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Method for adjusting and controlling structure and shape of calcium oxalate crystals by using colibacillus secretion

A technology of Escherichia coli and crystal structure, applied in the field of biochemistry, can solve the problem that there is no ideal method for stone prevention, and achieve the effect of consistent shape, high conversion rate and simple steps

Inactive Publication Date: 2015-02-18
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the formation mechanism of stones has not been fully elucidated so far, and there is no ideal method for the prevention of stones.

Method used

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  • Method for adjusting and controlling structure and shape of calcium oxalate crystals by using colibacillus secretion
  • Method for adjusting and controlling structure and shape of calcium oxalate crystals by using colibacillus secretion
  • Method for adjusting and controlling structure and shape of calcium oxalate crystals by using colibacillus secretion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Preparation of E. coli secretions:

[0026] Pick a single colony of Escherichia coli TOP10 preserved at 4°C, inoculate it in LB liquid medium, place it in a oscillating water bath with a constant temperature of 37.5°C and shake it at 110 rpm for 24 hours to obtain the original bacterial liquid, and centrifuge the original bacterial liquid at 4000 rpm to obtain the secretion of E. coli were stored at 4°C for later use.

[0027] Preparation of non-calculus calcium oxalate dihydrate crystals:

[0028] Prepare 100mM calcium chloride and 5mM sodium oxalate solutions respectively. Take 40ml of calcium chloride solution with a concentration of 100mM in a 100ml Erlenmeyer flask, add 10ml of Escherichia coli secretion, then add 40ml of sodium oxalate with a concentration of 5mM into the Erlenmeyer flask, shake it and place it in a constant temperature incubator at 37.5°C 24 hours. The supernatant was discarded, and the bottom crystals were washed twice with water and ethanol ...

Embodiment 2

[0030] The preparation of Escherichia coli secretion was the same as in Example 1.

[0031] Prepare 100mM calcium chloride and 5mM sodium oxalate solutions respectively. Take 40ml of calcium chloride solution with a concentration of 100mM in a 100ml Erlenmeyer flask, add 1ml of Escherichia coli secretion, and then add 40ml of sodium oxalate with a concentration of 5mM into the Erlenmeyer flask. After shaking, place it in a constant temperature incubator at 37.5°C for 24 hours. The supernatant was discarded, and the crystals at the bottom were washed with water and ethanol twice respectively. The resulting sample ( Figure 4 ) placed in an evaporating dish and dried at room temperature and then stored in a sample tube. The measured percentage of calcium oxalate dihydrate is 15%, and the rest is calcium oxalate monohydrate crystals, indicating that a very small amount of Escherichia coli secretion has a control effect on the crystal form of calcium oxalate.

Embodiment 3

[0033] The preparation of Escherichia coli secretion was the same as in Example 1.

[0034] Prepare 100mM calcium chloride and 5mM sodium oxalate solutions respectively. Take 40ml of calcium chloride solution with a concentration of 100mM in a 100ml Erlenmeyer flask, add 5ml of Escherichia coli secretion, and then add 40ml of sodium oxalate with a concentration of 5mM into the Erlenmeyer flask. After shaking, place it in a constant temperature incubator at 37.5°C for 24 hours. The supernatant was discarded, and the crystals at the bottom were washed with water and ethanol twice respectively. The resulting sample ( Figure 5 ) placed in an evaporating dish and dried at room temperature and then stored in a sample tube. The measured percentage of calcium oxalate dihydrate was 43.4%, and the rest was calcium oxalate monohydrate crystals, indicating that the crystallization ratio of calcium oxalate dihydrate increased with the increase in the amount of E. coli secretion.

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Abstract

The invention relates to a method for adjusting and controlling the structure and shape of calcium oxalate crystals by using colibacillus secretion. The method comprises the following steps of: mixing the colibacillus secretion with a matrix solution of which the ratio of [Ca2<+>] to [C2O4<2->] is (19.5-20.5):1, wherein the volume ratio is 1:(8-80); performing constant temperature incubation, and staying overnight; and washing and drying to obtain pure acalculous type calcium oxalate dehydrate crystals or a mixture of the calcium oxalate dehydrate crystals and calcium oxalate monohydrate crystals, wherein the colibacillus secretion is prepared through the steps of inoculating TOP10 single colonies of colibacillus to an LB liquid culture medium, performing low-speed oscillation on the culture medium at a constant temperature, staying the culture medium overnight, and performing high-speed centrifugation on the original bacterium liquid to obtain the colibacillus secretion. The method has the advantages that the calcium oxalate dehydrate crystals formed through colibacillus secretion adjustment and control have the characteristics of uniform size and consistent shape, the content of the calcium oxalate dehydrate crystals can be adjusted according to different additions, and the preparation condition is simple and non-hazardous, so that the method can be widely used in various fields.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to a new method for regulating the crystal structure and morphology of calcium oxalate. Background technique [0002] Urinary calculus is a common and frequently-occurring disease worldwide. Humans have known calculus for hundreds of years, and its incidence has been on the rise in recent years. Calculus is the product of pathological mineralization in organisms, which belongs to abnormal mineralization. The abnormal biomineralization process involves many disciplines such as medicine, biology, chemistry, crystallography, materials science and mineralogy, and thus has attracted widespread attention and great attention from experts in these fields. At present, it is mainly to study the composition of stones, detection methods, treatment options, and the interaction with macromolecules. Most stones can mix two or more components. Under the physiological pH value of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C55/07C07C51/41C12P1/04C12R1/19
Inventor 吴庆生赵振武
Owner TONGJI UNIV
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