Method for biosynthesis of nano-selenium by virtue of bacillus licheniformis and application of nano-selenium

A Bacillus licheniformis, biosynthesis technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, applications, etc., can solve problems such as human hazards, and achieve environmentally friendly, high yield, and remarkable effects

Active Publication Date: 2017-03-08
江西省水投富硒产业发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Excessive selenium can also

Method used

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  • Method for biosynthesis of nano-selenium by virtue of bacillus licheniformis and application of nano-selenium
  • Method for biosynthesis of nano-selenium by virtue of bacillus licheniformis and application of nano-selenium
  • Method for biosynthesis of nano-selenium by virtue of bacillus licheniformis and application of nano-selenium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Isolation, purification and identification of Bacillus licheniformis S13

[0049] 1. Isolation and purification of strain S13

[0050] A strain S13 that can tolerate higher concentrations of selenite and selenate was isolated from the soil.

[0051] 2. Identification of strain S13

[0052] 1.2.1 PCR amplification of 16S rRNA and gyrB gene sequences and sequencing:

[0053] Inoculate strain S13 on LB solid medium and culture for 24h, take 0.2mL sterilized PCR tube, add 10μL ddH 2 O, a sterile toothpick picked a single colony into the PCR tube and stirred to mix.

[0054] 1.2.2 Construct the PCR reaction system:

[0055] 16S rRNA: Using 8F (5'-CGGGATCCAGAGTTTGATCCTGGCTCAGAACGAACGCT-3') and 1506R (5'-CGGGATCCTACGGCTACCTTGTTACGACTTCACCCC-3') as primers, the 16S rRNA gene sequence was obtained by PCR amplification. The PCR reaction system is: ddH 2 O, 18.5 μL; 10×Buffer, 2.5 μL, dNTP Mix, 2 μL; primer 8F, 0.5 μL; primer 1506R, 0.5 μL; bacterial solution, 0.5 ...

Embodiment 2

[0060] Embodiment 2 Bacillus licheniformis S13 is to the tolerance concentration of selenite

[0061] Prepare solid LB with different concentrations of selenium-containing medium (each liter medium contains 10g of NaCl, 10g of tryptone, 5g of yeast extract, 15g of agar, and 1L of deionized water), and autoclave at 121°C for 20min; prepare a selenite mother solution, Sterilize by filtration, add selenite solution, make the content of selenite in the culture medium be 0mM, 10mM, 25mM, 35mM, 55mM, 70mM, 80mM respectively.

[0062] Pick a single colony of the S13 strain and inoculate it in LB liquid medium for 8 hours (150rpm, 37°C), and take the above bacterial solution and dilute it to OD 600 =0.8 of the mother liquor for later use; the mother liquor was diluted to 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 , respectively drop 2.5 μL of bacterial solutions of different concentrations on the selenium-containing plate, each concentration has 6 replicates, culture at 37°C for 48 hours,...

Embodiment 3

[0064] Embodiment 3 Bacillus licheniformis S13 is to the tolerance concentration of selenate

[0065] Prepare solid LB with different concentrations of selenium-containing medium (each liter medium contains 10g of NaCl, 10g of tryptone, 5g of yeast extract, 15g of agar, and 1L of deionized water), and autoclave at 121°C for 20min; prepare selenate mother liquor, filter Sterilize, add selenate solution, make the selenate content in the culture medium 0mM, 50mM, 100mM, 150mM, 400mM, 500mM, 600mM respectively.

[0066] Pick a single colony of the S13 strain and inoculate it in LB liquid medium for 8 hours (150rpm, 37°C), and take the above bacterial solution and dilute it to OD 600 =0.8 mother liquor, then dilute the mother liquor to 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , respectively drop bacterial liquid on different concentrations of selenium-containing plates, each concentration was replicated three times, cultivated at 37°C for 48 hours, and observed the growth of colo...

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Abstract

The invention provides a method for biosynthesis of nano-selenium by virtue of bacillus licheniformis and an application of the nano-selenium. According to the method and the application provided by the invention, the bacillus licheniformis S13, which is tolerant to selenite and selenate at relatively high concentrations, is separated from soil and the biological nano-selenium is synthesized by virtue of the strain S13; the biological nano-selenium is separated and purified, so that the mass preparation of the biological nano-selenium is achieved; and the nano-selenium can be applied to fertilizer, feed and selenium-enriched functional food processing as well as to health care products and medicine products. The nano-selenium, which is prepared by virtue of a biological fermentation process, has the characteristics of being environment-friendly, high in yield, safe and efficient and the like; and the produced biological nano-selenium, when applied or fed in the form of selenium-enriched fertilizer or selenium-enriched feed, can significantly enhance the selenium-enriching effect of crops, melons, vegetables, meat, milk and eggs.

Description

technical field [0001] The invention relates to the technical field of microbiology and biological nano-selenium preparation, in particular to a method for biosynthesizing nano-selenium by using bacillus licheniformis and its application. Background technique [0002] Selenium (Selenium, Se) element is one of the essential trace elements in many organisms, and it is an essential component of various selenium-containing enzyme proteins such as thioreductase, deiodinase, and glutathione peroxidase in organisms, and participates in Various metabolic pathways in the human body. Studies have found that selenium deficiency in the human body can lead to various diseases and increase the risk of cancer. my country is rich in selenium resources, but the distribution of selenium in nature is extremely uneven, resulting in selenium deficiency in more than two-thirds of my country. The Chinese Nutrition Society and the FAO / WHO / IAEA Joint Expert Committee determined that the appropriat...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P3/00A23L33/16A23K20/20A61K33/04C05D9/02A61P35/00C12R1/10
CPCA23V2002/00A61K33/04C05D9/02C12N1/20C12P3/00C12N1/205C12R2001/10A23V2250/1626
Inventor 郭岩彬李奎李柯赵桂慎吴文良谢斌陈志蓥王昊朱燕云
Owner 江西省水投富硒产业发展有限公司
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