CN108384738B - Pseudomonas aeruginosa, screening method thereof and application thereof in straw lactic acid fermentation - Google Patents
Pseudomonas aeruginosa, screening method thereof and application thereof in straw lactic acid fermentation Download PDFInfo
- Publication number
- CN108384738B CN108384738B CN201810399264.7A CN201810399264A CN108384738B CN 108384738 B CN108384738 B CN 108384738B CN 201810399264 A CN201810399264 A CN 201810399264A CN 108384738 B CN108384738 B CN 108384738B
- Authority
- CN
- China
- Prior art keywords
- pseudomonas aeruginosa
- lactic acid
- fermentation
- straw
- straws
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/38—Pseudomonas
- C12R2001/385—Pseudomonas aeruginosa
-
- 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
- C12N1/205—Bacterial isolates
-
- 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
- C12N1/02—Separating microorganisms from their culture media
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/56—Lactic acid
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides pseudomonas aeruginosa with a preservation number of CCTCC NO: m2018169; the invention also provides a screening method of the pseudomonas aeruginosa strain and application of the pseudomonas aeruginosa strain in straw lactic acid fermentation; the pseudomonas aeruginosa is obtained by screening aerobic tank activated sludge of a municipal sewage treatment plant, can efficiently degrade various typical products generated in the pretreatment process of lignin, is cultured for 36 hours at 37 ℃, and has degradation rates of coumaric acid, p-hydroxybenzaldehyde, vanillin, ferulic acid and syringaldehyde which are all substrates of 100 percent; the pseudomonas aeruginosa is used for straw lactic acid fermentation, the typical products can be prevented from inhibiting microbial metabolism during straw lactic acid fermentation, so that the conversion rate of lactic acid to straw is improved, the fermentation period is short, fermentation is finished after 48-72h of culture, plant straw is used as a fermentation raw material, waste is changed into valuable, environmental pollution is reduced, and the method has positive significance for ensuring grain safety.
Description
Technical Field
The invention relates to the fields of microbiology and straw lactic acid fermentation, in particular to pseudomonas aeruginosa, a screening method thereof and application of the pseudomonas aeruginosa in straw lactic acid fermentation.
Background
China is a big agricultural country, and the yield of crop straws exceeds 8 hundred million tons every year. Crop straws are mainly used for returning to fields, ensiling and yellow silage, methane, biochar, biomass fuel, biological energy and the like, but the straws are slowly decomposed by directly returning to the fields and the quality of cultivated land is not obviously improved; as silage or yellow silage, the production period is long, and the feed conversion rate is not high; the biogas produced by straw fermentation can produce a large amount of biogas slurry and biogas residues, and the severe haze is caused by the massive incineration of the straws. In summary, the crop straw resources are utilized in large quantities, but still have the problems of low utilization rate, secondary pollution and the like.
The method converts crop straw resources into products such as lactic acid and the like through microbial fermentation, and is a new way for realizing the efficient utilization of the straw resources. Lactic acid is a hydroxycarboxylic acid that is widely used in the food industry (preservatives, flavoring agents), the pharmaceutical industry (ointments, lotions, injections) and the leather industry (acidifiers, calcium scavengers). Are widely used in the food industry as souring agents, preservatives and reducing agents.
The industrial production of lactic acid mainly uses microbial fermentation and uses grain crops such as corn and the like as raw materials, so that the price of the lactic acid is high, and meanwhile, the lactic acid poses potential threats to national grain safety. Cellulose and hemicellulose in the main components of the straws can be converted into hexose and pentose through enzymolysis, and the theoretical conversion rate of lactic acid to sugar is 100 percent in the homolactic fermentation process of microorganisms. Therefore, the method for producing the lactic acid by using the crop straws as the raw material can realize the high-efficiency utilization of straw resources, improve the economic value of the straws, open up a new idea for the production of the lactic acid, provide wider and cheaper raw materials, reduce the production cost of the lactic acid, expand the application field of the lactic acid and achieve two purposes at one stroke.
Comparison document 1: the invention discloses a method for producing lactic acid by fermenting plant straw cellulose with carbon source for fermentation after partial enzymolysis, which is a patent with the application number of 'CN 201010609278.0' and the name of 'a lactic acid bacteria fermentation method', wherein the sugar concentration of the plant straw cellulose after partial enzymolysis is 1-10 wt%, the partial enzymolysis is to carry out enzymolysis on the plant straw cellulose with cellulase for 12-72 hours at 45-55 ℃, so that fermentation liquor contains enzymolysis sugar and cellulose without enzymolysis, and the lactic acid can be produced by fermentation while enzymolysis. Compared with the prior art, the production period is shortened from 136 hours to 64 hours, the inhibition effect of high-concentration glucose on lactic acid fermentation of lactic acid bacteria is removed, and the fermentation level is improved from 3.6 wt% to 6.8 wt%. The plant straw is used as the fermentation raw material, so that the waste is changed into valuable, the environmental pollution is reduced, and the method has positive significance for ensuring the grain safety. However, the conversion rate of the lactic acid to the straws is not high, the crop straws can be utilized by the microorganisms after being pretreated, and the straws can generate a plurality of byproducts in the pretreatment process to inhibit the metabolism of the microorganisms, wherein the main sources of the byproducts are the decomposition of lignin and hemicellulose, including micromolecular organic acids, furans, phenols and the like. Most microorganisms cannot utilize lignin, and the lignin and partial degradation products thereof can adsorb cellulase in the fermentation process to form coprecipitation, so that the activity of the lignin is reduced to loss; the lignin is decomposed to generate phenolic compounds such as ferulic acid, coniferyl aldehyde and the like, the fluidity of microbial cell membranes is improved, a large amount of potassium ions are lost, the cell membranes are damaged, the metabolism of microbes is inhibited, and therefore the conversion rate of lactic acid on straws is influenced.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a pseudomonas aeruginosa strain, a screening method thereof and application thereof in straw lactic acid fermentation, wherein the pseudomonas aeruginosa strain is screened from aerobic tank activated sludge of a municipal sewage treatment plant, can efficiently degrade various typical products generated in the pretreatment process of lignin, and avoids the typical products from inhibiting microbial metabolism during straw lactic acid fermentation, so that the conversion rate of lactic acid on straw is improved, the fermentation period is short, plant straw is used as a fermentation raw material, waste is changed into valuable, environmental pollution is reduced, and positive significance is brought to food safety guarantee.
One of the purposes of the invention is to provide a strain of Pseudomonas aeruginosa, namely Pseudomonas aeruginosa (Pseudomonas aeruginosa) FL02, which is screened from aerobic tank activated sludge of a municipal sewage treatment plant.
The 16S rRNA sequence of the strain FL02 is shown in SEQ ID NO. 1, and the 16S rRNA gene of the strain is amplified by PCR using primers 27F (5'-AGAGTT TGATCCTGGCTCAG-3') and 1492R (5'-TAC GGCTACCTTGTTACGACTT-3'). Sequence analysis and homology comparison of 16S rRNA sequence in GenBank shows 99% similarity with Pseudomonas aeruginosa by aligning strain FL 02. In conclusion, through morphological identification, physiological and biochemical experiments and 16S rRNA sequence analysis and homology comparison of strains, the strain Pseudomonas aeruginosa FL02 is determined to be Pseudomonas aeruginosa.
Based on the above information, strain FL02 was identified as Pseudomonas aeruginosa (Pseudomonas aeruginosa), which is now deposited in the chinese collection of type cultures (CCTCC), address: in the Wuhan university school of Wuhan 299 in the Wuchang area of Wuhan city, Hubei province, the preservation center of Wuhan university, the postcode: 430072, preservation number CCTCCNO: m2018169, date of deposit 2018, 3 months and 30 days.
The invention also aims to provide a screening method of the pseudomonas aeruginosa, which comprises the following steps:
step 1, adding an enrichment culture medium into a culture bottle, sequentially adding sludge of a secondary sedimentation tank of a water treatment plant and soil near a water outlet of a printing and dyeing plant, sealing, and performing shaking culture to obtain an enrichment culture solution, wherein the enrichment culture medium comprises vanillin, p-hydroxybenzaldehyde, syringaldehyde and yeast extract powder;
step 2, filling a screening culture medium in a culture bottle, adding the enrichment culture solution obtained in the step 1, sealing, and performing shaking culture to obtain a screening seed solution, wherein the screening culture medium comprises vanillin, p-hydroxybenzaldehyde, syringaldehyde, guaiacol, ferulic acid, coumaric acid, yeast extract powder, MgSO (MgSO) as well as vanillin, p-hydroxybenzaldehyde, syringaldehyde, guaiacol, ferulic acid, coumaric acid, yeast extract powder4 7H2O、CaCl2、(NH4)2SO4、K2HPO4NaCl and FeCl3；
Step 3, coating the screened seed liquid obtained in the step 2 on a solid screening culture medium plate after gradient dilution, and picking out a well grown single colony after standing culture;
and 4, preserving the single colony obtained in the step 3, and performing 16S rDNA identification to obtain the result of pseudomonas aeruginosa.
The invention also aims to provide application of the pseudomonas aeruginosa strain in straw lactic acid fermentation.
As a specific implementation mode of the application of the pseudomonas aeruginosa in straw lactic acid fermentation, the method comprises the following steps:
step S1, pretreatment of corn straws: weighing corn straw in aluminum foilAdding strong ammonia water into the bag, rapidly sealing, and repeatedly kneading the aluminum foil bag to make NH3Fully contacting with the straws, and standing for more than three days at normal temperature to obtain ammonia-treated straws;
step S2, fermenting straw with lactic acid: and (3) filling the pretreated corn straws obtained in the step (S1) in a culture bottle, inoculating pseudomonas aeruginosa condensed in the culture bottle and pseudomonas aeruginosa in the step (1), adding composite cellulase as a substrate, and standing, fermenting and culturing to obtain a lactic acid finished product.
The invention has the following beneficial effects:
1. the pseudomonas aeruginosa provided by the invention can efficiently degrade various typical products such as p-hydroxybenzaldehyde, coumaric acid, vanillin, ferulic acid and syringaldehyde generated in the pretreatment process of lignin, and the pseudomonas aeruginosa FL02 can be cultured for 36 hours at the temperature of 37 ℃ at 150r/min by using the compound as a unique carbon source, so that the degradation rate of each substrate, i.e. coumaric acid, p-hydroxybenzaldehyde, vanillin, ferulic acid and syringaldehyde reaches 100%.
2. The pseudomonas aeruginosa provided by the invention can efficiently degrade various typical products generated in the pretreatment process of lignin, and the typical products are prevented from inhibiting microbial metabolism during straw lactic acid fermentation, so that the conversion rate of lactic acid to straw is improved, and the conversion rate of lactic acid to straw in a straw lactic acid fermentation experiment is as high as 65%;
3. the application of the pseudomonas aeruginosa in straw lactic acid fermentation provided by the invention has a short fermentation period, the various typical products can be completely degraded after being cultured for 36 hours at 37 ℃, and the fermentation is finished after being cultured for 48-72 hours;
4. the invention utilizes plant straws as fermentation raw materials, changes waste into valuable, reduces environmental pollution and has positive significance for ensuring grain safety.
The preservation date of the strain is 2018, 3 and 30 months, and the preservation number is CCTCC NO: m2018169, classified and named as Pseudomonas aeruginosa, the name of the preservation unit is China Center for Type Culture Collection (CCTCC), and the address is the university school of Wuhan with eight paths 299 in Wuchang district, Wuhan city, Hubei province.
Detailed Description
EXAMPLE 1 screening of Strain FL02
1. Adding 90ml of enrichment medium (1g/L of vanillin, 1g/L of p-hydroxybenzaldehyde, 1g/L of syringaldehyde and 1g/L of yeast extract powder, pH7.0) into a 250ml triangular flask, respectively adding sludge of a secondary sedimentation tank of a water treatment plant and 10g of soil near a water outlet of a printing and dyeing mill, sealing, placing the mixture in a shaking table at 37 ℃, and performing shaking culture at 150r/min for 24 hours to obtain an enrichment culture solution.
2. 100ml of screening medium (1g/L vanillin +1g/L p-hydroxybenzaldehyde +1g/L syringaldehyde +1g/L guaiacol +1g/L ferulic acid +1g/L coumaric acid +0.5g/L yeast extract powder +0.2g/L MgSO 2. sup.4 7H2O,0.1g/L CaCl2,1g/L(NH4)2SO4,1g/L K2HPO4,0.1g/L NaCl,0.02g/L FeCl3pH7.0), adding 1ml of enrichment culture solution respectively, sealing, placing in a shaking table at 37 ℃, and performing shaking culture at 150r/min for 48h to obtain screening seed solution.
3. And (3) diluting the screened seed solution in a gradient manner, coating the diluted seed solution on a solid screening culture medium plate, performing static culture at 37 ℃ for 48 hours, and picking a single colony to obtain the strain FL 02.
Example 2 identification of Strain FL02
The 16S rRNA sequence of the strain FL02 is shown in SEQ ID NO. 1, and the 16S rRNA gene of the strain is amplified by PCR using primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGCTACCTTGTTACGACTT-3'). Sequence analysis and homology comparison of the 16S rRNA sequence in GenBank showed 99% similarity to Pseudomonas aeruginosa by aligning strain FL 02. The strain FL02 is subjected to morphological identification, physiological and biochemical experiments, 16S rRNA sequence analysis and homology comparison, and the similarity of the strain FL02 and Pseudomonas aeruginosa is 99 percent by comparison, so that the strain Pseudomonas aeruginosa FL02 is determined to be Pseudomonas aeruginosa.
The microbiological characteristics of strain FL02 are as follows: the cells are elongated and vary in length, and may be arranged in a club-like or linear form, in pairs or in short chains. One end of the thallus is provided with a single whip hair, the movement and activity of bacteria can be observed under a dark field microscope or a phase contrast microscope, the thallus is slender and has different lengths, and sometimes the thallus is arranged in a ball-rod shape or a linear shape in pairs or short chains. One end of the thallus has single penis hair, no spore and no capsule. Large, flat, moist, metallic luster, blue-green, transparent hemolytic ring colony and ginger flavor are formed on the blood plate.
The physicochemical characteristics of strain FL02 are as follows: gram-negative bacteria, oxidase positive, green pus producing, agar yellow-green staining, as shown in table 1.
TABLE 1
Example 3 degradation of coumaric acid, p-hydroxybenzaldehyde, vanillin, ferulic acid, syringaldehyde by Strain FL02
Detecting the degradation or utilization condition of the Pseudomonas aeruginosa strain (Pseudomonas aeruginosa) FL02 obtained by screening of the invention on coumaric acid, p-hydroxybenzaldehyde, vanillin, ferulic acid and syringaldehyde.
The specific test process is as follows: 5 parts of Pseudomonas aeruginosa strain (Pseudomonas aeruginosa) FL02 obtained by screening in equal amount are respectively added into culture media 1-5, wherein the culture media 1-5 respectively contain 0.5g/L yeast extract powder and 0.2g/L MgSO4.7H2O,0.1g/L CaCl2,1g/L(NH4)2SO4,1g/L K2HPO4,0.1g/L NaCl,0.02g/L FeCl3And the culture medium 1-5 respectively contains 1g/L vanillin, 1g/L p-hydroxybenzaldehyde, 1g/L syringaldehyde, 1g/L coumaric acid and 1g/L ferulic acid, and the Pseudomonas aeruginosa FL02 can be cultured for 36 hours at the temperature of 37 ℃ at 150r/min by using the compounds as unique carbon sources, and the result shows that the Pseudomonas aeruginosa strain (Pseudomonas aeruginosa) FL02 can grow by using the substrate as a carbon source. In addition, after 36h of culture, sampling and detecting the content of each substrate, the result shows that the Pseudomonas aeruginosa strain (Pseudomonas aeruginosa) FL02 can completely degrade each substrate, and the degradation rate of each substrate, namely coumaric acid, p-hydroxybenzaldehyde, vanillin, ferulic acid and syringaldehyde, can reach 100%.
Example 4 straw lactic acid fermentation experiment for detecting conversion rate of lactic acid to straw
1. The corn straw pretreatment method comprises the following steps: weighing 80g of corn straw in an aluminum foil bag, adding 20ml of concentrated ammonia water with the volume concentration of 25%, quickly sealing, and repeatedly kneading the aluminum foil bag to ensure that NH is generated3Fully contacting with the straws, and standing for more than three days at normal temperature to obtain the ammonia-treated straws.
2. Straw lactic acid fermentation experiment: two portions of pretreated corn straws with the concentration of 50g/L are taken and filled in a 250ml triangular flask with 100ml of pretreated corn straws, 1ml of bacillus coagulans FL209 and 1ml of pseudomonas aeruginosa FL02 are inoculated at the same time to serve as an experimental group, only 1ml of bacillus coagulans FL209 is inoculated to serve as a control group, and the experimental group and the control group are both added with 10FPU/g (substrate) of composite cellulase. Standing at 50 deg.C, fermenting, and adjusting pH once every 12 hr to 6.0-6.5. After culturing for 48-72h, or the pH is not changed any more, the fermentation is finished. The contents of lactic acid, acetic acid and total phenols in each experimental group are detected, the conversion rate is calculated, and the experimental group and the control group repeat the experiment for 3 times. As in table 2 below.
TABLE 2
The experimental results show that compared with a control group, the lactic acid concentrations of the experimental groups 1-3 are obviously improved, the yield of products inhibiting microbial metabolism, such as acetic acid, total phenols and the like, is increased, the microbial metabolism of typical products during straw lactic acid fermentation is prevented from being inhibited, the conversion rate of lactic acid to straw is increased, and the conversion rate of lactic acid to straw in the straw lactic acid fermentation experiment is as high as 65%.
The application of the pseudomonas aeruginosa in straw lactic acid fermentation provided by the invention has a short fermentation period, the various typical products can be completely degraded after being cultured for 36 hours at 37 ℃, and the fermentation is finished after being cultured for 48-72 hours; the invention utilizes plant straws as fermentation raw materials, changes waste into valuable, reduces environmental pollution and has positive significance for ensuring grain safety.
Wherein the deposit number of Bacillus coagulans FL209 is as follows: CCTCC No.2015773, specifically the patent with the application number of "CN 201610044993.1", entitled "Bacillus coagulans and its culture method and application in preparing biological organic fertilizer".
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Sequence listing
<110> university of agriculture in Huazhong
<120> pseudomonas aeruginosa strain, screening method thereof and application thereof in straw lactic acid fermentation
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1387
<212> DNA
<213> Pseudomonas aeruginosa (Pseudomonas aeruginosa)
<400> 1
gtcgagcgga tgaagggagc ttgctcctgg attcagcggc ggacgggtga gtaatgccta 60
ggaatctgcc tggtagtggg ggataacgtc cggaaacggg cgctaatacc gcatacgtcc 120
tgagggagaa agtgggggat cttcggacct cacgctatca gatgagccta ggtcggatta 180
gctagttggt ggggtaaagg cctaccaagg cgacgatccg taactggtct gagaggatga 240
tcagtcacac tggaactgag acacggtcca gactcctacg ggaggcagca gtggggaata 300
ttggacaatg ggcgaaagcc tgatccagcc atgccgcgtg tgtgaagaag gtcttcggat 360
tgtaaagcac tttaagttgg gaggaagggc agtaagttaa taccttgctg ttttgacgtt 420
accaacagaa taagcaccgg ctaacttcgt gccagcagcc gcggtaatac gaagggtgca 480
agcgttaatc ggaattactg ggcgtaaagc gcgcgtaggt ggttcagcaa gttggatgtg 540
aaatccccgg gctcaacctg ggaactgcat ccaaaactac tgagctagag tacggtagag 600
ggtggtggaa tttcctgtgt agcggtgaaa tgcgtagata taggaaggaa caccagtggc 660
gaaggcgacc acctggactg atactgacac tgaggtgcga aagcgtgggg agcaaacagg 720
attagatacc ctggtagtcc acgccgtaaa cgatgtcgac tagccgttgg gatccttgag 780
atcttagtgg cgcagctaac gcgataagtc gaccgcctgg ggagtacggc cgcaaggtta 840
aaactcaaat gaattgacgg gggcccgcac aagcggtgga gcatgtggtt taattcgaag 900
caacgcgaag aaccttacct ggccttgaca tgctgagaac tttccagaga tggattggtg 960
ccttcgggaa ctcagacaca ggtgctgcat ggctgtcgtc agctcgtgtc gtgagatgtt 1020
gggttaagtc ccgtaacgag cgcaaccctt gtccttagtt accagcacct cgggtgggca 1080
ctctaaggag actgccggtg acaaaccgga ggaaggtggg gatgacgtca agtcatcatg 1140
gcccttacgg ccagggctac acacgtgcta caatggtcgg tacaaagggt tgccaagccg 1200
cgaggtggag ctaatcccat aaaaccgatc gtagtccgga tcgcagtctg caactcgact 1260
gcgtgaagtc ggaatcgcta gtaatcgtga atcagaatgt cacggtgaat acgttcccgg 1320
gccttgtaca caccgcccgt cacaccatgg gagtgggttg ctccagaagt agctagtcta 1380
accgcaa 1387
Claims (5)
1. Pseudomonas aeruginosa (Pseudomonas aeruginosa) FL02, which is preserved in China center for type culture Collection with the preservation number of CCTCCNO: and M2018169.
2. The use of Pseudomonas aeruginosa (Pseudomonas aeruginosa) FL02 in straw lactic acid fermentation as claimed in claim 1.
3. The use according to claim 2, comprising the steps of:
step S1, pretreatment of corn straws: weighing corn stalk, putting into aluminum foil bag, adding strong ammonia water, rapidly sealing, and repeatedly kneading aluminum foil bag to obtain NH3Fully contacting with the straws, and standing for more than three days at normal temperature to obtain ammonia-treated straws;
step S2, fermenting straw with lactic acid: the pretreated corn straws obtained in the step S1 are filled in a culture bottle, simultaneously Bacillus coagulans (Bacillus coagulans) FL209 and Pseudomonas aeruginosa (Pseudomonas aeruginosa) FL02 as claimed in claim 1 are inoculated, and a finished lactic acid product is obtained after standing fermentation culture by adding composite cellulase as a substrate;
wherein, the bacillus coagulans FL209 is preserved in China center for type culture Collection with the preservation number of: CCTCC No. 2015773.
4. The use of claim 3, wherein in step S1, 80g of corn stalks are weighed into an aluminum foil bag, and 20ml of concentrated ammonia water with a volume concentration of 25% is added.
5. The use of claim 3, wherein in step S2, 100ml of pretreated corn stalks with a concentration of 50g/L are placed in a 250ml triangular flask, 1ml of Bacillus coagulans FL209 and 1ml of Pseudomonas aeruginosa FL02 are inoculated simultaneously, 10FPU/g of composite cellulase is added, standing fermentation is carried out at 50 ℃, pH is adjusted once every 12h, pH is controlled to be 6.0-6.5, and after cultivation for 84h or when pH is not changed any more, the fermentation is finished to obtain the finished lactic acid product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810399264.7A CN108384738B (en) | 2018-04-28 | 2018-04-28 | Pseudomonas aeruginosa, screening method thereof and application thereof in straw lactic acid fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810399264.7A CN108384738B (en) | 2018-04-28 | 2018-04-28 | Pseudomonas aeruginosa, screening method thereof and application thereof in straw lactic acid fermentation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108384738A CN108384738A (en) | 2018-08-10 |
| CN108384738B true CN108384738B (en) | 2021-05-25 |
Family
ID=63065067
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810399264.7A Active CN108384738B (en) | 2018-04-28 | 2018-04-28 | Pseudomonas aeruginosa, screening method thereof and application thereof in straw lactic acid fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108384738B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112080450B (en) * | 2020-09-30 | 2022-04-19 | 内蒙古农业大学 | Straw degrading bacteria and separation screening method |
| CN116536209B (en) * | 2023-05-15 | 2024-04-19 | 四川大学 | Pseudomonas azotoformans YF-58 for high yield of guaiacol and application thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8080403B2 (en) * | 2002-03-21 | 2011-12-20 | Council Of Scientific And Industrial Research | Biological process for color reduction of pulp and paper effluent |
| CN101857847A (en) * | 2010-05-06 | 2010-10-13 | 合肥工业大学 | Pseudomonas aeruginosa strain separating, purifying and domesticating method and use |
| CN106520615A (en) * | 2016-11-16 | 2017-03-22 | 河南农业大学 | Composite microbial system for quickly degrading maize straw and preparation and pretreatment method thereof |
-
2018
- 2018-04-28 CN CN201810399264.7A patent/CN108384738B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN108384738A (en) | 2018-08-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Bioaugmentation with an acetate-type fermentation bacterium Acetobacteroides hydrogenigenes improves methane production from corn straw | |
| Lu et al. | Characteristics of hydrogen and methane production from cornstalks by an augmented two-or three-stage anaerobic fermentation process | |
| CN101981199A (en) | Methods for the conversion of plant materials into fuels and chemicals by sequential action of two microorganisms | |
| US20070207531A1 (en) | Methods and bacterial strains for producing hydrogen from biomass | |
| CN108676755B (en) | Microbial liquid fertilizer containing bacillus and preparation method and application thereof | |
| EP3464555A1 (en) | Process for edible filamentous fungal cultivation and its integration in conventional sugar to ethanol production | |
| CN101519639A (en) | Paenibacillus polymyxa for preventing and treating plant fungal diseases and production method thereof | |
| CN108384738B (en) | Pseudomonas aeruginosa, screening method thereof and application thereof in straw lactic acid fermentation | |
| Didak Ljubas et al. | Production of different biochemicals by Paenibacillus polymyxa DSM 742 from pretreated brewers’ spent grains | |
| Atitallah et al. | Efficient bioethanol production from date palm (Phoenix dactylifera L.) sap by a newly isolated Saccharomyces cerevisiae X19G2 | |
| Sharma et al. | Effect of phenolic compounds on hydrogen production from municipal solid waste | |
| CN113046262B (en) | Stenotrophomonas-cauiu-1 and uses thereof | |
| Akponah et al. | Bio-ethanol production from cassava effluent using Zymomonas mobilis and Saccharomyces cerevisiae isolated from rafia palm (Elaesis guineesi) SAP | |
| CN112625948A (en) | Bacillus tequilensis S1 with nitrogen fixation function and application thereof in compost | |
| CN113337407B (en) | Mucor circinelloides CAULIU-FUNGUS-1 and application thereof | |
| CN112813010B (en) | Bacillus-CAULIU-1 and application thereof | |
| CN105349442B (en) | A kind of plum surprise saccharomycete and its application | |
| CN105385611B (en) | A kind of plum surprise saccharomycete and its application | |
| Denchev et al. | Biohydrogen production from lignocellulosic waste with anaerobic bacteria | |
| CN111394273B (en) | Bacillus beiLeisi CZ1 capable of producing fulvic acid and application thereof in secondary fermentation of livestock and poultry manure | |
| CN111100802B (en) | Enterococcus faecalis and application thereof | |
| CN113174343A (en) | Microbial flora for producing lactic acid by utilizing lignocellulose and fermentation method | |
| CN110734868A (en) | Bacillus coagulans RBE4-4 isolate with high lactic acid production capacity and application thereof | |
| KR101173468B1 (en) | Enterobacter sp?ES392 KACC 91568P and method of producing hydrogen by using the same | |
| CN108913616B (en) | Method for preparing biological bacterial fertilizer by utilizing tobacco leaf residues |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |