CN107541465B - In-situ enrichment method and screening method of native strain producing pullulanase - Google Patents
In-situ enrichment method and screening method of native strain producing pullulanase Download PDFInfo
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Abstract
The invention relates to an in-situ enrichment method and a screening method of an indigenous strain producing pullulanase. The in-situ enrichment method of the indigenous strains capable of producing the pullulanase comprises the following steps: and embedding the substrate adhered with the amylopectin into soil, taking out the substrate after enrichment, and collecting the soil on the surface of the substrate to obtain the strain-enriched soil. The screening method further comprises the following steps: and (3) uniformly mixing the strain-enriched soil with sterile water, centrifuging, taking the upper suspension for gradient dilution, taking the gradient dilution liquid to coat a separation flat plate for culture, and identifying after culture. The method is mainly characterized in that amylopectin is used as an initial induction substrate, the native strains for producing the pullulanase are subjected to in-situ enrichment in soil, the number of populations for producing the pullulanase can be increased under the original ecological environment condition, particularly, weak populations with the pullulanase activity can be proliferated to a certain extent, and the detection rate of target strains is further improved.
Description
Technical Field
The invention relates to an in-situ enrichment method and a screening method of an indigenous strain producing pullulanase, belonging to the technical field of bacterial enrichment screening.
Background
Microorganisms are an important part of the biological resources and material circulation in the nature, and it is widely believed that the types of microorganisms recognized by people are only 1% -10% of the actual stock, and people even consider that the types of microorganisms are less than 0.1%, so that the screening of wild microbial resources is still a long-term and arduous work. Starch is a polysaccharide compound which is greatly consumed in life, natural starch is composed of 10% -30% of amylose and 70% -90% of amylopectin, and the utilization rate of starch is mainly determined by the utilization rate of amylopectin. The alpha-1, 6 glycosidic bond contained in the amylopectin is difficult to hydrolyze, which becomes a bottleneck problem in the efficient utilization of the starch, while the pullulanase exclusively hydrolyzes the alpha-1, 6 glycosidic bond in the starch.
The screening method of the pullulanase-producing strain mainly adopts plate culture at present, and although the screening method has exploration and improvement measures in the aspects of increasing material diversity, optimizing components of a culture medium and the like in the screening process, the screening of the pullulanase-producing strain does not obtain substantial breakthrough. In the prior art, methods for screening and identifying by directly taking soil are commonly adopted, such as separation and identification of a pullulanase-producing strain in flue-cured tobacco rhizosphere soil and enzyme-producing activity (Hunan agricultural science, Zhujinfeng, Sunjiao and the like); or taking soil for indoor enrichment, and then screening and identifying. The traditional enrichment method of the native pullulanase-producing strain can also generally separate a few strains, but the problems are obvious, and the defects are concentrated in that: (1) the separation method is too programmed and single, the repetition rate of the separated strains is extremely high, the diversity is poor, and innovative research results are provided; (2) enrichment culture performed for a certain time before isolation culture is obviously stronger in pertinence and slightly better in effect than enrichment-free culture, but in-vitro enrichment also has the natural defect that the enrichment environment and the natural environment of a sample are different in nature. Any microorganism has a habitat on which it depends, which naturally leads to changes in population composition, especially for some vulnerable or rare populations, which are more likely to be lost during isolation by improper handling.
As the detection rate of the pullulanase-producing strains in the prior art is not high, even if individual pullulanase-producing strains can be screened, the diversity of the strains is far from abundant, the repeated screening condition is prominent, the biological activity is generally low, and most industrial development and application potentials are not high. Increasing the diversity of wild strains producing pullulanase and obtaining strains with high activity remain important research targets that people are eagerly to realize.
Disclosure of Invention
The invention aims to provide an in-situ enrichment method of an indigenous strain producing pullulanase, which can improve the screened diversity and the detection rate of the indigenous strain producing pullulanase.
The invention also provides a screening method of the indigenous strains for producing the pullulanase by adopting the in-situ enrichment method.
In order to achieve the purpose, the invention adopts the technical scheme that:
an in-situ enrichment method of an indigenous strain producing pullulanase, comprising the following steps: and embedding the substrate adhered with the amylopectin into soil, taking out the substrate after enrichment, and collecting the soil on the surface of the substrate to obtain the strain-enriched soil.
Preferably, the time for enriching the buried soil is 8-12 d.
In the technical scheme of the invention, the method is mainly characterized in that amylopectin is used as an initial induction substrate to perform in-situ enrichment in soil on the native strains producing the pullulanase, so that the number of the population producing the pullulanase can be increased under the original ecological environment condition, particularly weak populations with the pullulanase activity can be proliferated to a certain extent, and the detection rate of target native strains is further improved. The basic principle of the technical innovation of the invention is as follows: the soil on which the microorganisms live is an extremely complex special environment which cannot realize artificial hundred percent fitting, and the enrichment of specific microorganism populations in the original soil environment conforms to the principle of coevolution of microorganisms and the soil environment; the pullulan contains alpha-1, 6 glycosidic bonds, and native strains producing the pullulanase are easier to enrich by taking the pullulan as a substrate under the original ecological environment condition, so that the diversity of the strains producing the pullulanase is increased.
Preferably, the lining is absorbent cotton. In the in-situ enrichment process, the absorbent cotton is used as a substrate of the amylopectin, so that the opportunity of contacting the amylopectin in unit area with microorganisms can be increased, the absorbent cotton is not easy to rot in a short period, has no toxic or side effect, and is convenient for accurately collecting the enriched soil.
The lining material of the invention can also be selected from lining carriers meeting the conditions in the field (the principle of the selection of the lining material is natural, breathable, not easy to rot, does not change the physicochemical environment of soil in a short period, and can be sterilized at high temperature).
The preparation of the substrate adhered with the amylopectin comprises the following steps:
1) dissolving 15-20g of amylopectin in 50mL of sterile water to obtain an amylopectin solution;
2) 4-6g of sterilized lining material is soaked in the amylopectin solution and fully adsorbed to obtain the lining material adhered with the amylopectin.
The concrete operation of burying soil is as follows: digging a pit with a selected depth in soil in a sampling environment, spraying sterile water into the pit, taking the liner adhered with amylopectin, finishing the liner into a flat cuboid, flatly placing the flat cuboid at the bottom of the pit, and covering absorbent cotton with in-situ soil during digging the pit.
The concrete operation of burying soil is as follows: digging a pit hole with the depth of 5-10cm in the soil of the sampling environment, spraying 150mL of sterile water into the pit hole, taking the lining adhered with the amylopectin, finishing the lining into a flat rectangular shape, flatly placing the flat rectangular shape at the bottom of the pit, and covering the absorbent cotton with in-situ soil during digging the pit hole. The environment and the depth of the cotton wool soaked with the amylopectin and placed in the soil in the in-situ enrichment process can be diversified according to the purpose and the intention of the test, so that the blindness, the contingency and the low efficiency of sampling in the traditional screening test are overcome to a certain extent, and the test becomes more controllable and efficient.
Preferably, the lining material is taken out, and the soil on the surface of the lining material is collected after the excess soil is shaken off. Since the lining is buried in the soil for a relatively long time, the lining is taken out with lumpy soil, and the soil attached with the lumpy soil is not in contact with the absorbent cotton, so that the pullulanase-producing strains are few, and the soil can be taken out from the surface of the absorbent cotton after shaking the soil.
The screening method of the indigenous strains producing the pullulanase by adopting the in-situ enrichment method comprises the following steps: and (3) uniformly mixing the strain-enriched soil with sterile water, centrifuging, taking the upper suspension for gradient dilution, taking the gradient dilution liquid to coat a separation flat plate for culture, and identifying after culture.
Preferably, the strain-enriched soil is evenly mixed with sterile water and then centrifuged, the upper suspension is taken for 10-fold gradient dilution, and 10 times of the upper suspension is taken-4、10-5、10-6And coating a separation plate with the gradient diluent for culture, and identifying after culture.
The screening method of the indigenous strains for producing the pullulanase comprises the following steps: weighing 5g of enriched soil, adding the soil into a conical flask filled with 45mL of sterile water and glass beads, and shaking at room temperature at 150r/minStanding for 5min for 30min, taking the upper layer bacterial suspension, performing gradient dilution by 10 times, and taking 10-4、10-5、10-6And coating a separation plate with the gradient diluent for culture, and identifying after culture. The gradient dilution plate coating can avoid the phenomenon that strains are too dense and difficult to distinguish due to too many strains. Wherein the glass beads are used for assisting in shaking and dissolving, and are added with a proper amount.
The identification is carried out by dropwise adding Lu's iodine solution, and the strain with hydrolysis ring is pullulanase positive strain. The culture is carried out for 36-60h at constant temperature of 30 ℃ by inversion culture. The separation plate is a bacterial culture medium plate added with pullulan.
The in-situ enrichment is the inevitable trend of separation and screening of indigenous microbial resources, the technical scheme of the invention has the advantages of clear purpose, reliable effect and convenient operation of the preparation process. Can be used for the enrichment of indigenous bacteria and can also be popularized to the enrichment screening work of microorganism resources such as actinomycetes, fungi and the like.
Drawings
FIG. 1 is a schematic view of the hydrolysis loop of the selected strain in test example 1.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
Example 1
The in-situ enrichment method of the indigenous strains producing the pullulanase in the embodiment comprises the following steps:
A. substrate treatment
Taking 5g of sterilized absorbent cotton from a clean bench, putting the sterilized absorbent cotton into a sterilized beaker filled with 15g of amylopectin and 50mL of sterile water, turning over the absorbent cotton to enable the amylopectin to penetrate into and adhere to the absorbent cotton as much as possible, and preserving the pretreated absorbent cotton blocks in an aseptic manner.
B. In-situ enrichment by placing soil
Digging a pit with the depth of 5cm in the soil of the sampling environment (the soil of the wasteland on the two sides of the Luoyang suburb Luoye river), spraying 100mL of sterile water into the pit, taking out the absorbent cotton blocks treated in the step A, arranging the cotton blocks into a square shape (taking the non-breaking as the standard), flatly placing the square shape at the bottom of the pit, and covering the absorbent cotton with in-situ soil when the pit is dug. After 8d of in-situ enrichment, the absorbent cotton blocks are carefully taken out, and after the excess soil is shaken off, the surface soil of the absorbent cotton is collected in a super clean bench to be used as a soil sample cultured by a dilution smearing plate.
The screening method of the indigenous strains for producing the pullulanase in the embodiment comprises the following steps:
weighing 5g of the enriched soil, adding into a conical flask filled with 45mL of sterile water and a small amount of glass beads, shaking at room temperature at 150r/min for 30min, standing for 5min, taking the upper layer bacterial suspension, performing gradient dilution by 10 times, and taking 10-4、10-5、10-6The gradient dilutions were plated on separate plates (pullulan as substrate) for incubation. And (3) carrying out inverted culture at constant temperature of 30 ℃ for 48 h. And dripping Lu's iodine solution to detect whether the bacterial colony has a hydrolysis ring, wherein the bacterial colony with the hydrolysis ring is a bacterial strain capable of producing pullulanase.
The medium formulation in the separation plate was: 10-12g/L of glutinous rice starch, 8g/L of peptone, 2g/L of yeast powder and MgSO 24·7H2O 0.5g/L，K2HPO41g/L, agar 18g/L, pH 7.0.
Example 2
The in-situ enrichment method of the indigenous strains producing the pullulanase in the embodiment comprises the following steps:
A. substrate treatment
Taking 5g of sterilized absorbent cotton from a clean bench, putting the sterilized absorbent cotton into a sterilized beaker filled with 20g of amylopectin and 50mL of sterile water, turning over the absorbent cotton to enable the amylopectin to penetrate into and adhere to the absorbent cotton as much as possible, and preserving the pretreated absorbent cotton blocks in an aseptic manner.
B. In-situ enrichment by placing soil
Digging a 10cm deep pit in the soil of the sampling environment (the soil of the plough layer of the corn field in the suburb of Luoyang), spraying 150mL of sterile water into the pit, taking out the absorbent cotton blocks treated in the step A, arranging the cotton blocks into a square shape (taking the non-breaking as the standard), flatly placing the cotton blocks at the bottom of the pit, and covering the absorbent cotton with in-situ soil during digging the pit. After 12d of in-situ enrichment, the absorbent cotton blocks are carefully taken out, and after the excess soil is shaken off, the surface soil of the absorbent cotton is collected in a super clean bench to be used as a soil sample cultured by a dilution smearing plate.
The screening method of the indigenous strains for producing the pullulanase in the embodiment comprises the following steps:
weighing 5g of enriched soil, adding into a conical flask filled with 45mL of sterile water and a small amount of glass beads, shaking at 180r/min at room temperature for 30min, standing for 10min, taking the upper layer bacterial suspension, performing gradient dilution by 10 times, and taking 10-4、10-5、10-6The gradient dilutions were plated on separate plates (pullulan as substrate) for incubation. And (3) carrying out inverted culture at constant temperature of 30 ℃ for 48 h. And dripping Lu's iodine solution to detect whether the bacterial colony has a hydrolysis ring, wherein the bacterial colony with the hydrolysis ring is a bacterial strain capable of producing pullulanase.
The medium formulation in the separation plate was: 10-12g/L of glutinous rice starch, 8g/L of peptone, 2g/L of yeast powder and MgSO 24·7H2O 0.5g/L，K2HPO41g/L, agar 18g/L, pH 7.0.
Comparative example 1
The screening method of the indigenous strains for producing the pullulanase in the comparative example comprises the following steps:
1) collecting samples: reclaiming the same soil of the barren lands of the Luoyang rural areas Luohai in the example 1 to a laboratory;
2) the enrichment method comprises the following steps: and (4) enriching by using an indoor shaking table.
Taking 10g of soil sample, soaking the soil sample in 100mL of sterile normal saline for lh, shaking up, sucking 1.0mL of the soil sample into 50mL of enrichment medium (10-12 g/L of glutinous rice starch, 8g/L of peptone, 2g/L of yeast powder and MgSO 2)4·7H2O 0.5g/L，K2HPO41g/L, pH 7.0), culturing at room temperature or 32 deg.C for 18-20 h, performing gradient dilution 10 times, and taking 10-4-10-6And (5) coating a separation plate with the gradient diluent for culture.
The medium formulation in the separation plate was: 10-12g/L of glutinous rice starch, 8g/L of peptone, 2g/L of yeast powder and MgSO 24·7H2O 0.5g/L，K2HPO41g/L, agar 18g/L, pH 7.0.
Comparative example 2
The screening method of the indigenous strains for producing the pullulanase in the comparative example comprises the following steps:
1) collecting samples: the same luoyang suburb corn field plough layer soil as in example 2 was harvested to the laboratory;
2) the enrichment method comprises the following steps: and (4) enriching by using an indoor shaking table.
Taking 10g of soil sample, soaking the soil sample in 100mL of sterile normal saline for lh, shaking up, sucking 1.0mL of the soil sample into 50mL of enrichment medium (10-12 g/L of glutinous rice starch, 8g/L of peptone, 2g/L of yeast powder and MgSO 2)4·7H2O 0.5g/L，K2HPO41g/L, pH 7.0), culturing at room temperature or 32 deg.C for 18-20 h, performing gradient dilution 10 times, and taking 10-4-10-6And (5) coating a separation plate with the gradient diluent for culture.
The medium formulation in the separation plate was: 10-12g/L of glutinous rice starch, 8g/L of peptone, 2g/L of yeast powder and MgSO 24·7H2O 0.5g/L，K2HPO41g/L, agar 18g/L, pH 7.0.
Test example 1
Taking soil of barren lands on both sides of Luo river in Luoyang suburban area as an example, the method for screening the pullulanase-producing indigenous strains in example 1 and the comparative example was used to isolate the strains.
The method of example 1 was set up for 3 replicates, and 0.1mL of each dilution gradient was plated on 3 plates and statistically analyzed for the number of colonies with distinct characteristics. The colony statistics are shown in Table 2, and the screening results of example 1 are shown in FIG. 1-A; the screening results of the comparative example are shown in fig. 1-C, and the experimental results show that the colony number with hydrolysis circle obtained by separating and screening the pullulanase-producing indigenous strains by using the technology of the invention is obviously more than that of the control group.
Marking and purifying the identified strain, and identifying culture medium with pullulan (the formula is 1.5g/L of pullulan, 8g/L of peptone, 2g/L of yeast powder and MgSO4·7H2O 0.5g/L，K2HPO41g/L, agar 18g/L, pH 7.0. ) The degradation effect of the strains on pullulan is identified, the identification result of the screened strain of example 1 is shown in figure 1-B, and the identification result of the screened strain of comparative example 1 is shown in figure 1-D. The figure shows that the bacterial strain obtained by screening by the screening method has better degradation effect on pullulan. This shows that the technical method of the invention is effective and feasible, and is a product worthy of popularization and applicationA pullulanase indigenous strain separation and screening technology.
TABLE 1 separation effect of native strain of pullulanase after in-situ enrichment
| Is provided with | Number of colonies with distinguishing characteristics | Number of colonies with hydrolysis Ring |
| Comparative example | 3 | 2 |
| Example 1 (repeat 1) | 8 | 4 |
| Example 1 (repeat 2) | 11 | 5 |
| Example 1 (repeat 3) | 9 | 5 |
Test example 2
Taking the plough layer soil of the corn field in the suburb of Luoyang as an example, the screening method of the indigenous strains for producing the pullulanase in the example 2 and the comparative example is adopted to separate the strains.
The screening method of example 2 was set up in 3 replicates, and 0.1mL of each dilution gradient was used to coat 3 plates and the number of colonies with distinct characteristics was statistically analyzed. The 16S rDNA of the isolated strain having pullulanase activity was amplified using universal primers for bacteria (F: 5'-AGAGTTTGATCCTGGCTCAG-3', R: 5 '-AAGGAGGTGWTCCARCC-3'), the amplified product was sequenced, and the species of the strain was identified by analysis based on the sequencing results, with the results shown in Table 2.
TABLE 2 separation effect of native strain of pullulanase after in-situ enrichment
As can be seen from Table 2, from the colonies isolated by the method of the present invention, 5 strains belonging to different genera were identified; comparison example method the pullulanase-active strains isolated were classified into two genera. Further illustrates that the technical method of the invention is effective and feasible and has obvious advantages.
<110> university of Henan science and technology
<120> in-situ enrichment method and screening method of indigenous strains producing pullulanase
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<212> DNA
<213> Artificial sequence
<221> 16S rDNA forward primer sequence
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agagtttgat cctggctcag 20
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<213> Artificial sequence
<221> 16S rDNA reverse primer sequence
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aaggaggtgw tccarcc 17
Claims (4)
1. The screening method of the native strain for producing the pullulanase is characterized by comprising the following steps of: the method comprises the following steps: embedding the substrate adhered with the amylopectin into soil, taking out the substrate after enrichment, shaking off redundant soil, and collecting the soil on the surface of the substrate to obtain strain-enriched soil; uniformly mixing the strain-enriched soil with sterile water, centrifuging, taking the upper suspension for gradient dilution, taking the gradient dilution liquid to coat a separation flat plate for culture, and identifying after culture;
the lining is absorbent cotton;
the time for enriching the buried soil is 8-12 d;
the preparation of the substrate adhered with the amylopectin comprises the following steps:
1) dissolving amylopectin in sterile water to obtain amylopectin solution;
2) soaking sterilized liner in amylopectin solution, and adsorbing completely;
the concrete operation of burying soil is as follows: digging a pit in the soil of the sampling environment, spraying sterile water into the pit, taking the lining adhered with the amylopectin, placing the lining at the bottom of the pit, and covering the absorbent cotton with in-situ soil during digging the pit.
2. The method for screening an indigenous strain capable of producing pullulanase according to claim 1, wherein: the identification is carried out by dropwise adding Lu's iodine solution, and the strain with hydrolysis ring is pullulanase positive strain.
3. The method for screening an indigenous strain capable of producing pullulanase according to claim 1, wherein: the culture is carried out for 36-60h at constant temperature of 30 ℃ by inversion culture.
4. The method for screening an indigenous strain producing pullulanase according to claim 1, 2 or 3, wherein: the separation plate is a bacterial culture medium plate added with pullulan.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101886056A (en) * | 2010-07-02 | 2010-11-17 | 北京桑德环保集团有限公司 | Method for separating anaerobic ammonia oxidizing bacteria |
| CN103255079A (en) * | 2013-03-13 | 2013-08-21 | 江南大学 | Heatproof acidic pullulanase production strain and its screening method |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101886056A (en) * | 2010-07-02 | 2010-11-17 | 北京桑德环保集团有限公司 | Method for separating anaerobic ammonia oxidizing bacteria |
| CN103255079A (en) * | 2013-03-13 | 2013-08-21 | 江南大学 | Heatproof acidic pullulanase production strain and its screening method |
Non-Patent Citations (2)
| Title |
|---|
| Accounting for inherent variability of growth in microbial risk assessment;Marks,H M.等;《International Journal of Food Microbiology》;20051231;第100卷(第1-3期);第275-287页 * |
| 多孔介质中微生物生长行为和传输过程的数学模型研究进展;刚洪泽等;《化学与生物工程》;20090603;第26卷(第4期);第1-6页 * |
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