CN104818235A - Enterobacter ludwigii and application thereof - Google Patents

Enterobacter ludwigii and application thereof Download PDF

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Publication number
CN104818235A
CN104818235A CN201510256197.XA CN201510256197A CN104818235A CN 104818235 A CN104818235 A CN 104818235A CN 201510256197 A CN201510256197 A CN 201510256197A CN 104818235 A CN104818235 A CN 104818235A
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beta
enzyme
mannase
fermentation
test
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蔡俊
杨苗
王常高
杜馨
林建国
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Hubei University of Technology
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Hubei University of Technology
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts

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Abstract

The invention discloses enterobacter ludwigii and application thereof. A bacterial strain is enterobacter ludwigii MY271 collected in China Center for Type Culture Collection (CCTCC) in March 31, 2015, and has a collection number of CCTCC M2015182. According to the bred bacterial strain disclosed by the invention, beta-mannanase can be produced with high yield by using konjaku flour, manno-oligosaccharides can also be obtained, and the obtained manno-oligosaccharides have very good biological regulation functions, can be used for effectively reducing the cholesterol level of a human body, reducing blood sugar and promoting the growth of bifidobacteria in intestinal tracts, and are good food additives.

Description

A kind of Lu Shi enterobacteria and application thereof
Technical field
The invention belongs to microbial technology field, utilize Rhizoma amorphophalli powder to produce the stronger Lu Shi enterobacteria (Enterobacter ludwigii) of 'beta '-mannase ability and application thereof in particular to a strain.
Background technology
Hemicellulose is the assorted poly-polysaccharide of the second largest class of nature, and wherein beta-mannase is a kind of main hemicellulose.Beta-mannase is extensively present in locust bean gum, guar gum, konjaku powder and other plant polysaccharide.The saccharan (beta-mannase, glucomannan, polygalactomannan) that 'beta '-mannase (β-mannanase, EC 3.2.1.78) can be hydrolyzed containing β-Isosorbide-5-Nitrae seminose glycosidic bond on main chain is manna oligosaccharide.The manna oligosaccharide obtained has good biological regulation function, and the growth that effectively can reduce body's cholesterol level, reduce blood sugar, promote bifidus bacillus in enteron aisle is good foodstuff additive.This enzyme is also widely used in all many-sides such as feed, papermaking, textile printing and dyeing, oil production and biological study.As in paper industry, the hemicellulose degrading enzymes such as 'beta '-mannase and zytase class is collaborative to be used, and pulp treatment, obviously can improve papery, reduces the environmental pollution that chlorine floats and alkaline extraction brings; 'beta '-mannase is the biological gel breaker of high-quality of oil well petroleum fracturing liquid; In fodder industry, be used as feed enzyme additive, play the effect eliminating antinutritional factor and the nutritive value improving feed.
Take microorganism as the enzyme source of 'beta '-mannase, there is easy to prepare and that enzymic activity is high advantage, because enzyme procurement cost is low, facilitate the Study and appliance to this enzyme.Lin etc. obtain 'beta '-mannase by aspergillus niger (Aspergillus niger NCH189) liquid state fermentation, and product enzyme level is 27.4U/mL.Hossain etc. carry out liquid state fermentation with genus bacillus (Bacillus sp.KK01), and producing enzyme level can reach 90U/mL.Ferreira etc. are that carbon source is cultivated thermophile bacteria (Trichoderma harzianum strainT4) and produced the outer 'beta '-mannase of born of the same parents with wheat bran.The people such as Li Jianfang, Wu Minchen, by means such as screening and ultraviolet mutagenesises, screen the excellent acidic beta-mannase superior strain of a strain (Aspergil-lus usamii YL-01-78) from the bacterial strain that its laboratory is preserved.Yang Youhui etc. utilize Congo red plate screening model to screen from bud pole bacterium to obtain the strain excellent producing 'beta '-mannase.The people such as Luo Qiang, Sun Qiling, by the method for ion implantation and ultraviolet compounded mutagenesis, screen the bacillus subtilis (Bacillus subtilis) that 'beta '-mannase is produced in a strain.
Through consulting domestic and international prior art, temporarily not reporting and using Lu Shi enterobacteria to utilize Rhizoma amorphophalli powder to produce the document of 'beta '-mannase.
Summary of the invention
The object of the present invention is to provide a kind of Lu Shi enterobacteria (Enterobacter ludwigii) and application thereof of producing 'beta '-mannase.
In order to realize object of the present invention, contriver carries out strain improvement research repeatedly by lot of experiments, and from the soil of konjaku planting base, Zhuxi, Shiyan, finally screen the bacterial strain Lu Shi enterobacteria (Enterobacter ludwigii) that 'beta '-mannase is produced in a strain, by carrying out culture condition and medium optimization to it, improve the product enzyme level of this bacterial strain, for the suitability for industrialized production of 'beta '-mannase and application provide scientific basis.
Particularly, present invention obtains a strain and produce the stronger Lu Shi enterobacteria (Enterobacter ludwigii) of 'beta '-mannase ability, by its called after Lu Shi enterobacteria (Enterobacter ludwigii) MY271, be preserved in China typical culture collection center on March 31st, 2015, preservation address is Wuhan, China, and deposit number is: CCTCC NO:M 2015182.
Lu Shi enterobacteria (Enterobacter ludwigii) MY271 of the high yield 'beta '-mannase that the present invention relates to has following characteristic sum characteristic:
Morphological feature: bacterium colony is rounded, oyster white, smooth surface, neat in edge.
Physiological property: utilize glucose, fructose, rhamnosyl, sucrose, maltose, N.F,USP MANNITOL, seminose, semi-lactosi, L-arabinose, inositol, lactose, sorbyl alcohol and D-raffinose etc. as carbon source.
Metabolic characteristic: metabolic process can produce 'beta '-mannase.
Optimum fermentation culture conditions is: fermentation time 48h, temperature 31 DEG C, inoculum size 9%, initial pH7.0, liquid amount 50mL/250mL, medium component: Rhizoma amorphophalli powder 14g/L, peptone 18g/L, KH 2pO 40.5g/L, Na 2hPO 40.7g/L, MgSO 40.01g/L, ZnSO 40.15g/L.
Lu Shi enterobacteria (Enterobacter ludwigii) MY271 of the high yield 'beta '-mannase that the present invention relates to obtains as follows: gather soil sample 4 parts from konjaku planting base, Zhuxi, Shiyan, Hubei.Taking 5g mixed soil sample in every increment product joins in stroke-physiological saline solution, at 30 DEG C, on the constant-temperature table of 160r/min, fully vibration 2h prepares bacteria suspension, then gets 2mL soil supension and joins enrichment medium (Rhizoma amorphophalli powder 5.0g/L, peptone 5.0g/L, KH 2pO 41.0g/L, MgSO 40.1g/L, pH nature 0.1MPa sterilizing 20min), 30 DEG C, shaking culture 48h under 160r/min.Enrichment culture gained bacterium liquid carries out gradient dilution, and the bacterium liquid 100 μ L got after suitably dilution coats primary dcreening operation plate culture medium (Rhizoma amorphophalli powder 5.0g/L, NH 4cl 5.0g/L, KH 2pO 41.0g/L, MgSO 40.1g/L, agar 20g/L, pH nature 0.1MPa sterilizing 20min), be inverted in 30 DEG C of incubators and cultivate 48h, each dilution gradient do 3 times parallel.After strain culturing 48h, appropriate Congo red dye liquor is added in flat board, leave standstill 10min, observe size and the transparency of yellow hydrolysis circle, select that hydrolytic circle is large and transparency is good bacterial strain is stored in test tube slant substratum (peptone 10g/L, extractum carnis 3.0g/L, NaCl 5.0g/L, agar 20g/L, natural pH, 0.1MPa sterilizing 20min), test in order to shake flask fermentation.By shake flask fermentation (Rhizoma amorphophalli powder 5.0g/L, peptone 5.0g/L, KH 2pO 41.0g/L, MgSO 40.1g/L, pH nature, 0.1MPa sterilizing 20min) test, the fermentation broth enzyme of a strain Lu Shi enterobacteria is wherein lived the highest, by its called after Lu Shi enterobacteria (Enterobacter ludwigii) MY271, be preserved in China typical culture collection center on March 31st, 2015, deposit number is: CCTCC M 2015182 simultaneously.
Compared with prior art, Lu Shi enterobacteria (Enterobacter ludwigii) the MY271 tool that the present invention obtains has the following advantages and marked improvement:
(1) bacterial strain of institute of the present invention seed selection can utilize Rhizoma amorphophalli powder to produce 'beta '-mannase, and in the rear gained fermented liquid of fermentation, the enzyme work of 'beta '-mannase is up to 62.76U/ml.
(2) manna oligosaccharide that beta-mannase enzyme liberating mannosans obtains has good biological regulation function, and the growth that effectively can reduce body's cholesterol level, reduce blood sugar, promote bifidus bacillus in enteron aisle is good foodstuff additive.
Accompanying drawing explanation
To be inoculum size live on enzyme Fig. 1 affects column diagram;
To be liquid amount live on enzyme Fig. 2 affects column diagram;
To be initial pH live on enzyme Fig. 3 affects column diagram;
To be leavening temperature live on enzyme Fig. 4 affects column diagram;
To be fermentation period live on enzyme Fig. 5 affects column diagram;
To be carbon source live on enzyme Fig. 6 affects column diagram;
To be carbon source addition live on enzyme Fig. 7 affects column diagram;
To be nitrogenous source live on enzyme Fig. 8 affects column diagram;
To be nitrogenous source addition live on enzyme Fig. 9 affects column diagram;
To be inorganic salt live on enzyme Figure 10 affects column diagram;
Figure 11 is the optimization test result column diagram of biphosphate potassium application rate;
Figure 12 is the optimization test result column diagram of peptone consumption;
Figure 13 is the optimization test result column diagram of sodium-chlor consumption;
Figure 14 is the optimization test result column diagram of zinc sulfate consumption.
Embodiment
Form is described in further detail foregoing of the present invention again by the following examples, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following embodiment, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
It should be noted that, the term enzyme in the present invention is lived and is defined as: per minute hydrolysis beta-mannase produces enzyme amount needed for 1 μm of ol reducing sugar and is defined as an enzyme activity unit (U).
The preparation method of crude enzyme liquid is: get the bacterium liquid of certain volume in centrifuge tube, 8000rpm is centrifugal, and 15min gets supernatant liquor, after dilution suitable multiple, is then crude enzyme liquid.
The measuring method that 'beta '-mannase enzyme is lived is: get 1.5mL0.5% konjaku purifying powder solution in 25mL colorimetric cylinder, add 0.5mL crude enzyme liquid in 55 DEG C of water-bath reaction 10min, add 3mLDNS solution, boil 5min in boiling water bath after, be cooled to room temperature immediately, add distilled water and complement to 25mL, this is experimental group; Control group replaces crude enzyme liquid with boiling inactivator liquid.Light absorption value is measured at 540nm place.
The training systern of embodiment one: Lu Shi enterobacteria MY271
(1) bacterial strain: Lu Shi enterobacteria MY271CCTCC M 2015182
(2) method steps:
Fermention medium is initial medium (Rhizoma amorphophalli powder 5.0g/L, peptone 5.0g/L, KH 2pO 41.0g/L, MgSO 40.1g/L).Initial fermentation condition is: natural pH, inoculum size 5%, liquid amount 50mL/250mL, 30 DEG C, 160r/min shake flask fermentation 48h.
Adopt the inoculum size of single factor experiment investigation Lu Shi enterobacteria MY271 bacterial strain, liquid amount, initial pH, leavening temperature, fermentation period on the impact of beta-mannase production of enzyme.
A. the optimization test of inoculum size
Other fermentation condition is initial fermentation condition, when inoculum size is respectively 3%, 5%, 7%, 9%, 11%, 13% and 15%, investigates the impact that inoculum size Dui Lushi enterobacteria MY271 produces 'beta '-mannase.
As shown in Figure 1, when inoculum size is 9%, enzyme is lived the highest, is 4.11U/mL.Therefore select inoculum size to be 9%.
B. the optimization test of fermentation shake flask liquid amount
Fermentation inoculation measures the optimal result that above-mentioned test is determined, other fermentation condition is initial fermentation condition, adopt 250mL shaking flask dress liquid, when shaking flask liquid amount is respectively 25mL, 50mL, 75mL, 100mL, 125mL, investigate the impact that liquid amount Dui Lushi enterobacteria MY271 produces 'beta '-mannase.
As shown in Figure 2, when shaking flask liquid amount is 50mL/250mL, enzyme is lived and is reached maximum value 4.79U/mL.Therefore select liquid amount to be 50mL/250mL.
C. the optimization test of initial pH
Fermentation inoculum size and liquid amount get the optimal result that above-mentioned test is determined, other fermentation condition is initial fermentation condition, adjust initial pH and be respectively 4.0,4.5,5.0,5.5,6.0,6.5,7.0,7.5,8.0 and 8.5, shake flask fermentation measures 'beta '-mannase enzyme after cultivating and lives, and investigates the impact that pH Dui Lushi enterobacteria MY271 produces 'beta '-mannase.
As shown in Figure 3, when the initial pH that ferments is 7.0, enzyme is lived and is reached maximum value 7.0U/mL.Therefore select initial pH to be 7.0.
D. the optimization test of leavening temperature
Fermentation inoculum size, liquid amount and initial pH get the optimal result that above-mentioned test is determined, other fermentation condition is initial fermentation condition, measure 'beta '-mannase enzyme after carrying out fermentation culture respectively in 25 DEG C, 28 DEG C, 31 DEG C, 34 DEG C, 37 DEG C and 40 DEG C of shaking tables to live, investigate the impact that temperature Dui Lushi enterobacteria MY271 produces 'beta '-mannase.
Known by Fig. 4, when leavening temperature is 31 DEG C, enzyme is lived the highest, is 7.63U/mL.Therefore select leavening temperature to be 31 DEG C.
E. the optimization test of fermentation period
Fermentation inoculum size, liquid amount, initial pH and temperature get the optimal result that above-mentioned test is determined, other fermentation condition is initial fermentation condition, live every 12h sampling and measuring 'beta '-mannase enzyme, determine that Lu Shi enterobacteria MY271 produces the optimum fermentation period of 'beta '-mannase.
Known by Fig. 5, as fermentation 48h, enzyme is lived and is reached maximum value 7.94U/mL.Therefore select fermentation period to be 48h.
The medium component optimization of embodiment two: Lu Shi enterobacteria MY271
(1) bacterial strain: the bacterial classification Lu Shi enterobacteria MY271 that embodiment one filters out
(2) method steps:
Culture condition is fermentation time 4h, temperature 31 DEG C, inoculum size 9%, initial pH7.0, liquid amount 50mL/250mL.
A. carbon source is selected and addition test
With Rhizoma amorphophalli powder, locust bean gum, sucrose, D-MANNOSE, PEARLITOL 25C, Zulkovsky starch, maltose, glucose, D-Fructose, alpha-lactose, Xylo-Mucine, D-wood sugar, molasses for carbon source variable, addition is 5g/L, and other compositions are initial incubation based component.
As shown in Figure 6, when taking Rhizoma amorphophalli powder as carbon source, bacterial strain MY271 induces product enzyme effect best, and enzyme is lived as 8.01U/mL, and locust bean gum takes second place, and therefore selects Rhizoma amorphophalli powder to be optimum carbon source.
Choosing Rhizoma amorphophalli powder according to upper experiment is optimum carbon source, and research addition is respectively the impact that 5g/L, 6g/L, 7g/L, 8g/L, 9g/L, 10g/L, 11g/L and 12g/L Dui Lushi enterobacteria MY271 produces 'beta '-mannase.
Known by Fig. 7, when Rhizoma amorphophalli powder content is 8g/L, enzyme is lived the highest, is 17.73U/mL.Therefore select Rhizoma amorphophalli powder addition to be 8g/L.
B. nitrogenous source Selection experiment
Carbon source and addition thereof use above-mentioned test-results, with peptone, yeast powder, extractum carnis, yeast extract, corn steep liquor, wheat bran leach liquor, soyflour, ammonium sulfate, ammonium nitrate, ammonium chloride, diammonium hydrogen citrate, Secondary ammonium phosphate, primary ammonium phosphate, Pidolidone for nitrogenous source variable, addition is 5g/L, and other compositions are initial incubation based component.
Known by Fig. 8, when selecting peptone, enzyme is lived the highest, is 17.06U/mL, therefore selects peptone to be optimum nitrogen source.
Choosing peptone according to above-mentioned test is optimum nitrogenous source, and research addition is respectively the impact that 3g/L, 5g/L, 7g/L, 9g/L, 11g/L, 13g/L, 15g/L, 17g/L, 19g/L, 21g/L and 23g/L Dui Lushi enterobacteria MY271 produces 'beta '-mannase.
As shown in Figure 9, when peptone content is 19g/L, enzyme is lived and is reached maximum value 38.42U/mL.Therefore select peptone addition to be 19g/L.
C. the selection of inorganic salt
Carbon source, nitrogenous source and addition thereof use above-mentioned test-results, study its Dui Lushi enterobacteria MY271 produce the impact of 'beta '-mannase with dipotassium hydrogen phosphate, potassium primary phosphate, Sodium phosphate dibasic, SODIUM PHOSPHATE, MONOBASIC, sodium-chlor, magnesium sulfate, calcium sulfate, barium sulfate, copper sulfate, ferrous sulfate, manganous sulfate, zinc sulfate for inorganic salt variable.
As shown in Figure 10, zinc sulfate, sodium-chlor, magnesium sulfate, Sodium phosphate dibasic, potassium primary phosphate can promote that Lu Shi enterobacteria MY271 produces 'beta '-mannase preferably, therefore select zinc sulfate, sodium-chlor, magnesium sulfate, Sodium phosphate dibasic, potassium primary phosphate to carry out follow-up test as inorganic salt.
D.Plackett-Burman tests
With Rhizoma amorphophalli powder, peptone, potassium primary phosphate, Sodium phosphate dibasic, zinc sulfate, sodium-chlor and magnesium sulfate be Plackett-Burman test, test design as table 1, to determine significant factors.
Table 1 Plackett-Burman experimental factor and level
Table 2 Plackett-Burman test design and result
The regression analysis of table 3 Plackett-Burman test design
Table 4 Plackett-Burman designs analysis of variance table
Significant at 5％level,*represent the significant factor
Getting fiducial interval is 95%, is significant factors, and has the data of 99.42% can explain this test by table 3 and 4 known Rhizoma amorphophalli powders, Sodium phosphate dibasic and magnesium sulfate.
E. steepest climbing and the test of minimum addition
According to Plackett-Burman test, do steepest hill climbing test with significant factors Rhizoma amorphophalli powder, Sodium phosphate dibasic and magnesium sulfate, significant factors peptone, sodium-chlor, potassium primary phosphate and zinc sulfate do not do the test of minimum addition.
The design of table 5 steepest hill climbing test and result
As shown in Table 5, when the consumption of Rhizoma amorphophalli powder, Sodium phosphate dibasic and magnesium sulfate is respectively 14g/L, 0.8g/L, 0.05g/L, enzyme is lived and is reached maximum value 49.750U/mL.Therefore select the consumption of Rhizoma amorphophalli powder, Sodium phosphate dibasic and magnesium sulfate to be respectively 14g/L, 0.8g/L, 0.05g/L as central composite design central point.
When other medium component consumption is constant, biphosphate potassium application rate is respectively 0g/L, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, measures 'beta '-mannase enzyme and lives, the results are shown in Figure 11.
As shown in Figure 11, when biphosphate potassium application rate is 0.5g/L, enzyme is lived and is reached maximum value 51.17U/mL.Therefore select biphosphate potassium application rate to be 0.5g/L.
When other medium component consumption is constant, peptone consumption is respectively 14g/L, 15g/L, 16g/L, 17g/L, 18g/L, 19g/L, measures 'beta '-mannase enzyme and lives, the results are shown in Figure 12.
As shown in Figure 12, when peptone consumption is 18g/L, enzyme is lived and is reached maximum value 52.34U/mL.Therefore select peptone consumption to be 18g/L.
When other medium component consumption is constant, sodium-chlor consumption is respectively 0g/L, 0.05g/L, 0.1g/L, 0.15g/L, 0.2g/L, measures 'beta '-mannase enzyme and lives, the results are shown in Figure 13.
As shown in Figure 13, along with the increase of sodium chloride content, 'beta '-mannase enzyme is lived and is substantially tended to balance, and for saving cost, therefore selective chlorination sodium consumption is 0g/L.
When other medium component consumption is constant, zinc sulfate consumption is respectively 0g/L, 0.05g/L, 0.1g/L, 0.15g/L, 0.2g/L, measures 'beta '-mannase enzyme and lives, the results are shown in Figure 14.
As shown in Figure 14, when zinc sulfate consumption is 0.15g/L, enzyme is lived and is reached maximum value 55.03U/mL.Therefore select zinc sulfate consumption to be 0.15g/L.
F. central composite design (Central composite design, CCD)
3 kinds of remarkably influenced 'beta '-mannase enzymes medium component alive is obtained, i.e. Rhizoma amorphophalli powder, Sodium phosphate dibasic and magnesium sulfate through PB design screening.With the concentration (g/L) of this 3 remarkable kind factor for independent variable(s), the enzyme (U/mL) alive of 'beta '-mannase is response value, carries out the central composite design test of 3 factor 5 levels, in table 6.
The central composite design experimental factor of table 6 and level
Table 7 Central Composite test design and result
The analysis of table 8 second order polynomial regression
SS＝Sum of Squares，DF＝Degrees of Freedom，Significant at 5％level，*represent the significant factor.
According to table 7 testing data, (U/mL) is lived for response quautity Y with 'beta '-mannase, Rhizoma amorphophalli powder (g/L), Sodium phosphate dibasic (g/L), magnesium sulfate (g/L) are independent variable(s) A, B, C, utilize Design-Expert8.0.5 software to carry out regression analysis, can obtain second order empirical model is:
Y＝60.08+1.14A+0.88B-2.24C+2.85AB-1.76AC+2.06BC-3.26A 2-1.96B 2-0.91C 2
Known on the online data regression equation of 93.06% by table 8, illustrate that this regression equation has certain interpretability to experimental result.
Example three: fermentation proof test
Lu Shi enterobacteria MY271 take culture condition as fermentation time 48h, temperature 31 DEG C, inoculum size 9%, initial pH7.0, liquid amount 50mL/250mL, medium component Rhizoma amorphophalli powder 14g/L, peptone 18g/L, KH 2pO 40.5g/L, Na 2hPO 40.7g/L, MgSO 40.01g/L, ZnSO 40.15g/L carries out fermentation proof test.
The culture condition drawn according to the test design optimization of embodiment one and two and substratum composition ferment and to obtain 'beta '-mannase enzyme theoretical value 63.02U/mL alive, for verifying accuracy and the validity of this model, fermentation test is carried out under the combination of prediction best medium, repeat 3 tests, after actual verification, 'beta '-mannase enzyme mean value alive is 62.76U/mL.This model visible can be lived by the rear 'beta '-mannase enzyme of predict fermentation.

Claims (4)

1. Lu Shi enterobacteria (Enterobacter ludwigii) MY271, its Patent Deposit Designation is CCTCC M2015182.
2. Lu Shi enterobacteria MY271 according to claim 1 ferments and produces the technique of 'beta '-mannase.
3. Lu Shi enterobacteria MY271 according to claim 1 is utilizing the application in Rhizoma amorphophalli powder production 'beta '-mannase.
4. Lu Shi enterobacteria MY271 according to claim 1 is utilizing the application in Rhizoma amorphophalli powder production manna oligosaccharide.
CN201510256197.XA 2015-05-18 2015-05-18 Enterobacter ludwigii and application thereof Pending CN104818235A (en)

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Publication number Priority date Publication date Assignee Title
CN107629980A (en) * 2017-09-30 2018-01-26 中南民族大学 The Ludwig enterobacteria SXZ N5 bacterial strains and purposes of one plant of production huperzine and Huperzine B
CN112961807A (en) * 2021-03-30 2021-06-15 中国科学院成都生物研究所 Microbial composition and application thereof in promoting germination and growth of highland barley seeds
CN114921387A (en) * 2022-07-01 2022-08-19 中国农业科学院农田灌溉研究所 Enterobacter ludwigii with algae-lysing activity and preparation method of permeable microspheres of enterobacter ludwigii

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Cited By (4)

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Publication number Priority date Publication date Assignee Title
CN107629980A (en) * 2017-09-30 2018-01-26 中南民族大学 The Ludwig enterobacteria SXZ N5 bacterial strains and purposes of one plant of production huperzine and Huperzine B
CN112961807A (en) * 2021-03-30 2021-06-15 中国科学院成都生物研究所 Microbial composition and application thereof in promoting germination and growth of highland barley seeds
CN112961807B (en) * 2021-03-30 2023-01-20 中国科学院成都生物研究所 Microbial composition and application thereof in promoting germination and growth of highland barley seeds
CN114921387A (en) * 2022-07-01 2022-08-19 中国农业科学院农田灌溉研究所 Enterobacter ludwigii with algae-lysing activity and preparation method of permeable microspheres of enterobacter ludwigii

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