclastic adaptive capacity to environment strong, produce the efficient product ammonia ore-leaching bacteria that the ammonia ability is outstanding, for the leaching of the difficult copper mine of high alkalinity provides a kind of novel method.

Technical scheme of the present invention is: the ammonia ore-leaching bacteria is produced in a strain, this bacterium belong to produce the alkali Providence ( providencia sp), classification is by name: Providence providencia sp,name is called JAT-1, and at China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, in registration and preservation on June 13rd, 2012, preserving number is: CGMCC No.6214.

This bacterium is analyzed (as Fig. 2) and bacterial classification homology analysis (as Fig. 3) through gramstaining (as Fig. 1) and the relevant bacterial strain phylogenetic tree based on 16S rDNA gene order, be accredited as belong to product alkali Providence kind, Providencia ( providencia sp).JAT-1 with providencia sp. Sam 130-9Ahomology is up to 99%.

Method for cultivation of bacteria of the present invention, it is that 2.0 ~ 4.0 g/L Sodium phosphate dibasics and concentration are that 0.1 ~ 0.5 g/L sal epsom is as substratum, without somatomedin that the method adopts the carbon source that concentration is 5 ~ 25g/L, the nitrogenous source that concentration is 5 ~ 20g/L, potassium primary phosphate, the concentration that concentration is 1.0 ~ 3.0 g/L.Described carbon source is Trisodium Citrate or glucose, and nitrogenous source is urea.Culture condition is: 20 ~ 40 ℃ of temperature, pH value 5.0 ~ 11.0, aerobic or amphimicrobian.

The application of bacterium of the present invention, will produce ammonium bacteria shaking culture in substratum and, to logarithmic phase, with moistening red litmus paper, be suspended from the cultivation bottleneck, and it is blue that test paper becomes rapidly, illustrates that Production by Bacteria ammonia is good.The preparation fresh culture, inoculation logarithmic phase bacterium is as leaching agent.The product ammonium bacteria carries out in the 250mL erlenmeyer flask the leaching experiment of alkaline copper mine.Under 20 ~ 40 ℃ of temperature, ore pulp liquid-solid ratio 4:1 ~ 9:1, bacterium initial inoculation concentration 10 ~ 30% conditions, leach certain copper mine, leaching yield reaches 33.26% ~ 39.86%.

The application of this bacterial strain will be conducive to expand the bacterial leaching field, enrich the ore-leaching bacteria kind, improve traditional ammonia soaks the problems such as the cost of existence is large, energy consumption is high, environmental pollution is serious.

The accompanying drawing explanation

Fig. 1 is for producing the gramstaining schematic diagram of ammonia ore-leaching bacteria JAT-1.

Fig. 2 sets schematic diagram for producing ammonia ore-leaching bacteria JAT-1 to the phylogenetic evolution of relevant bacterial strain.

Fig. 3 for produce ammonia ore-leaching bacteria JAT-1 with providencia sp. Sam 130-9Athe 16S rDNA gene order comparison schematic diagram of bacterial strain.

Embodiment

Following examples are used for the present invention is described, but are not used for limiting the scope of the invention.

Embodiment mono-:

Adopt urea medium to carry out enrichment culture to bacterium in soil, carry out plate isolation after clearly obtaining efficient bacterium, obtain pure strain.Medium component is: Trisodium Citrate 5g/L, urea 5g/L, potassium primary phosphate 1.0 g/L, Sodium phosphate dibasic 2.0 g/L, sal epsom 0.1 g/L.Under 5.0,20 ℃ of conditions of pH value, the constant temperature aerobic is cultivated 48 hours, obtains breeding bacterium liquid.Under 20 ℃ of temperature, ore pulp liquid-solid ratio 4:1, bacterium initial inoculation concentration 10% condition, leach certain copper mine, after 144h, copper leaches and reaches 33.26%.

Embodiment bis-:

Adopt urea medium to carry out enrichment culture to bacterium in soil, carry out plate isolation after clearly obtaining efficient bacterium, obtain pure strain.Medium component is: glucose 5g/L, urea 10g/L, potassium primary phosphate 2.0g/L, Sodium phosphate dibasic 3.0 g/L, sal epsom 0.3 g/L.Under 7.0,20 ℃ of conditions of pH value, the constant temperature aerobic is cultivated 48 hours, obtains breeding bacterium liquid.Under 20 ℃ of temperature, ore pulp liquid-solid ratio 6:1, bacterium initial inoculation concentration 20% condition, leach certain copper mine, after 144h, copper leaches and reaches 38.17%.

Embodiment tri-:

Adopt urea medium to carry out enrichment culture to bacterium in soil, carry out plate isolation after clearly obtaining efficient bacterium, obtain pure strain.Medium component is: Trisodium Citrate 25g/L, urea 10g/L, potassium primary phosphate 2.0 g/L, Sodium phosphate dibasic 3.0 g/L, sal epsom 0.4 g/L.Culture condition is constant temperature anaerobism under 8.0,30 ℃ of conditions of pH value.The bacterial growth lag phase is shorter, and logarithmic phase about 24 ~ 48 hours, then enters stationary phase greatly.Under 30 ℃ of temperature, ore pulp liquid-solid ratio 7:1, bacterium initial inoculation concentration 30% condition, leach certain copper mine, after 144h, copper leaches and reaches 36.7%.

Embodiment tetra-:

Adopt urea medium to carry out enrichment culture to bacterium in soil, carry out plate isolation after clearly obtaining efficient bacterium, obtain pure strain.Medium component is: glucose 25g/L, urea 20g/L, potassium primary phosphate 3.0 g/L, Sodium phosphate dibasic 4.0 g/L, sal epsom 0.5 g/L.Under 11.0,40 ℃ of conditions of pH value, the constant temperature anaerobism is cultivated 48 hours, obtains breeding bacterium liquid.Under 40 ℃ of temperature, ore pulp liquid-solid ratio 9:1, bacterium initial inoculation concentration 30% condition, leach certain copper mine, after 144h, copper leaches and reaches 39.86%.

Claims

1. the ammonia ore-leaching bacteria is produced in a strain, it is characterized in that, this bacterium belong to produce the alkali Providence ( providencia sp), name is called JAT-1, and in Chinese common micro-organisms culture presevation administrative center registration and preservation, preserving number is: CGMCC 6214.

2. a method of cultivating product ammonia ore-leaching bacteria claimed in claim 1, it is characterized in that: the medium component of the method is: the potassium primary phosphate that the nitrogenous source that the carbon source that concentration is 5 ~ 25g/L, concentration are 5 ~ 20g/L, concentration are 1.0 ~ 3.0 g/L, concentration are that 2.0 ~ 4.0 g/L Sodium phosphate dibasics and concentration are 0.1 ~ 0.5 g/L sal epsom, without somatomedin, wherein, described carbon source is Trisodium Citrate or glucose, nitrogenous source is urea

Culture condition is: 20 ~ 40 ℃ of temperature, pH value 5.0 ~ 11.0, aerobic or amphimicrobian.

3. the application of a product ammonia ore-leaching bacteria claimed in claim 1 in the microbial leaching field.