CN117187108A - Streptomyces chromogenes and application thereof - Google Patents
Streptomyces chromogenes and application thereof Download PDFInfo
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- CN117187108A CN117187108A CN202310919388.4A CN202310919388A CN117187108A CN 117187108 A CN117187108 A CN 117187108A CN 202310919388 A CN202310919388 A CN 202310919388A CN 117187108 A CN117187108 A CN 117187108A
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Abstract
The invention belongs to the field of microorganisms, and particularly relates to streptomyces pigmentary and application thereof. The invention provides a Streptomyces sp BG-R1 deposited in the microorganism strain collection of Guangdong province at month 31 of 2023 with the deposit number: GDMCC No:63510. the streptomycete BG-R1 water-soluble mauve pigment has good coloring effect on silk textiles as a natural pigment, does not need to be extracted by a chemical reagent, has simple dyeing step, does not need to add a dyeing auxiliary agent, is easy to color and high in color fastness, overcomes the difficulty of light sensitivity of microbial pigments, and is safe and nontoxic.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to streptomyces pigmentary and application thereof.
Background
The coloring matter is a raw material imparting a certain color. The choice of commodity often depends on the feel of sight, touch, smell, etc., and pigments are an important part of vision, so that whether pigments are used properly or not can also play a role in the quality of products. Synthetic pigment: the pigment obtained from chemical synthesis is called synthetic pigment; inorganic pigment: common inorganic pigments are ferric oxide, carbon black, chromium oxide green and the like, which have good light resistance and are insoluble in water; natural pigment: the common natural pigments include carmine, cochineal, chlorophyll, curcumin, and bilirubin.
Natural pigments are derived from natural substances, mainly extracted from plant tissues, and also include some pigments from animals and microorganisms. Most plant pigments have no side effect and high safety. Most plant pigments are anthocyanin compounds, carotenoid compounds and flavonoid compounds, are bioactive substances, and are functional effective components in plant medicines and health-care foods. The plant pigment has natural color tone, can increase the color tone and is similar to the natural color tone, and is natural beauty. The plant pigment has a small content in plant body, and is difficult to separate and purify, and peculiar smell can be generated when some coexists exist, so that the production cost is higher than that of the synthetic pigment. In addition, most plant pigments have poor dyeing force, are not easy to dye uniformly, and are fresh and bright without synthetic pigments. And the textile needs to be washed, and has more requirements on friction fastness, soaping fastness and light fastness.
Microbial pigment is a secondary metabolite of microorganism, and has various colors such as red, orange, yellow, green, cyan, purple, black, brown, etc. Microbial pigments can be classified into water-soluble and water-insoluble pigments. Compared with other natural dyes, the microbial pigment has short production period and low cost, and is easier for industrial production. The method is to culture microbe in liquid culture medium to metabolize great amount of pigment, and to obtain pigment solution through separation, extraction and concentration. Patent CN 112375791B discloses a method for preparing and dyeing streptomyces spectabilis haematochrome, which comprises the following steps: (1) culturing and fermenting; (2) ultrasonic extraction; (3) extraction; (4) purifying. Compared with other natural dyes, the Streptomyces spectabilis haematochrome has short production period and low cost, and is easy for industrial production; the fermentation medium is optimized, the phenomenon of pilling does not occur when the streptomyces spectabilis is cultivated, and the yield is higher; the dyeing liquid prepared from the Streptomyces spectabilis haematochrome is high in dye-uptake, and the soaping color fastness and the rubbing color fastness of the dyed silk fabric are both of grade 4 and accord with the dyeing standard of the silk fabric. The patent CN 110983816A discloses an application of actinomycin as a dye in printing and dyeing textile materials such as terylene, chinlon, cotton, silk and the like by using actinomycin obtained from marine plants as a natural pigment, and has good dyeing effect and good antibacterial effect; especially has good dyeing effect on silk fabrics, the dry rubbing fastness is 4-5 grade, the wet rubbing fastness is 4 grade, the soaping fastness is 4 grade, actinomycin is applied to dyed materials, the raw materials are easy to obtain, the cost is low, and the actinomycin has good application prospect in industry as a functional dye due to antibacterial activity. However, the extraction of the pigment requires an organic solvent and a dyeing auxiliary agent in the dyeing process, and the actual green and safe performance is not realized.
The extraction liquid dyeing method has the advantages of wide application range, easy industrial production, complex extraction process and higher cost; meanwhile, a large amount of organic solvents are needed to produce a large amount of industrial sewage. In contrast to non-water-soluble pigments, water-soluble pigments can be secreted directly outside the cell, and when fermented using a liquid medium, pigment molecules are mostly in the fermentation broth, and pigment solutions can be obtained by centrifugation and filtration. The operation is simple and convenient, no organic solvent is needed, and industrial wastewater is not generated. The separated pigment solution can be directly used for dyeing the fabric. Therefore, development of more microorganisms producing water-soluble pigments, and high pigment-coloring ability and color fastness are required.
Disclosure of Invention
In order to solve the above problems, the present invention provides a Streptomyces sp BG-R1 deposited at the microorganism strain collection in Guangdong province at 2023, month 05 and 31 under the deposit number: GDMCCNo:63510. the streptomycete BG-R1 water-soluble mauve pigment has good coloring effect on silk textiles as a natural pigment, does not need to be extracted by a chemical reagent, has simple dyeing steps, does not need to add a dyeing auxiliary agent, is easy to color, has high color fastness, and is safe and nontoxic.
In one aspect, the invention provides a Streptomyces sp BG-R1 deposited at the microorganism strain collection, cantonese province, at month 31 of 2023 under accession number: GDMCC No:63510.
on the other hand, the invention provides a fermentation method of the streptomyces BG-R1, which comprises the step of inoculating the streptomyces BG-R1 into a culture solution of Gao's first.
Specifically, the formula of the culture solution of Gao's No. one can be soluble starch 15-25g/L and KNO 3 0.5-1.5g/L，K 2 HPO 4 0.2-0.8g/L，MgSO 4 ·7H 2 O 0.2-0.8g/L，NaCl 0.2-0.8g/L，FeSO 4 ·7H 2 O 0.01-0.02g/L。
Preferably, the formula of the culture solution of Gao's No. one is soluble starch 20g/L and KNO 3 1g/L，K 2 HPO 4 0.5g/L，MgSO 4 ·7H 2 O 0.5g/L，NaCl 0.5g/L，FeSO 4 ·7H 2 O 0.01g/L。
On the other hand, the invention also provides a preparation method of the pigment solution, which comprises the following steps:
(1) Inoculating the streptomycete BG-R1 into the culture solution of the first Gao's strain for culture;
(2) Centrifuging and filtering to obtain supernatant.
Specifically, the culture temperature in step (1) may be 25 to 30℃and the culture time may be 5 to 10 days.
Preferably, the incubation temperature in step (1) is 28℃and the incubation time is 7d.
Specifically, the pigment solution is a mauve pigment solution.
In yet another aspect, the present invention provides a pigment solution prepared by the aforementioned preparation method.
In yet another aspect, the present invention provides a method of dyeing a fabric comprising immersing the fabric in the pigment solution described above.
Specifically, the pigment solution is a mauve pigment solution.
Specifically, the fabric is silk fabric.
Specifically, the fabric dyeing method also comprises the steps of heat preservation dyeing after temperature rising, cleaning and drying.
Specifically, the bath ratio of step (1) may be 1:20-40.
Preferably, the bath ratio is 1:35.
Further, the post-heating thermal-insulation dyeing of the bacterial cells comprises two heating steps.
Preferably, the temperature is raised to 55-65 ℃ for the first time, and the temperature is kept for dyeing for 25-35min; heating to 75-85 ℃ for the first time, and preserving heat and dyeing for 15-25min.
Further preferably, the temperature is raised to 60 ℃ for the first time, and the dyeing is carried out for 30 minutes; and heating to 80 ℃ for the second time, and preserving heat and dyeing for 20min.
In still another aspect, the invention provides application of the streptomycete BG-R1 or pigment solution in textile and leather printing and dyeing industries.
Specifically, the spinning includes, but is not limited to: cashmere, wool, silk, cotton, chemical fibre or modal.
In particular, the leather includes, but is not limited to: cow leather, sheep leather, PVC leather, vegetable fiber synthetic leather or mycelium leather.
In yet another aspect, the invention provides the use of the aforementioned Streptomyces BG-R1 or pigment solution in cosmetics.
Specifically, the cosmetic includes, but is not limited to: hair dye, lipstick or foundation.
In yet another aspect, the invention provides the use of the aforementioned Streptomyces BG-R1 or pigment solutions for the preparation of pigments or paints.
Specifically, the coating includes, but is not limited to: liquid, powder or high solids coatings.
The invention has the technical effects that:
(1) The streptomycete BG-R1 has short growth cycle, strong reproductive capacity and more purplish red pigment secretion;
(2) The pigment separation and extraction are simple, the extraction of chemical reagents is not needed, the dyeing step is simple, and the addition of dyeing auxiliary agents is not needed;
(3) The color fastness is high;
(4) Is safe and nontoxic.
Preservation description:
strain name: streptomyces sp BG-R1;
preservation number: GDMCC No:63510;
taxonomic name: streptomyces sp;
preservation time: 2023, 05, 31;
preservation unit: the collection of microorganism strains in Guangdong province;
preservation address: guangzhou city first middle road No. 100 college No. 59 building 5.
Drawings
FIG. 1 is a photograph of Streptomyces BG-R1 colonies cultured for 7d.
FIG. 2 is a photograph of fermentation broth of Streptomyces BG-R1 cultured for 7d.
FIG. 3 shows the dyeing of the purplish red pigment extracted from Streptomyces BG-R1 after fermentation in silk.
FIG. 4 shows the dyeing of the purplish red pigment extracted from fermentation of Streptomyces BG-R1 in different fabrics.
Detailed Description
The present invention will be described in further detail with reference to the following examples, which are not intended to limit the present invention, but are merely illustrative of the present invention. The experimental methods used in the following examples are not specifically described, but the experimental methods in which specific conditions are not specified in the examples are generally carried out under conventional conditions, and the materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.
Example 1 isolation and identification of strains
Soil sample: yunnan forest soil.
Isolation medium: the culture medium of Gaoshi No.1 comprises the following specific formula: soluble starch (20 g/L), KNO 3 (1g/L)，K 2 HPO 4 (0.5g/L)，MgSO 4 ·7H 2 O(0.5g/L)，NaCl(0.5g/L)，FeSO 4 ·7H 2 O (0.01 g/L), agar (20 g/L), deionized water (deionized water) is added to a volume of 1L, and pH=7.4-7.6. Sterilizing with steam at 121deg.C for 20min.
Fermentation medium: the culture medium of Gaoshi No.1 comprises the following specific formula: soluble starch (20 g/L), KNO 3 (1g/L)，K 2 HPO 4 (0.5g/L)，MgSO 4 ·7H 2 O(0.5g/L)，NaCl(0.5g/L)，FeSO 4 ·7H 2 O (0.01 g/L) was added with deionized water to a constant volume of 1L and pH=7.4-7.6. Sterilizing with steam at 121deg.C for 20min.
1.1 isolation of strains
Spreading the collected soil sample in sterile culture dish, naturally drying at 25-28deg.C for 7-14d, and grinding. Firstly, placing a soil sample into a 70 ℃ oven for 1 hour to kill bacteria, then weighing 5g of the soil sample, placing the soil sample into a 150mL triangular flask containing 15-20 small glass beads which are sterilized, adding 45mL of sterile water, and shaking the triangular flask at 30 ℃ for 20 minutes; subsequently, the dilution was carried out with sterile water with a dilution gradient of 10 for the experiment -2 、10 -3 、10 -4 And 10 -5 A total of 4. 100 mu L of soil dilutions with different dilution gradients are respectively and uniformly coated in the separation culture medium by a coating rod, and are cultured for 10-14d at 30 ℃ to check the pigment production condition of the bacterial colony. The single colony for producing pigment is selected, separated and purified, and then is preserved in 20% glycerol at low temperature.
1.2 morphological observations
On the isolation medium, hyphae grew well. The aerial hyphae are off-white, and the surfaces of the fungus coating are raised; spiral spore filaments, oval or long rod shape, and regular two ends; the mycelium in the basal body is mauve, and the culture medium contains mauve secretory pigment.
1.3 identification of species
The isolated strain was sent to the engineering bioengineering (Shanghai) Co., ltd for identification. Bacterial 16S rRNA universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGCTACCTTGTTACGACTT-3') were used. After PCR, the sequence is analyzed in 3 segments, the 16s rRNA full-length sequence is spliced, then the comparison analysis is carried out in a NCBI database, and the sequence similarity with the Streptomyces lilacinus (Streptomyces lavendulae) BP39 and Streptomyces aspen (Streptomyces yangpuensis) CM253 sequences in the database is highest. The 16S rRNA base sequence is shown in SEQ ID NO.1, and the strain is identified as Streptomyces sp and designated as Streptomyces BG-R1. The microorganism strain is deposited in the Guangdong province microorganism strain collection at the month 31 of 2023 with the deposit number: GDMCC No:63510.
example 2
2.1 Streptomyces sp. BG-R1 secretes pigments for dyeing silk, the specific steps are as follows:
(1) Fermentation culture and pigment preparation
Inoculating Streptomyces sp BG-R1 into the isolated culture medium, and culturing at 28deg.C for 5-7 days to obtain spores; inoculating the spores into a fermentation medium; the amount of fermentation medium in a 250 mL Erlenmeyer flask was 100mL. The culture was carried out at a constant temperature of 28℃with shaking at a shaker speed of 160rpm for 7d. After the fermentation broth was centrifuged (8000 g,10 min), the supernatant was sterile filtered, and the filtrate was a pigment solution.
(2) Dyeing of fabric
The pigment solution is used for dyeing silk. 40X 40cm silk is treated according to a bath ratio of 1:35 is soaked in the pigment solution, the temperature is raised to be 3 ℃ per minute, the temperature is raised to be 60 ℃ and then the dyeing is carried out for 30min, the temperature is raised to be 80 ℃ and the dyeing is continued for 20min. Taking out, cleaning with water, and oven drying at 60deg.C. The dyed silk sample is shown in fig. 3.
2.2 color fastness detection
And (3) feeding the dyed silk to a middle spinning standard (Shenzhen) detection limited company for detecting the washing fastness, the acid perspiration fastness, the alkali perspiration fastness, the friction fastness, the washing fastness, the light fastness and the non-chlorine bleaching fastness. The detection results are shown in Table 1.
TABLE 1
The results show that the red pigment produced by Streptomyces sp.BG-R1 has very good coloring effect on silk. The color fastness to water reaches 4 levels, the rubbing color fastness reaches 4-5 levels, the color fastness to washing reaches 3-4 levels, the color fastness to non-chlorine bleaching reaches 4 levels, the color fastness to light reaches 4 levels, and the requirements of silk clothing are met. In particular, the red pigment has good light-proof capability, and overcomes the difficulty of sensitivity of microbial pigment to light.
Example 3
The dyeing of Streptomyces sp. BG-R1 secretory pigments with different fabrics is performed as follows:
(1) Fermentation culture and pigment preparation
As in example 2.
(2) Dyeing of fabric
The pigment solution is used for dyeing different fabrics, including silk, cashmere, mohair, cotton cloth, modal, nylon and terylene. A fabric of about 5 x 5cm size was treated at a bath ratio of 1:30 is soaked in pigment solution, the temperature is raised to 3 ℃ per minute, the temperature is raised to 60 ℃ and then the dyeing is carried out for 30min, the temperature is raised to 80 ℃ and the dyeing is continued for 20min. Taking out, cleaning with water, oven drying at 60deg.C, and photographing.
As shown in FIG. 4, the dye solution prepared from Streptomyces BG-R1 has certain dyeing capability on different types of fabrics. Wherein, the dye has good coloring effect on protein fibers, silk, cashmere and mohair; the color of the chemical fiber such as nylon and terylene is lighter; for cellulose fiber cotton and modal, the coloring effect is poor, the color is light, and other auxiliary agents may be needed to improve the coloring effect.
Example 4
Streptomyces sp. BG-R1 secretes pigments for silk dyeing, comprising the following steps:
(1) Fermentation culture and pigment preparation
As in example 2.
(2) Dyeing of fabric
The pigment solution was used for dyeing silk. 40X 40cm silk is treated according to a bath ratio of 1:25 are soaked in pigment solution, the temperature is raised to 3 ℃ per minute, the temperature is raised to 55 ℃ and then the dyeing is carried out for 25 minutes, the temperature is raised to 75 ℃ and the dyeing is continued for 15 minutes. Taking out, cleaning with water, and oven drying at 60deg.C.
The color fastness test was the same as in example 2. The detection results are shown in Table 2.
TABLE 2
The above results show that the dyeing composition still has better coloring effect and color fastness under the conditions of reducing the bath ratio, dyeing temperature and dyeing time. The color fastness to water is 3, the color fastness to acid and alkali perspiration is 3, the color fastness to rubbing is 4-5 grade, the color fastness to washing is 3 grade, the color fastness to non-chlorine bleaching is 4 grade, and the color fastness to light is 3 grade.
Claims (10)
1. Streptomyces sp BG-R1 deposited at the microorganism strain collection in Guangdong province at month 31 of 2023 under the accession number: GDMCC No:63510.
2. the fermentation method of streptomyces BG-R1 of claim 1, comprising inoculating streptomyces BG-R1 to culture broth No. one gao; the culture of Gao's No. oneThe formula of the liquid is as follows: soluble starch 15-25g/L KNO 3 0.5-1.5g/L，K 2 HPO 4 0.2-0.8g/L，MgSO 4 ·7H 2 O 0.2-0.8g/L，NaCl 0.2-0.8g/L，FeSO 4 ·7H 2 O 0.01-0.02g/L。
3. A method for preparing a pigment solution, comprising the steps of:
(1) Inoculating the streptomycete BG-R1 of claim 1 into the culture solution of the first Gao's strain of claim 2 for culturing;
(2) Centrifuging and filtering to obtain supernatant.
4. A pigment solution prepared by the method of claim 3.
5. A method of dyeing a fabric comprising immersing the fabric in the pigment solution of claim 4.
6. The method for dyeing fabrics according to claim 5, wherein the fabrics are silk fabrics and the bath ratio is 1:20-40.
7. The method for dyeing fabrics according to claim 5, further comprising the steps of heating, dyeing with heat, washing and drying.
8. The method of dyeing fabrics according to claim 7, wherein the post-warming, thermal-insulation dyeing comprises two warming steps.
9. The method for dyeing fabrics according to claim 8, wherein the temperature is raised to 55-65 ℃ for the first time, and the fabric is dyed for 25-35min with heat preservation; heating to 75-85 ℃ for the second time, and keeping the temperature for dyeing for 15-25min.
10. Use of streptomycete BG-R1 of claim 1 or the pigment solution of claim 4 in textile, leather printing and dyeing.
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