CN104551001B - A kind of alternaric bacteria Y1309-1 application in preparing nanometer gold - Google Patents
A kind of alternaric bacteria Y1309-1 application in preparing nanometer gold Download PDFInfo
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- CN104551001B CN104551001B CN201410819613.8A CN201410819613A CN104551001B CN 104551001 B CN104551001 B CN 104551001B CN 201410819613 A CN201410819613 A CN 201410819613A CN 104551001 B CN104551001 B CN 104551001B
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Abstract
The invention discloses a kind of alternaric bacteria Y1309-1 application in preparing nanometer gold.The present invention adopts the alternaric bacteria Y1309-1 nanogold particle prepared, and its normal temperature and pressure that reacts on preparing nanometer gold carries out, and reaction cost is low, and reacted collecting cells is convenient, it is easy to amplify preparative-scale.Nanogold particle prepared by alternaric bacteria Y1309-1 of the present invention can to relevant industries containing heavy metal waste liquid and soil carry out repair and regeneration.
Description
Technical field
The present invention relates to biological technical field, be specifically related to a kind of alternaric bacteria Y1309-1 application in preparing nanometer gold.
Background technology
Nanometer gold not only possesses the general character such as skin effect, quantum effect, small-size effect and the macro quanta tunnel effect etc. of nanoparticle, also there is the physical and chemical performance of oneself uniqueness, be widely used in fields such as nano-device, biomedicine, oncotherapy, cell imaging and pharmaceutical carriers.
The biosynthesis of nano metal refers to and utilizes microorganism or plant to synthesize nano metal at normal temperatures and pressures, it is not necessary to uses toxic chemical raw material or does not produce toxic by-products, being the eco-friendly nano material synthetic method of a kind of green.Nano-particle prepared by biological synthesis method has narrow size distribution, stability height, good biocompatibility, productivity height, low cost and other advantages, has a wide range of applications potentiality in fields such as nanometer biotechnology, medicine and catalysis.
More existing results of study show, microorganism can synthesize the bioactive substance that the reason of metal nano material is its generation and include protein, reducing sugar, reductive glutathione etc. and as gold, cadmium, palladium, platinum etc. carry out enrichment, metal ion can be reduced into nano material, or two metal ion species in system are reduced into Nanoalloy, and nano metal is stably played an important role by biomolecule.Therefore, the alternaric bacteria in the present invention also has the ability that gold, cadmium, palladium, platinum plasma reduction are assembled into nanoparticle or alloy.
Summary of the invention
Goal of the invention: the deficiency existed for prior art, it is an object of the invention to provide a kind of alternaric bacteria Y1309-1 application in preparing nanometer gold.Prepare the method mild condition of nanometer gold, it is not necessary to using multiple chemical reagent, reaction cost is low, environmental friendliness.
Technical scheme: for realizing the purpose of foregoing invention, the technical solution used in the present invention is: a kind of alternaric bacteria Y1309-1 application in preparing nanometer gold.
Above-mentioned application, described alternaric bacteria Y1309-1, its Classification And Nomenclature is alternaric bacteria Y1309-1(Alternariasp.Y1309-1), this bacterial strain is preserved in China typical culture collection center on July 3rd, 2014, and deposit number is CCTCCNo:M2014311.Preservation address: Wuhan University's preservation center, Wuchang District, Wuhan City, Hubei Province (the first affiliated primary school of Wuhan University opposite).
This alternaric bacteria bacterium colony is white just, fades to taupe, and mycelia is sparse, substrate pitchy.Conidiophore single, different in size, the raw not branch in top, conidium has a barrier film in length and breadth, the shape of falling club, avette.
The screening technique of above-mentioned alternaric bacteria Y1309-1 is as follows:
1) take 35 fungal strains being isolatable from Southeast China University's campus soil of Laboratories Accession, cultivate 3-5 days in 28 DEG C on PDA solid medium respectively；
2) 35 strain strains are inoculated in 100mL potato dextrose broth respectively, in 28 DEG C, 150rpm cultivate within 2 days, obtain thalline；
3) collect thalline and wash with sterilized water, 20g wet thallus is added in 100ml sterile deionized water, in 28 DEG C, pH7,150rpm cultivate 48h；
4) it is centrifuged off the thalline in step 3), collects supernatant.The chlorauric acid solution of final concentration 0.5-2.0mmol/L is added in the supernatant of step 4), in 28 DEG C, 150rpm react 3-48h, judge whether to generate nanometer gold (generate the solution of nanometer gold be purple to darkviolet) according to the color of solution after reaction, the system reacted is repeated screening experiment and the nanometer gold generated is confirmed；
5) thalline in step 3) is collected, after sterilized water washs, 20g wet thallus is added in the aseptic chlorauric acid solution of 100ml final concentration 0.5-2.0mmol/L, in 28 DEG C, pH7,150rpm react 3-48h, judge in its born of the same parents, whether to generate nanometer gold (generate the cell of nanometer gold be purple to darkviolet) according to the color of thalline after reaction, the system reacted is repeated screening experiment and the nanometer gold generated is confirmed；The bacterial strain that can generate nanometer gold that screening obtains is chosen a strain preservation called after alternaric bacteria Y1309-1.
Described alternaric bacteria Y1309-1 prepares specifically comprising the following steps that of nanometer gold and is cultivated after 4 days in 28 DEG C on PDA solid medium by alternaric bacteria Y1309-1, is inoculated in 100mL potato dextrose broth, in 28 DEG C, 150rpm cultivates and obtains thalline in 2 days；Collect thalline, after sterilized water washs, 20g wet thallus added in the aseptic chlorauric acid solution of 100ml concentration 0.5-2.0mmol/L, in 28 DEG C, pH7,150rpm react after 6h, broken thalline, gather in the crops nanogold particle.
Above-mentioned nanogold particle particle diameter is 30 ~ 40nm.
Beneficial effect: compared with prior art have the advantage that the present invention adopts the alternaric bacteria Y1309-1 nanogold particle prepared, its normal temperature and pressure that reacts on preparing nanometer gold carries out, reaction cost is low, and reacted collecting cells is convenient, it is easy to amplify preparative-scale.Nanogold particle prepared by alternaric bacteria Y1309-1 of the present invention can to relevant industries containing heavy metal waste liquid and soil carry out repair and regeneration.
Accompanying drawing explanation
Fig. 1 is the design sketch of the alternaric bacteria Y1309-1 of embodiment 1 gold nano grain prepared；
Fig. 2 is the stereoscan photograph of gold nano grain prepared by the alternaric bacteria of embodiment 1；
Fig. 3 is the EDS spectrogram of gold nano grain prepared by the alternaric bacteria of embodiment 1；
Fig. 4 is the colibacillary design sketch of suppression of embodiment 2；
Fig. 5 is the design sketch suppressing golden yellow staphylococcus of embodiment 2.
Detailed description of the invention
Below in conjunction with accompanying drawing be embodied as further illustrating present disclosure, but should not be construed as limitation of the present invention.Without departing from the spirit and substance of the case in the present invention, the amendment that the inventive method, step or condition are made or replacement, belong to the scope of the present invention.
Embodiment 1: the condition of culture of alternaric bacteria Y1309-1
Alternaric bacteria Y1309-1 is PDA solid medium (Rhizoma Solani tuber osi 20%, glucose 2%, deionized water 1000mL, pH is natural, and 121 DEG C of sterilizing 30min are stand-by) in 28 DEG C of cultivations, liquid medium within (PDA solid medium: Rhizoma Solani tuber osi 20%, glucose 2%, agar 1.5%, deionized water 1000mL, pH are natural, and 121 DEG C of sterilizing 30min are stand-by) in 28 DEG C, 150rpm cultivates.
Embodiment 2 alternaric bacteria Y1309-1 prepares the concrete steps of nanometer gold
Alternaric bacteria Y1309-1 is cultivated after 4 days in 28 DEG C on PDA solid medium, is inoculated in 100mL potato dextrose broth, in 28 DEG C, 150rpm cultivates 2 days；Collect thalline, after sterilized water washs, 20g wet thallus added in the aseptic chlorauric acid solution of 100ml (concentration 0.5-2.0mmol/L), in 28 DEG C, after pH7,150rpm react 6h, broken thalline, gather in the crops nanogold particle.
Accompanying drawing 1 is left for adding the thalline before gold chloride, right for adding the darkviolet thalline having wrapped up nanometer gold after gold chloride reaction 6h, illustrates alternaric bacteria can in born of the same parents quickly synthesizing nano golden.
The transmission electron microscope photo of the nanogold particle prepared in accompanying drawing 2 and accompanying drawing 3 respectively rod method mycetocyte and EDS collection of illustrative plates, it is possible to clearly indicate the nanogold particle generated in born of the same parents.
The gold nano grain synthesized by embodiment 3 present invention bacteriostasis to escherichia coli and staphylococcus aureus
Escherichia coli and staphylococcus aureus are coated solid medium (Carnis Bovis seu Bubali cream 0.5%, NaCl0.5%, peptone 1%, pH7.6 ~ 7.8) by step 1；
Step 2 places a sterilizing filter paper on solid medium, and it adds the gold nano grain solution 5 μ l prepared by embodiment 2；
Escherichia coli and staphylococcus aureus are cultivated 24h in 37 DEG C by step 3, measure its antibacterial circle diameter respectively and average, determining that escherichia coli antibacterial circle diameter under nanometer gold effect is 1.21cm, staphylococcus aureus antibacterial circle diameter under nanometer gold effect is 1.42cm.
Accompanying drawing 4 and accompanying drawing 5 be nanometer gold synthesized by embodiment 2 to escherichia coli and silver Staphylococcus aureus inhibition zone size, illustrate that nanometer gold prepared by alternaric bacteria is all inhibited to gram negative bacteria and gram positive bacteria.
Claims (3)
1. the alternaric bacteria Y1309-1 application in preparing nanometer gold, it is characterised in that described alternaric bacteria Y1309-1, its Classification And Nomenclature is alternaric bacteria Y1309-1(Alternariasp.Y1309-1), this bacterial strain is preserved in China typical culture collection center on July 3rd, 2014, and deposit number is CCTCCNo:M2014311.
2. application according to claim 1, it is characterized in that, described alternaric bacteria Y1309-1 prepares specifically comprising the following steps that of nanometer gold and is cultivated after 4 days in 28 DEG C on PDA solid medium by alternaric bacteria Y1309-1, be inoculated in 100mL potato dextrose broth, in 28 DEG C, 150rpm cultivate within 2 days, obtain thalline；Collect thalline, after sterilized water washs, 20g wet thallus added in the aseptic chlorauric acid solution of 100ml concentration 0.5-2.0mmol/L, in 28 DEG C, pH7,150rpm react after 6h, broken thalline, gather in the crops nanogold particle.
3. application according to claim 2, it is characterised in that described nanogold particle particle diameter is 30 ~ 40nm.
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| KR100557314B1 (en) * | 2004-11-08 | 2006-03-06 | (주)바이오드림스 | Nano-silicasilver and method for the preparation thereof |
| CN1854735A (en) * | 2005-04-19 | 2006-11-01 | 林远 | Fluid cell equipment-microcarrier clinical diagnosis chip |
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| KR100557314B1 (en) * | 2004-11-08 | 2006-03-06 | (주)바이오드림스 | Nano-silicasilver and method for the preparation thereof |
| CN1854735A (en) * | 2005-04-19 | 2006-11-01 | 林远 | Fluid cell equipment-microcarrier clinical diagnosis chip |
Non-Patent Citations (1)
| Title |
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| "Fungus-mediated synthesis of silver nanoparticles and their activity";Monali Gajbhiye等;《Nanomedicine: Nanotechnology, Biology, and Medicine》;20091231(第5期);全文 * |
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