CN109709334A - A method of analysis oil plant dregs of rice mineral acts in fermentation of bacillus production iraq subtilis actinomycin A - Google Patents
A method of analysis oil plant dregs of rice mineral acts in fermentation of bacillus production iraq subtilis actinomycin A Download PDFInfo
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- CN109709334A CN109709334A CN201811417088.1A CN201811417088A CN109709334A CN 109709334 A CN109709334 A CN 109709334A CN 201811417088 A CN201811417088 A CN 201811417088A CN 109709334 A CN109709334 A CN 109709334A
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Abstract
The invention belongs to field of microbial fermentation, disclose a kind of method that analysis oil plant dregs of rice mineral acts in fermentation of bacillus production iraq subtilis actinomycin A, respectively using the extraction albumen of the oil plant dregs of rice or the oil plant dregs of rice as nitrogen source, using glucose as carbon source, respectively under the conditions of adding or not adding two kinds of inorganic salts, it is mixed with water as culture medium, protein production iraq subtilis actinomycin A is extracted using the bacillus amyloliquefaciens fermentation oil plant dregs of rice or the oil plant dregs of rice after sterilizing, records the yield of iraq subtilis actinomycin A respectively；Compare in the first step when extracting albumen as nitrogen source using the oil plant dregs of rice or the oil plant dregs of rice respectively, the yield of corresponding iraq subtilis actinomycin A, analysis oil plant dregs of rice mineral produces the effect in iraq subtilis actinomycin A in fermentation of bacillus.As a result prove: oil plant dregs of rice mineral, which produces iraq subtilis actinomycin A in fermentation of bacillus, has facilitation, can exercise inorganic salt action in microbiological culture media.
Description
Technical field
The present invention relates to microbial fermentation and analysis field, in particular to analysis oil plant dregs of rice etc. contain the agricultural and sideline of different component
Effect of the product mineral in other microbial metabolic products.
Background technique
The oil plant dregs of rice are the by-products after the processing such as rapeseed, sesame, are rich in a variety of nutriments, especially protein content
About 35%~45%, therefore nitrogen source is used frequently as microbial fermentation.But also containing many other in the oil plant dregs of rice
Substance, such as cellulose, minerals, phenolic substances, grease, mineral due to can not separation and Extraction, to being at present
There are no the methods that analysis oil plant dregs of rice mineral acts in microbial fermentation.
Summary of the invention
In order to solve at present to be asking for the method acted in microbial fermentation there are no analysis oil plant dregs of rice mineral
Topic, the present invention provides the sides that a kind of analysis oil plant dregs of rice mineral acts in fermentation of bacillus production iraq subtilis actinomycin A
Method, the discovery oil plant dregs of rice act not only as nitrogen source use, and minerals therein equally also can replace needed for microbial fermentation
Inorganic salts.
The present invention be solve the problems, such as it is set forth above used by technical solution are as follows:
The method that analysis oil plant dregs of rice mineral acts in fermentation of bacillus production iraq subtilis actinomycin A, the method
Are as follows:
The first step, respectively using the extraction albumen of the oil plant dregs of rice or the oil plant dregs of rice as nitrogen source, using glucose as carbon source, point
Not under the conditions of adding or not adding two kinds of inorganic salts, is mixed with water as culture medium, bacillus amyloliquefaciens are utilized after sterilizing
Ferment the oil plant dregs of rice or oil plant dregs of rice extraction protein production iraq subtilis actinomycin A, records the yield of iraq subtilis actinomycin A respectively；
Second step compares in the first step when extracting albumen as nitrogen source using the oil plant dregs of rice or the oil plant dregs of rice respectively, and corresponding she is withered
The yield of lavendustin A, effect of the analysis oil plant dregs of rice mineral in fermentation of bacillus production iraq subtilis actinomycin A.
According to the above scheme, respectively using the extraction albumen of the oil plant dregs of rice or the oil plant dregs of rice as nitrogen source when, it is effective in the two
Protein content is equal, so as to comparative analysis.Wherein, the concentration of the oil plant dregs of rice in the medium is 90-150g/L, and the oil plant dregs of rice extract
The concentration of albumen in the medium is 40-60g/L.
According to the above scheme, the carbon source can be glucose, molasses, glycerol, sucrose or cornstarch etc., and carbon source is being trained
Supporting concentration in base is 40-80g/L.
According to the above scheme, the inorganic salts mainly select the inorganic salts of the elements such as potassium, phosphorus, magnesium, manganese.In the present invention preferably without
Machine salt is K2HPO4·3H2O、MgSO4·7H2O、MnSO4·H2The mixture of O, concentration is most preferably three in the medium
0.5g/L K2HPO4·3H2O,0.5g/L MgSO4·7H2O,0.005g/L MnSO4·H2O。
According to the above scheme, the condition of the sterilizing are as follows: 121 DEG C sterilizing 20-30 minutes.
According to the above scheme, the bacillus amyloliquefaciens CX-20 (Bacillus amyloliquefaciens CX-20),
It is preserved in China typical culture collection center on November 14th, 2018, abbreviation CCTCC, preservation address: China, Wuhan,
Wuhan University, deposit number are CCTCC NO:M 2018794.
According to the above scheme, the additive amount of the bacillus amyloliquefaciens is the 4-6% of culture medium total volume.
According to the above scheme, the condition of the fermentation are as follows: be stirred and ferment at 28-37 DEG C using airlift fermentor
36-72 hours, it is during which not necessarily to feed supplement, without change fermentation condition.
According to the above scheme, the oil plant dregs of rice mainly include rapeseed meal, dregs of beans or peanut meal etc..
According to the above scheme, the method that the oil plant cake protein extracts is preferably alkali extraction-acid precipitation.
Compared with prior art, the beneficial effects of the present invention are:
(1) the present invention provides a kind of analysis oil plant dregs of rice minerals in fermentation of bacillus production iraq subtilis actinomycin A
The method of effect, although up to the present there are no from the oil plant dregs of rice extract minerals method, can not directly, independent analysis this
The effect of a little minerals, but the present invention analyzes the effect of oil plant dregs of rice mineral from the negative in two ways, one is logical
It crosses and adds additional inorganic salts and analyse whether there is inhibiting effect as nitrogen source to the oil plant dregs of rice, after all in microbial fermentation, culture
It can inhibit microbial fermentation containing excessive inorganic salts in base, and then influence the yield of microbial metabolic products；Another kind is logical
Albumen in the extraction oil plant dregs of rice is crossed, most of minerals in the oil plant dregs of rice are removed, then in the case where not adding inorganic salts, analysis
Whether the oil plant dregs of rice can have negative effect to microbial secondary metabolite when extracting albumen as nitrogen source, because in microbial fermentation
In, it equally can also inhibit microbial fermentation when very few containing inorganic salts amount in culture medium, and then influence microbial metabolic products
Yield.
(2) present invention goes out oil plant dregs of rice mineral in fermentation of bacillus production iturin by Technical Analysis
Effect in A, i.e. the oil plant dregs of rice act not only as nitrogen source use, and minerals therein equally also can replace microbial fermentation institute
The inorganic salts needed, although most of researcher thinks that the minerals in the oil plant dregs of rice have effect, up to the present not
Positive evidence shows effect of these minerals for microbial fermentation, therefore the present invention is to promoting the oil plant dregs of rice in microbial fermentation
Play a significant role in terms of the understanding of value.
Detailed description of the invention
Fig. 1 is that different nitrogen sources, (rapeseed meal, rapeseed meal extract albumen, albumen to bacillus amyloliquefaciens of embodiment of the present invention CX-20
Peptone, yeast powder and ammonium nitrate) in addition or influence in the case where do not add inorganic salts to Iturin A fermentation.
Wherein, each label meaning is as follows: 1, adding inorganic salts in 90g/L culture medium；2, nothing is not added in 90g/L rapeseed meal culture medium
Machine salt；3, add inorganic salts in 40g/L rapeseed protein culture medium；Inorganic salts are not added in 4:40g/L rapeseed protein culture medium； 5,
Add inorganic salts in 22g/L peptone culture medium；6, inorganic salts are not added in 22g/L peptone culture medium；7,30g/L yeast powder culture
Add inorganic salts in base；8, inorganic salts are not added in 30g/L yeast powder culture medium；9, add inorganic salts in 8g/L ammonium nitrate culture medium；10,
Inorganic salts are not added in 8g/L ammonium nitrate culture medium.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment and attached drawing content that the present invention is furture elucidated, but this
Invention is not limited solely to the following examples.
In following embodiments, used bacillus amyloliquefaciens CX-20 (Bacillus amyloliquefaciens
CX-20), it is preserved in China typical culture collection center on November 14th, 2018, abbreviation CCTCC, preservation address: in
State, Wuhan, Wuhan University, deposit number are CCTCC NO:M 2018794.
Embodiment
A method of analysis rapeseed meal mineral produces iraq subtilis actinomycin A effect in fermentation of bacillus, and this method is specific
Are as follows:
(1) rapeseed meal, glucose and water are mixed, finally make the concentration 90g/L of rapeseed meal in water, glucose it is dense
Degree is 80g/L, is adding or do not adding inorganic salts (0.5g/L K2HPO4·3H2O,0.5g/L MgSO4·7H2O,0.005g/L
MnSO4·H2O) in two kinds of culture mediums, 121 DEG C sterilize 20 minutes；After cooling, it is added thereto according to the 5% of fermentating liquid volume
Then bacillus amyloliquefaciens CX-20 carries out shake culture under conditions of 28 DEG C, 220rpm, be during which not necessarily to feed supplement, without
Change fermentation condition；Fermentation 72 hours measures the content of iraq subtilis actinomycin A in fermentation liquid.
(2) albumen in rapeseed meal is extracted using the method that alkali soluble acid sinks, rapeseed meal is extracted into albumen, glucose and water and is mixed
It closes, rapeseed meal is finally made to extract the concentration 40g/L (protein content with 90g/L rapeseed meal in above-mentioned steps (1) of albumen in water
It is equal), the concentration of glucose is 80g/L, is adding or do not adding inorganic salts (0.5g/L K2HPO4·3H2O,0.5g/L
MgSO4·7H2O,0.005g/L MnSO4·H2O) in two kinds of culture mediums, 121 DEG C sterilize 20 minutes；After cooling, according to fermentation
Bacillus amyloliquefaciens CX-20 is added in liquid product 5% thereto, then carries out shake culture under conditions of 28 DEG C, 220rpm,
Period is not necessarily to feed supplement, without change fermentation condition；Fermentation 72 hours measures the content of iraq subtilis actinomycin A in fermentation liquid.
Comparative example
Have an impact to further judge whether inorganic salts produce iraq subtilis actinomycin A to fermentation of bacillus, the present invention is same
Other three kinds of nitrogen sources, including peptone (animal origin), yeast powder (plant origin) and ammonium nitrate (inorganic nitrogen) are analyzed, most
Under suitable concentration (respectively 22g/L, 30g/L, 8g/L), influence whether addition to iraq subtilis actinomycin A yield of inorganic salts.It is right
Ratio other conditions are identical as embodiment, the difference is that only that the concentration of nitrogen source and nitrogen source is different.
As shown in Figure 1, relative to addition inorganic salts in the case where Iturin A yield, the case where not adding inorganic salts
Under, the yield of Iturin A is all substantially reduced in these three nitrogen sources of comparative example: the wherein culture medium in peptone as nitrogen source
In, the yield of Iturin A is reduced to 0.05g/L by 1.07g/L；In culture medium of the yeast powder as nitrogen source, Iturin
The yield of A is reduced to 0.23g/L by 0.56g/L；In culture medium of the ammonium nitrate as nitrogen source, the yield of Iturin A by
0.05g/L is reduced to 0g/L.
When similarly, using rapeseed meal extraction albumen as nitrogen source, inorganic salts do not add the yield for equally also making Iturin A
0.24g/L is reduced to by 1.48 g/L.But when using rapeseed meal as nitrogen source, the yield of Iturin A when adding inorganic salts
For 1.15 g/L, the yield of Iturin A has been increased to 1.25g/L instead when without adding inorganic salts.
When therefore, using rapeseed meal as nitrogen source, under conditions of not adding inorganic salts, bacillus amyloliquefaciens produce her withered grass bacterium
The amount of plain A is higher；And when rapeseed meal extracts albumen as nitrogen source, in the case where not adding inorganic salts, bacillus amyloliquefaciens
The amount for producing iraq subtilis actinomycin A significantly reduces；The two comprehensive results obviously prove that rapeseed meal mineral is produced in fermentation of bacillus
Iraq subtilis actinomycin A has facilitation, can exercise inorganic salt action in microbiological culture media.Not only using this method of the present invention
The effect that rapeseed meal mineral produces iraq subtilis actinomycin A in fermentation of bacillus can be analyzed, other oil meals etc. can also be analyzed
Effect of the agricultural byproduct mineral in other microbial metabolic products containing different component.
The above is only a preferred embodiment of the present invention, it is noted that come for those of ordinary skill in the art
It says, without departing from the concept of the premise of the invention, several modifications and variations can also be made, these belong to of the invention
Protection scope.
Claims (10)
1. the method that analysis oil plant dregs of rice mineral acts in fermentation of bacillus production iraq subtilis actinomycin A, it is characterised in that
The method are as follows:
The first step is existed using the extraction albumen of the oil plant dregs of rice or the oil plant dregs of rice as nitrogen source using glucose as carbon source respectively respectively
Under the conditions of adding or not adding two kinds of inorganic salts, is mixed with water as culture medium, fermented after sterilizing using bacillus amyloliquefaciens
The oil plant dregs of rice or the oil plant dregs of rice extract protein production iraq subtilis actinomycin A, record the yield of iraq subtilis actinomycin A respectively；
Second step compares in the first step when extracting albumen as nitrogen source using the oil plant dregs of rice or the oil plant dregs of rice respectively, her corresponding withered grass bacterium
The yield of plain A, effect of the analysis oil plant dregs of rice mineral in fermentation of bacillus production iraq subtilis actinomycin A.
2. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that effective in the two when respectively using the extraction albumen of the oil plant dregs of rice or the oil plant dregs of rice as nitrogen source
Protein content is equal.
3. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that the concentration of the oil plant dregs of rice in the medium is 90-150 g/L, and the oil plant dregs of rice extract albumen in culture medium
In concentration be 40-60 g/L.
4. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that concentration is 40-80 g/L to carbon source in the medium.
5. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that the inorganic salts are mainly selected from the inorganic salts of potassium, phosphorus, magnesium, manganese.
6. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that the inorganic salts are K2HPO4·3H2O、 MgSO4·7H2O、MnSO4·H2The mixture of O, three
Concentration is 0.5 g/L K to person in the medium2HPO4·3H2O, 0.5 g/L MgSO4·7H2O, 0.005 g/L MnSO4·
H2O。
7. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that the bacillus amyloliquefaciens are CX-20, are preserved in Chinese Typical Representative on November 14th, 2018
Culture collection, abbreviation CCTCC, preservation address: China, Wuhan, Wuhan University, deposit number are CCTCC NO:M
2018794。
8. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that the additive amount of the bacillus amyloliquefaciens is the 4-6% of culture medium total volume.
9. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that the condition of the fermentation are as follows: be stirred and ferment at 28-37 DEG C using airlift fermentor
36-72 hours, it is during which not necessarily to feed supplement, without change fermentation condition.
10. analysis oil plant dregs of rice mineral according to claim 1 is made in fermentation of bacillus production iraq subtilis actinomycin A
Method, it is characterised in that the oil plant dregs of rice mainly include rapeseed meal, dregs of beans or peanut meal；The carbon source can be grape
Sugar, molasses, glycerol, sucrose or cornstarch.
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| CN114807274A (en) * | 2022-04-11 | 2022-07-29 | 海南康植肽生物科技有限公司 | Efficient preparation method and application of bacillomycin D |
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