CN117187129A - Enterobacter cloacae for producing DDP-IV inhibitor and application thereof - Google Patents
Enterobacter cloacae for producing DDP-IV inhibitor and application thereof Download PDFInfo
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses enterobacter cloacae producing a DDP-IV inhibitor and application thereof, and relates to a strain and application thereof. The enterobacter cloacae producing the DDP-IV inhibitor is enterobacter cloacae (Enterobacter cloacae) HB47, and is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.28084. The enterobacter cloacae (Enterobacter cloacae) HB47 is a strain with both hypoglycemic and siderophore production. The enterobacter cloacae (Enterobacter cloacae) HB47 of the invention is added into a rice field of a rice-crab comprehensive planting and breeding mode, so that the feed digestion utilization rate of procambarus clarkia can be improved, and the antibacterial effect can be produced.
Description
Technical Field
The invention relates to a strain and application thereof.
Background
The rice and shrimp co-cropping mode is one of typical modes of comprehensive rice field cultivation, and forms a mutually beneficial symbiotic composite ecological system by rice planting and procambarus clarkii (commonly called as crayfish) cultivation, wherein rice grows in water, and shrimps are arranged at the downstream of rice, so that space can be saved, one-water dual-purpose, one-field double-harvest, grain stabilizing and synergy can be realized, and the environment is protected. According to the report of the development of the Procambrus clarkii industry in China (2021), the Procambrus clarkii breeding industry in 2021 has a yield value of about 748.38 hundred million yuan, which is increased by 11.4 percent in the same ratio; wherein, the cultivation area of the procambarus clarkia paddy field is about 126.14 ten thousand hm 2 The culture yield is 20.623 hundred million kg, which respectively accounts for 86.6 percent and 86.2 percent of the total culture area and the total culture yield of procambarus clarkia, and respectively accounts for 49.2 percent and 63.4 percent of the total comprehensive planting and culture area and the total yield of national rice and fish. The efficient mode of rice and shrimp co-cropping ecological planting and raising developed in the hidden river city of Hubei province is more praised as a successful model of modern agricultural development.
However, the rice and shrimp co-cropping mode also has double-edged property, and due to long-term flooding and high underground water level, secondary submerged cultivation of the rice field soil is often caused, and the rice field soil becomes cold-soaked fields and muddy fields.
Disclosure of Invention
The invention provides enterobacter cloacae producing a DDP-IV inhibitor and application thereof.
The enterobacter cloacae producing the DDP-IV inhibitor is enterobacter cloacae (Enterobacter cloacae) HB47, and is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.28084.
The invention also discloses the application of the enterobacter cloacae producing the DDP-IV inhibitor in the rice and shrimp co-cropping mode.
The invention adds the enterobacter cloacae (Enterobacter cloacae) HB47 bacterial liquid producing the DDP-IV inhibitor into a rice field with a rice and shrimp co-cropping mode to promote the growth of rice and procambarus clarkia.
The bacterial colony of the enterobacter cloacae (Enterobacter cloacae) HB47 on the LB solid culture medium is circular, yellowish and opaque, the bacterial colony is stuck on the surface of the culture medium, the center is slightly convex, the bacterial colony is more moist, the edge is neat, and the bacterial colony is gram-negative bacteria after gram staining.
Enterobacter cloacae (Enterobacter cloacae) HB47 can strongly produce siderophores with high iron chelating ability, and the Su value of siderophores in 37 ℃ environment is 68%. Under the iron deficiency condition, the enterobacter cloacae (Enterobacter cloacae) HB47 can produce a siderophore with higher affinity, and the siderophore can absorb and utilize iron in the environment so as to act on the siderophore and animals and plants, and inhibit the growth and reproduction of pathogenic bacteria when competing limited iron nutrition with pathogenic bacteria, thereby achieving the biological control effect on procambarus clarkia and rice, improving the immunity defense capability of procambarus clarkia and reducing the proportion of pathogenic bacteria in the intestinal flora of procambarus clarkia.
The diameter D of a hydrolysis circle of the lactobacillus cloacae (Enterobacter cloacae) HB47 on the SKM solid culture medium is 6.04mm, the diameter D of a colony is 3.86mm, and the D/D is 1.56; the yield of DDP-IV inhibitor was as high as 31.2% in 37 ℃. The enterobacter cloacae (Enterobacter cloacae) HB47 can produce a DDP-IV inhibitor, can control the blood sugar of eriocheir sinensis, and can better absorb sugar substances in feed, especially some non-starch polysaccharide; further can improve the digestion utilization rate of the feed and enhance the growth performance.
The enterobacter cloacae (Enterobacter cloacae) HB47 is a strain with both hypoglycemic and siderophore production. The enterobacter cloacae (Enterobacter cloacae) HB47 of the invention is added into a rice field of a rice-crab comprehensive planting and breeding mode, so that the feed digestion utilization rate of procambarus clarkia can be improved, and the antibacterial effect can be produced. The enterobacter cloacae (Enterobacter cloacae) HB47 can form obvious inorganic phosphorus-soluble ring on Meng Jinna inorganic phosphorus culture medium, the diameter D of the inorganic phosphorus-soluble ring is 10.07mm, the colony diameter D is 8.85mm, and the D/D is 1.14; the fertilizer has a certain function of dissolving inorganic phosphorus, can dissolve inorganic phosphorus in soil, can decompose the inorganic phosphorus which is difficult to convert and utilize in the soil into soluble phosphorus to be utilized by rice plants, and can promote the growth of rice.
The reduction of the pH at 37 ℃ has little influence on the number of HB47 spores of enterobacter cloacae (Enterobacter cloacae) and has stronger acid resistance; the concentration of bile salts has a great influence on the spore number of the HB47 of the enterobacter cloacae (Enterobacter cloacae), and when the concentration of the bile salts is increased to 1.5%, the survival rate of HB47 spores is less than 80%; enterobacter cloacae (Enterobacter cloacae) HB47 has extremely high tolerance to acidity and has relatively high tolerance to bile salts. Therefore, enterobacter cloacae (Enterobacter cloacae) HB47 can survive and act in the intestine of procambarus clarkia. The enterobacter cloacae (Enterobacter cloacae) HB47 strain is safe and reliable, does not cause death of procambarus clarkia, compared with a control group, the feed coefficient of the enterobacter cloacae (Enterobacter cloacae) HB47 group is obviously lower than that of the control group, the weight gain rate and the specific growth rate of a treatment group are obviously higher than those of the control group, and the utilization rate of the feed can be obviously improved by the enterobacter cloacae (Enterobacter cloacae) HB47.
The enterobacter cloacae (Enterobacter cloacae) HB47 can greatly improve the plant height and fresh weight of rice seedlings, and slightly improve the root length and stem thickness of the rice seedlings, thereby being very beneficial to the growth of rice plants.
The use of enterobacter cloacae (Enterobacter cloacae) HB47 can improve the availability of soil nutrients, increase the yield of rice and play a role in improving the fertilizer for the submerged rice soil.
Enterobacter cloacae (Enterobacter cloacae) HB47 is Enterobacter cloacae and belongs to the genus Enterobacter (Enterobacter Hormaeche and Edwards, 1960); the culture medium is preserved in China general microbiological culture Collection center (CGMCC), the preservation address is 1 # 3 of North West Lu of the Korean area of Beijing, the preservation number is 28084 of CGMCC, and the preservation date is 2023, 8 months and 18 days.
Drawings
FIG. 1 is a graph showing the results of an experiment in which Enterobacter cloacae (Enterobacter cloacae) HB47 is subjected to inorganic phosphorus lysis on Meng Jinna inorganic phosphorus bacteria medium;
FIG. 2 is a graph showing the results of a siderophore screening assay of Enterobacter cloacae (Enterobacter cloacae) HB47 on CAS solid detection media;
FIG. 3 is a graph showing the results of a hydrolysis test of milk protein from Enterobacter cloacae (Enterobacter cloacae) HB47 on SKM medium;
FIG. 4 is a phylogenetic tree constructed from Enterobacter cloacae (Enterobacter cloacae) HB 47;
FIG. 5 is a graph showing the comparative effect of E.cloacae (Pseudomonas glycinae) HB47 strain on pot growth of rice seedlings in example 2.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other.
The first embodiment is as follows: the enterobacter cloacae producing the DDP-IV inhibitor is enterobacter cloacae (Enterobacter cloacae) HB47, and is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.28084.
Selecting a gram of hula aquatic products test field of Heilongjiang aquatic products research institute of China academy of aquatic products in 2023 and 4The method comprises the steps of randomly selecting 300 healthy procambarus clarkii (25.0+/-1.87) g, filling air and water, bagging, transporting to the university of northeast agricultural animal science and technology college, selecting 20 procambarus clarkii with normal body color and active reaction, and anesthetizing by using MS-222 anesthetic (250 mg/L). Wiping the surface of procambarus clarkia with absolute ethanol, dissecting with sterilized scissors and tweezers in an ultra-clean workbench, taking out the whole intestinal tract of procambarus clarkia, gently squeezing out the content, placing into a conical flask containing glass beads and 50mL of sterile water, oscillating at 180r/min for 30min at room temperature, performing gradient dilution, and collecting 10 -3 、10 -4 、10 -5 100 mu L of the gradient is coated on an LB solid culture medium plate, each gradient is repeated for 3 times, and the mixture is placed at 28 ℃ for culturing for 24-48 hours. After 48h of culture, strains with different shapes are selected for separation and purification, wherein the strain HB47 is obtained.
Strain HB47 was purified and inoculated with sterile toothpicks onto plates of Meng Jinna inorganic phosphorus bacteria medium. Each gradient was repeated 3 times and incubated at 28℃for 24-48 h. The colony diameter D and the diameter D of the inorganic phosphate ring were measured by the crisscross method, and D/D was calculated by forming an apparent inorganic phosphate ring around the colony of strain HB47 (as shown in FIG. 1). The diameter D of the inorganic phosphate ring of strain HB47 was 10.07mm, the colony diameter D was 8.85mm, and D/D was 1.14. The strain HB47 has a certain function of decomposing inorganic phosphorus.
Then, the purified strain HB47 was transferred to LB plate for 24 hours for reactivation, then single colony was picked up by sterilized toothpick and transferred to chrome azure S (Chromeazrol S, CAS) solid detection medium, and inverted culture was carried out at 37℃for 2-3D, and it was observed that a distinct color-changing ring was formed around the colony of strain HB47 (as shown in FIG. 2), colony diameter D and color-changing ring diameter D were measured by the crisscross method, the color-changing ring diameter D of strain HB47 was 19.98mm, colony diameter D was 5.02mm, and D/D was 3.98, indicating that strain HB47 has a siderophore with extremely strong Gao Tieao binding ability.
Strain HB47 was further tested:
(1) Inoculating the activated strain HB47 lawn in an iron-limiting SA liquid culture medium, and carrying out shake cultivation for 48 hours at 37 ℃;
(2) Transferring the bacterial suspension to be tested, which grows for 48 hours, into a sterilized 10mL centrifuge tube, and centrifuging at 13000rpm for 15min;
(3) Transferring the supernatant into a test tube treated by concentrated hydrochloric acid, adding a certain amount of freshly prepared CAS detection solution to make the volume ratio of the supernatant to the detection solution be 1:1, fully mixing uniformly, and standing at room temperature for 1h;
(4) Measuring an absorbance value (A) at a wavelength of 630nm, taking double distilled water as a control for zeroing, taking the absorbance value (Ar) at the wavelength of 630nm after the unvaccinated SA limited iron culture medium and the detection liquid which are measured by the same method are mixed as a reference value, and expressing the activity unit of the siderophore by the following formula:
Su≈(Ar-As)/Ar×100；
wherein: su is the siderophore content; ar is the OD value of the SA limited iron culture medium and the supernatant of the detection liquid which are not inoculated; as is the OD of the culture supernatant. (when the unit of siderophore activity is less than 10, it is generally considered negative, and the mixture of siderophore and detection liquid has no color change.)
As shown by test, the Su value of the strain HB47 for producing the siderophore at 37 ℃ is 68%, which proves that the strain has extremely strong siderophore producing capacity.
Strain HB47 was plated on SKM (2% nonfat dry milk medium) to form transparent circles, and the colony of strain HB47 formed distinct hydrolytic circles (as shown in fig. 3) around the colony, with the hydrolytic circle diameter D of strain HB47 being 6.04mm, the colony diameter D being 3.86mm, and D/D being 1.56. The strain HB47 bacterial liquid cultivated for 2 generations is centrifugated for 15min under the condition of 8000r/min, and the supernatant liquid is removed to leave bacterial mud. The cells were resuspended in PBS and the absorbance of the cell solution was adjusted to 1.0 by washing 3 times with 0.1mol/L sterile phosphate buffer solution (PBS, pH 6.8). Culturing at 37deg.C for 24 hr, centrifuging at 4deg.C and 8000r/min for 15min, filtering supernatant with 0.22pm water-based filter membrane bacterial filter to obtain cell metabolite (CFS), and preserving at-80deg.C. Further test: 25. Mu.L, 1.6mmol/L glycyl-prolyl-p-nitroaniline and 25. Mu.L CFS were accurately added dropwise to a 96-well plate. The reaction was stopped by adding 50. Mu.L of DDP-IV (0.01U/mL) at 37℃for 15min, continuing the reaction at 37℃for 1h, and finally adding 100. Mu.L of 1mol/L sodium acetate buffer (pH=4.0), and the absorbance of the reaction solution was measured at 405nm using an ELISA reader. DDP-IV inhibition rate calculation formula:
A sample to be measured : 25. Mu.L of sample +25. Mu.LGly-pro-phy +50. Mu.LDDP-IV +100. Mu.L of sodium acetate;
A sample blank : 25. Mu.L of sample +50. Mu.L of LTris-HCl +25. Mu.L of LGly-pro-phy +100. Mu.L of sodium acetate;
A negative control :25 mu LTris-HCl+25 mu LGly-pro-phy+50 mu LDDP-IV+100 mu L sodium acetate;
A negative blank :75 mu LTris-HCl+25 mu LGly-pro-phy+100 mu L sodium acetate.
The productivity of DDP-IV inhibitor at 37℃was as high as 31.2% for strain HB47, indicating that strain HB47 has a certain ability to produce DDP-IV inhibitor.
Strain HB47 stress resistance detection:
acid resistance detection: strain HB47 was inoculated into LB liquid medium at pH 2.0, 2.5 and 3.0 in an inoculum size of 2% (v/v), and the viable count was measured after plating by pipetting the bacterial liquids for 0, 1, 2 and 3 hours, respectively, and culturing at 37℃for 24 hours.
And (3) detecting the bile salt resistance: the activated strain HB47 was diluted in sterile physiological saline at a multiple ratio, a suitable dilution gradient was selected and 1mL of the dilution was aspirated and placed in a sterilized dish. Plates were then poured with LB solid medium containing 0.3% and 1.0% sodium taurocholate, while MRS solid medium without sodium taurocholate was used as a control group, cultured at 37℃for 48 hours, colony counts were performed, and the survival rate of the strain was calculated.
As shown in Table 1, the decrease in pH had a smaller effect on the number of spores of strain HB47, and the survival rate of spores of strain HB47 was 87.5% at pH 2.0, and the acid resistance was stronger. The concentration of bile salts has a large influence on the spore number of the strain HB47, and when the concentration of bile salts is increased to 1.5%, the spore survival rate of the strain HB47 is as low as 79.5%. The strain HB47 has extremely high tolerance to acidity and strong tolerance to bile salts.
TABLE 1
Physiological and biochemical identification of strain HB 47: the strain HB47 after preservation was streaked on a solid LB medium plate in three regions, single colonies were isolated and their morphology was described, and gram staining and physiological biochemical identification of the strain were performed according to the general bacterial System identification handbook.
Bacterial strain HB47 has a round colony shape, is yellowish and opaque, is stuck on the surface of a culture medium, has slightly raised center, is relatively moist and neat in edge, and is gram-negative bacteria after gram staining. The physiological and biochemical indexes of a part of strain HB47 are shown in Table 2. According to the description of physiological characteristics of Enterobacter from the Bojie's Manual of bacteria identification, strain HB47 has the same characteristics as physiological biochemistry of a model species of Enterobacter cloacae (Enterobacter cloacae), and from each physiological biochemistry it is inferred that strain HB47 may be Enterobacter cloacae (Enterobacter cloacae).
TABLE 2
16S rRNA identification of strain HB 47: and (3) selecting a bacterial genome DNA extraction kit of Beijing Soxhaust biological technology company, and extracting, separating and purifying the strain DNA. The bacterial universal primer 27F/1492R is adopted for PCR amplification, and the PCR amplification system is a 25 mu L system: 10 Xbuffer 2.5. Mu.L, taq enzyme 0.5. Mu.L, primer 27F 0.5. Mu.L, primer 142R 0.5. Mu.L, DNA template 1. Mu.L, ddH 2 O20. Mu.L. The reaction procedure is set to 95 ℃ for 5min of pre-denaturation; denaturation at 94℃for 50s, annealing at 56℃for 30s, extension at 72℃for 1.5min, cycle times for 30 times, extension at 72℃for 10min again, and preservation at 4 ℃. The PCR amplified product was sent to RuiBiotech company for sequencing. Sequencing results of strain 16S rRNA were aligned by NCBI database and phylogenetic tree was constructed (as shown in FIG. 4). BLAST ratios in NCBIThe similarity of the 16S rRNA gene sequence of the strain HB47 was found to be 99% with Enterobacter cloacae (Enterobacter cloacae). From the phylogenetic tree of strain HB47, it can be seen that the smallest branch of strain HB47 is located at the same site as Enterobacter cloacae (Enterobacter cloacae) (NR 118568.1), the evolutionary distance is closer, and the comprehensive physiological and biochemical index identifies the HB47 strain as Enterobacter cloacae (Enterobacter cloacae).
Example 1
Effect of enterobacter cloacae (Enterobacter cloacae) HB47 on the growth performance of procambarus clarkia.
The procambarus clarkia culture test is carried out in a circulating water tank of a laboratory of the institute of the aquatic products of the black longjiang of the academy of aquatic products of China, and procambarus clarkia used for the test is all sourced from a hula aquatic test field of the institute of the aquatic products of the black longjiang of the academy of aquatic products of China. Selecting 200 tails of procambarus clarkia with initial weight of 25.0+ -1.87 g propagated in the current year, placing in a cement pond with water depth of 4.3m×1.5m×1.0m (10 cm), and temporarily culturing with a large amount of herba Sagittariae as shielding material for 1 week to adapt to test environment, and feeding control group fodder during temporary culturing periodPuffed feed special for shrimp feed). After temporary rearing, selecting 120 tails of procambarus clarkii with similar weight, better activity and no damage on the body surface, randomly dividing the tails into 2 groups (a control group and a treatment group), wherein each group is 3 parallel, each parallel 20 tails are placed in 6 circulating water rearing boxes with the height of water body of 10cm and a large amount of water grass as a shielding object, tap water with aeration time of more than 24 hours is used as test water body, an oxygenation pump is adopted for 24 hours for aeration during the test, and the daily consumption is 08:00 and 16:00 feeding.
The treatment group was fed with 1.5X10 mix 6 CFU/mL of Enterobacter cloacae (Enterobacter cloacae) HB47 bacterial liquid The special puffed feed for shrimp feed is characterized in that before feeding, bacterial liquid is fully mixed with the feed (the feed-liquid ratio is 1:0.1), and the bacterial liquid is fully adsorbed on the feed. Control group feedingMaterial (/ ->Puffed feed special for shrimp feed); feeding for 2 times a day, apparent feeding, sucking at the bottom of the day for 1 time, changing water amount about 1/2, water temperature (23+ -1) deg.C during test, dissolved oxygen (6+ -0.5) mg/L, pH value of 7.1+ -0.5, and cultivation period of 7d. Observing and recording whether the appendages are complete or not at 09:00 a day, and calculating the mortality and the stump rate; weighing each treated procambarus clarkia after the cultivation is finished, and calculating the average weight, the weight growth rate and the feed coefficient, wherein the specific calculation formula is as follows:
body weight gain = [ (W) 1 -W 0 )/W 0 ]×100％
Mortality = D/Z x 100%
Stump rate = C/Z x 100%
Feed coefficient=w 2 /(W 1 -W 0 )×100％
Wherein W is 0 For initial weight, W 1 For final body weight, W 2 For feed consumption, D is the death number of each treatment in 24 hours, C is the shrimp mantissa with appendage loss in each treatment in 24 hours, and Z is the mantissa of procambarus clarkia put in each treatment.
As shown in Table 3, the results of the experiments show that the mortality and the amputation rate of procambarus clarkia are not significantly different among groups, and the results show that the Enterobacter cloacae (Enterobacter cloacae) HB47 does not influence the mortality and the amputation rate of procambarus clarkia, and the strain is safe and reliable. The feed coefficient of the treatment group is obviously lower than that of the control group, and the weight gain rate of the treatment group is obviously higher than that of the control group, which indicates that the enterobacter cloacae (Enterobacter cloacae) HB47 can obviously improve the feed utilization rate.
TABLE 3 Table 3
Example 2
Inoculating Enterobacter cloacae (Enterobacter cloacae) HB47 strain into 50mL LB liquid medium, shake culturing at 28deg.C and 180r/min overnight, and sterilizing with sterile waterDiluting the culture solution to a thallus concentration of 1×10 5 CFU/mL was used as bacterial suspension for further use. 150 seeds of Longjing 31 rice with plump and consistent grains are selected, soaked in warm water at 55 ℃ for 18 hours, and then are grouped and respectively placed in 1 multiplied by 10 5 CFU·mL -1 Soaking enterobacter cloacae (Enterobacter cloacae) HB47 bacterial suspension or clear water for 5 hours; the seeds after soaking are washed 3 times by distilled water, then are evenly placed in a culture dish (diameter is 10 cm) filled with 3 layers of filter paper, and then are respectively sprayed with 5mL of the bacterial suspension (the seeds are soaked by adopting enterobacter cloacae HB47 bacterial suspension before) or clear water (the seeds are soaked by adopting clear water before) on the surfaces of the seeds. After accelerating germination at 28 ℃, sowing the seedlings in 15kg paddy field soil transplanting pots, taking 30 seedlings from each pot after 28 days, and measuring root length, plant height, stem thickness and fresh weight of the seedlings, wherein the seedlings are repeated for 3 times.
The experimental results are shown in Table 4 and FIG. 5, and the Enterobacter cloacae (Enterobacter cloacae) HB47 has obvious growth promoting effect on rice under the potting condition. As can be seen from Table 4, the root length, plant height, stem thickness and fresh weight of the rice plants subjected to the seed soaking treatment by the Enterobacter cloacae (Enterobacter cloacae) HB47 are respectively increased by 4.76%, 49.81%, 6.34% and 30.19%, so that the Enterobacter cloacae (Enterobacter cloacae) HB47 can greatly improve the plant height and fresh weight of the rice seedlings, and the root length and stem thickness of the rice seedlings are slightly improved, thereby being very beneficial to the growth of the rice plants.
TABLE 4 Table 4
In this example, the mass concentration of corn starch in the culture medium of Enterobacter cloacae (Enterobacter cloacae) HB47 bacterial liquid was 1%, the mass concentration of beef extract was 1.5%, the mass concentration of calcium carbonate was 0.3%, and the mass concentration of dipotassium hydrogen phosphate was 0.02%.
The seed solution of Enterobacter cloacae (Enterobacter cloacae) HB47 is inoculated to the culture medium of the Enterobacter cloacae (Enterobacter cloacae) HB47 strain solution at the inoculum size of 3 percent, and then the culture is carried out for 24 hours under the conditions of 37 ℃ and 200r/min and pH 7, and the number of viable bacteria in the HB47 strain solution of the Enterobacter cloacae (Enterobacter cloacae) reaches 7.97×10 8 CFU/mL. The number of viable bacteria before the optimization is 5.46 multiplied by 10 7 CFU/mL (non-optimized pre-culture medium is bean sprout juice culture medium, bean sprout juice culture medium: 100mL of bean sprout juice, 10g of sucrose, (NH) 4 ) 2 SO 4 2g，NaCl 0.4g，ZnSO 4 0.08g, distilled water to 1000mL, pH 7, and autoclaved at 115℃for 30 min).
Claims (2)
1. A strain of enterobacter cloacae producing DDP-IV inhibitor is enterobacter cloacae (Enterobacter cloacae) HB47, and is preserved in China general microbiological culture Collection center with a preservation number of CGMCC No.28084.
2. Use of enterobacter cloacae producing a DDP-IV inhibitor according to claim 1 in a rice shrimp co-operation mode.
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