CN113403353B - Culture medium and method for producing undecyl prodigiosin - Google Patents
Culture medium and method for producing undecyl prodigiosin Download PDFInfo
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Abstract
The invention discloses a culture medium and a method for producing undecyl prodigiosin, and the method takes collagen polypeptide as a precursor source for fermentation production of undecyl prodigiosin, and relates to the technical field of biomass conversion and microbial fermentation. The weight average molecular weight of the collagen polypeptide is 0.2-20 k Da, the collagen polypeptide with the weight average molecular weight can promote the growth of streptomyces, induce and stimulate the production of undecyl prodigiosin, greatly shorten the fermentation time and improve the fermentation efficiency. The collagen polypeptide is more suitable for producing undecyl prodigiosin by fermentation of streptomycete than soybean meal and malt extract powder. The content of precursor amino acid in the meat protein polypeptides of rabbits, cows, chickens and ducks is higher than that of soybean meal and malt extract powder and lower than that of collagen polypeptide, so that although the 4 meat protein polypeptides can produce undecyl prodigiosin, compared with the collagen polypeptide, the fermentation time is greatly prolonged, and the yield is relatively low. Therefore, the collagen polypeptide has good application prospect in the fermentation production of the undecyl prodigiosin.
Description
Technical Field
The invention belongs to the technical field of biomass conversion and microbial fermentation, and particularly relates to a culture medium and a method for producing undecyl prodigiosin.
Background
Undecyl Prodigiosin (UP) is an antibiotic with multiple biological activities such as antibiosis, antitumor and immunosuppression, and has huge application prospects in the pharmaceutical industry, so the Undecyl Prodigiosin (UP) is widely concerned by people. UP may be derived from Streptomyces coelicolor: (Streptomyces coelicolor) Streptomyces lividans (A)Streptomyces coelescens) And Serratia marcescens: (Serratia marcescens) Synthesized by fermentation. WhileS. marcescensIs a conditional pathogen, has relatively low safety, and can cause urinary tract and lung infection, septicemia and other problems when the immunity of a human body is low. Therefore, UP is currently produced mainly by fermentation of streptomyces microorganisms.
Based on a specific production strain, the addition of the UP precursor is a method which is simple to operate and flexible for improving the UP production efficiency, and is very important for the UP production. Biosynthetic precursors of UP mainly include Proline (Pro), Glycine (Gly), Serine (Ser), acetyl-coa and malonyl-coa. Acetyl coenzyme A and malonyl coenzyme A are widely available and can be easily obtained from common substrates. UP and the structures of precursor amino acids and intermediates thereof are shown in figure 1, Pro, Gly and other precursor amino acids are very important for synthesis of UP, but the content of the precursor amino acids in conventional fermentation substrates is limited, and the cost for industrial production of UP is high by directly adding free amino acids, so that new UP precursor amino acid resources are urgently needed to be searched and developed.
Disclosure of Invention
The invention aims to provide a culture medium for producing undecyl prodigiosin, and aims to provide abundant nutrient substances and precursor amino acids for growth of microorganisms and production of undecyl prodigiosin by utilizing collagen polypeptides so as to develop precursor resources for synthesizing undecyl prodigiosin, improve the yield of undecyl prodigiosin and improve the fermentation efficiency.
The invention also aims to provide a method for producing undecyl prodigiosin by utilizing collagen polypeptide fermentation, develop the characteristic biotransformation function of the collagen polypeptide, expand the application of the collagen polypeptide in the field of antibiotic production and realize the resource utilization and high-value utilization of the collagen polypeptide.
The technical problem to be solved by the invention is realized by adopting the following technical scheme.
In a first aspect, the invention adopts the technical scheme that: a culture medium for the production of undecyl prodigiosin, the medium comprising collagen polypeptides having a weight average molecular weight of 0.2-20 k Da. Preferably, the collagen polypeptide has a weight average molecular weight of 0.3k Da to 5k Da.
Further, the addition amount of the collagen polypeptide in the culture medium is 0.1-80 g/L.
Further, the collagen polypeptide is prepared by hydrolyzing collagen by at least one of enzymatic hydrolysis, alkaline hydrolysis, acid hydrolysis or high-temperature hydrolysis.
Further, the inorganic salt component in the medium comprises at least one of potassium salt, sodium salt and magnesium salt.
Further, the potassium salt is at least one selected from the group consisting of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium chloride and potassium sulfate; the sodium salt is selected from at least one of sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride and sodium sulfate; the magnesium salt is selected from at least one of magnesium hydrogen phosphate, magnesium chloride and magnesium sulfate.
In a second aspect, the invention adopts the technical scheme that: a method for producing undecyl prodigiosin by utilizing collagen polypeptide fermentation, which comprises the step of inoculating streptomyces with the capability of producing undecyl prodigiosin into the culture medium of any one of the first aspect to carry out the production fermentation of the undecyl prodigiosin.
Further, the streptomyces having an undecyl prodigiosin producing ability comprises streptomyces coelicolorStreptomyces coelicolorAnd/or Streptomyces lividansStreptomyces coelescens。
Further, the inoculation amount of the streptomyces is 0.5% -10%.
Further, the fermentation temperature is 26-34 ℃, the pH is 6-9, and the rotation speed is 150-250 rpm.
Further, the method also comprises a step of separating fermentation liquor obtained by fermentation production after the production and fermentation of the undecyl prodigiosin.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a culture medium for producing undecyl prodigiosin, which comprises collagen polypeptides with the weight-average molecular weight of 0.2-20 k Da. The collagen polypeptide has special amino acid composition, contains a large amount of undecyl prodigiosin precursor amino acid, can simplify the components of a fermentation medium, and reduces the production cost of the antibiotic. The embodiment of the invention provides a method for producing undecyl prodigiosin by utilizing collagen polypeptide fermentation, which comprises the step of inoculating streptomyces with the capability of producing undecyl prodigiosin into the culture medium containing the collagen polypeptide to carry out the fermentation production of the antibiotic. The inventor creatively discovers that: the collagen polypeptide with the weight-average molecular weight can promote the growth of microorganisms, improve the yield of the undecyl prodigiosin, greatly shorten the fermentation time and improve the fermentation efficiency of the undecyl prodigiosin.
It should be noted that the collagen polypeptide provided in the embodiments of the present invention can increase the yield of undecyl prodigiosin, mainly because the collagen polypeptide not only can provide abundant nutrition for the growth of microorganisms, but also can provide necessary precursor amino acids for the production of undecyl prodigiosin. The collagen polypeptide contains a large amount of precursor amino acids (glycine, proline and serine) of the undecyl prodigiosin, and can induce and stimulate the microbial fermentation to produce the undecyl prodigiosin, thereby improving the production efficiency of the antibiotic.
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In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention, and therefore should not be considered as limiting the scope, and those skilled in the art can also obtain other related drawings based on the drawings without inventive efforts.
FIG. 1 is the structures of UP and its precursor amino acids and intermediates;
FIG. 2 shows the phenotypic characteristics and cell morphology of Streptomyces SLL-523;
FIG. 3 shows the effect of collagen polypeptide on fermentation of Streptomyces CGMCC 4.7172 to produce undecyl prodigiosin;
FIG. 4 is the content of undecyl prodigiosin precursor amino acids in collagen polypeptides and 2 plant biomass;
FIG. 5 shows the effect of collagen polypeptide on fermentation of Streptomyces SLL-523 to produce undecyl prodigiosin;
FIG. 6 is a graph showing the effect of collagen polypeptides on the intracellular precursor amino acid content of Streptomyces SLL-523;
FIG. 7 shows the effect of collagen polypeptide and 4 meat protein polypeptides on fermentation of Streptomyces CGMCC 4.7172 to produce undecyl prodigiosin;
FIG. 8 shows the content of undecyl prodigiosin precursor amino acids in 4 meat protein polypeptides.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used are not indicated by the manufacturer, and are all conventional products available commercially.
It is understood that the operating conditions for the fermentation culture may be further varied depending on the microorganism having the ability to produce undecylprodigiosin, so as to achieve the optimum culture effect. The choice of these operating conditions is within the routine discretion of the skilled person.
The following provides a specific description of the culture medium and method for producing undecylprodigiosin according to the examples of the present invention.
The embodiment of the invention provides a culture medium for producing undecyl prodigiosin, which comprises collagen polypeptide, wherein the weight average molecular weight of the collagen polypeptide is 0.2-20 k Da, and preferably, the weight average molecular weight of the collagen polypeptide is 0.3-5 k Da. Collagen has the characteristics of reproducibility, easy acquisition and special amino acid composition, but is not easy to be absorbed and utilized by cells due to compact structure, larger molecular weight and lower solubility. Compared with collagen and gelatin, the collagen polypeptide has the excellent properties of lower relative viscosity, smaller molecular weight, higher free amino group content, larger solubility and the like.
The inventors have creatively discovered that a collagen polypeptide with a particular amino acid composition can serve as the sole carbon and nitrogen source for streptomyces growth and antibiotic production. The undecyl prodigiosin is produced by adopting the fermentation of the collagen polypeptide with the weight-average molecular weight range of 0.2-20 k Da, and continuous tests and researches find that the collagen polypeptide with the weight-average molecular weight range can obviously promote the growth of microorganisms and improve the yield of undecyl prodigiosin when being used for the fermentation of the microorganisms undecyl prodigiosin. Notably, the collagen polypeptides provided in the examples of the present application can also shorten the fermentation time and improve the fermentation efficiency.
The collagen polypeptide provided by the embodiment of the invention is derived from collagen which is one of the largest renewable animal biomass resources in the world. The collagen is widely present in skin, bone, tendon, ligament, blood vessel and other parts of animals, and can be conveniently extracted by adopting a conventional method. Collagen polypeptide is a hydrolysate of collagen, and is more easily utilized by microorganisms. The collagen polypeptide has special amino acid composition and contains a large amount of glycine, proline and serine. In fact, undecyl prodigiosin is an important antibiotic with a three-pyrrole ring backbone structure, each pyrrole ring contains a N from glycine, proline and serine, respectively, and thus these three amino acids are essential precursors for the biosynthesis of undecyl prodigiosin. In particular, proline, as the initial precursor of undecyl prodigiosin, contributes its pyrrole ring into the tripyrrole backbone structure of undecyl prodigiosin. Therefore, the collagen polypeptide can provide abundant precursor amino acids for the production of the undecyl prodigiosin.
The collagen polypeptide is prepared by at least one hydrolysis method of acid hydrolysis, alkali hydrolysis, high-temperature hydrolysis and enzyme hydrolysis; preferably, the collagen polypeptide is prepared by hydrolyzing collagen by an enzymatic hydrolysis method. Generally, any conventional hydrolysis method is suitable for preparing the collagen polypeptide.
It is necessary to supplement the processes of acid hydrolysis, alkaline hydrolysis, high temperature hydrolysis and enzymatic hydrolysis to the existing well-established processes, which can be referred to in the prior art.
It should be added that the collagen polypeptide may be obtained from commercially available products, and is not limited to the preparation method mentioned in the examples of the present invention.
The features and properties of the present invention are described in further detail below with reference to examples.
In the following examples, the streptomyces biomass was measured by dry cell method, the yield of undecyl prodigiosin was measured by high performance liquid chromatography, and the content of specific amino acids in various biomass and the content of intracellular amino acids in streptomyces were measured by amino acid analyzer.
In the following examples, the collagen polypeptide is commercially available, and the hydrolysis method is not specifically limited, butThe weight average molecular weight of the collagen polypeptide is required to satisfy the requirement. Streptomyces (I), (II)Streptomyces coelicolor CGMCC 4.7172) purchased from China general microbiological culture Collection center. Streptomyces (I), (II)Streptomyces coelescens SLL-523), screening and preservation of Sichuan university college of light industry science and engineering, wherein the 16s rDNA sequence of the streptomycete SLL-523 is shown as a sequence table SEQ ID NO: 1.
Example 1
Testing collagen polypeptide pairsS. coelicolorThe growth effect is specifically as follows: (1) control group: liquid culture medium: 4 g/L glucose, 4 g/L yeast extract powder, 10 g/L malt extract powder, 0.05 g/L K 2 HPO 4 ,8.0 g/L NaH 2 PO 4 ,0.02 g/L MgCl 2 . (2) Experimental groups: collagen polypeptides with the weight average molecular weight of 10 kDa of 30 g/L, 40 g/L and 50 g/L are respectively added into a control group culture medium, and are numbered as an experimental group 1-1, an experimental group 1-2 and an experimental group 1-3. To be provided withS. coelicolor CGMCC 4.7172 is experimental strain, and is prepared by activating and proliferating CGMCC 4.7172 strain by 8% (8%)v/v) The inoculation amount of (A) is inoculated into each culture medium, the fermentation temperature is 30 ℃, the rotating speed is 200 r/min, and the initial pH is 6.0. The fermentation time was 3 d, and the dry cell weight of each medium at 2d and 3 d was measured, and the results are shown in Table 1.
TABLE 1 Effect of collagen Polypeptides on the growth of CGMCC 4.7172 Strain
As can be seen from Table 1, the CGMCC 4.7172 strain has a significantly higher dry cell weight in the experimental group with collagen polypeptide added than in the control group without collagen polypeptide added, and the dry cell weight increases with the addition of collagen polypeptide. Therefore, the collagen polypeptide can promote the growth of the strain.
Example 2
Testing collagen polypeptide pairsS. coelescensThe growth effect is specifically as follows: (1) control group: 4 g/L glucose, 4 g/L yeast extract powder, 10 g/L malt extract powder, 0.25 g/L KH 2 PO 4 ,0.15 g/L NaCl,2.0 g/L MgSO 4 . (2) Experimental groups: collagen polypeptides with the weight average molecular weight of 15 kDa and the numbers of experimental group 2-1, experimental group 2-2 and experimental group 2-3 are respectively added into the culture medium of the control group, wherein the collagen polypeptides are 1 g/L, 10 g/L and 20 g/L. To be provided withS. coelescens SLL-523 is an experimental strain, and the phenotypic characteristics and cell morphology of the strain are shown in figure 2. After activation and proliferation of SLL-523 strain, 2%, (v/v) The inoculation amount of the strain is inoculated into each culture medium, the fermentation temperature is 34 ℃, the rotating speed is 180 r/min, and the initial pH is 8.0. The fermentation time was 5 days, and the dry cell weight of each medium was measured, and the results are shown in Table 2.
As can be seen from FIG. 2, after the strain SLL-523 was cultured on a plate medium for a certain period of time, the surface started to become dusty, spores began to be produced, and the color of the spores changed from white to gray as time passed. The hypha of the SLL-523 strain is observed by SEM, obvious aerial hypha and spore filament can be seen, the spores are arranged in a chain shape, the length of the spores is 1-2 mu m, and the hypha has the typical hypha morphological characteristics of streptomyces. Uploading the 16S rDNA sequence of the strain to NCBI and EzbioCluod databases for Blast comparison analysis, and finding out that the strain with the highest homology with the strain isStreptomyces coelescensDSM 40421 (AF503496), with 100.00% similarity.
TABLE 2 Effect of collagen Polypeptides on the growth of SLL-523 Strain
As shown in Table 2, the SLL-523 strain showed a significantly higher dry cell weight in the collagen polypeptide-containing test group than in the control group to which no collagen polypeptide was added, and the dry cell weight increased with the increase in the amount of collagen polypeptide added. It can be seen that the collagen polypeptide can also promote the growth of the strain.
Example 3
The influence of the collagen polypeptide on the production of undecyl prodigiosin by fermenting streptomyces CGMCC 4.7172 is tested, and the method specifically comprises the following steps: (1) control group: minimal Medium (MM): 0.5 g/L KH 2 PO 4 ,1.5 g/L Na 2 HPO 4 ,1.0 g/L NaCl,0.1 g/L MgSO 4 . (2) Experimental group: 40 g/L of soybean meal, malt extract powder and collagen polypeptide with weight average molecular weight of 3k Da, which are numbered as 4SP, 4ME and 4CP culture medium, are added into MM culture medium respectively. Activating and proliferating strain CGMCC 4.7172 with CGMCC 4.7172 as experimental strain, centrifuging to remove supernatant, washing with sterile physiological saline, and suspending in an amount of 1% (in)v/v) The inoculation amount of (2) is inoculated into each culture medium, the fermentation temperature is 32 ℃, the rotating speed is 220 r/min, and the initial pH is 7.5. The fermentation time was 2d, the yield of undecyl prodigiosin in each medium was measured, and the contents of proline, glycine and serine in 3 kinds of biomass, soybean meal, malt extract meal and collagen polypeptide were analyzed, and the results are shown in fig. 3 and 4.
As can be seen from FIG. 3, the concentration of undecyl prodigiosin after 2d fermentation of the strain CGMCC 4.7172 in MM medium with collagen polypeptides introduced therein was over 500. mu.g/L, while the presence of undecyl prodigiosin was not detected in 4SP and 4ME medium.
As can be seen from FIG. 4, the content of 3 undecyl prodigiosin precursor amino acids, proline, glycine and serine, in the collagen polypeptide was significantly higher than that of the soybean meal and the malt extract meal. Therefore, the collagen polypeptide rich in precursor amino acid is more suitable for producing the undecyl prodigiosin by the fermentation of the strain CGMCC 4.7172 than soybean meal and malt extract powder.
Example 4
The method is used for testing the influence of the collagen polypeptide on the fermentation of the streptomyces SLL-523 strain to produce the undecyl prodigiosin, and comprises the following steps: (1) control group: MM medium. (2) Experimental groups: 20 g/L malt extract powder and collagen polypeptide with weight average molecular weight of 2k Da, which are respectively encoded as 2ME and 2CP culture medium, are added into MM culture medium. Activating and proliferating SLL-523 strain as experimental strain, centrifuging to remove supernatant, washing with sterile physiological saline, and suspending the strain at a ratio of 5% (5%)v/v) The inoculation amount of the strain is inoculated into each culture medium, the fermentation temperature is 28 ℃, the rotating speed is 250 r/min, and the initial pH is 7.0. The fermentation time was 1 d, the yield and dry cell weight of undecyl prodigiosin in each medium were measured, and the contents of intracellular proline, glycine and serine were analyzed, and the results are shown in FIGS. 5 and 6. Intracellular amino acid content is expressed in% Dry Cell Weight (DCW)Shown in the figure.
As shown in FIG. 5, the SLL-523 strain had a dry cell weight of 0.31 g/L after being cultured in MM medium for 1 day, while the dry cell weight was increased to 0.71 g/L and 1.19 g/L after addition of malt extract and collagen polypeptide, respectively. Therefore, the collagen polypeptide can promote the growth of cells. And 2CP Medium had an undecyl prodigiosin concentration of about 600. mu.g/L at 1 d, while the presence of UP was not detected in both MM and 2 ME. This indicates that the collagen polypeptide can increase the yield of undecyl prodigiosin of SLL-523 strain.
As can be seen from FIG. 6, the contents of intracellular proline, glycine and serine of SLL-523 strain in the 2CP medium were all higher than those in the 2ME and MM media. Therefore, the collagen polypeptide can greatly improve the content of the intracellular undecyl prodigiosin precursor amino acid, thereby inducing and stimulating the bacterial strain to produce undecyl prodigiosin.
Example 5
Comparing the effect of the collagen polypeptide and the 4 meat protein polypeptides on the production of undecyl prodigiosin by fermenting streptomyces CGMCC 4.7172 strain, the method comprises the following steps: rabbit, bovine, Chicken and Duck meat protein polypeptides and collagen polypeptides with the weight average molecular weight of 5k Da, which are respectively numbered as 2Rabbit, 2Beef, 2Chicken, 2Duck and 2CP culture media, are respectively added into the MM culture medium by 20 g/L. Using CGMCC 4.7172 strain as experimental strain, activating and proliferating CGMCC 4.7172 strain, centrifuging to remove supernatant, washing with sterile physiological saline, and suspending the strain in an amount of 4% (in weight percent)v/v) The inoculum size of (2) is inoculated into each culture medium, the fermentation temperature is 29 ℃, the rotating speed is 240 r/min, and the initial pH is 8.5. The fermentation time was 16 d, the yield of undecyl prodigiosin in each medium was measured, and the proline, glycine and serine contents of each meat protein polypeptide were analyzed, and the results are shown in fig. 7 and fig. 8.
As can be seen from FIG. 7, the strain CGMCC 4.7172 detected undecyl prodigiosin after 13 days of fermentation in 2Rabbit, 2Beef, 2Chicken and 2Duck media, and the antibiotic could be detected by 2 days of culture in 2CP media. It can be seen that these four meat protein polypeptides have a later onset of undecyl prodigiosin production and a lower yield than the collagen polypeptides.
As can be seen from fig. 8, these 4 meat protein polypeptides do not differ much in serine content, but have relatively less proline and glycine content, compared to the collagen polypeptide. This indicates that the biosynthesis of undecyl prodigiosin mainly depends on the availability of amino acids in the substrate, and the special amino acid composition of the collagen polypeptide makes the collagen polypeptide more favorable for promoting the streptomyces to produce undecyl prodigiosin than the 4 meat protein polypeptides.
Example 6
The influence of the addition of the collagen polypeptide on the fermentation performance of the streptomyces CGMCC 4.7172 strain is as follows: (1) control group medium: 5 g/L glucose, 5 g/L yeast extract powder, 10 g/L malt extract powder, 2.0 g/L KCl, 0.5 g/L Na 2 SO 4 ,1.0 g/L MgHPO 4 . (2) Experimental group culture medium: 0.5 g/L of collagen polypeptide was added to the culture medium of the experimental group to obtain experimental group 6-1. After activation and proliferation of the CGMCC 4.7172 strain, the supernatant is centrifuged, and the bacteria are washed and suspended by sterile normal saline to obtain a seed solution, wherein the specific fermentation conditions and experimental results are shown in Table 3.
TABLE 3 influence of the addition of collagen polypeptide on the fermentation Performance of Streptomyces CGMCC 4.7172 Strain
As shown in Table 3, the addition of collagen polypeptide can improve the yield of undecyl prodigiosin of CGMCC 4.7172 strain. Meanwhile, after the collagen polypeptide is introduced, the pH value is increased to 9.00 from 8.57 of a control group, and therefore, the pH value of fermentation liquor can be increased by the collagen polypeptide. This indicates that the collagen polypeptide can indeed alter the fermentation environment of streptomyces, thereby affecting the biosynthesis of antibiotics.
Example 7
The influence of the addition of the collagen polypeptide on the fermentation performance of the streptomyces CGMCC 4.7172 strain is as follows: (1) control group medium: 5.0 g/L K 2 SO 4 ,4.0 g/L Na 3 PO 4 ,0.5 g/L MgCl 2 . (2) Experimental group culture medium: adding 5.0 g/L of 20k Da collagen polypeptide into the experimental group culture medium,the experimental group 7-1 was obtained. After activation and proliferation of the CGMCC 4.7172 strain, the supernatant is centrifuged, and the bacteria are washed and suspended by sterile normal saline to obtain a seed solution, wherein the specific fermentation conditions and experimental results are shown in Table 4.
TABLE 4 influence of the addition of collagen polypeptide on the fermentation Performance of Streptomyces CGMCC 4.7172 Strain
As can be seen from Table 4, when collagen polypeptide with a weight average molecular weight of 20 kDa was introduced into an inorganic salt medium containing no carbon source and no nitrogen source, 70.78. mu.g/L of undecyl prodigiosin could be detected at 5 days.
In conclusion, the method for producing undecyl prodigiosin by utilizing collagen polypeptide fermentation provided by the invention comprises the collagen polypeptide with the weight-average molecular weight of 0.2-20 k Da. The inventor creatively discovers that: the collagen polypeptide can provide abundant nutrition for the growth of microorganisms, and also can provide a large amount of precursor amino acids for the production of the undecyl prodigiosin, so that the undecyl prodigiosin is produced by inducing and stimulating the fermentation of the microorganisms, the yield is improved, the fermentation time is shortened, and the fermentation efficiency is improved.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
<110> Sichuan university
<120> culture medium and method for producing undecyl prodigiosin
<160> 1
<210> 1
<211> 1434
<212> DNA
<213> Streptomyces sp
<400> 1
tgcttacaca tgcagtcgaa cgatgaacca cttcggtggg gattagtggc gaacgggtga 60
gtaacacgtg ggcaatctgc ccttcactct gggacaagcc ctggaaacgg ggtctaatac 120
cggatactga ccctcgcagg catctgcgag gttcgaaagc tccggcggtg aaggatgagc 180
ccgcggccta tcagcttgtt ggtgaggtaa tggctcacca aggcgacgac gggtagccgg 240
cctgagaggg cgaccggcca cactgggact gagacacggc ccagactcct acgggaggca 300
gcagtgggga atattgcaca atgggcgaaa gcctgatgca gcgacgccgc gtgagggatg 360
acggccttcg ggttgtaaac ctctttcagc agggaagaag cgaaagtgac ggtacctgca 420
gaagaagcgc cggctaacta cgtgccagca gccgcggtaa tacgtagggc gcaagcgttg 480
tccggaatta ttgggcgtaa agagctcgta ggcggcttgt cacgtcggtt gtgaaagccc 540
ggggcttaac cccgggtctg cagtcgatac gggcaggcta gagttcggta ggggagatcg 600
gaattcctgg tgtagcggtg aaatgcgcag atatcaggag gaacaccggt ggcgaaggcg 660
gatctctggg ccgatactga cgctgaggag cgaaagcgtc gggagcgaac aggattagat 720
accctggtag tccacgccgt aaacggtggg cactaggtgt gggcaacatt ccacgttgtc 780
cgtgccgcag ctaacgcatt aagtgccccg cctggggagt acggccgcaa ggctaaaact 840
caaaggaatt gacgggggcc cgcacaagcg gcggagcatc tggcttaatt cgacgcaacg 900
cgaagaacct taccaaggct tgacatacgc cggaaagcat cagagatggt gccccccttg 960
tggtcggtgt acaggtggtg catggctgtc gtcagctcgt gtcgtgagat gttgggttaa 1020
gtcccgcaac gagcgcaacc cttgtcccgt gttgccagca agcccttcgg ggtgttgggg 1080
actcacggga gaccgccggg gtcaactcgg aggaaggtgg ggacgacgtc aagtcatcat 1140
gccccttatg tcttgggctg cacacgtgct acaatggccg gtacaatgag ctgcgatacc 1200
gcaaggtgga gcgaatctca aaaagccggt ctcagttcgg attggggtct gcaactcgac 1260
cccatgaagt cggagtcgct agtaatcgca gatcagcatt gctgcggtga atacgttccc 1320
gggccttgta cacaccgccc gtcacgtcac gaaagtcggt aacacccgaa gccggtggcc 1380
caaccccttg tgggagggag ctgtcgaagg tgggactggc gatggacgaa gtcg 1434
Claims (10)
1. A culture medium for the production of undecyl prodigiosin, characterized in that it comprises collagen polypeptides having a weight average molecular weight of between 0.2k Da and 20k Da.
2. An undecyl prodigiosin producing medium according to claim 1, wherein said collagen polypeptides have a weight average molecular weight ranging from 0.3k Da to 5k Da.
3. The undecylprodigiosin production medium according to claim 1 or 2, wherein said collagen polypeptide is added in an amount of 0.1 to 80 g/L in the medium.
4. The undecylprodigiosin producing medium according to claim 1 or 2, wherein said collagen polypeptide is obtained by hydrolyzing collagen by at least one of enzymatic hydrolysis, alkaline hydrolysis, acid hydrolysis and high-temperature hydrolysis.
5. A culture medium for producing undecylprodigiosin according to claim 3, wherein said collagen polypeptide is obtained by hydrolyzing collagen by at least one of enzymatic hydrolysis, alkaline hydrolysis, acid hydrolysis and high-temperature hydrolysis.
6. A culture medium for producing undecylprodigiosin according to claim 1, wherein said inorganic salt component in said culture medium comprises at least one of potassium salt, sodium salt and magnesium salt.
7. A method for producing undecyl prodigiosin, characterized in that it comprises the step of introducing Streptomyces coelicolorStreptomyces coelicolorAnd/or Streptomyces lividansStreptomyces coelescensInoculating the culture medium of any one of claims 1-6 to produce undecyl prodigiosin by fermentation.
8. A method of producing undecyl prodigiosin according to claim 7, wherein said streptomyces is inoculated in an amount ranging from 0.5% to 10%.
9. The method of producing undecylprodigiosin according to claim 7, wherein said fermentation temperature is 26-34 ℃, pH is 6-9, and the rotation speed is 150-.
10. A method of producing undecyl prodigiosin according to claim 7 further comprising the step of isolating the fermentation broth resulting from said production fermentation after said undecyl prodigiosin production fermentation.
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