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Efficient calcium mineralizing bacterium and application thereof

A kind of mineralizing bacteria and high-efficiency technology, applied in the field of microorganisms, can solve the problems of destroying cultural relics, easily clogging stone cultural relics, permeability limitation, etc., and achieve the effect of strong calcium mineralization performance

Pending Publication Date: 2020-08-28
INST OF SOIL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The permeability of general chemical reinforcements has certain restrictions, and it is easy to block the pores of stone cultural relics, so that the internal moisture cannot come out, which may further damage the cultural relics; while bacteria can penetrate into the interior of stone cultural relics to the maximum extent, Induced calcium carbonate crystallization does not block the pores of the stone

Method used

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  • Efficient calcium mineralizing bacterium and application thereof
  • Efficient calcium mineralizing bacterium and application thereof
  • Efficient calcium mineralizing bacterium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The strain is isolated from EM strains, and the specific method is as follows: add 1 g of EM strains to 9 mL of sterile water, oscillate and shake well, and gradiently dilute to 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 A total of 6 concentrations, spread the dilution on the MRS plate (MRS solid medium composition: K 2 HPO 4 2.0g / L, MgSO 4 ·7H 2 O 0.2g / L, FeSO 4 •7H 2 O 0.03g / L, ammonium citrate 2.0 g / L, sodium acetate 5.0 g / L, glucose 20.0 g / L, peptone 4.0 g / L, beef extract 4.0 g / L, yeast extract 2.0 g / L, agar 20 g / L, pH=6.5-7.0), incubate at 30°C for 2-3 days, until colonies grow out, pick colonies of different shapes and streak them on fresh MRS medium for 3 times to obtain pure bacteria. The obtained strains were inoculated into MRS liquid medium, and cultured on a shaker at 30°C and 180 r / min for 24 hours. The culture medium was mixed with 80 wt.% glycerol 1:1, and stored in a -80°C refrigerator.

Embodiment 2

[0019] Amplification and sequencing primers:

[0020] Forward primer: 27F 5'-AGAGTTTGATCCTGGCTCAG-3'

[0021] Reverse primer: 1492R 5'-GGTTACCTTGTTACGACTT-3'

[0022] The total DNA of the EMCa1 strain was extracted, and the 16s rDNA gene of the strain was amplified by PCR with primers 27F and 1492R, and sequenced with primers 27F and 1492R. The 16s rDNA gene sequence of the strain is shown in Seq ID No:1.

[0023] On http: / / blast.ncbi.nlm.nih.gov / Blast.cgi, the 16srDNA gene sequence of EMCa1 was compared with the known sequence of GenBank with BLAST software. The results showed that the sequence homology between the strain EMCa1 and the 16S rDNA gene (Bacillus cereus strain C5 16S ribosomal RNA gene, partial sequence) of Bacillus cereus C5 reached 100%, which proved that the strain EMCa1 belonged to the genus Bacillus cereus.

Embodiment 3

[0025] Add 20 g / L coral sand to 100 mL MRS medium, sterilize at 115°C for 30 minutes, and inoculate the strain EMCa1 at a ratio of 1%, and use the medium without bacteria as a control, at 30°C, After culturing on a shaker at 180 r / min for 20 days, the supernatant was taken to measure the concentration of calcium ions in the solution with an inductively coupled plasma emission spectrometer, and the coral sand was taken out to dry and analyzed by XRD with an X-ray diffractometer. Results The concentration of calcium ions in the control solution without bacteria was 36.9 mg / L, and the proportion of calcite in coral sand was 2%, while the treatment with EMCa1, the concentration of calcium ions in the solution was 7.19 mg / L, and the proportion of calcite in coral sand was 7.19 mg / L. The ratio is 20%. It can be seen that the strain EMCa1 converts calcium ions in the solution into calcium carbonate.

[0026] With the same method, after inserting other bacterial strains at the same ti...

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Abstract

The invention discloses an efficient calcium mineralizing bacterium and an application thereof, and belongs to the field of microorganisms. The bacterium disclosed by the invention is bacillus cereusEMCa1. The strain is in a short rod shape, and is in a short or long chain, the size of the bacterium is about (1.0-1.2) x (3.0-5.0) [mu]m, the strain is a gram staining positive bacterium, white circular bacterial colonies appear after the strain is cultured for 24 h on an LB agar plate culture medium at 30 DEG C, a diameter is about 5-7 mm, the edge is irregular, and the bacterial colony surfaceis like wax. The induced calcium carbonate deposition characteristic of the strain is as follows: after 20 days of culture, the strain can convert free calcium ions in a coral sand solution into a calcium carbonate precipitate, and in a calcium carbonate deposition experiment, the concentration of calcium ions is reduced from 36.9 mg / L to 7.19 mg / L, and the proportion of calcite in coral sand isincreased from 2% to 20%. According to the efficient calcium mineralizing bacterium and the application thereof, calcium ions can be mineralized to generate calcium carbonate, so that the efficient calcium mineralizing bacterium is applied to the restoration of stone cultural relics.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a high-efficiency calcium mineralization bacterium and its application. Background technique [0002] my country is rich in resources of stone relics, and calcium-containing rocks are the most commonly used matrix materials in a large number of stone relics left over from history. Stone cultural relics are affected by factors such as ambient atmosphere, acid rain, organisms, and changes in surface temperature and humidity, which lead to weathering and decomposition of stone components. At present, inorganic or organic polymer materials are mostly used as protection methods to strengthen stone cultural relics. However, due to the mismatch between the mechanical properties of polymer materials and the original cultural relic base material and the aging of protective materials, cultural relics often cause discoloration, hard shell formation and The peeling of the surface...

Claims

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Application Information

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IPC IPC(8): C12N1/20C04B41/00C12R1/085
CPCC04B41/00C12R2001/085C12N1/205
Inventor 王一明杨晓倩林先贵
Owner INST OF SOIL SCI CHINESE ACAD OF SCI
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