CN113151073B - Giraldii annulata producing strain and purification preparation and application of pigment thereof - Google Patents
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Abstract
A prodigiosin-producing ring strain and purification preparation and application of pigments thereof relate to the fields of microbial fermentation technology and active product separation and identification. The prodigiosin-producing strain is Zooshikella sp.SZ-14, and the accession number of the collection center is as follows: GDMCC No.61474. And (3) coating a strain SZ-14 plate, carrying out streak separation, selecting a single colony, culturing, then transferring, and culturing for 48 hours to obtain a strain culture solution. Centrifuging to obtain precipitate, and extracting with ethanol and ethyl acetate step by step to obtain crude extract of pigment. The crude extract is eluted constantly by a silica gel column with 200-300 meshes and then is prepared by high performance liquid chromatography, and then the pure cyclic prodigiosin can be obtained. The prepared prodigiosin can efficiently kill various harmful algae, has huge potential for developing algicide agents, and has wide application prospect in the aspect of treating algal bloom by a biological method.
Description
Technical Field
The invention relates to the fields of microbial fermentation technology and active product separation and identification, in particular to a prodigiosin-producing ring strain and purification preparation and application of pigments thereof.
Background
Prodigiosins (prodigiosines) are natural red pigments containing three pyrrole ring structures, and the structures of the prodigines (prodigiines) usually contain a methoxy pyrrole ring skeleton, and comprise prodigiosines (prodigiosines) and homologs and analogs thereof. Prodigiosin, also known as prodigiosin, is known as 2-methyl-3-pentyl-6-methoxypsoridine, and is the most common pigment in prodigiosinines. The molecular formula is C 20 H 25 N 3 O (M = 323) is hardly soluble in water, and is easily soluble in organic solvents such as methanol, ethanol, ethyl acetate, acetone, chloroform, and the like. Under the acidic condition, the prodigiosin is red or mauve, and has a special absorption peak at 530-540 nm; under alkaline or neutral conditions, the prodigiosin is yellow or orange yellow and has a special absorption peak at 460-480 nm. The current research shows that prodigiosin has multiple biological activities of antibiosis, tumor resistance, immunosuppression and the like, and has great medicine research and development potential. Meanwhile, the compound has wide attention in the aspects of printing and dyeing, algal bloom prevention and control and the like, and is a natural compound with a great application prospect.
Depending on the number of carbon atoms in the pyrrolidine ring side chain alkyl group and whether cyclization occurs, prodigiosin-like substances can be roughly classified into two main classes, linear (Linear) and Cyclic (Cyclic). Common linear species are prodigiosin and Undecylprodigiosin (C) 25 H 35 N 3 O), the corresponding ring type is ring prodigiosin (C) 20 H 23 N 3 O) and undecyl prodigiosin (also known as streptorubicin B, C) 25 H 33 N 3 O). Prodigiosin-like substances can be produced by a variety of microorganisms including Serratia (Serratia), streptomyces (Streptomyces), vibrio (Vibrio), heiella (Hahella) and Pseudomonas alteromonas (Pseudomonas).
At present, the production and fermentation of Prodigiosin mainly take Serratia marcescens Marcescensp. The fundamental reasons are that the varieties of the strains capable of synthesizing natural cyclic prodigiosin reported at present are few, the varieties of the prodigiosin generated by the strains capable of synthesizing are complex, the ratio of the cyclic prodigiosin is small, and the yield is low. Therefore, the research and development in the field can be greatly promoted by exploring and expanding the types of prodigiosin-producing strains, separating and screening the high-yield strains of the cyclic prodigiosin and establishing an extraction and purification method of the cyclic prodigiosin.
Disclosure of Invention
The first purpose of the invention is to provide a prodigiosin-producing strain.
The second purpose of the invention is to provide a method for preparing the cyclic prodigiosin by purifying the cyclic prodigiosin-producing strain.
The third purpose of the invention is to provide the application of the cyclic prodigiosin in the treatment of harmful algal bloom.
The prodigiosin-producing strain is Zooshikella sp.SZ-14, which has been preserved in Guangdong province culture collection center at 20 days 02 and 2021, and the address is as follows: guangzhou city, first furious Zhonglu No. 100 large yard No. 59 building No. 5, zip code: 510070, accession number of the collection: GDMCC No.61474.
The method for preparing the cyclic prodigiosin by purifying the cyclic prodigiosin producing strain comprises the following steps:
1) Inoculating the strain Zooshikella sp.SZ-14 into a Zobell 2216E liquid culture medium, and culturing the strain in a dark environment by a shaking table to obtain a seed solution;
2) Transferring the seed liquid into a Zobell 2216E liquid culture medium, and culturing by shaking tables in a dark place to obtain a strain culture;
3) Centrifuging the strain culture, collecting precipitate, adding anhydrous ethanol, and performing oscillation and light-proof extraction; centrifuging the mixture, taking the supernatant extract, and performing rotary evaporation to obtain an ethanol crude extract;
4) Adding ethyl acetate into the crude ethanol extract, and oscillating and extracting in a dark place; centrifuging the mixture, taking the supernatant extract, and performing rotary evaporation to obtain a pigment crude extract;
5) Dissolving the crude extract of pigment in ethyl acetate, adding silica gel powder, adsorbing, mixing, rotary evaporating, and pouring out dry powder;
6) Soaking silica gel powder in n-hexane, ultrasonically removing bubbles, and draining and pouring into a chromatographic column;
7) Silica gel column chromatography: loading the dry powder sample in the step 5) into a chromatographic column by adopting a dry method, eluting until all red substances flow out, combining and collecting the deep red and brownish red components A, and performing rotary evaporation to dryness;
8) Dissolving the component A in methanol, and separating and purifying the prodigiosin by using a preparative high performance liquid chromatograph;
9) Collecting the substance with the liquid chromatography retention time of 12-15min, and rotationally evaporating methanol in the solution; adding equal volume of ethyl acetate into the residual water phase, and oscillating and extracting in a dark place; centrifuging the mixed solution to obtain an upper ethyl acetate phase, performing rotary evaporation to dryness, and dissolving with methanol to obtain the prodigiosin.
In the step 1), the conditions of the shaking table light-proof culture can be 28-37 ℃, 100-200rpm and 16-24 h.
In the step 2), the inoculation amount of the seed liquid can be 1-5%; the conditions of shaking table light-proof culture can be 28-37 ℃, 100-200rpm and 24-48 h.
In the step 3), the centrifugation condition can be 6000 to 10,000g for 10 to 15min; the oscillating light-proof extraction can be carried out for 2 to 6 hours at the temperature of 4 ℃; the temperature of the rotary evaporation to dryness can be 50-60 ℃.
In the step 4), the oscillating light-proof extraction can be carried out for 2 to 6 hours at the temperature of 4 ℃; the centrifugation condition can be 6000-10,000g, 10-15 min; the temperature of the rotary evaporation to dryness can be 40-45 ℃.
In the step 5), the ratio of the pigment crude extract to the ethyl acetate to the silica gel powder can be 40-55 mg: 4.5-5 mL: 1g; preferably, the ratio of the pigment crude extract to the ethyl acetate to the silica gel powder is 50 mg: 5 mL: 1g; the fineness of the silica gel powder can be 200-300 meshes; the temperature of the rotary evaporation to dryness can be 40-45 ℃.
In the step 6), the fineness of the silica gel powder can be 200-300 meshes; the ratio of the silica gel powder to the n-hexane can be 25 g: 50-100 mL; the time for removing bubbles by ultrasonic can be 10-15 min.
In step 7), the elution may be performed in the following order: (a) eluting with 100-200 mL of n-hexane; (b) Ethyl acetate/methanol (v/v/v) = 10: 4: 1; the temperature of the rotary evaporation to dryness can be 40-45 ℃.
In step 8), the conditions for separation and purification by preparative high performance liquid chromatography may be: (a) preparing a column: sunfire TM Prep C18Column (10X 100mm,5 μm; waters, ireland); (b) mobile phase: 55% methanol solution (methanol: ddH) 2 O = 55: 45, v/v,0.035% trifluoroacetic acid); (c) flow rate: 5.0mL/min; (d) detecting wavelength: 535nm;
in the step 9), the temperature of the methanol in the rotary evaporation solution can be 50-60 ℃; the time for oscillating light-proof extraction can be 2-6 h; the centrifugation condition can be 6000-10,000g, 10-15 min; the temperature of the re-rotary evaporation to dryness can be 40-45 ℃.
The molecular formula of the cyclic prodigiosin is C 20 H 23 ON 3 The molecular weight is 321.
Experiments show that the prepared cyclic prodigiosin has obvious algae killing effect on 19 kinds of water bloom and red tide algae (see table 1). The prodigiosin can be applied to the treatment of harmful algal blooms.
The invention discloses a cyclic prodigiosin (cyclopropodigiosin) producing strain, and an extraction preparation method and application of the cyclic prodigiosin. The strain is Zooshikella sp.SZ-14, the strain SZ-14 is subjected to plate coating, streaking separation, single colony culture is picked and then is transferred, and the strain culture solution is obtained after culture for 48 hours. Centrifuging to obtain precipitate, and extracting with ethanol and ethyl acetate step by step to obtain crude extract of pigment. The crude extract is eluted constantly by a silica gel column (200-300 meshes), and then is prepared by high performance liquid chromatography, so that the pure cyclic prodigiosin can be obtained. The prepared cyazosin can efficiently kill various harmful algae, has huge potential for developing algicide and has wide application prospect in the aspect of treating algal bloom by a biological method.
Drawings
FIG. 1 is a morphological diagram of the prodigiosin-producing bacterium Zooshikella sp. Wherein, A is the colony morphology of a flat plate, B is the morphology after 48h of shake flask culture, and C is the scanning electron microscope image of the strain.
FIG. 2 is a phylogenetic tree constructed by the strain Zooshikella sp.SZ-14 based on the 16S rRNA gene sequence.
FIG. 3 is a liquid chromatogram of the preparation of cyclic prodigiosin.
FIG. 4 is a liquid chromatogram and TIC chart of the prepared and concentrated cyclic prodigiosin.
FIG. 5 is a first mass MS chart of prodigiosin
FIG. 6 is a secondary mass MS/MS plot of cyclic prodigiosin.
Figure 7 is the ionic fragment fragmentation structure of cyclic prodigiosin.
FIG. 8 is a nuclear magnetic resonance-hydrogen spectrum of cyclic prodigiosin.
FIG. 9 is a nuclear magnetic resonance-carbon spectrum of prodigiosin cyclic form.
Detailed Description
The following examples will further illustrate the present invention with reference to the accompanying drawings.
The embodiment of the invention provides a prodigiosin-like substance high-yield strain and a method for preparing cyclopropodiosis in a cyclic prodigiosin by purifying the strain. The determination of the algae killing spectrum shows that the prodigiosin has strong application value in the prevention and treatment of harmful algal blooms.
1. Separation, screening and identification of strains
(1) Taking mangrove wetland soil samples in national natural protection areas of northern Lun estuary in Guangxi province, urban defense and harbor city, using 10 times of sterile seawater for gradient dilution, after shaking and uniform mixing, respectively taking 0.2mL of the diluted soil samples to be coated on a Zobell 2216E solid plate (5 g/L of bactopeptone, 1g/L of yeast extract, 0.1g/L of ferric phosphate, 15g/L of agar powder, constant volume of aged seawater to be 1L, pH 7.2), and culturing for 7d in a dark place at 30 ℃;
(2) Selecting different colonies according to differences of colony morphology, size, color, etc., inoculating in 20mL Zobell 2216E liquid culture medium, placing in a shaking table (30 ℃,150 rpm), and culturing for 3d in a dark place;
(3) Re-streaking the culture on a Zobell 2216E solid plate, and culturing at 30 ℃ in the dark for 2-3 days;
(4) Repeating the step (3) for multiple times until a pure culture is obtained;
(5) Morphological characteristics of the cyclic prodigiosin-producing strain SZ-14 are shown in FIG. 1. The genome of the strain SZ-14 is extracted, 16S rRNA gene amplification is carried out, and PCR products are connected through a T vector and then sent to Shanghai Biotech for sequencing. The sequences were then primed and uploaded to the Korean EzBioCloud website (https:// www.ezbiocloud.net /) for alignment analysis, and the MEGA X software was used to construct phylogenetic trees, the results of which are shown in FIG. 2. Through comparison and identification, the strain SZ-14 and Zoohikella marina JC333 T The similarity of this strain is highest, 99.79%, so strain SZ-14 belongs to the genus Zooshikela, designated Zooshikela sp.SZ-14, whose 16S rRNA sequence is uploaded to the NCBI database under the GenBank accession number MK911731.
2. Cultivation of the Strain
(1) Inoculating the strain Zooshikella sp.SZ-14 into 20mL Zobell 2216E liquid culture medium, placing the strain in a shaking table (30 ℃,150 rpm), and culturing for 24h in a dark place to obtain seed liquid;
(2) Transferring the seed solution to 500mL of Zobell 2216E liquid culture medium in a volume ratio of 1%, placing the culture medium in a shaking table (30 ℃,150 rpm), and culturing for 48h in a dark place to obtain a strain culture.
3. Preparation of crude pigment extract
(1) The above culture was centrifuged (10,000g, 10 min); adding 20mL of absolute ethyl alcohol into the precipitate, and oscillating and extracting for 2 hours at 4 ℃ in a dark place;
(2) The mixture was centrifuged (10,000g, 10min); taking the supernatant extract, and rotary evaporating at 52 ℃ to dryness to obtain an ethanol crude extract;
(3) Adding 20mL of ethyl acetate into the crude ethanol extract, and oscillating and extracting for 2 hours in a dark place at 4 ℃;
(4) The mixture was centrifuged (10,000g, 10min); taking the supernatant extract, and rotary evaporating at 42 deg.C to obtain pigment crude extract.
4. Purification preparation of cyclic prodigiosin
(1) Dissolving 50mg of the crude extract of the pigment in 5mL of ethyl acetate, adding 1g of silica gel powder (200-300 meshes) for uniformly adsorbing, drying by distillation at 42 ℃, and pouring out dry powder for later use;
(2) Filling a silica gel column: weighing 25g of silica gel powder (200-300 meshes), soaking the silica gel powder in n-hexane (50-100 mL), ultrasonically removing bubbles for 10min, and draining the silica gel powder into a chromatographic column (30 mm multiplied by 400 mm) by using a glass rod;
(3) Silica gel column chromatography: and (2) adopting a dry method for loading, adding the dry powder sample in the step (1) into a chromatographic column, and eluting according to the following sequence: (a) elution with n-hexane is 100mL; (b) Continuously eluting with n-hexane, ethyl acetate, methanol (v/v/v) = 10: 4: 1 until red substance completely flows out;
(4) Mixing and collecting dark red and brownish red component A, and rotary evaporating at 42 deg.C;
(5) Dissolving component A in 1mL methanol, and separating and purifying prodigiosin cycloprodiosin by using preparative high performance liquid chromatography (SHIMADZU, SIL-AP20, japan). Wherein (a) preparing a column: sunfireTMPrep C18Column (10X 100mm,5 μm; waters, ireland); (b) mobile phase: 55% methanol solution (methanol: ddH) 2 O = 55: 45, v/v,0.035% trifluoroacetic acid); (c) flow rate: 5.0mL/min; (d) detecting wavelength: 535nm;
(6) Collecting substance with liquid chromatography retention time of 12-15min (see FIG. 3), and rotary evaporating methanol in the solution at 52 deg.C;
(6) Adding equal volume of ethyl acetate into the residual water phase, and oscillating and extracting for 2 hours in a dark place at 4 ℃;
(7) The mixture was centrifuged (10,000g, 10min); taking the upper ethyl acetate phase, and rotationally evaporating at 42 ℃ to dryness to obtain the cyclopropodiosis in the prodigiosin (the purity of the sample is shown in figure 4). FIG. 5 is a first-order mass spectrum MS diagram of prepared and purified prodigiosin obtained by high-resolution LC-MS (Waters 2767, USA). FIG. 6 is a second-order mass spectrum MS/MS diagram of the detection of the cyclic prodigiosin by a Q-active high-resolution liquid chromatograph-mass spectrometer (Thermo, USA). FIG. 7 shows the ion fragment breaking structure of the cyclic prodigiosin. FIGS. 8 and 9 are the hydrogen and carbon spectra, respectively, as identified by a nuclear magnetic resonance spectrometer (Bruker AV600, switzerland).
5. Determination of algae-killing Effect
(1) The blue algae is purchased from aquatic organism research institute of Chinese academy of sciences, the culture medium used by the algae is BG11 culture medium, and the culture conditions are as follows: the illumination period is 12/12h (light/dark), and the illumination intensity is 50 mu mol photons m -2 s -1 The temperature is 25 +/-1 ℃. The other algae species except the blue algae are provided by an important laboratory algae species library of offshore marine environmental science and national center of Xiamen university, the culture medium used by the algae species is f/2 culture medium, and the culture conditions are as follows: the light period is 12/12h (light/dark), and the light intensity is 100 mu mol photons m -2 s -1 The temperature is 20 +/-1 ℃;
(2) Culturing the algae cells to a logarithmic growth phase, and subpackaging to 48-hole cell culture plates with 1mL per hole;
(3) Adding the prepared cyclic prodigiosin into a pore plate, adding the same amount of DMSO into a control group, measuring the spontaneous fluorescence value of chlorophyll of algae cells by using a microplate reader at regular intervals, wherein the calculation formula of the algae killing rate is as follows: algae removal rate (%) = (F) t -F 0 )/ F 0 X 100%, wherein, F 0 The autofluorescence value of chlorophyll of algae cells is measured in 0 h; f t Chlorophyll autofluorescence of algal cells treated with t h.
(4) As shown in Table 1, the extracted prodigiosin has a remarkable algicidal effect on various algal blooms and red tide algae species, and the algicidal activity is higher than that of prodigiosin on Chlorella, zostera marina, platymonas subcordiformis, thalassilia ensii, aphanizomenon pterocarpum, phaeodactylum tricornutum, zostera sphaerica and Praenophyta robusta.
FIG. 1 is a morphological diagram of the prodigiosin-producing bacterium Zooshikella sp. As can be seen from FIG. 1, colonies of the strain Zooshikella sp.SZ-14 are dark red with a green metallic luster; the culture solution is bright red and mixed with red secretion; the cells are spiral rods with length of 5-15 μm and width of 0.3-0.7 μm, without flagella and surface smooth and without folds.
FIG. 2 is a phylogenetic tree constructed by the strain Zooshikella sp.SZ-14 based on the 16S rRNA gene sequence. As can be seen from FIG. 2, the strain SZ-14 belongs to the genus Zooshikela, hahellaceae, which is currently only three strains of bacteria and has been reported to produce prodigiosin-like substances.
FIG. 3 is a liquid chromatogram of the preparation of cyclic prodigiosin. As can be seen from the figure, the time of appearance peak of the cyclic prodigiosin is 12-15min when the liquid chromatogram is prepared.
FIG. 4 is a liquid chromatogram and TIC chart of the prepared and concentrated annular prodigiosin. As can be seen from the figure, the prepared cyclic prodigiosin has an absorption peak at 535nm, and the peak type is single, and the sample is relatively pure.
Figures 5 and 6 are mass spectra of cyclic prodigiosin. As can be seen from FIGS. 5 and 6, the molecular addition ion value ([ M + H ] of prodigiosin cyclate](+) is 322.45, its molecular weight is 321, and its molecular formula may be calculated as C 20 H 23 ON 3 。
Figure 7 is the structural formula of cyclic prodigiosin. The dashed arrows in the figure identify the fracture sites where fracture fragment signals (including m/z307, m/z290, m/z175, m/z160, etc.) detectable by secondary mass spectrometry are located.
FIGS. 8 and 9 are the hydrogen and carbon nuclear magnetic resonance spectra of the cyclic prodigiosin.
TABLE 1
The invention provides a prodigiosin substance high-yield strain and a method for purifying and preparing a cyclic prodigiosin by using the same. The cyclic prodigiosin and the prodigiosin are two different compounds, and the algae killing effect of the cyclic prodigiosin is better than that of the prodigiosin, see table 1. At present, no report on the algae killing activity of the prodigiosin. The determination of the algicidal spectrum shows that the prodigiosin has strong application value in the prevention and treatment of harmful algal blooms.
Claims (6)
1. A prodigiosin-producing bacterium, characterized in that it is Zooshikella sp.SZ-14, which has been deposited at the Guangdong province Collection center on 20 D.02/2021, with the accession numbers: GDMCC No.61474.
2. A process for the purification of prodigiosin-producing bacteria to produce prodigiosin-containing ring as claimed in claim 1 comprising the steps of:
1) Inoculating the strain Zooshikella sp.SZ-14 into a Zobell 2216E liquid culture medium, and culturing the strain in a dark environment by a shaking table to obtain a seed solution;
2) Transferring the seed liquid into a Zobell 2216E liquid culture medium, and culturing by shaking tables in a dark place to obtain a strain culture; the inoculation amount of the seed liquid is 1-5%; the conditions of shaking table light-proof culture are 28-37 ℃, 100-200rpm and 24-48 h;
3) Centrifuging the strain culture, collecting precipitate, adding anhydrous ethanol, and performing oscillation and light-proof extraction; centrifuging the mixture, taking the supernatant extract, and performing rotary evaporation to obtain an ethanol crude extract; the centrifugation condition is 6000-10,000g, 10-15 min; the oscillating light-proof extraction is carried out for 2 to 6 hours at the temperature of 4 ℃; the temperature for rotary evaporation to dryness is 50-60 ℃;
4) Adding ethyl acetate into the crude ethanol extract, and oscillating and extracting in a dark place; centrifuging the mixture, taking the supernatant extract, and performing rotary evaporation to obtain a pigment crude extract; the oscillating light-proof extraction is carried out for 2-6 h at 4 ℃; the centrifugation condition is 6000-10,000g, 10-15 min; the temperature for rotary evaporation to dryness is 40-45 ℃;
5) Dissolving the crude extract of pigment in ethyl acetate, adding silica gel powder, adsorbing, mixing, rotary evaporating to dryness, and pouring out dry powder; the ratio of the pigment crude extract to the ethyl acetate to the silica gel powder is 40-55 mg: 4.5-5 mL: 1g; the fineness of the silica gel powder is 200-300 meshes; the temperature for rotary evaporation to dryness is 40-45 ℃;
6) Soaking silica gel powder in n-hexane, ultrasonically removing bubbles, draining, and pouring into a chromatographic column; the fineness of the silica gel powder is 200-300 meshes; the ratio of the silica gel powder to the n-hexane is 25 g: 50-100 mL; the time for removing bubbles by ultrasonic is 10-15 min;
7) Silica gel column chromatography: loading the dry powder sample in the step 5) into a chromatographic column by adopting a dry method, eluting until all red substances flow out, combining and collecting the deep red and brownish red components A, and performing rotary evaporation to dryness; the elution is carried out in the following order: (a) eluting with 100-200 mL of n-hexane; (b) The volume ratio of n-hexane to ethyl acetate to methanol is 10: 4: 1; the temperature for rotary evaporation to dryness is 40-45 ℃;
8) Dissolving the component A in methanol, and separating and purifying the prodigiosin by using a preparative high performance liquid chromatograph;
9) Collecting the substance with the liquid chromatography retention time of 12-15min, and rotationally evaporating methanol in the solution; adding equal volume of ethyl acetate into the residual water phase, and oscillating and extracting in a dark place; centrifuging the mixed solution to obtain an upper ethyl acetate phase, performing rotary evaporation to dryness, and dissolving with methanol to obtain the prodigiosin.
3. The method for preparing cyclic prodigiosin according to claim 2, wherein in the step 1), the conditions of shaking table light-shielding culture are 28-37 ℃, 100-200rpm and 16-24 h.
4. The method for preparing cyclic prodigiosin according to claim 2, wherein in step 4), the ratio of the pigment crude extract, ethyl acetate and silica gel powder is 50 mg: 5 mL: 1g.
5. A method for preparing cyclic prodigiosin according to claim 2, wherein in the step 8), the conditions for separation and purification by preparative high performance liquid chromatography are as follows: (a) preparing a column: sunfire TM Prep C18Column, 10X 100mm,5 μm; waters, ireland; (b) mobile phase: 55% methanol solution of ddH 2 O = 55: 45, v/v,0.035% trifluoroacetic acid; (c) flow rate: 5.0mL/min; (d) detecting wavelength: 535nm.
6. A method for preparing cyclic prodigiosin according to claim 2, wherein in the step 9), the temperature of methanol in the solution obtained by rotary evaporation to dryness is 50-60 ℃; the time for oscillating and light-proof extraction is 2-6 h; the centrifugation condition is 6000 to 10,000g for 10 to 15min; the temperature for rotary evaporation to dryness is 40-45 ℃.
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| CN111088210A (en) * | 2020-01-21 | 2020-05-01 | 泉州师范学院 | Strain for producing prodigiosin pigment and prodigiosin pigment synthetic gene cluster |
| CN111621458A (en) * | 2020-06-30 | 2020-09-04 | 江南大学 | BVG90_11450 gene-deleted serratia marcescens engineering bacterium |
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| CN102888351A (en) * | 2012-09-03 | 2013-01-23 | 广东药学院 | Prodigiosin high-producing strain and production method thereof |
| CN111088210A (en) * | 2020-01-21 | 2020-05-01 | 泉州师范学院 | Strain for producing prodigiosin pigment and prodigiosin pigment synthetic gene cluster |
| CN111621458A (en) * | 2020-06-30 | 2020-09-04 | 江南大学 | BVG90_11450 gene-deleted serratia marcescens engineering bacterium |
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| Jong Suk Lee等.Exceptional Production of both Prodigiosin and Cycloprodigiosin as Major Metabolic Constituents by a Novel Marine Bacterium, Zooshikella rubidus S1-1.《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》.2011,第77卷(第14期),第4967-4973页. * |
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