Paenibacillus NGMCC 1.200843 with phosphate-solubilizing and starch-solubilizing effects from rhinoceros excrement and application of paenibacillus NGMCC 1.200843

Abstract

The invention discloses a strain of paenibacillus NGMCC 1.200843 from rhinoceros feces, and belongs to a new discovery in the technical field of microorganisms. The strain is identified as a new species of paenibacillus, the number of the strain is NGMCC 1.200843 = CGMCC 1.64763T, and the strain is registered and preserved in the China General Microbiological Culture Collection Center. The strain can form a larger transparent circle on a specific culture medium, which indicates that the strain has stronger phosphate solubilizing and starch solubilizing capabilities. Under the current conditions that the soil fertility is reduced and the fertilizer is abused, the strain provides a high-quality strain resource for developing a microbial fertilizer with an organophosphorus degradation function, and is beneficial to promoting the development of the microbial fertilizer. The produced hydrolase has wide application potential in the industrial fields of food, fermentation, pharmacy and the like. Especially, amylase can hydrolyze starch to generate various products, has important significance on the biotechnology industry, and is applied to multiple industries such as food, fermentation, spinning and papermaking. In a word, the invention provides a new resource for development of microbial fertilizers and industrial enzyme preparations.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a spore-forming bacterium NGMCC 1.200843 with the functions of dissolving phosphate and starch, and an application thereof. Background Art

[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), inorganic phosphorus bound to aluminum / iron (in acidic soils), and inorganic phosphorus bound to calcium (in alkaline soils). However, only 0.1% of the phosphorus in cultivated land worldwide can be absorbed by plant roots. In response to this extreme deficiency, the production and application of large amounts of phosphorus fertilizers have increased. In fact, plants have low phosphorus use efficiency under phosphorus stress or even when phosphorus is sufficient, and long-term application of phosphorus fertilizers has adverse effects on the environment, and phosphorus fertilizers are expensive. Phosphorus solubilizing bacteria (PSB) can convert insoluble phosphorus in the soil into available phosphorus that can be directly absorbed by plants, thereby improving plant phosphorus absorption and utilization. These phosphorus solubilizing microorganisms have different potentials for phosphorus solubilization, which mainly depends on the solubilization mechanism they adopt, molecular genetics, and their ability to release phosphorus in the soil. Screening highly efficient phosphorus solubilizing microorganisms and developing them into phosphorus solubilizing bacterial agents for inoculation, or directly applying them to the soil, can activate insoluble phosphates 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 genus Paenibacillus are endospore-forming Gram-negative bacteria, facultative anaerobes or obligate aerobes, with a DNA G+C content of 39-59 mol% and motile via peritrichous flagella. Bacteria belonging to the genus Paenibacillus have been isolated from various environments, and many of these species are associated with humans, animals, plants, and the environment. Most of them are found in soil, often associated with plant roots, and these rhizosphere bacteria promote plant growth and can be used in agriculture. Paenibacillus-derived antimicrobials also have applications in medicine, including polymyxins and fusaricides. 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. Nevertheless, most of these spore-forming bacteria remain underexplored with potential for biotechnological applications.

[0004] To this end, the present invention aims to provide a spore-forming bacterium NGMCC 1.200843 having the functions of solubilizing phosphate and starch and its application.

[0005] On the other hand, the present invention also provides the use of the spore-forming bacteria NGMCC 1.200843 and its metabolites in degrading organic phosphorus and the use of the prepared biological microbial agent in agricultural production.

[0006] On the other hand, the present invention also provides the use of the spore-forming bacteria NGMCC 1.200843 and its metabolites in hydrolyzing starch, and the use of the secreted hydrolytic enzymes or the prepared biological agents in the chemical, medical, feed or food industries.

[0007] 3. The spore-forming bacteria according to claim 1

[0008] The difficulty and significance of the technical problem solved by the present invention are:

[0009] The spore-forming bacteria NGMCC 1.200843 of the present invention is isolated and screened from rhino feces. The bacteria can produce phosphatase that can degrade organic phosphorus, and play an important role in promoting plant root absorption and maintaining soil fertility in the agricultural production process. In addition, the bacteria can produce hydrolytic enzymes that can degrade starch, which is crucial in converting starch into oligosaccharides, and has potential applications in many industrial processes such as food, fermentation and pharmaceutical industries. Studying the bacteria is conducive to obtaining new spore-forming bacteria resources with potential functions from the special environment of animal intestines.

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

[0011] 1. The spore-forming bacteria NGMCC 1.200843 provided by the present invention is Gram-negative, rod-shaped, and facultatively aerobic. The 16S rRNA sequence of the bacteria was determined by PCR technology, and the nucleotide sequence was compared with the nucleotide sequence in Genbank by Blast. The results showed that NGMCC 1.200843 was similar to Paenibacillus lautus NBRC 15380. T The 16SrRNA sequence of NGMCC 1.200843 was the highest, with an identity value of 98.91%. The whole genome sequencing analysis of NGMCC 1.200843 and DNA-DNA molecular hybridization experiments showed that NGMCC 1.200843 and Paenibacillus lautus NBRC 15380 T The hybridization ratio was 68.50%, which was lower than the threshold of 70%; the average nucleotide identity (ANI) results showed that NGMCC 1.200843 and Paenibacillus lautus NBRC 15380 TThe ANI value of NGMCC 1.200843 was 95.76%, which was lower than the threshold of 97%. Phenotypic characteristics and phylogenetic studies showed that NGMCC 1.200843 was identified as a new species of Paenibacillus, with a genome size of 7,228,608 bp, 6,681 genes, and a GC content of 49.69 mol%.

[0012] 2. The spore-forming bacteria NGMCC 1.200843 of the present invention has phosphatase that can degrade organic phosphorus and produces hydrolases that can degrade starch, and both show clear transparent circles in the plate test. Its phosphorus-solubilizing activity has broad development space as a microbial fertilizer in promoting plant root absorption and maintaining soil fertility in agricultural production. It secretes hydrolases that can degrade starch and convert starch into oligosaccharides, and has good development and application prospects in many industrial processes such as food, fermentation and pharmaceutical industries. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 This is a diagram of the shape of the mGAM colonies on the plate of NGMCC 1.200843 in the embodiment of the present invention;

[0014] Figure 2 is a scanning electron microscope image of NGMCC 1.200843 in an embodiment of the present invention;

[0015] Figure 3 It is a phylogenetic relationship diagram of NGMCC 1.200843 and the main group model strains under the Paenibacillus family constructed based on the 16S rRNA gene sequence in the embodiment of the present invention;

[0016] Figure 4 It is a phylogenetic relationship diagram of the genome of NGMCC 1.200843 and the main group model strains under the Paenibacillus family constructed based on the core genome in the embodiment of the present invention;

[0017] Figure 5 It is a genome functional gene analysis diagram of NGMCC 1.200843 based on KEGG, COG, Cazy and Go databases in an embodiment of the present invention;

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

[0019] Figure 7 This is a graph showing the results of a phosphate solubilization test of NGMCC 1.200843 in an embodiment of the present invention when lecithin is used as the sole phosphorus source.

[0020] Figure 8 This is a graph showing the results of a starch hydrolysis test of NGMCC 1.200843 in a medium containing 1% soluble starch in an embodiment of the present invention. DETAILED DESCRIPTION

[0021] The following describes the preferred embodiments of the present invention with reference to the drawings in the specification, so that the technical content is clearer and easier to understand. The present invention can be embodied in many different forms of embodiments, and the protection scope of the present invention is not limited to the embodiments mentioned in the text.

[0022] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme of the present invention will be further described in detail below in combination with the embodiments of the present invention and the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0023] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present invention can be combined with each other. The present invention will be described in detail below in conjunction with the embodiments.

[0024] Example:

[0025] The embodiment of the present invention provides a Paenibacillus, belonging to the Paenibacillus family and the Paenibacillus genus, identified as: NGMCC 1.200843, registered and deposited in the China General Microbial Culture Collection Center, with a patent deposit number of CGMCC No: 27644, and a deposit date of 2023-06-15. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The Paenibacillus NGMCC 1.200843 of the present invention is isolated and screened from rhinoceros feces.

[0026] Morphological observation of a new species of spore-forming fungi

[0027] 1.1 Strain source

[0028] Fresh feces of male rhinoceros. Take fresh feces of male rhinoceros, put the sample into an anaerobic jar after collection, and immediately transfer it to an anaerobic glove box. In the glove box, take about 1g of fecal sample and dissolve it in 10ml of sterile anaerobic phosphate buffer (PBS; pH 7) containing 1% cysteine, and filter it through a cell sieve (70μm and 40μm) to remove a large amount of fecal residue. Take 1ml from the prepared stock solution and further treat it according to 3 different methods: (Ⅰ) treatment with 70% ethanol, (Ⅱ) incubation at 65℃ for 30min, and (Ⅲ) incubation at 80℃ for 10min. (IV) is the stock solution. Dilute the 4 different treatment solutions 10 times with sterile anaerobic PBS to obtain 10 -1 ~10 -7The fecal solution was diluted in a gradient and spread on yeast extract casein hydrolyzed fatty acid medium (YCFA) and modified anaerobic medium (MGAM) agar plates supplemented with sterile sheep blood (5%) and rumen fluid (10%), respectively. -6 The isolates were isolated on MGAM agar plates after 1 / 2 dilution and subjected to phenotypic, physiological and phylogenetic analyses. The isolates were stored at -80°C in the form of glycerol suspension (20%, w / v).

[0029] Preparation method of modified Qifu anaerobic medium (MGAM) agar plate: 15.0g of peptone, 10.0g of tryptone, 3.0g of soy peptone, 5.0g of yeast extract powder, 2.0g of beef powder, 13.5g of digested serum powder, 1.2g of beef liver extract powder, 3.0g of glucose, 2.5g of potassium dihydrogen phosphate, 3.0g of sodium chloride, 0.3g of soluble starch, 0.3g of L-cysteine, 0.15g of sodium thioglycolate, 17.0g of agar powder, add water to adjust the total amount to 1000ml. Sterilize with high pressure steam at 121℃ for 15 minutes, cool to room temperature, add 1ml of sterile 0.1% vitamin K1 solution, 2.5g of hemin chloride, slowly add 50ml of sterile defibrinated sheep blood and 100ml of clarified bovine rumen fluid and pour on the plate to prepare MGAM agar plate.

[0030] Yeast extract casein hydrolyzed fatty acid medium (YCFA) agar plate preparation method: cysteine ​​1.0g, tryptone 10.0g, yeast extract 2.5g, sodium bicarbonate 4.0g, dipotassium hydrogen phosphate 0.45g, potassium dihydrogen phosphate 0.45g, sodium chloride 0.9g, heme 0.01g, magnesium sulfate heptahydrate 0.09g, calcium chloride 0.09g, agar powder 15.0g, add water to adjust the total amount to 1000ml. Sterilize with high pressure steam at 121°C for 15 minutes, cool to room temperature, add filter-sterilized reagent II (resazurin 1.0 mg, biotin 10.0 μg, cobalamin 10.0 μg, p-aminobenzoic acid 30.0 μg, folic acid 50.0 μg, pyridoxal 150.0 μg) and reagent III (thiamine 0.5 μg, riboflavin 0.5 μg, glucose 25 mM), then slowly add 50 ml of sterile defibrinated sheep blood and 100 ml of clarified bovine rumen fluid and pour into the plate to make YCFA agar plates.

[0031] 1.2 Morphological observation

[0032] The isolated and purified new species of Bacillus spores were transferred to MGAM agar plates and cultured in a biochemical incubator at 37°C for 24 hours. NGMCC 1.200843 formed milky white, low-convex, smooth, round colonies with a diameter of 1.0 to 2.0 mm after being cultured at 37°C for 24 hours on MGAM agar plates ( Figure 1). The bacteria showed typical characteristics of spore-forming fungi under electron microscope: a single bacterium was in the shape of a rod with two pointed ends, with a diameter of about 0.40-0.50 μm and a length of about 3.50-6.50 μm, without spores or flagella ( Figure 2 ).

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

[0034] Table 1 Sodium chloride tolerance results of NGMCC 1.200843

[0035]

[0036] NGMCC 1.200843 grows when cultured in a medium containing 0% to 2% sodium chloride, but does not grow when cultured in a medium containing more than 3% sodium chloride.

[0037] 1.4 NGMCC 1.200843 Temperature Tolerance Test (Table 2)

[0038] Table 2 Temperature tolerance results of NGMCC 1.200843

[0039]

[0040] NGMCC 1.200843 grew when cultured at 20℃~37℃, but did not grow at 4℃, 10℃, 45℃, and 50℃.

[0041] 1.5 NGMCC 1.200843 acid-base tolerance test (Table 3)

[0042] Table 3 Acid and alkali tolerance results of NGMCC 1.200843

[0043]

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

[0045] From the morphological aspect, it can be preliminarily determined that NGMCC 1.200843 belongs to the family Paenibacillus and the genus Paenibacillus. However, it is not possible to determine which species of the family Paenibacillus and the genus Paenibacillus it belongs to based on the colony morphology and microscopic observation alone. In order to further determine its taxonomic status, it is necessary to conduct biochemical and molecular biological analysis.

[0046] 2 Biochemical experimental identification

[0047] 2.1 API 50CH biochemical reaction identification

[0048] Inoculate the purified bacteria onto a modified Qifu anaerobic medium (MGAM) sheep blood agar plate and place it in a biochemical incubator at 37°C for 24 hours. Then pick enough colonies with a cotton swab or inoculation loop and add them to the suspension to make a high-concentration bacterial suspension (S). Add the high-concentration bacterial suspension (S) to 0.85% NaCl (5ml) to prepare a turbidity of 2.0McF McF turbidity bacterial suspension, and record the number of drops n. Open the 50CHB medium ampoule, add 2n drops of the above bacterial solution for inoculation, and then divide the bacterial suspension into each reaction well, 65μl per well, shake gently to mix, cover the reaction cover, and place it in a biochemical incubator at 37°C for 24h and 48h to interpret the results.

[0049] 2.2 API ZYM biochemical reaction identification

[0050] The purified bacteria were inoculated into the modified Qifu anaerobic medium (MGAM) sheep blood agar plate and placed in a biochemical incubator at 37°C for 24 hours. Then, a cotton swab or inoculation loop was used to pick up enough colonies into a 2ml API suspension medium ampoule to prepare a McFarland turbidity bacterial suspension with a turbidity of 5.0-6.0 McF. The bacterial suspension was then dispensed into each reaction well, 65 μl per well, gently shaken and mixed, covered with a reaction cover, placed in a biochemical incubator at 37°C for 4 hours, and then one drop of ZYM-A and ZYM-B was added to each cup and the results were observed, as shown in Table 4.

[0051] 2.3 API 20E biochemical reaction identification

[0052] The purified bacteria were inoculated into the modified Qifu anaerobic medium (MGAM) sheep blood agar plate and placed in a biochemical incubator at 37°C for 24 hours. Then, a cotton swab or an inoculation loop was used to pick up enough colonies into a 5ml 0.85% ampoule to prepare a turbidity of 0.5McF McF turbidity bacterial suspension, which was then dispensed into each reaction well, 65μl per well, and gently shaken to mix. The wells for ADH, LDC, ODC, H2S, and URE reactions were covered with mineral oil, covered with a reaction cover, and placed in a biochemical incubator at 37°C for 24 hours, and the results were observed.

[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] Use a sterile tip or a sterile toothpick to pick up a single colony of a suspected new species (preliminary judgment based on colony size, morphology, and color) as a PCR reaction template, and use primers 27F-1492R (27F: 5′-AGAGTTTGATCMTGGCTCAG-3′, 1492R: 5′-GGYTACCTTGTTACGACTT-3′) for colony PCR amplification. The amplified product was sent to Guangzhou Tianyi Huiyuan Biotechnology Co., Ltd. for sequencing. The sequencing results were placed in GenBank and compared with known nucleic acid sequences in GenBank by BLAST. The results showed that NGMCC 1.200843 and NBRC 15380 T The identity value of the nucleic acid sequence was 98.91%. Based on the principle that identities ≥ 97% are considered to be the same species, NGMCC 1.200843 was preliminarily determined to be a suspected new species of the genus Bacillus.

[0060] SeqMan and DNA star software were used to splice the obtained sequences, perform forward and reverse detection, and remove the vector and primer sequences. Then, MAGA 7.0 software was used to construct a phylogenetic tree using the maximum likelihood method, and finally the generated phylogenetic tree was tested using Bootstrap. T As an outgroup. Figure 4 As shown in the figure, it can be seen that NGMCC 1.200843 is in an independent evolutionary branch on the cluster tree, which is different from Paenibacillus lautus NBRC 15380. T , Paenibacillus glucanolyticus DSM 5162 T andPaenibacillus qingshengiiS1-9 T Clustered into a cluster, its Bootstrap support rate is 50% ( Figure 3 ), showing a close relationship. The identity values ​​of NGMCC 1.200843 and other known species of Paenibacillus family in Blast comparison were all lower than 98%, indicating a distant relationship. The study showed that NGMCC 1.200843 was supported as a new species in Paenibacillus at the 16S rRNA classification level.

[0061] 4 Genomics analysis

[0062] After obtaining the pure culture of NGMCC 1.200843, the present invention carried out whole genome sequencing, performed predictive analysis on the basic characteristics of its genome, and conducted genome comparative analysis between NGMCC 1.200843 and other known species of the genus Bacillus.

[0063] 4.1 Whole genome sequencing and assembly

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

[0065] 4.2 Analysis of basic genome characteristics

[0066] GeneMarkS (http: / / topaz.gatech.edu / ) software was used to predict the CDS of coding genes, and tRNAscan-SE and rRNAmmer software were used to predict tRNAs and rRNAs in the genome. IslandPath-DIOMB software was used to predict gene islands. In this study, the whole genome of NGMCC 1.200843 isolated from fresh rhino feces was sequenced using the third-generation sequencing platform Pacific Biosciences (PacBio). The full length of the NGMCC 1.200843 genome is 7,228,608bp, with a G+C content of 49.69mol%, encoding 6,681 genes ( Figure 5 ).

[0067] 5. Genomic DNA-DNA hybridization

[0068] The whole genome sequence of NGMCC 1.200843 was hybridized online with the representative sequences of different species of Paenibacillus that can be retrieved in the GenBank library using DNA-DNA hybridization online software (genome and genome distance calculator, GGDC). The sequences with DNA-DNA hybridization similarity greater than 70% were classified as 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%, while the hybridization ratios of other known closely related Paenibacillus species were between 21.10% and 27.50%, indicating that NGMCC1.200843 did not belong to any known species and was a new species of the genus Paenibacillus.

[0070] Table 5. Genome sequence consistency and sequence hybridization results of Bacillus sp. NGMCC 1.200843 and different known Bacillus sp. species.

[0071]

[0072] 6 Phosphate solubilization activity

[0073] Phosphate-solubilizing bacterial culture medium was used, and 0.2 g / l of lecithin was added to the culture medium as the only phosphorus source. T Inoculate in 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. Use a vernier caliper to measure the diameter of the transparent zone (halo) around the bacterial growth and the diameter of the colony. Measure three times in parallel and calculate the average value. The transparent zone around the growing colony indicates phosphate dissolution. The calculation formula "Phosphorus solubility index = (halo diameter + colony diameter) / colony diameter" is used as the phosphorus solubility index. See the results. Figure 7 The specific analysis results are shown in Table 6.

[0074] Preparation method of agar plate for phosphate-solubilizing bacterial culture medium (organic phosphorus culture medium): 10.0g glucose, 0.5g ammonium sulfate, 0.5g yeast extract powder, 0.3g sodium chloride, 0.3g potassium chloride, 0.3g magnesium sulfate, 0.03g ferrous sulfate, 0.03g manganese sulfate, 0.2g lecithin, 1.0g calcium carbonate, 17.0g agar powder, add water to adjust the total amount to 1000ml, 121℃, high-pressure steam sterilization for 15 minutes.

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

[0076]

[0077] From the antibacterial activity analysis results in Table 6, it can be seen that the NGMCC 1.200843 strain of the present invention shows a clear halo band around its colony during the decomposition of organic phosphorus ( Figure 7 , B), In the negative control, no clear zone was observed around the E. coli colonies ( Figure 7, A). The colony diameter of strain NGMCC 1.200843 is 5.29±0.34mm, the transparent zone diameter is 29.04±1.9mm, and it has a high phosphate solubility rate, with a PSI of 4.46±0.30. The results show that strain NGMCC 1.200843 has a certain ability to decompose organic phosphorus, indicating that it has the characteristics of phosphate-solubilizing bacteria. Therefore, the strain of the present invention has potential application prospects in agriculture, biomedicine or chemical industry.

[0078] 7. Starch degrading activity

[0079] The amylase activity of strain NGMCC 1.200843 was identified using a soluble starch medium containing 1% soluble starch. The strain NGMCC 1.200843 was inoculated in the soluble starch medium, and Escherichia coli L-7 was inoculated in the same way as a negative control. All plates were cultured at 37°C for 2 days, and the formation of transparent circles was observed. In order to detect hydrolyzed starch, Lugol's iodine solution was used to add iodine solution to the culture of strain NGMCC 1.200843 and submerge it, and starch hydrolysis was observed by color change. A negative result is the formation of a dark blue, purple or black complex. A positive result is the color of the brown-red iodine solution, and the result is shown in Figure 7 .

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

[0081] Figure 8 The results of the study on the hydrolysis of starch by strain NGMCC 1.200843 grown in a basal medium containing 1% soluble starch are shown. A clear transparent band was observed around the colonies of strain NGMCC 1.200843. When exposed to an iodine solution, the colonies changed from milky white to reddish brown. This color change indicates that when the amylopectin that makes up the soluble starch is hydrolyzed, the first dextrin formed 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 Escherichia coli colonies, and the bacterial cells appeared black after the addition of iodine solution, indicating that the starch was not hydrolyzed. The above results indicate that strain NGMCC 1.200843 is able to hydrolyze soluble starch and secrete related extracellular hydrolases. These enzymes are usually α-amylase and oligo-1,6-glucosidase. These enzymes hydrolyze starch by breaking the glycosidic bonds between glucose subunits and allow starch hydrolysis products to enter the cells.

[0082] The above specific embodiments are merely explanations of the present invention and are not limitations of the present invention. After reading this specification, those skilled in the art may make modifications to the embodiments without any creative contribution as needed. However, such modifications are protected by the patent law as long as they are within the scope of the claims of the present invention.

Claims

1. A strain of Paenibacilus with phosphate and starch solubilizing effects, identified as a new species of Paenibacilus, with an isolation number of NGMCC 1.200843=CGMCC 1.64763T. The Paenibacilus is deposited and registered in the China General Microbiological Culture Collection Center, with a patent deposit number of CGMCC No: 27644; the 16s ribosomal DNA sequence of NGMCC 1.200843 is shown in SEQ ID NO.1, and the genome GenBank access number is: JBHDJG000000000.

2. Use of the spore-forming bacteria NGMCC 1.200843 and its metabolites as described in claim 1 in the degradation of organic phosphorus and use of the prepared biological agent in agricultural production.

3. Use of the spore-forming bacteria NGMCC 1.200843 and its metabolites as claimed in claim 1 in hydrolyzing starch, and use of the secreted hydrolytic enzymes or prepared biological agents in chemical, medical, feed or food industries, etc.

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