CN106635879B - The cultural method of spatial plant lactobacillus LCT-LP1 and application - Google Patents
The cultural method of spatial plant lactobacillus LCT-LP1 and application Download PDFInfo
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- CN106635879B CN106635879B CN201610940382.5A CN201610940382A CN106635879B CN 106635879 B CN106635879 B CN 106635879B CN 201610940382 A CN201610940382 A CN 201610940382A CN 106635879 B CN106635879 B CN 106635879B
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- 238000000034 method Methods 0.000 title claims abstract description 18
- 241000186660 Lactobacillus Species 0.000 title claims abstract description 9
- 229940039696 lactobacillus Drugs 0.000 title claims abstract description 9
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 141
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 141
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 141
- 239000001963 growth medium Substances 0.000 claims abstract description 22
- 244000005700 microbiome Species 0.000 claims abstract description 17
- 230000002906 microbiologic effect Effects 0.000 claims abstract description 10
- 230000002265 prevention Effects 0.000 claims abstract description 5
- 229910001069 Ti alloy Inorganic materials 0.000 claims description 32
- 230000007797 corrosion Effects 0.000 claims description 23
- 238000005260 corrosion Methods 0.000 claims description 23
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- 239000010935 stainless steel Substances 0.000 claims description 19
- 239000000463 material Substances 0.000 claims description 6
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- 239000007836 KH2PO4 Substances 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- 235000015278 beef Nutrition 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 5
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- 241000193830 Bacillus <bacterium> Species 0.000 claims 1
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
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- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
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- FFUAGWLWBBFQJT-UHFFFAOYSA-N hexamethyldisilazane Chemical compound C[Si](C)(C)N[Si](C)(C)C FFUAGWLWBBFQJT-UHFFFAOYSA-N 0.000 description 2
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- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 244000283207 Indigofera tinctoria Species 0.000 description 1
- 244000164595 Lactobacillus plantarum subsp plantarum Species 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
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- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
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- JJWKPURADFRFRB-UHFFFAOYSA-N carbonyl sulfide Chemical compound O=C=S JJWKPURADFRFRB-UHFFFAOYSA-N 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
Abstract
The invention discloses the cultural method of spatial plant lactobacillus LCT-LP1 a kind of and applications.The present invention provides a kind of methods for cultivating lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1, include the following steps: using lactobacillus plantarum described in the culture of lactobacillus plantarum culture medium；The deposit number of lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 is CGMCC NO.12578.The present invention facilitates the mechanism and prevention and control measure research that microorganism corrodes space flight equipment in long-term space closed cabin, help to deepen to the understanding of microbiological hazards in manned spacecraft technological reserve related to reinforcement, help to explore significantly more efficient manned spacecraft microbiological comprehensive control measure, provides technical support for the development and operation of manned spacecraft.
Description
Technical field
The present invention relates to microorganism and field of biotechnology, and in particular to a kind of training of spatial plant lactobacillus LCT-LP1
Support methods and applications.
Background technique
As mankind's space exploration is movable increasingly frequently, cosmic space becomes a key areas of mankind's activity.
There is specific microbiologic populations in space closed cabin.Although each article all have passed through strict sterilization, microorganism still can
Enter space with spacefarer's body, human secretion, aerospace parts etc., and forms specified microorganisms in spacecraft closed in space
Group both includes harmless micro-organisms, also includes some pathogenic and corrosivity microorganism." peace " number ten spaces Yu Nianlai of operation
234 kinds of microorganisms are detected in standing.Particular surroundings factor in space closed cabin, including microgravity, low-intensity magnetic field and particle radiation
Deng, so that microbes generate variation, cause their requirements to living environment very low, is more easier to grow and breed, it is certain
The corrosivity of microorganism can enhance.The mass propagation of corrosivity microorganism in space closed cabin, so that various space materials are gradually
Corrosion is generated, accelerates the degradation consume of space flight device, it would be possible to cause the equipment in space station to run failure, influence spacecraft
Long-term normal operation and its in-orbit service life, or even very big threat will also result in flight safety.Spacecraft service condition is disliked
Bad, time in orbit is long, and repairs condition relative deficiency, once microbiologic(al) corrosion equipment fault occurs, loses inestimable.It is external
The in-orbit experience of manned spacecraft and the PRELIMINARY RESULTS of domestic ground research show that microbes seriously threaten space equipment safety.
Carry out the mechanism and prevention and control measure research that microorganism corrodes space flight equipment in long-term space closed cabin, deepens to manned spacecraft
The understanding of middle microbiological hazards technological reserve related to reinforcement is explored significantly more efficient manned spacecraft microbiological comprehensive control and is arranged
It applies, provides technical support for the development and operation of manned spacecraft.
Summary of the invention
The object of the present invention is to provide the cultural method of spatial plant lactobacillus LCT-LP1 a kind of and applications.
The present invention provides it is a kind of cultivate lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 method,
Include the following steps: using lactobacillus plantarum (Lactobacillus plantarum) described in the culture of lactobacillus plantarum culture medium
LCT-LP1。
The present invention also protects a kind of method for preparing lactobacillus plantarum LCT-LP1 culture, includes the following steps: using plant
Object lactobacillus culture medium culture lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1.
Solute and its concentration in the lactobacillus plantarum culture medium in any description above lactobacillus plantarum culture medium
It is as follows: peptone 0.5-1.5g/100ml, beef extract 0.5-1.5g/100ml, yeast extract 0.4-0.6g/100ml, KH2PO40.1-
0.3g/100ml, trisodium citrate 0.1-0.3g/100ml, sodium acetate 0.1-0.3g/100ml, glucose 1-3g/100ml,
Tween80 0.05-0.15mL/100ml、MgSO4·7H2O 0.048-0.068g/100ml、MnSO4·4H2O0.015-
0.035g/100ml。
The solute and its concentration in the lactobacillus plantarum culture medium concretely peptone 1g/100ml, beef
Cream 1g/100ml, yeast extract 0.5g/100ml, KH2PO40.2g/100ml, trisodium citrate 0.2g/100ml, sodium acetate 0.2g/
100ml, glucose 2g/100ml, Tween 800.1mL/100ml, MgSO4·7H2O0.058g/100ml、MnSO4·
4H2O0.025g/100ml。
The solvent of any description above lactobacillus plantarum culture medium is water.
The pH of any description above lactobacillus plantarum culture medium is 6.2-6.4.
Lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1, in preservation on the 01st in 06 month in 2016
In China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number, deposit number is CGMCC NO.12578.Lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 CGMCC
NO.12578 is referred to as lactobacillus plantarum LCT-LP1.
The present invention also protects lactobacillus plantarum LCT-LP1 in microorganisms to the application in space flight equipment corrosion mechanism.
The present invention also protects application of the lactobacillus plantarum LCT-LP1 in the research of prevention and control microbiologic(al) corrosion space flight equipment.
The present invention also protects lactobacillus plantarum LCT-LP1 researching and developing the application in novel space flight corrosion-resistant material.
The present invention also protects lactobacillus plantarum LCT-LP1 answering in the model that preparation microorganism corrodes space flight equipment
With.
The present invention also protects application of the lactobacillus plantarum LCT-LP1 in the product of preparation corrosion space flight equipment.
The present invention provides one kind to have corrosive spatial plant lactobacillus LCT-LP1 to common space flight high molecular material
Cultural method and application.The experimental results showed that lactobacillus plantarum LCT-LP1 space flight group, simulated microgravity group and ground
1g gravity control group bacterial strain can corrode titanium alloy and stainless steel, but under the conditions of space flight and under the conditions of simulated microgravity more
It is easy to form biomembrane in metal material surface, the corrosive power of titanium alloy and stainless steel is increased compared with ground 1g gravity control group
By force.The present invention facilitates the mechanism and prevention and control measure research that microorganism corrodes space flight equipment in long-term space closed cabin, helps
In in-depth to the understanding of microbiological hazards in manned spacecraft technological reserve related to reinforcement, help to explore significantly more efficient load
People's spacecraft microbiological comprehensive control measure provide technical support for the development and operation of manned spacecraft.
Detailed description of the invention
Fig. 1 is the scanning electron microscope aspect graph of lactobacillus plantarum LCT-LP1.
Fig. 2 is the lawn aspect graph of lactobacillus plantarum LCT-LP1.
Fig. 3 is the single colonie aspect graph of lactobacillus plantarum LCT-LP1.
Fig. 4 is lactobacillus plantarum LCT-LP1 complete genome sequence spectrogram.
Fig. 5 is the COG functional analysis of lactobacillus plantarum LCT-LP1.
Fig. 6 is the GO annotation of gene function of lactobacillus plantarum LCT-LP1.
The KEGG metabolic pathway that Fig. 7 is lactobacillus plantarum LCT-LP1 is analyzed.
The biofilm formation that Fig. 8 is lactobacillus plantarum LCT-LP1 is analyzed.
Fig. 9 is that lactobacillus plantarum LCT-LP1 is analyzed in the biofilm formation of TC4 titanium alloy surface.
Figure 10 is that lactobacillus plantarum LCT-LP1 is analyzed in the biofilm formation of 304 stainless steel surfaces.
Figure 11 is sem analysis of the lactobacillus plantarum LCT-LP1 to TC4 titanium alloy corrosive power.
Figure 12 is sem analysis of the lactobacillus plantarum LCT-LP1 to 304 Corrosion of Stainless Steel abilities.
Figure 13 is that lactobacillus plantarum LCT-LP1 analyzes the AFM of metal erosion ability.
Figure 14 is that lactobacillus plantarum LCT-LP1 analyzes the EDS of 304 Corrosion of Stainless Steel abilities.
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly
What routine biochemistry reagent shop was commercially available.Quantitative test in following embodiment is respectively provided with three repeated experiments, as a result makes even
Mean value.
Lactobacillus plantarum is placed in different experiments environment in following embodiment to test, is divided into according to experimental situation difference
Space flight group, ground 1g gravity control group and simulated microgravity group；
The experimental situation of space flight group are as follows: space flight condition, i.e. Shenzhou spacecraft space carrying environment；
The experimental situation of ground 1g gravity control group are as follows: ground 1g gravity condition, i.e. ground stand environment；
The experimental situation of simulated microgravity group are as follows: under ground simulation microgravity condition, i.e., with gyrator 30 turns/min mould
Quasi- microgravity environment.
Lactobacillus plantarum culture medium: peptone 10g, beef extract 10g, yeast extract 5g, KH2PO42g, trisodium citrate 2g, second
Sour sodium 2g, glucose 20g, Tween 801mL, MgSO4·7H2O0.58g、MnSO4·4H2O0.25g, distilled water are settled to 1L,
PH 6.2-6.4,121 DEG C of sterilizing 15min.
The acquisition of embodiment 1, lactobacillus plantarum LCT-LP1
One, strain acquires
To the acquisition of microorganism has been carried out inside " divine boat ten " Manned reentry capsule, bacteria culture media is selected to carry out sample
Primary dcreening operation, then the different single colonie of picking colony form carries out scribing line culture, after being purified to single bacterial strain, is named as bacterial strain LCT-
LP1。
Two, Morphological Identification
The bacterial strain LCT-LP1 that step 1 is obtained carries out Gram's staining, and bacterial strain LCT-LP1 is gram sun as the result is shown
Property, brevibacterium.The single colonie of bacterial strain LCT-LP1 is seeded to the growthform for observing bacterium colony on plate, and is seen with scanning electron microscope
Examine its microscopic morphology.Scanning electron microscopic observation thallus is in rod-short or spherical shape arrangement (see Fig. 1).Bacterium colony on soft agar is thin
Close, color is white (see Fig. 2), and colonial morphology is larger sometimes, there is raised (see Fig. 3).
Three, 16S rDNA is identified
The 16S rDNA of one bacterial strain LCT-LP1 of detecting step, sequencing result is as shown in the sequence 1 of sequence table.16s rDNA
Qualification result shows bacterial strain LCT-LP1 and lactobacillus plantarum Lactobacillus plantarum subsp.plantarum
The similitude of ATCC 14917 (T) is 99.74%.
It is identified by morphology and 16S rDNA, can determine that bacterial strain LCT-LP1 belongs to lactobacillus plantarum, therefore again will
It is named as lactobacillus plantarum LCT-LP1.
Four, the preservation of lactobacillus plantarum LCT-LP1
Lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 was preserved in China on 06 01st, 2016
Microbiological Culture Collection administration committee common micro-organisms center, deposit number are CGMCC NO.12578.Lactobacillus plantarum
(Lactobacillus plantarum) LCT-LP1 CGMCC NO.12578 is referred to as lactobacillus plantarum LCT-LP1.
Five, the Whole genome analysis of lactobacillus plantarum LCT-LP1
Paired-End sequencing is carried out to lactobacillus plantarum LCT-LP1 using high-throughput Solexa sequencing technologies.
Large fragment genomic DNA is interrupted at random using ultrasonic method Covaris and generates a series of DNA fragmentations；Then it uses
T4DNA Polymerase, Klenow DNA Polymerase and T4PNK repair Cheng Pingmo for the cohesive end to be formed is interrupted
End, then by 3 ' ends plus base " A ", enable DNA fragmentation to hold the special joint with " T " base to connect with 3 ', use electrophoresis
The target fragment connection product that need to be recycled is selected, the DNA fragmentation that round pcr amplification both ends have connector is reused；Establish 300bp
Small fragment library.The initial data obtained to sequencing is filtered and counts；With SOAP denovo V1.05 software and
To treated, reads data assemble SOAPaligner/soap2 software.And genome and gene regions coverage are carried out
Evaluation；Gene order is obtained from assembling result using Glimmer3.0 software and compares the sequence of gene and each database
It is right, obtain corresponding functional annotation information.
By the Whole genome analysis to lactobacillus plantarum LCT-LP1, whole genome sequence total length is 3,293,
854bp, G/C content 44.53%, including 3133 coded sequences and 92 rna genes (complete genome sequence map is as shown in Figure 4),
COG database compares to be annotated to 20 groups of COG function classifications (Fig. 5) altogether, while detailed GO annotation of gene function has been carried out to it
(Fig. 6) and KEGG metabolic pathway analyze (Fig. 7).
The biofilm formation experiment of embodiment 2, lactobacillus plantarum LCT-LP1
1, the lactobacillus plantarum culture medium of 50mL sterilizing aseptically, is respectively added into each 100mL indigo plant lid bottle, so
Accessing lactobacillus plantarum LCT-LP1 afterwards, (initial concentration of the lactobacillus plantarum LCT-LP1 in cultivating system is 104A/m1), 37
DEG C culture 2 days.
2, the lactobacillus plantarum LCT-LP1 culture that step 1 obtains is transferred in 96 orifice plates and carries out 37 DEG C of constant temperature cultures
For 24 hours, its biofilm formational situation in the orifice plate is observed, the bacterium colony in orifice plate is then selected and is transferred in teat glass and trained
It supports and compares, observe and compare the stabilization formational situation of its biofilm again.
Three groups of processing are arranged: in first group of processing, the condition of culture of step 1 and step 2 is the culture of space flight group
Condition；In second group, the condition of culture of step 1 and step 2 is the condition of culture of ground 1g gravity control group；In third group,
The condition of culture of step 1 and step 2 is the condition of culture of simulated microgravity group.
As a result as shown in Figure 8.In Fig. 8, M18-981 and M18-1016 are simulated microgravity group bacterium colony, and T18-685 is space
Flight crew bacterium colony, D18-390, D18-391 and D18-392 are ground 1g gravity control group bacterium colony, and three arrows are space in figure
The bacterium colony of 3 formation biomembranes of flight crew.The result shows that forming the bacterium colony of biofilm in orifice plate under the conditions of space flight
At most, the bacterium colony followed by under the conditions of simulated microgravity.The bacterium colony for forming biofilm passes through switching test tube amplification passage training
It supports, discovery can still form stable biofilm.The result shows that lactobacillus plantarum LCT-LP1 is under the conditions of space flight
Be more prone to produce biomembrane, followed by under the conditions of simulated microgravity, this show lactobacillus plantarum LCT-LP1 in space sealed compartment or
Biofilm formation probability increases under the conditions of simulated microgravity.
The corrosion test of embodiment 3, lactobacillus plantarum LCT-LP1
One, metallic test test piece is prepared
1, metal material TC4 titanium alloy (Ti-6A1-4V, (alpha+beta) alloy) and 304 stainless steels (OCr18Ni9) are chosen, are added
Work at cross-sectional diameter be 10mm, a length of 4mm cylindrical sample, successively with granularity be 400#, 800#, 1200#, 2000# gold
The polishing of phase sand paper.
2, by the 304 stainless steel samples diamond paste polishing treatment after step 1 polishing.
3, by after step 1 polishing TC4 titanium alloy sample iron oxide and oxidation chromium solution polish, acetone removes sample
Then surface grease corrodes 1-2min with 10% (mass fraction) sodium hydrate aqueous solution at 50-60 DEG C, to remove Surface Oxygen
Change layer.
4, after the processing of step 2 and step 3,304 stainless steel samples and TC4 titanium alloy sample are rinsed with tap water
Afterwards, it then immerses in 30% (volume fraction) aqueous solution of nitric acid and purifies 2-6min, be finally dipped in 70-90 DEG C of distilled water and clean, blow
It is added to 15min in 75% (volume fraction) ethanol water after dry to sterilize, is put into sterile batch cultur ware, mark good number,
It is air-dried in aseptic operating platform, it is spare.
Two, corrosion experiment
Strain to be tested are as follows: lactobacillus plantarum LCT-LP1, (the ATCC number: 14917) of lactobacillus plantarum 14917.
1, using 37 DEG C of culture strains to be tested of lactobacillus plantarum culture medium, obtaining concentration is 108The bacteria suspension of CFU/mL.
2, the TC4 titanium alloy test button of step 1 preparation and 304 stainless steel metal samples are added to 50mL's respectively
20mL lactobacillus plantarum culture medium is added in anaerobism bottle for anaerobism bottle, first with the air in high pure nitrogen stripping culture medium, then plus
The bacteria suspension for entering 1mL step 1, in 37 DEG C of constant incubator cultures.During the cultivation process, continous way experiment is carried out, respectively in reality
It tests when proceed to the 7th, 14,21 day, analyzes the sample, evaluate the corrosion behavior of metal material.
Three groups of processing are set, and in first group of processing, the condition of culture of step 1 and step 2 is the culture of space flight group
Condition；In second group, the condition of culture of step 1 and step 2 is the condition of culture of ground 1g gravity control group；In third group,
The condition of culture of step 1 and step 2 is the condition of culture of simulated microgravity group.
(1) surface biological film and etch state sem analysis.
When experiment proceeds to the 14th day, sample is placed in culture dish, delays along wall and adds 2.5% glutaraldehyde fixer, by sample
Flood at least 5mm, fixed 4h.Sample, distilled water cleaning sample film 3 times, each 15min are taken out from fixer.Successively use body
The ethanol water that fraction is 30%, 50%, 70%, 85% and 95% is dehydrated, and every grade of 15-20min is dehydrated with dehydrated alcohol
2 times, each 15-20min.Sample is put into new culture dish, pours into hexamethyldisilazane (sample is flooded at least 5mm)
3min is placed, ventilation is dry.After drying then sample is observed towards metal spraying processing is carried out above using SEM.
As a result as shown in Figure 9 and Figure 10.Fig. 9 is point of the TC4 titanium alloy test button of lactobacillus plantarum LCT-LP1 processing
Analyse result figure.Figure 10 is the analysis result figure of 304 stainless steel metal samples of lactobacillus plantarum LCT-LP1 processing.In two kinds of gold
Belonging in the testing of materials, apparent biofilm formation is not observed in the ground lactobacillus plantarum LCT-LP1 1g gravity control sample,
And lactobacillus plantarum LCT-LP1 space flight group sample forms macroscopic biomembrane, lactobacillus plantarum LCT-LP1 simulation
Microgravity group also observes that macroscopic biomembrane.The result shows that under the conditions of space flight and under the conditions of simulated microgravity
Lactobacillus plantarum is more stronger than ground 1g gravity control group in the ability that titanium alloy and stainless steel surface form biomembrane.Control strain
Biomembrane is formed under the conditions of 14917 space flight of lactobacillus plantarum and in titanium alloy and stainless steel surface under the conditions of simulated microgravity
Ability compared with the 1g gravity control group of ground without significant difference, can not observe apparent biofilm formation.
(2) it when experiment proceeds to the 7th, 14,21 day, is seen with QUANTA FEG250 FEI scanning electron microscope
Examine sample corrosion product pattern.
As a result as is illustrated by figs. 11 and 12.Figure 11 is the TC4 titanium alloy test button of lactobacillus plantarum LCT-LP1 processing
Observe result figure.Figure 12 is the observation result figure of 304 stainless steel metal samples of lactobacillus plantarum LCT-LP1 processing.At two kinds
In metal material test, lactobacillus plantarum LCT-LP1 space flight group sample metal surface is compared with the ground lactobacillus plantarum LCT-LP1
The lactobacillus plantarum of 1g gravity control sample aggregation is more, and surface corrosion region is also more, lactobacillus plantarum LCT-LP1 simulation
The lactobacillus plantarum number and surface corrosion region area of microgravity group sample metal surface aggregation and space flight group situation without
Significant difference is all larger than the ground lactobacillus plantarum LCT-LP1 1g gravity control sample.Control strain lactobacillus plantarum 14917
The lactobacillus plantarum number of sample metal surface aggregation and surface corrosion area under the conditions of space flight and under the conditions of simulated microgravity
Domain area and ground 1g gravity control group are without significant difference.
(3) when experiment proceeds to the 7th, 14,21 day, with Dimension Icon Veeco atomic force microscope pair
Sample is observed, and using standard silicon probe, carries out contact type scanning with the force constant of 0.06N/m.As a result as shown in figure 13, figure
13A is the observation result figure of the TC4 titanium alloy test button of lactobacillus plantarum LCT-LP1 processing.Figure 13 B is lactobacillus plantarum
The observation result figure of 304 stainless steel metal samples of LCT-LP1 processing.Ra is profile arithmetic average error in figure, reflects metal
Sample shows roughness, and the unit of coordinate is nm in figure.The result shows that lactobacillus plantarum LCT-LP1 is to 304 in each group processing
Stainless steel metal sample or TC4 titanium alloy test button all have corrosivity, and as the extension extent of corrosion of time aggravates.
14th day, the increase degree of stainless steel surface roughness was lower than titanium alloy.
Lactobacillus plantarum LCT-LP1 space flight group: compared with the 0th day, the 14th day titanium alloy surface roughness Ra value
Increase to 2.5 times.
Lactobacillus plantarum LCT-LP1 simulated microgravity group: compared with the 0th day, the 14th day titanium alloy surface roughness Ra
Value increases to 2.5 times.
The ground lactobacillus plantarum LCT-LP1 1g gravity control group: compared with the 0th day, the 14th day titanium alloy surface is coarse
Degree Ra value increases to 2.3 times.
14917 space flight group of lactobacillus plantarum: compared with the 0th day, the 14th day titanium alloy surface roughness Ra value increases
Add to 2.0 times.
14917 simulated microgravity group of lactobacillus plantarum: compared with the 0th day, the 14th day titanium alloy surface roughness Ra value
Increase to 1.9 times.
14917 ground 1g gravity control group of lactobacillus plantarum: compared with the 0th day, the 14th day titanium alloy surface roughness
Ra value increases to 1.9 times.
21st day, stainless steel surface roughness sharply increased, by more serious biological corrosion.
Simulated microgravity group lactobacillus plantarum LCT-LP1 to the corrosive power of titanium alloy and stainless steel and space flight group without
Significant difference, titanium alloy Ra is 10.8nm at the 14th day, and stainless steel Ra is 12.5nm at the 21st day, is all larger than ground 1g gravity
Control group.Control strain lactobacillus plantarum 14917 is under the conditions of space flight condition and simulated microgravity to titanium alloy and stainless
The corrosive power and ground 1g gravity control group of steel are without significant difference.The result shows that space flight group and simulated microgravity tissue culture
Lactobacillus plantarum LCT-LP1 is all larger than ground 1g gravity control group to the corrosive power of titanium alloy and stainless steel under the conditions of supporting.
(4) when experiment proceeds to the 7th, 14,21 day, in energy spectrum analysis (EDS) network analysis corrosion sample
Elemental composition.As a result as shown in figure 14, Figure 14 is elemental composition analysis knot in the corrosion sample of lactobacillus plantarum LCT-LP1 processing
Fruit.EDS analysis shows: at 14 days, surface carbon element content significantly improved space flight group sample, oxygen element amount increase
To 2.1%, increase significantly than ground 1g gravity control group, 21 days results also further detect that carbon and oxygen element increase
Significantly.This shows that the vital movement of lactobacillus plantarum LCT-LP1 under the conditions of space flight is more strong, illustrates metal material surface
Biological corrosion is more serious.
Show lactobacillus plantarum LCT-LP1 in space flight group, simulated microgravity group and ground 1g weight based on the above results
Titanium alloy and stainless steel can be corroded under the conditions of power control group, but more held under the conditions of space flight and under the conditions of simulated microgravity
Biomembrane easily is formed in metal material surface, the corrosive power of titanium alloy and stainless steel is enhanced compared with ground 1g gravity control group.
<110>Fuller gives birth suddenly object engineering science and technology (Beijing) Co., Ltd
<120>cultural method of spatial plant lactobacillus LCT-LP1 and application
<130> GNCYXMN161621
<160> 1
<210> 1
<211> 1530
<212> DNA
<213>lactobacillus plantarum (Lactobacillus plantarum)
<400> 1
tagagtttga tcctggctca ggacgaacgc tggcggcgtg cctaatacat gcaagtcgaa 60
cgaactctgg tattgattgg tgcttgcatc atgatttaca tttgagtgag tggcgaactg 120
gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata acacctggaa acagatgcta 180
ataccgcata acaacttgga ccgcatggtc cgagcttgaa agatggcttc ggctatcact 240
tttggatggt cccgcggcgt attagctaga tggtggggta acggctcacc atggcaatga 300
tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg cccaaactcc 360
tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg agcaacgccg 420
cgtgagtgaa gaagggtttc ggctcgtaaa actctgttgt taaagaagaa catatctgag 480
agtaactgtt caggtattga cggtatttaa ccagaaagcc acggctaact acgtgccagc 540
agccgcggta atacgtaggt ggcaagcgtt gtccggattt attgggcgta aagcgagcgc 600
aggcggtttt ttaagtctga tgtgaaagcc ttcggctcaa ccgaagaagt gcatcggaaa 660
ctgggaagct tgagtgcaga agaggacagt ggaactccat gtgtagcggt gaaatgcgta 720
gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg acgctgaggc 780
tcgaaagtat gggtagcaaa caggattaga taccctggta gtccataccg taaacgatga 840
atgctaagtg ttggggggtt tccgcccttc agtgctgcag ctaacgcatt aagcattccg 900
cctggggagt acggccgcaa ggctgaaact caaaggaatt gacgggggcc cgcacaagcg 960
gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct tgacatacta 1020
tgcaaatcta agagattaga cgttcccttc ggggacatgg atacaggtgg tgcatggttg 1080
tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa cccttattat 1140
cagttgccag cattaagttg ggcactctgg tgagactgcc ggtgacaaac cggaggaagg 1200
tggggatgac gtcaaatcat catgcccctt gtgacctggg ctacacacgt gctacaatgg 1260
atggtacagc gagttgcgaa ctcgcgagag taagctaatc tcttaaagcc attctcagtt 1320
cggattgtag gctgcaactc gcctacatga agtcggaatc gctagtaatc gcggatcagc 1380
atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc atgagagttt 1440
gtaacaccca aagtcggtgg ggtaaccttt taggaaccag ccgcctaagg tgggacagat 1500
gattagggtg aagtcgtaac aaggtaacca 1530
Claims (9)
1. a kind of culture lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 method, include the following steps:
Using lactobacillus plantarum described in lactobacillus plantarum culture medium culture (Lactobacillus plantarum) LCT-LP1；The plant
Solute in object lactobacillus culture medium and its concentration in the lactobacillus plantarum culture medium are as follows: peptone 0.5-1.5g/
100ml, beef extract 0.5-1.5g/100ml, yeast extract 0.4-0.6g/100ml, KH2PO40.1-0.3g/100ml, citric acid three
Sodium 0.1-0.3g/100ml, sodium acetate 0.1-0.3g/100ml, 80 0.05-0.15mL/ of glucose 1-3g/100ml, Tween
100ml、MgSO4·7H2O 0.048-0.068g/100ml、MnSO4·4H2O 0.015-0.035g/100ml；The plant cream
Bacillus (Lactobacillus plantarum) LCT-LP1 deposit number be CGMCC NO.12578.
2. a kind of method for preparing lactobacillus plantarum LCT-LP1 culture includes the following steps: using lactobacillus plantarum culture medium
Culture lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1；Solute in the lactobacillus plantarum culture medium and
Its concentration in the lactobacillus plantarum culture medium is as follows: peptone 0.5-1.5g/100ml, beef extract 0.5-1.5g/
100ml, yeast extract 0.4-0.6g/100ml, KH2PO40.1-0.3g/100ml, trisodium citrate 0.1-0.3g/100ml, acetic acid
Sodium 0.1-0.3g/100ml, 80 0.05-0.15 mL/100ml of glucose 1-3g/100ml, Tween, MgSO4·7H2O
0.048-0.068g/100ml、MnSO4·4H2O 0.015-0.035g/100ml；The lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 deposit number be CGMCC NO.12578.
3. method according to claim 1 or 2, it is characterised in that: the solvent of the lactobacillus plantarum culture medium is water.
4. method according to claim 1 or 2, it is characterised in that: the pH of the lactobacillus plantarum culture medium is 6.2-6.4.
5. lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 in microorganisms to space flight equipment etching machine
Application in reason；The lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 deposit number be CGMCC
NO.12578；The space flight equipment is titanium alloy and/or stainless steel.
6. lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 prevention and control microbiologic(al) corrosion space flight equipment study
In application；The lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 deposit number be CGMCC
NO.12578；The space flight equipment is titanium alloy and/or stainless steel.
7. lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 is in researching and developing novel space flight corrosion-resistant material
Using；The lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 deposit number be CGMCC
NO.12578；The material is titanium alloy and/or stainless steel.
8. lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 corrodes space flight equipment in preparation microorganism
Application in model；The lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 deposit number be CGMCC
NO.12578；The space flight equipment is titanium alloy and/or stainless steel.
9. lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 preparation corrosion space flight equipment product in
Using；The lactobacillus plantarum (Lactobacillus plantarum) LCT-LP1 deposit number be CGMCC
NO.12578；The space flight equipment is titanium alloy and/or stainless steel.
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