CN108277184A - Produce the bacillus and its preparation method and application of algin catenase - Google Patents
Produce the bacillus and its preparation method and application of algin catenase Download PDFInfo
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Abstract
The deposit number of a kind of bacillus, the bacillus (sp.W003) is：CGMCC NO.15010.The deposit number of a kind of bacillus, the bacillus (sp.W024) is：CGMCC NO.15011.A method of algin catenase is prepared using above-mentioned bacillus, is included the following steps：The bacillus is activated, the bacterium solution after activation is applied on slant medium；Bacillus on slant medium described in picking is inoculated in the container equipped with culture medium and is cultivated, and is 25 35 DEG C in temperature, cultivates 12 16h to the logarithm middle and later periods on the shaking table that rotating speed is 150 230r/min, obtain seed liquor；The seed liquor is inoculated in the fermentation tank equipped with enzymatic production culture medium, in 25 35 DEG C of temperature, rotating speed is 150 230r/min, and pH ferments 24 days under the conditions of being 78, obtains algin oligosaccharide zymotic fluid.Also provide a kind of bacillus production algin catenase, fermenting and producing algin oligosaccharide culture medium in application.The above-mentioned produced algin degradation enzyme vigor of bacillus is high, and fermentation efficiency is high.
Description
Technical field
The present invention relates to microorganism field, more particularly to bacillus of production algin catenase and preparation method thereof and
Using.
Background technology
Algin oligosaccharide is that algin is smaller through the degree of polymerization obtained from hydrolysis and the low molecular weight fraction of good water solubility.By
There are a variety of physiological activity in algin oligosaccharide, such as anticancer, anti-freezing, lipid-loweringing, immunological regulation and anti-aging effects, you can as guarantor
Health food also can be used as medicine or fungicide, therefore can be widely used in the fields such as food, medicine, agricultural and cosmetics
In, there is huge application and Development volue.
The preparation method for obtaining algin oligosaccharide at present mainly has chemical degradation method and enzymatic isolation method, chemical degradation method to there is production
The shortcomings of object is inhomogenous, poor repeatability, big environmental pollution degree, and enzymatic isolation method is because efficient, specificity is strong, reaction condition temperature
Advantages become main direction of studying in recent years with action time is short etc., and algin catenase derives from a wealth of sources in nature, mesh
Before, it has been reported that multiple-microorganism can produce algin catenase, such as Alteromonad (Alteromonas), Pseudoalteromonas
(Pseudoalteromonas), vibrio marinopraesens (Vibrio) etc..However, many of these microorganisms are pathogenic bacteria, such as ocean
Vibrios can make mammalian infections cause septicemia, cause algin catenase enzyme activity low, and fermentation efficiency is low, thus limits
The application of this quasi-microorganism.
Invention content
Based on this, it is necessary to provide that a kind of enzyme activity is high, the bacillus of the high production algin catenase of fermentation efficiency and
Preparation method and application.
The deposit number of a kind of bacillus, the bacillus (sp.W003) is：CGMCC NO.15010.
The deposit number of a kind of bacillus, the bacillus (sp.W024) is：CGMCC NO.15011.
A kind of application of bacillus as described above in producing algin catenase.
A kind of application for the culture medium producing algin oligosaccharide using fermentation of bacillus as described above, the culture medium
In carbon source be sodium alginate, Kelp Powder or kelp paste.
A method of algin catenase is prepared using bacillus as described above, is included the following steps：
The bacillus is activated, the bacterium solution after activation is applied on slant medium；
Bacillus on slant medium described in picking is inoculated in the container equipped with culture medium and is cultivated,
Temperature is 25-35 DEG C, and rotating speed is that culture 12-16h obtains seed liquor to the logarithm middle and later periods on the shaking table of 150-230r/min；
The seed liquor is inoculated in the fermentation tank equipped with enzymatic production culture medium, in 25-35 DEG C of temperature, rotating speed 150-
230r/min, pH ferment 2-4 days under the conditions of being 7-8, obtain algin oligosaccharide zymotic fluid.
The inoculum concentration of the seed liquor in the fermentation tank is 5%-15% in one of the embodiments,.
The inclined-plane culture based formulas is sodium alginate 10.0-15.0g/L, ammonium sulfate in one of the embodiments,
3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-4.0g/L, ferrous sulfate 0.01-0.1g/L and agar
15.0-20.0g/L。
The culture medium formula is in one of the embodiments,：
Sodium alginate 10.0-15.0g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-
4.0g/L and ferrous sulfate 0.01-0.1g/L；
Or Kelp Powder 20.0-40.0g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-
4.0g/L and ferrous sulfate 0.01-0.1g/L；
Or kelp paste 200-500g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-
4.0g/L and ferrous sulfate 0.01-0.1g/L.
In being formulated in one of the embodiments, to the culture medium plus metal ion, the metal ion add
It is 0.05%-1.0% to add quality percent by volume.
The metal ion is potassium ion, magnesium ion or iron ion in one of the embodiments,.
The produced degradable algins of algin degrading enzyme of above-mentioned bacillus sp.W003 and sp.W024 generate brown alga
Glue oligosaccharides, to mass produce algin oligosaccharide.The produced algin degradation enzyme vigor of bacterial strain uses therefor is high, fermentation efficiency
Height is the algin degrading enzyme production bacterial strain of great potential.
Description of the drawings
Fig. 1 is that the bacillus of an embodiment prepares the method flow schematic diagram of algin catenase；
Fig. 2 be an embodiment bacillus (sp.W003) after cultivate 48h, using Gram iodine solution to tablet dye
Color obtains the schematic diagram of dissolving circle；
Fig. 3 be an embodiment bacillus (sp.W024) after cultivate 48h, using Gram iodine solution to tablet dye
Color obtains the schematic diagram of dissolving circle；
Fig. 4 is embodiment bacillus (sp.W003) Gram's staining figure；
Fig. 5 is embodiment bacillus (sp.W024) Gram's staining figure；
Fig. 6 is the electron microscope of an embodiment bacillus (sp.W003)；
Fig. 7 is the electron microscope of an embodiment bacillus (sp.W024)；
Fig. 8 is the gene development tree graph of an embodiment bacillus (sp.W003)；
Fig. 9 is the gene development tree graph of an embodiment bacillus (sp.W024)；
Specific implementation mode
With reference to embodiment, to a kind of bacillus and its preparation method and application of production algin catenase make into
The detailed description of one step.
The bacillus (sp.W003) of one embodiment, preservation time：2017-12-04, preservation address：China, Beijing
City Chaoyang District, Institute of Microorganism, Academia Sinica, deposit number：CGMCC NO.15010.In one embodiment, gemma bar
Bacterium (sp.W003) form be in rodlike, Gram-negative, on inclined-plane cultivate 48h bacterium colony be in shallow white, round dimpling,
Surface is smooth, lawn moistens, bacterium colony is smaller, and transparent circle is apparent, atrichia and sporogenesis.In one embodiment, gemma bar
The optimum temperature of bacterium (sp.W003) is 30 DEG C, optimal pH 7.5.
The bacillus (sp.W024) of one embodiment, preservation time：2017-12-04, preservation address：China, Beijing
City Chaoyang District, Institute of Microorganism, Academia Sinica, deposit number：CGMCC NO.15011.In one embodiment, gemma bar
Bacterium (sp.W024) is in rod-short, and gram staining is negative, and the colonial morphology that 48h is cultivated on inclined-plane is similar to sp.W003,
In；The optimum temperature of bacillus (sp.W024) is 35 DEG C, optimal pH 7.0.
Above-mentioned production bacillus (sp.W003) and bacillus (sp.W024) nutritional requirement are simple, and fermentation time is short；Institute
Enzyme activity with bacillus (sp.W003) and the algin degrading enzyme of bacillus (sp.W024) production is higher；Bacillus used
(sp.W003) and (sp.W024) generate algin degrading enzyme can degrade algin production algin oligosaccharide.
Bacillus (sp.W003) of the present invention and the produced algin degrading enzyme of bacillus (sp.W024) can
Degradation algin, generates algin oligosaccharide, to mass produce algin oligosaccharide.The produced algin degrading enzyme of bacterial strain uses therefor
Enzyme activity is high, and fermentation efficiency is high, is the algin degrading enzyme production bacterial strain of great potential.
Above-mentioned bacillus can apply in the culture medium for producing algin catenase, fermenting and producing algin oligosaccharide.
In one embodiment, the carbon source in culture medium can be sodium alginate, Kelp Powder or kelp paste.
Referring to Fig. 2, a kind of method that algin catenase is prepared using bacillus as described above, including walk as follows
Suddenly：
S110, bacillus is activated, the bacterium solution after activation is applied on slant medium；
In one embodiment, inclined-plane culture based formulas be sodium alginate 10.0-15.0g/L, ammonium sulfate 3.0-8.0g/L,
Magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-4.0g/L, ferrous sulfate 0.01-0.1g/L and agar 15.0-20.0g/L.
Bacillus on S120, picking slant medium, which is inoculated in the container equipped with culture medium, to be cultivated,
It it is 25-35 DEG C in temperature, rotating speed is that culture 12-16h obtains seed liquor to the logarithm middle and later periods on the shaking table of 150-230r/min；
In one embodiment, culture medium formula can be：Sodium alginate 10.0-15.0g/L, ammonium sulfate 3.0-
8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-4.0g/L and ferrous sulfate 0.01-0.1g/L；Or Kelp Powder
20.0-40.0g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-4.0g/L and ferrous sulfate
0.01-0.1g/L；Or kelp paste 200-500g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate
1.0-4.0g/L and ferrous sulfate 0.01-0.1g/L.
S130, it is inoculated with seed liquor in the fermentation tank equipped with enzymatic production culture medium, in 25-35 DEG C of temperature, rotating speed is
150-230r/min, pH ferment 2-4 days under the conditions of being 7-8, obtain algin oligosaccharide zymotic fluid.
In one embodiment, in step s 130, the inoculum concentration of the seed liquor in fermentation tank can be 5%-15%.
In one embodiment, in step s 130, further include into culture medium formula plus metal ion, wherein gold
The addition quality percent by volume for belonging to ion is 0.05%-1.0%.
In one embodiment, metal ion can be potassium ion, magnesium ion or iron ion.
In one embodiment, further include the steps that algin oligosaccharide zymotic fluid product testing.Algin oligosaccharide is sent out
Zymotic fluid centrifuges 5-20min at 8000-12000r/min, collects supernatant, and enzyme activity and HPLC are measured with ultraviolet absorption method
Measure algin oligosaccharide concentration.
Embodiment
Embodiment 1
The screening of bacillus (sp.W003)：
(1) enrichment culture：By kelp, rotten 14d obtains the kelp sample that rots in water, and picking samples are placed in fresh enrichment
In culture medium for 24 hours in 30 DEG C of cultures.
(2) it isolates and purifies：By the bacterium solution after culture with different extension rates 10-1、10-2、10-3、10-4、10-5According to coating
Flat band method is coated on plating medium, and the single bacterium colony of the apparent dissolving circle of selection formation on tablet, uses after 37 DEG C of culture 48h
Conventional microbiological separation method is isolated and purified repeatedly, until obtaining pure bacillus (sp.W003).
Embodiment 2
The screening of bacillus (sp.W024)：
(1) enrichment culture：By kelp, rotten 14d obtains the kelp sample that rots in water, and picking samples are placed in fresh enrichment
In culture medium 48h is cultivated in 30 DEG C.
(2) it isolates and purifies：By the bacterium solution after culture with different extension rates 10-1、10-2、10-3、10-4、10-5According to coating
Flat band method is coated on plating medium, and the single bacterium colony of the apparent dissolving circle of selection formation on tablet, uses after 37 DEG C of culture 48h
Conventional microbiological separation method is isolated and purified repeatedly, until obtaining pure bacillus (sp.W024).
Wherein, the component and proportioning of above-mentioned enrichment culture and the culture medium arrived used in the process of isolating and purifying are as follows：
Enriched medium：Peptone 5.0g/L, yeast extract 10.0g/L, seawater configuration.
Plating medium：Sodium alginate 10.0g/L, ammonium sulfate 5.0g/L, magnesium sulfate 2.0g/L, dipotassium hydrogen phosphate 2.0g/
L, ferrous sulfate 0.01g/L, agar 20.0g/L, tap water configuration.
Embodiment 3
The identification of bacillus (sp.W003) and bacillus (sp.W024)
Bacillus (sp.W003) and (sp.W024) of high yield algin catenase are obtained by screening, bacterial strain uses therefor exists
After growing 48h on primary dcreening operation culture medium, bacillus (sp.W003) periphery of bacterial colonies can form apparent dissolving circle as shown in Fig. 2, gemma
Bacillus (sp.W003) form is in rodlike, Gram-negative (Fig. 4).The bacterium colony that 48h is cultivated on inclined-plane is in shallow white, circle
Shape dimpling, surface is smooth, lawn moistens, bacterium colony is smaller, and transparent circle is apparent, atrichia and sporogenesis.In one embodiment,
The optimum temperature of bacillus (sp.W003) is 30 DEG C, optimal pH 7.5.
The bacillus (sp.W024) of one embodiment, preservation time：2017-12-04, preservation address：China, Beijing
City Chaoyang District, Institute of Microorganism, Academia Sinica, deposit number：CGMCC NO.15011.In one embodiment, gemma bar
It is as shown in Figure 3 that bacterium (sp.W024) periphery of bacterial colonies can form apparent dissolving circle.Bacillus (sp.W024) is in rod-short, gram
Negative staining (Fig. 5).The colonial morphology that 48h is cultivated on inclined-plane is similar to sp.W003, wherein bacillus (sp.W024)
Optimum temperature is 35 DEG C, optimal pH 7.0.
It is bacillus according to thalli morphology and molecular biology identification result (Fig. 8, Fig. 9) Preliminary Identification bacterial strain uses therefor
Belong to, and is named as bacillus (sp.W003) and bacillus (sp.W024).
Embodiment 4
The measurement of bacillus (sp.W003) and bacillus (sp.W024) supernatant of bacteria solution liquid enzymatic activity：
Bacillus (sp.W003) and bacillus (sp.W024) on picking slant medium are inoculated in are equipped with respectively
In the container of culture medium, cultivation temperature is respectively 30 DEG C and 35 DEG C, is placed on the shaking table that rotating speed is 200r/min and cultivates
48h obtains the zymotic fluid of bacterial strain；Fermented supernatant fluid after above-mentioned 200 μ L degermings is mixed with 1.8mL liquid culture mediums,
30 DEG C of incubators are placed in, culture is for 24 hours.After culture, mixture is placed in 100 DEG C of boiling water bath 10min, enzyme is made to inactivate, stopped
Enzyme digestion reaction.Then, the absorbance value with UV spectrophotometer measuring mixed liquor at 235nm.Enzyme activity unit is defined as
Absorbance per minute often adds 0.1 to be an enzyme activity unit (EU), and it is respectively 70.0U/mg and 63.8U/mg to measure enzyme activity.
The component and proportioning of the culture medium arrived used in the process of above-mentioned enzyme assay are as follows：
Culture medium：Sodium alginate 10.0-15.0g/L or Kelp Powder 20.0-40.0g/L or kelp paste 200-500g/
L, ammonium sulfate 5.0g/L, magnesium sulfate 2.0g/L, dipotassium hydrogen phosphate 2.0g/L, ferrous sulfate 0.01g/L.
Embodiment 5
Algin catenase is prepared using bacillus (sp.W003) and bacillus (sp.W024)
By on slant medium bacillus (sp.W003) and bacillus (sp.W024) respectively be inoculated in equipped with production
In the container of enzyme culture medium, cultivation temperature is 30 DEG C, after being placed on the shaking table that rotating speed is 200r/min in culture 16h to logarithm
Phase obtains the seed liquor of bacterial strain.The seed liquor of inoculation 10%, fermentation condition control in the shaking flask equipped with 10.0g/L sodium alginates
System ferments 2 days in 30 DEG C, shaking speed 200r/min of temperature, obtains algin oligosaccharide zymotic fluid, wherein bacillus
(sp.W003) and the produced algin oligosaccharide concentration of bacillus (sp.W024) is respectively 2.70g/L and 2.18g/L.
Embodiment 6
Algin catenase is prepared using bacillus (sp.W003) and bacillus (sp.W024)
By on slant medium bacillus (sp.W003) and bacillus (sp.W024) respectively be inoculated in equipped with production
In the container of enzyme culture medium, cultivation temperature is 30 DEG C, after being placed on the shaking table that rotating speed is 200r/min in culture 16h to logarithm
Phase obtains the seed liquor of bacterial strain.The seed liquor of inoculation 10%, fermentation condition in the 5L fermentation tanks equipped with 30.0g/L Kelp Powders
Control ferments 4 days in 7-8 in 30 DEG C of temperature, speed of agitator 200r/min, pH control, obtains algin oligosaccharide zymotic fluid,
Middle bacillus (sp.W003) and the produced algin oligosaccharide concentration of bacillus (sp.W024) are respectively 3.73g/L and 3.20g/
L。
Several embodiments of the invention above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
Cannot the limitation to the scope of the claims of the present invention therefore be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the guarantor of the present invention
Protect range.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
1. a kind of bacillus, which is characterized in that the bacillus sp.W003 is preserved in Chinese common micro-
Biological inoculum preservation administrative center, address：China, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode：
100101, deposit number is：CGMCC NO.15010.
2. a kind of bacillus, which is characterized in that the bacillus sp.W024 is preserved in Chinese common micro-
Biological inoculum preservation administrative center, address：China, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode：
100101, deposit number is：CGMCC NO.15011.
3. a kind of application of bacillus as claimed in claim 1 or 2 in producing algin catenase.
4. a kind of application for the culture medium producing algin oligosaccharide using fermentation of bacillus as claimed in claim 1 or 2, special
Sign is that the carbon source in the culture medium is sodium alginate, Kelp Powder or kelp paste.
5. a kind of method preparing algin catenase using bacillus as claimed in claim 1 or 2, which is characterized in that packet
Include following steps：
The bacillus is activated, the bacterium solution after activation is applied on slant medium；
Bacillus on slant medium described in picking is inoculated in the container equipped with culture medium and is cultivated, in temperature
It it is 25-35 DEG C, rotating speed is that culture 12-16h obtains seed liquor to the logarithm middle and later periods on the shaking table of 150-230r/min；
The seed liquor is inoculated in the fermentation tank equipped with enzymatic production culture medium, in 25-35 DEG C of temperature, rotating speed 150-
230r/min, pH ferment 2-4 days under the conditions of being 7-8, obtain algin oligosaccharide zymotic fluid.
6. according to the method described in claim 5, it is characterized in that, the inoculum concentration of the seed liquor in the fermentation tank is 5%-
15%.
7. according to the method described in claim 5, it is characterized in that, the inclined-plane culture based formulas is sodium alginate 10.0-
15.0g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-4.0g/L, ferrous sulfate 0.01-
0.1g/L and agar 15.0-20.0g/L.
8. according to the method described in claim 5, it is characterized in that, the culture medium formula is：
Sodium alginate 10.0-15.0g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-
4.0g/L and ferrous sulfate 0.01-0.1g/L；
Or Kelp Powder 20.0-40.0g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-
4.0g/L and ferrous sulfate 0.01-0.1g/L；
Or kelp paste 200-500g/L, ammonium sulfate 3.0-8.0g/L, magnesium sulfate 0.5-3.0g/L, dipotassium hydrogen phosphate 1.0-4.0g/L
And ferrous sulfate 0.01-0.1g/L.
9. according to the method described in claim 8, it is characterized in that, adding metal ion, institute in being formulated to the culture medium
The addition quality percent by volume for stating metal ion is 0.05%-1.0%.
10. according to the method described in claim 9, it is characterized in that, the metal ion be potassium ion, magnesium ion or iron from
Son.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108929859A (en) * | 2018-08-03 | 2018-12-04 | 中国热带农业科学院热带生物技术研究所 | One type bacterial strain of bacillus HB172198 and its application |
| CN109355238A (en) * | 2019-01-08 | 2019-02-19 | 中国科学院烟台海岸带研究所 | A kind of enterobacter cloacae strain and its application in degradation brown alga |
| CN111100825A (en) * | 2020-02-27 | 2020-05-05 | 中国科学院烟台海岸带研究所 | Bacillus and application thereof in industry |
| CN111100827A (en) * | 2020-02-27 | 2020-05-05 | 中国科学院烟台海岸带研究所 | Bacillus capable of producing high-activity alginate lyase and application thereof |
| CN111100829A (en) * | 2020-02-27 | 2020-05-05 | 中国科学院烟台海岸带研究所 | Bacterial strain capable of degrading specific fragment brown algae polysaccharide |
| CN112592914A (en) * | 2020-12-31 | 2021-04-02 | 青岛海大生物集团有限公司 | Special green alga polysaccharide lyase and production process thereof |
| CN112694995A (en) * | 2021-01-11 | 2021-04-23 | 山东省科学院生物研究所 | Alginate degrading bacterium Bacillus HZ-1 and application thereof in holothurian culture |
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| CN108929859A (en) * | 2018-08-03 | 2018-12-04 | 中国热带农业科学院热带生物技术研究所 | One type bacterial strain of bacillus HB172198 and its application |
| CN108929859B (en) * | 2018-08-03 | 2021-07-20 | 中国热带农业科学院热带生物技术研究所 | Bacillus-like strain HB172198 and application thereof |
| CN109355238A (en) * | 2019-01-08 | 2019-02-19 | 中国科学院烟台海岸带研究所 | A kind of enterobacter cloacae strain and its application in degradation brown alga |
| CN111100825A (en) * | 2020-02-27 | 2020-05-05 | 中国科学院烟台海岸带研究所 | Bacillus and application thereof in industry |
| CN111100827A (en) * | 2020-02-27 | 2020-05-05 | 中国科学院烟台海岸带研究所 | Bacillus capable of producing high-activity alginate lyase and application thereof |
| CN111100829A (en) * | 2020-02-27 | 2020-05-05 | 中国科学院烟台海岸带研究所 | Bacterial strain capable of degrading specific fragment brown algae polysaccharide |
| CN111100825B (en) * | 2020-02-27 | 2021-05-11 | 中国科学院烟台海岸带研究所 | Bacillus and application thereof in industry |
| CN111100829B (en) * | 2020-02-27 | 2021-07-13 | 中国科学院烟台海岸带研究所 | Bacterial strain capable of degrading specific fragment brown algae polysaccharide |
| CN112592914A (en) * | 2020-12-31 | 2021-04-02 | 青岛海大生物集团有限公司 | Special green alga polysaccharide lyase and production process thereof |
| CN112592914B (en) * | 2020-12-31 | 2024-05-28 | 青岛海大生物集团股份有限公司 | Special chlorella polysaccharide lyase and production process thereof |
| CN112694995A (en) * | 2021-01-11 | 2021-04-23 | 山东省科学院生物研究所 | Alginate degrading bacterium Bacillus HZ-1 and application thereof in holothurian culture |
| CN112694995B (en) * | 2021-01-11 | 2022-07-26 | 山东省科学院生物研究所 | Algin degrading bacterium Bacillus HZ-1 and application thereof in holothurian culture |
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