Bacillus-like strain ngmcc 1.200843 from rhinoceros feces having phosphorus and starch solubilizing action and use thereof

Abstract

The application discloses a Paenibacillus sp. NGMCC 1.200843 derived from rhinoceros feces, belongs to a new discovery in the technical field of microorganisms, and is identified as a new species of Paenibacillus, numbered as NGMCC 1.200843=CGMCC 1.64763T, and has been registered and preserved in the China General Microbiological Culture Collection Center. The strain can form a larger transparent circle on a specific culture medium, indicating that the strain has strong phosphorus and starch degradation capacity. Under the current situation of soil fertility decline and fertilizer abuse, the strain provides a high-quality strain resource for the development of microbial fertilizers with organic phosphorus degradation function, and helps to promote the development of microbial fertilizers. The hydrolytic enzymes produced by the strain have wide application potential in the industrial fields of food, fermentation and pharmaceuticals. In particular, amylase can hydrolyze starch to produce various products, and has important significance for the biotechnology industry, and the application range covers multiple industries such as food, fermentation, textile and papermaking. In a word, the application provides a new resource for the development of microbial fertilizers and industrial enzyme preparations.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbial technology, in particular to a Paenibacillus NGMCC 1.200843 with phosphorus and starch solubilizing effect and application thereof. BACKGROUND

[0002] Phosphorus (P) is the second most important element for plant growth and development. Phosphorus exists in three different forms, such as organic phosphorus (orthophosphate, phospholipids and nucleic acids), aluminum / iron-bound inorganic phosphorus (in acid soils), and calcium-bound inorganic phosphorus (in alkaline soils). However, only 0.1% of the phosphorus in the global cultivated land can be absorbed by plant roots. In order to cope with this extreme deficiency, the production and application of a large amount of phosphorus fertilizer has increased. In fact, plants have low phosphorus utilization efficiency under phosphorus stress or even when phosphorus is sufficient, and long-term application of phosphorus fertilizer has adverse effects on the environment, and the cost of phosphorus fertilizer is high. Phosphorus solubilizing bacteria (PSB) can convert insoluble phosphorus in the soil into available phosphorus that can be directly absorbed by plants, thereby improving the absorption and utilization of phosphorus by plants. These phosphorus solubilizing microorganisms have different potential for dissolving phosphorus, mainly depending on their adopted dissolution mechanism, molecular genetics, and their ability to release phosphorus in the soil. Screening of efficient phosphorus solubilizing microorganisms, developing them into phosphorus solubilizing bacterial agents for inoculation, or directly applying them to the soil, can activate insoluble phosphate in the soil, which has important theoretical and practical significance in agricultural production.

[0003] Paenibacillus was first proposed by Ash et al. in 1993, with Paenibacillus polymyxa as the model species. Most members of the Paenibacillus genus are endospore-forming, gram-negative bacteria, facultative anaerobes or obligate aerobes, with a DNA G+C content of 39-59 mol%, and move by peritrichal flagella. Bacteria belonging to the Paenibacillus genus have been isolated from various environments, and many species are related to humans, animals, plants and the environment. Most of them exist in soil, usually associated with plant roots, and these rhizosphere bacteria promote plant growth and can be used in agriculture. Paenibacillus-derived antibacterial agents also have applications in medicine, including polymyxins and fusaricidins. Other useful molecules include exopolysaccharides (EPS) and enzymes such as amylases, cellulases, hemicellulases, lipases, pectinases, oxygenases, dehydrogenases, lignin-modifying enzymes and mutagenic enzymes, which can be used in detergents, food and feed, textiles, paper, biofuels and healthcare. Despite this, most of these Paenibacillus have not been fully explored and have potential for biotechnological applications.

[0004] To this end, the present application aims to provide a Paenibacillus NGMCC 1.200843 with phosphorus and starch solubilizing effect and application thereof.

[0005] The application also provides an application of the Paenibacillus NGMCC 1.200843 and metabolites thereof in degrading organophosphorus and an application of a biological bacterial agent prepared by the Paenibacillus NGMCC 1.200843 in agricultural production.

[0006] The application also provides an application of the Paenibacillus NGMCC 1.200843 and metabolites thereof in hydrolyzing starch and an application of a secreted hydrolytic enzyme or a biological bacterial agent prepared by the Paenibacillus NGMCC 1.200843 in chemical industry, medicine, feed or food industry and the like.

[0007] 3. The Paenibacillus according to claim 1

[0008] The difficulty and significance of solving the technical problem of the application are as follows:

[0009] The Paenibacillus NGMCC 1.200843 is isolated and screened from rhinoceros feces. The Paenibacillus NGMCC 1.200843 can produce phosphatase with the function of degrading organophosphorus, and plays an important role in promoting plant root absorption and maintaining soil fertility in the process of agricultural production. In addition, the Paenibacillus NGMCC 1.200843 can produce hydrolytic enzyme with the function of degrading starch, and is important for the conversion of starch into oligosaccharide, and has potential application in many industrial processes such as food, fermentation and pharmaceutical industry. Research on the Paenibacillus NGMCC 1.200843 is conducive to obtaining new Paenibacillus resources with potential functions from the special environment of animal intestinal tract.

[0010] Compared with the prior art, the application has the following beneficial effects:

[0011] 1. The Paenibacillus NGMCC 1.200843 provided in the application is gram-stained negative, rod-shaped and facultative aerobic bacteria. The 16S rRNA sequence of the Paenibacillus NGMCC 1.200843 is determined by PCR technology, and is compared with the nucleotide sequence in Genbank by Blast comparison. The results show that the 16S rRNA sequence of the Paenibacillus NGMCC 1.200843 has the highest similarity with the 16S rRNA sequence of Paenibacillus lautus NBRC 15380 T , and the identity values are 98.91%. The whole genome sequencing analysis of the Paenibacillus NGMCC 1.200843, the DNA-DNA molecular hybridization experiment and the average nucleotide identity (ANI) result show that the hybridization ratio of the Paenibacillus NGMCC 1.200843 and the Paenibacillus lautus NBRC 15380 T is 68.50%, which is lower than the threshold value of 70%; the average nucleotide identity (ANI) result shows that the Paenibacillus NGMCC 1.200843 and the Paenibacillus lautus NBRC 15380 TThe ANI value of NGMCC 1.200843 is 95.76%, which is lower than the threshold value of 97%. The phenotype characteristics and phylogenetic studies show that NGMCC 1.200843 is identified as a new species of Paenibacillus genus, the genome size is 7,228,608 bp, contains 6,681 genes, and the GC content is 49.69 mol%.

[0012] 2、The Paenibacillus NGMCC 1.200843 of the present application has phosphatase for degrading organic phosphorus and produces hydrolytic enzyme for degrading starch, and both show clear transparent circles in plate tests, the phosphorus solubilizing activity as microbial fertilizer promotes plant root absorption and maintains soil fertility in extensive agricultural production, has broad development space, secretes starch-degrading hydrolytic enzyme, can convert starch into oligosaccharides, and has good development and application prospect in many industrial processes such as food, fermentation and pharmaceutical industry. BRIEF DESCRIPTION OF DRAWINGS

[0013] Figure 1 is the mGAM upper plate colony shape diagram of NGMCC 1.200843 in the embodiment of the present application;

[0014] Figure 2 is the scanning electron microscope diagram of NGMCC 1.200843 in the embodiment of the present application;

[0015] Figure 3 is the phylogenetic relationship diagram of NGMCC 1.200843 and the main group mode strains under Paenibacillaceae based on 16S rRNA gene sequence in the embodiment of the present application;

[0016] Figure 4 is the genomic phylogenetic relationship diagram of NGMCC 1.200843 and the main group mode strains under Paenibacillaceae based on core genome in the embodiment of the present application;

[0017] Figure 5 is the genomic function gene analysis diagram of NGMCC 1.200843 based on KEGG, COG, Cazy and Go databases in the embodiment of the present application;

[0018] Figure 6 is the polar lipid analysis diagram of NGMCC 1.200843 in the embodiment of the present application;

[0019] Figure 7 is the result diagram of NGMCC 1.200843 in the phosphorus-dissolving test containing lecithin as the only phosphorus source in the embodiment of the present application.

[0020] Figure 8 is the result diagram of NGMCC 1.200843 in the starch hydrolysis test containing 1% soluble starch medium in the embodiment of the present application. DETAILED DESCRIPTION

[0021] The technical solutions of the present application will be described below in more detail with reference to the accompanying drawings and preferred embodiments of the present application, so that the technical solutions of the present application can be more clearly understood and be convenient for understanding. The present application can be embodied in many different forms, and the protection scope of the present application is not limited to the embodiments described herein.

[0022] In order to make the person skilled in the art better understand the technical solutions of the present application, the technical solutions of the present application will be described in further detail below with reference to the embodiments of the present application and the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor should belong to the protection scope of the present application.

[0023] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict. The present application will be described in detail below with reference to the embodiments.

[0024] Embodiment:

[0025] The present application provides a kind of bacillospora, belongs to bacillospora, bacillospora is identified as: NGMCC 1.200843, is registered and preserved in China General Microbiological Culture Collection Center, and the patent preservation number is CGMCC No: 27644, and the preservation date is 2023-06-15. The address of preservation is Beijing City Chaoyang District Beichen West Road No. 1 Yard No. 3, Beijing City Chaoyang District Beichen West Road No. 1 Yard No. 3. The bacillospora NGMCC 1.200843 of the present application is isolated and screened from rhinoceros feces.

[0026] 1 bacillospora new species morphological observation

[0027] 1.1 strain source

[0028] Fresh feces of male rhinoceros. Fresh feces of male rhinoceros is taken, and the sample is collected and put into anaerobic tank, and then immediately transferred into anaerobic glove box. About 1g of feces sample is dissolved in 10ml of sterile anaerobic phosphate buffer (PBS; pH 7) containing 1% cysteine in the glove box, filtered through a cell sieve (70μm and 40μm), and a large amount of feces residue is removed. 1ml is taken from the prepared stock solution, and further treated according to three different methods: (I) treated with 70% ethanol, (II) incubated at 65℃ for 30min, (III) incubated at 80℃ for 10min. (IV) is the stock solution. The four different treatment solutions are diluted 10 times with sterile anaerobic PBS to obtain 10 -1 ~10 -7Diluted fecal solution was spread on yeast extract casein hydrolyzed fatty acid medium (YCFA) and modified Gifu anaerobic medium (MGAM) agar plates supplemented with sterile sheep blood (5%) and rumen fluid (10%), respectively. Strain NGMCC 1.200843 was isolated from 10 -6 fold dilutions on MGAM agar plates, and subjected to phenotypic, physiological and phylogenetic analyses. The isolates were stored as glycerol suspensions (20%, w / v) at -80°C.

[0029] Modified Gifu anaerobic medium (MGAM) agar plate preparation: tryptone 15.0 g, pancreatic casein peptone 10.0 g, soybean peptone 3.0 g, yeast extract powder 5.0 g, beef extract powder 2.0 g, digested serum powder 13.5 g, beef liver extract powder 1.2 g, glucose 3.0 g, potassium dihydrogen phosphate 2.5 g, sodium chloride 3.0 g, soluble starch 0.3 g, L-cysteine 0.3 g, sodium thioethanolate 0.15 g, agar powder 17.0 g, and water to make a total volume of 1000 ml. Autoclaved at 121°C for 15 min, cooled to room temperature, and then added with 1 ml of sterile 0.1% vitamin Kl solution, 2.5 g of hematin chloride, 50 ml of sterile defibrinated sheep blood, and 100 ml of clarified bovine rumen fluid, and poured into plates to prepare MGAM agar plates.

[0030] Yeast extract casein hydrolyzed fatty acid medium (YCFA) agar plate preparation: cysteine 1.0 g, tryptone 10.0 g, yeast extract 2.5 g, sodium bicarbonate 4.0 g, potassium phosphate dibasic 0.45 g, potassium phosphate monobasic 0.45 g, sodium chloride 0.9 g, hematin 0.01 g, magnesium sulfate heptahydrate 0.09 g, calcium chloride 0.09 g, and agar powder 15.0 g, and water to make a total volume of 1000 ml. Autoclaved at 121°C for 15 min, cooled to room temperature, and then added with filter-sterilized reagent two (1.0 mg of resazurin, 10.0 μg of biotin, 10.0 μg of cobalamin, 30.0 μg of p-aminobenzoic acid, 50.0 μg of folic acid, and 150.0 μg of pyridoxal) and reagent three (0.5 μg of thiamine, 0.5 μg of riboflavin, and 25 mM of glucose), and then slowly added with 50 ml of sterile defibrinated sheep blood and 100 ml of clarified bovine rumen fluid, and poured into plates to prepare YCFA agar plates.

[0031] 1.2 Morphological observation

[0032] The isolated and purified novel Paenibacillus-like species was transferred to MGAM agar plates and incubated at 37°C for 24 h in a biochemical incubator. NGMCC 1.200843 formed 1.0-2.0 mm in diameter, milky white, low convex, smooth, and round colonies on MGAM agar plates after incubation at 37°C for 24 h (Fig. 1). Figure 1). The electron microscope observation showed that the bacteria had typical characteristics of the genus Alicyclobacillus: single cell showed a rod shape with two pointed ends, the diameter was about 0.40-0.50 μm, the length was about 3.50-6.50 μm, no spores and no flagella Figure 2

[0033] 1.3 NGMCC 1.200843 sodium chloride tolerance experiment (Table 1)

[0034] Table 1 NGMCC 1.200843 sodium chloride tolerance results

[0035]

[0036] NGMCC 1.200843 grew in the medium containing 0%-2% sodium chloride, and did not grow in the medium containing more than 3% sodium chloride.

[0037] 1.4 NGMCC 1.200843 temperature tolerance experiment (Table 2)

[0038] Table 2 NGMCC 1.200843 temperature tolerance results

[0039]

[0040] NGMCC 1.200843 grew at 20°C-37°C, and did not grow at 4°C, 10°C, 45°C and 50°C.

[0041] 1.5 NGMCC 1.200843 acid and alkali tolerance experiment (Table 3)

[0042] Table 3 NGMCC 1.200843 acid and alkali tolerance results

[0043]

[0044] NGMCC 1.200843 grew at pH 6-8, and did not grow at pH 3-5 and pH 9-11.

[0045] The morphological identification can preliminarily determine that NGMCC 1.200843 belongs to the genus Alicyclobacillus. However, only the colony morphology and microscopic observation cannot determine which species of the genus Alicyclobacillus they belong to. In order to further determine the classification status, biochemical and molecular biology analysis is necessary.

[0046] 2 Biochemical experiment identification

[0047] 2.1 API 50CH biochemical reaction identification

[0048] ​The pure bacteria were inoculated into the modified Gifu anaerobic medium (MGAM) sheep blood agar plate, and incubated in a biochemical incubator at 37°C for 24 h. Then, enough colonies were picked up with a cotton swab or an inoculation loop and added into a suspension to prepare a high-concentration bacterial suspension (S). The high-concentration bacterial suspension (S) was added into 0.85% NaCl (5 ml) to prepare a bacterial suspension with a turbidity of 2.0 McF MacConkey turbidity, and the number of drops n was recorded. A 50 CHB medium ampoule was opened, 2n drops of the above bacterial solution were inoculated, and then the bacterial suspension was divided into each reaction well at 65 μl per well, mixed gently, covered with a reaction cover, and incubated in a biochemical incubator at 37°C for 24 h and 48 h for result interpretation.

[0049] 2.2 API ZYM biochemical reaction identification

[0050] The pure bacteria were inoculated into the modified Gifu anaerobic medium (MGAM) sheep blood agar plate, and incubated in a biochemical incubator at 37°C for 24 h. Then, enough colonies were picked up with a cotton swab or an inoculation loop and added into a suspension to prepare a high-concentration bacterial suspension (S). The high-concentration bacterial suspension (S) was added into 0.85% NaCl (5 ml) to prepare a bacterial suspension with a turbidity of 2.0 McF MacConkey turbidity, and the number of drops n was recorded. A 50 CHB medium ampoule was opened, 2n drops of the above bacterial solution were inoculated, and then the bacterial suspension was divided into each reaction well at 65 μl per well, mixed gently, covered with a reaction cover, and incubated in a biochemical incubator at 37°C for 24 h and 48 h for result interpretation.

[0051] 2.3 API 20E biochemical reaction identification

[0052] The pure bacteria were inoculated into the modified Gifu anaerobic medium (MGAM) sheep blood agar plate, and incubated in a biochemical incubator at 37°C for 24 h. Then, enough colonies were picked up with a cotton swab or an inoculation loop and added into a suspension to prepare a high-concentration bacterial suspension (S). The high-concentration bacterial suspension (S) was added into 0.85% NaCl (5 ml) to prepare a bacterial suspension with a turbidity of 2.0 McF MacConkey turbidity, and the number of drops n was recorded. A 50 CHB medium ampoule was opened, 2n drops of the above bacterial solution were inoculated, and then the bacterial suspension was divided into each reaction well at 65 μl per well, mixed gently, covered with a reaction cover, and incubated in a biochemical incubator at 37°C for 24 h and 48 h for result interpretation.

[0053] Table 4 Biochemical reaction results of NGMCC 1.200843

[0054]

[0055]

[0056]

[0057]

[0058] 3. 16S rRNA phylogenetic analysis of NGMCC 1.200843

[0059] Single colony of suspected new species (preliminary judged by colony size, shape, color) was picked up with sterile Tip head or sterilized toothpick as PCR reaction template, and primer 27F-1492R (27F: 5'-AGAGTTTGATCMTGGCTCAG-3', 1492R: 5'-GGYTACCTTGTTACGACTT-3') was used for colony PCR amplification. Amplification product was sent to Guangzhou Tianyi Huiyuan Biotechnology Co., Ltd. for sequencing. Sequencing results were put into GenBank and BLAST compared with known nucleic acid sequences in GenBank. It was found that the identity value of nucleic acid sequence of NGMCC 1.200843 and NBRC 15380 T was 98.91%. According to the principle of Identities≥97% to determine the same species, it was preliminarily determined that NGMCC 1.200843 belonged to a suspected new species of Paenibacillus.

[0060] The obtained sequences were spliced, forward and reverse detected, and vector and primer sequences were removed by using SeqMan software and DNA star software. Then, MAGA 7.0 software was used to construct a phylogenetic tree by using maximum likelihood method, and finally the generated phylogenetic tree was tested by using Bootstrap. Bacillus subtilis subsp. virginiana HUB-1-047 T was used as an outgroup. The results, as shown in Figure 4 , showed that NGMCC 1.200843 was in an independent evolutionary branch on the clustering tree, and Paenibacillus lautus NBRC 15380 T , Paenibacillus glucanolyticus DSM 5162 T and Paenibacillus qingshengii S1-9 T were clustered into a cluster, with a Bootstrap support rate of 50% Figure 3 , showing a close genetic relationship. The identity value of Blast comparison of NGMCC 1.200843 and other known species of Paenibacillus was lower than 98%, and the genetic relationship was far. The research showed that the classification level of 16S rRNA supported that NGMCC 1.200843 belonged to a new species of Paenibacillus.

[0061] 4. Genomic analysis

[0062] The present application obtains pure culture of NGMCC 1.200843, carries out whole genome sequencing, carries out prediction analysis on its genome basic characteristics, and carries out genome comparison analysis on NGMCC 1.200843 and other known species of Paenibacillus.

[0063] 4.1 Whole genome sequencing and assembly

[0064] The whole genome sequencing of NGMCC 1.200843 is completed by Beijing Nuoweizhuyuan Technology Co., Ltd. The single molecule real-time sequencing technology of the third generation sequencing platform Pacific Biosciences is used to determine the whole genome of NGMCC 1.200843, and the SMRT Analysis2.3.0 software package is used to filter the sequencing data for quality control, and the complete circular genome sequence is assembled.

[0065] 4.2 Genome basic characteristics analysis

[0066] The GeneMarkS (http: / / topaz.gatech.edu / ) software is used for CDS prediction of coding genes, and the tRNAscan-SE and rRNAmmer software is used for tRNAs and rRNA prediction in the genome. The IslandPath-DIOMB software is used for gene island prediction. In this study, the whole genome sequencing of NGMCC 1.200843 isolated from fresh feces of rhinoceros is carried out by the third generation sequencing platform Pacific Biosciences (PacBio). The whole genome of NGMCC 1.200843 is 7,228,608 bp, the G+C content is 49.69 mol%, and 6,681 genes are encoded Figure 5 ) in the genome.

[0067] 5 Genome DNA-DNA hybridization

[0068] The online software of DNA-DNA hybridization (genome and genome distance calculator, GGDC) is used for online genome hybridization of the whole genome sequence of NGMCC 1.200843 and the representative sequences of different species of Paenibacillus that can be retrieved in the current GenBank library. The DNA-DNA hybridization similarity greater than 70% is divided into the same species (see Table 5).

[0069] As can be seen from Table 5, NGMCC 1.200843 and Paenibacillus lautus NBRC 15380 TThe hybridization ratio was 68.50%, and the hybridization ratios of other known related Paenibacillus were between 21.10% and 27.50%, indicating that NGMCC1.200843 does not belong to any known species and is a new species of Paenibacillus.

[0070] Table 5 Consistency of genomic sequences and sequence hybridization results of Paenibacillus NGMCC 1.200843 and different known species of Paenibacillus.

[0071]

[0072] 6 Phosphorus solubilizing activity

[0073] The phosphorus-solubilizing bacterial culture medium was used, and lecithin 0.2 (g / l) was added as the only phosphorus source in the culture medium. The strain NGMCC1.200843 T was inoculated at the center of the plate. Escherichia coli L-7 was used as a negative control. All plates were incubated at 37°C for 7 days. The diameters of the transparent zone (halo) around the bacterial growth and the diameter of the colony were measured with a vernier caliper, and the average value was calculated by measuring three times in parallel. The transparent zone around the growing colony indicates phosphate dissolution. The formula "phosphorus dissolution index = (halo diameter + colony diameter) / colony diameter" was used as the phosphorus dissolution index. The results are shown in Figure 7 , and the specific analysis results are shown in Table 6.

[0074] The preparation method of the phosphorus-solubilizing bacterial culture medium (organic phosphorus culture medium) agar plate is as follows: glucose 10.0 g, ammonium sulfate 0.5 g, yeast extract powder 0.5 g, sodium chloride 0.3 g, potassium chloride 0.3 g, magnesium sulfate 0.3 g, ferrous sulfate 0.03 g, manganese sulfate 0.03 g, lecithin 0.2 g, calcium carbonate 1.0 g, agar powder 17.0 g, and water was added to a total amount of 1000 ml, and sterilized by high-pressure steam at 121°C for 15 minutes.

[0075] Table 6 Qualitative analysis of the phosphate release capacity of the strain NGMCC 1.200843

[0076]

[0077] As can be seen from the antibacterial activity analysis results in Table 6, the NGMCC 1.200843 strain of the present application showed a clear halo zone around the colony in the decomposition of organic phosphorus ( Figure 7 , B), and no transparent zone was observed around the Escherichia coli colony in the negative control ( Figure 7, A). The colony diameter of strain NGMCC 1.200843 was 5.29±0.34 mm, the transparent circle diameter was 29.04±1.9 mm, and the PSI was 4.46±0.30, which indicated that the strain had a high phosphorus solubilization rate. The results showed that the strain NGMCC 1.200843 had a certain ability to decompose organic phosphorus, indicating that it had the characteristics of phosphorus-solubilizing bacteria. Therefore, the strain of the present application has potential application prospects in the fields of agriculture, biological medicine or chemical industry.

[0078] 7 Amylolytic activity

[0079] The amylase activity of strain NGMCC 1.200843 was identified using soluble starch medium containing 1% soluble starch. Strain NGMCC 1.200843 was inoculated into soluble starch medium, and E. coli L-7 was inoculated as a negative control in the same way. All plates were incubated at 37°C for 2 days, and the production of transparent circles was observed. In order to detect the hydrolysis of starch, iodine solution was added to the culture of strain NGMCC 1.200843 and submerged, and the starch hydrolysis was observed by color change. The negative result was the formation of a dark blue, purple or black complex. The positive result was the color of the brownish red iodine solution, and the results are shown in Figure 7 .

[0080] Preparation method of soluble starch medium agar plate: soluble starch 10.0 g, dihydrogen phosphate 1.0 g, magnesium sulfate 1.0 g, sodium chloride 1.0 g, ammonium sulfate 2.0 g, calcium carbonate 2.0 g, ferrous sulfate 0.001 g, manganese chloride 0.001 g, zinc sulfate 0.001 g, agar powder 17.0 g, add water to a total of 1000 ml, 121°C, 15 minutes high pressure steam sterilization.

[0081] Figure 8 The results of the study on the hydrolysis of starch by strain NGMCC 1.200843 grown in the basic medium containing 1% soluble starch are shown. A clear transparent band was observed around the colony of strain NGMCC 1.200843. When exposed to iodine solution, the colony changed from cream to reddish brown. This color change indicates that during the hydrolysis of amylopectin, which constitutes soluble starch, the first formed dextrin is erythrodextrin, which changes color from blue to purple to reddish brown after the addition of iodine. In the control group, no transparent band was formed around the E. coli colony, and the bacterial cells turned black after the addition of iodine solution, indicating that the starch was not hydrolyzed. The above results show that strain NGMCC 1.200843 can hydrolyze soluble starch and secrete related extracellular hydrolytic enzymes. These enzymes are usually alpha-amylase and oligo-1,6-glucosidase. These enzymes hydrolyze starch by breaking the glycosidic bonds between glucose subunits and allow the starch hydrolysate to enter the cell.

[0082] The above specific embodiments are only an explanation of the present application, which is not a limitation of the present application, and the person skilled in the art can make a modification of the present embodiment without a creative contribution according to the need after reading the present specification, but as long as it is within the scope of the claims of the present application, it is protected by the patent law.

Claims

1. A new species of Paenibacillus with phosphate-solubilizing and starch-solubilizing properties has been identified, with isolation number NGMCC 1.200843 = CGMCC 1.64763T. This Paenibacillus strain is deposited at the China General Microbiological Culture Collection Center (CGMCC No. 27644). The 16S ribosomal DNA sequence of NGMCC 1.200843 is shown in SEQ ID NO.1, and its GenBank accession number is JBHDJG000000000.

2. The application of Bacillus subtilis NGMCC 1.200843 as described in claim 1 in the degradation of organophosphates.

3. The application of Bacillus subtilis NGMCC 1.200843 as described in claim 1 in the hydrolysis of starch.

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