CN113916820A - Method for rapidly determining content of total carotenoids in bacterial liquid - Google Patents
Method for rapidly determining content of total carotenoids in bacterial liquid Download PDFInfo
- Publication number
- CN113916820A CN113916820A CN202111057601.2A CN202111057601A CN113916820A CN 113916820 A CN113916820 A CN 113916820A CN 202111057601 A CN202111057601 A CN 202111057601A CN 113916820 A CN113916820 A CN 113916820A
- Authority
- CN
- China
- Prior art keywords
- bacterial liquid
- carotenoid content
- total carotenoid
- concentration
- bacterial
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000007788 liquid Substances 0.000 title claims abstract description 116
- 230000001580 bacterial effect Effects 0.000 title claims abstract description 113
- 235000021466 carotenoid Nutrition 0.000 title claims abstract description 107
- 150000001747 carotenoids Chemical class 0.000 title claims abstract description 107
- 238000000034 method Methods 0.000 title claims abstract description 50
- 241000894006 Bacteria Species 0.000 claims abstract description 53
- 238000012417 linear regression Methods 0.000 claims abstract description 30
- 230000003287 optical effect Effects 0.000 claims description 27
- 239000000725 suspension Substances 0.000 claims description 5
- 230000004071 biological effect Effects 0.000 abstract description 12
- 239000003960 organic solvent Substances 0.000 abstract description 10
- 238000001035 drying Methods 0.000 abstract description 5
- 230000007613 environmental effect Effects 0.000 abstract description 5
- 241001052560 Thallis Species 0.000 abstract description 2
- 238000012423 maintenance Methods 0.000 abstract description 2
- 238000012545 processing Methods 0.000 abstract description 2
- 241000797752 Erythrobacter pelagi Species 0.000 description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 238000007796 conventional method Methods 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 241001526679 Mimachlamys nobilis Species 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 238000000861 blow drying Methods 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000000643 oven drying Methods 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
Abstract
The invention relates to the technical field of biology, in particular to a method for rapidly determining the total carotenoid content in bacterial liquid. By constructing a standard curve of the concentration value of the bacterial liquid and the total carotenoid content in the bacterial liquid, a linear regression equation between the concentration value and the total carotenoid content can be established, and then the carotenoid content of the bacterial liquid at any concentration can be calculated by using the equation. Compared with the traditional method, the method has the advantages of less steps, no need of using an organic solvent, no need of drying thalli, no need of processing bacteria and capability of keeping the biological activity of the carotenoid. Therefore, the invention has the advantages of rapidness, convenience, environmental protection, low cost, maintenance of the biological activity of the carotenoid and the like.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a method for rapidly determining the content of total carotenoids in a bacterial liquid.
Background
Carotenoids (carotenoids) are a general term for a group of natural pigments that are widely found in nature and are mainly synthesized by plants, bacteria and fungi. The natural carotenoid can be used as a coloring agent and a food additive and also can be used as an antioxidant due to the characteristics of various biological activities, and has important biological functions, such as high efficiency quenching of singlet oxygen; enhancing the immune function of the organism; the bacteria can participate in various life activities in the bacterial body, and the adaptability of the bacteria to the environment can be improved. At present, carotenoids are widely applied to the fields of foods, cosmetics, health products and the like. With the pursuit of healthy life, the market demand of natural carotenoids is huge, and the global market is expected to reach $ 69 hundred million in 2026. Therefore, the development of a product rich in natural carotenoids is of great significance.
Bacteria are important sources for obtaining natural carotenoids for human beings, and obtaining natural carotenoids by culturing bacteria is one of the main approaches at present. The traditional method for determining the content of total carotenoids in bacterial liquid comprises the following steps: 1. sample preparation: the bacterial suspension was centrifuged at 5000g for 10 minutes to obtain bacterial cells. 2. Wall breaking treatment: the wall is broken by grinding, ultrasonic crushing, high-speed bead grinding, repeated freeze thawing and other methods. 3. Organic solvent extraction: extracting carotenoid in thallus with organic solvent such as methanol and acetone in shaking table at room temperature in dark condition until thallus turns white. 4. Obtaining a supernatant: centrifuging at 5000g for 10 min to obtain carotenoid-containing supernatant, oven drying and weighing thallus precipitate. 5. Determination of total carotenoid content: and detecting the absorbance of the supernatant by using an ultraviolet spectrophotometer at a wavelength of 480 nm. Finally, the total carotenoid content produced by the bacteria was calculated according to the following formula:
total carotenoid content (TCC,. mu.g/g) ═ A × D × V/(E × W)
Wherein A is the absorbance of the total carotenoid content at 480nm, D is the dilution ratio, V is the volume of the extract (mL), E is the extinction coefficient of the total carotenoid of 0.16, and W is the dry weight (g) of the cells.
The disadvantages of this method are: 1. the steps are complicated, the time consumption is long, and the whole process needs 12 hours. 2. Is not environment-friendly, and needs a large amount of organic solvents such as methanol, acetone and the like. 3. The biological activity of the carotenoid is destroyed, and the natural carotenoid has the biological activity characteristic and has stronger sensitivity to light, heat and the like. 4. The cost is high, for example, the organic solvent required for measuring the carotenoid content in 1L of bacterial liquid, methanol is 3-5 Yuan renmingbu, and acetone is 10-15 Yuan rengbu.
In view of the important role of bacteria in obtaining natural carotenoid, the method for determining the yield of the carotenoid produced by the bacteria, which is developed rapidly, simply, conveniently, environmentally and at low cost, has wide application prospect. However, no report has been made so far for rapidly detecting the total carotenoid content of bacteria by using the bacterial liquid concentration and the total carotenoid content to construct a linear regression equation. The method can rapidly detect the total carotenoid content in the bacterial liquid by utilizing a linear regression equation established by the bacterial liquid concentration and the total carotenoid content, and can solve the problems in the traditional method. Therefore, the invention has the advantages of rapidness, convenience, environmental protection, low cost, capability of keeping the biological activity of the carotenoid and the like.
Disclosure of Invention
The invention aims to overcome the defects of complicated steps, long time consumption, no environmental protection, high cost, damaged biological activity of carotenoid and the like in the determination of the total carotenoid content of bacterial liquid by using the traditional method, and simultaneously provides a rapid, simple, convenient, environment-friendly and low-cost determination method capable of keeping the biological activity of the carotenoid.
The principle of the invention lies in that the bacteria liquid concentration of bacteria under a certain wavelength and the optical density have a positive linear relation, and the bacteria liquid concentration can be rapidly measured by utilizing a spectrophotometer. We have found that: the bacteria liquid concentration and the carotenoid content have a positive linear relation, so that a linear regression equation between the bacteria liquid concentration and the total carotenoid content in the bacteria liquid is established by constructing a standard curve of the bacteria liquid concentration and the total carotenoid content in the bacteria liquid, and then the carotenoid content of the bacteria liquid at any concentration can be calculated by utilizing the equation. The method comprises the following steps: (1) obtaining the concentration of the carotenoid production bacteria solution; (2) establishing a linear regression equation for measuring the total carotenoid content of the bacterial liquid and the concentration of the bacterial liquid; (3) and obtaining the total carotenoid content of the bacterial liquid according to a linear regression equation. Compared with the traditional method, the method has the advantages of less steps, no need of using an organic solvent, no need of drying thalli, no need of processing bacteria and capability of keeping the biological activity of the carotenoid. Therefore, the invention has the advantages of rapidness, convenience, environmental protection, low cost, maintenance of the biological activity of the carotenoid and the like.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for rapidly determining the content of total carotenoids in a bacterial liquid comprises the following steps:
A. preparing standard bacterial liquid with different concentrations, measuring the optical density value of the standard bacterial liquid as X, drawing a standard curve by taking the concentration of the standard bacterial liquid as Y, and establishing a linear regression equation for rapidly measuring the concentration of the bacterial liquid: y ═ AX + B.
B. And (2) measuring the total carotenoid content of the standard bacterial liquid by adopting a traditional method as y, drawing a standard curve by taking the concentration of the standard bacterial liquid as x, and establishing a linear regression equation for rapidly measuring the total carotenoid content of the bacterial liquid: and y is ax + b.
C. And (4) obtaining an optical density value of the unknown bacterial liquid, substituting the optical density value into the linear regression equation established in the step A to obtain the concentration of the unknown bacterial liquid, and substituting the linear regression equation established in the step B to obtain the total carotenoid content of the unknown bacterial liquid.
Preferably, the bacterium is a bacterium capable of producing carotenoids.
Preferably, the optical density value is obtained using an ultraviolet spectrophotometer.
Preferably, in step a, the concentration of the bacterial liquid is obtained by using a plate colony counting method, and the bacterial liquid is configured into the standard bacterial liquid.
Preferably, the method further comprises the following steps:
D. checking: and D, obtaining the total carotenoid content of the unknown bacterial liquid by adopting a traditional method, wherein the total carotenoid content of the unknown bacterial liquid obtained in the step C has no significant difference compared with the total carotenoid content of the unknown bacterial liquid obtained in the step C.
The application of the method for rapidly determining the total carotenoid content in the bacterial liquid is characterized in that the total carotenoid content of any bacterial liquid is rapidly obtained.
After a double linear regression equation is established and verified to be correct, the total carotenoid content of the bacterial liquid of the same species can be quickly obtained by substituting the double linear regression equation as long as the optical density value of the bacterial liquid is directly obtained, and convenience is provided for relevant experiments and researches for producing the carotenoid by bacteria.
Compared with the prior art, the implementation of the invention has the following beneficial effects:
(1) is rapid and simple. The traditional method for determining the content of the total carotenoids in the bacteria needs the steps of centrifugation, wall breaking, extraction, drying and the like, the whole process needs 12 hours, and the method is tedious and time-consuming; the invention only needs two steps to obtain the result: firstly, obtaining the concentration of bacterial liquid. Secondly, obtaining the total carotenoid content of the bacterial liquid according to an equation; the whole process can be completed within a few minutes. Therefore, the invention is fast and simple.
(2) Is environment-friendly. The traditional method needs a large amount of organic solvents such as methanol, acetone and the like, and is not environment-friendly; the present invention does not require the use of organic solvents. Therefore, the invention is environment-friendly.
(3) The cost is low. The traditional method needs a large amount of organic solvents such as methanol, acetone and the like, and needs electric drying or nitrogen blow-drying, which costs a certain amount. The invention does not need to use organic solvent and does not need drying. Therefore, the cost is low.
(4) The bacterial activity and carotenoid bioactivity are maintained. The traditional method needs wall breaking and extraction treatment on bacteria, the bacteria die and carotenoid degradation is easy to lose activity. The invention does not need to treat bacteria, and can maintain the activity of the bacteria and the biological activity of the carotenoid.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and all other embodiments obtained by those skilled in the art without any inventive work are within the scope of the present invention.
In order to avoid redundancy of the application text, the present invention is described in examples of only Erythrobacter pelagi of the genus gibberellic of the family gibberellicaceae. It is a bacterium which is separated from the intestinal tract of chlamys nobilis 'south Australia golden shellfish' and can produce carotenoid. The bacterium is an orange, aerobic, rod-shaped gram-negative bacterium. Based on 16S rRNA and phylogenetic analysis, the similarity of the gene sequence of the 16S rRNA of the strain to Erythrobacter pelagi UST081027-248 is 99.2%.
Those skilled in the art will appreciate that similar experimental results will be obtained using other bacteria capable of producing carotenoids.
Example 1: the method is adopted to determine the total carotenoid content of the E.pelagi bacterial liquid at the temperature of 25 DEG C
Time: 5/6/2021, site: shantou university.
The operation steps are as follows:
e.pelagi bacterial liquid concentration rapid determination: the bacterial suspension concentration (per mL) was obtained by plate colony counting. Bacterial liquid is serially diluted into 6 bacterial liquids with different concentrations. The optical density values measured with a uv spectrophotometer λ 600nm wavelength were 0.037, 0.057, 0.102, 0.262, 0.399, 0.619. Corresponding plate colony counts (× 10)6pieces/mL) results were 21, 40, 45, 170, 290, 550. And drawing a standard curve and establishing a linear regression equation by taking the optical density value of the bacterial liquid as X and the counting result of the plate bacterial colony as Y: Y889.25X-32.754 (R)2＝0.9808)。
And (3) testing a standard curve linear regression equation: substituting the measured optical density values of the bacterial liquid of 0.117, 0.138 and 0.257 into the equation to obtain the bacterial concentration (multiplied by 10)6pieces/mL) was 71, 90, 196. Bacterial concentration obtained from actual plate colony count (× 10)6piece/mL) 40,50. 140 was analyzed by the T test (T test) of SPSS 19.0 statistical software to determine no significant difference (p) between the linear regression equation of the bacterial suspension and the plate colony counts>0.05)。
2. Establishing a linear regression equation of the concentration of the bacterial liquid and the total carotenoid content of the bacterial liquid:
diluting the E.pelagi bacterial liquid in 8 concentration gradients according to a certain proportion, accurately transferring 500 mu L of the diluted bacterial liquid, and measuring the optical density values of the E.pelagi bacterial liquid by an ultraviolet spectrophotometer to be 0.47, 0.598, 0.833, 1.062, 1.292, 1.309, 1.360 and 1.440 under the condition that the lambda is 600 nm. Obtaining the concentration (multiplied by 10) of the bacterial liquid according to the linear regression method of the optical density value and the concentration of the bacterial liquid6individual/mL) is: 385. 499, 708, 912, 1116, 1131, 1177, 1248.
Determining the total carotenoid content (mug/mL) in the bacterial liquid with different concentration gradients in the step 1 as 0.338, 0.430, 0.719, 0.828, 1.039, 1.040, 1.116 and 1.163 by a traditional method (as described in the background art), taking the total carotenoid content (mug/mL) as y, taking the bacterial liquid concentration obtained in the step 1 as x, drawing a standard curve, and establishing a linear regression equation for determining the total carotenoid content (mug/mL) of the E.pelagi bacterial liquid: y-0.0009 x-0.0063 (R)2＝0.9887)。
3. And (3) effect inspection: randomly taking a certain amount of bacteria liquid with different concentrations, measuring the optical density values of 0.469, 0.983, 1.185 and 1.316 by an ultraviolet spectrophotometer under the condition that the wavelength is 600nm, and substituting the measured optical density values into the equation in the step 1 to obtain the concentration of the bacteria liquid (multiplied by 10)6individual/mL) is: 384. 841, 1021, 1137. Substituting the obtained concentration into the equation in step 2 to obtain the total carotenoid content (μ g/mL) of the bacteria liquid at the concentration of 0.339, 0.751, 0.913 and 1.017. Whereas the total carotenoid content (μ g/mL) determined by the conventional method (as described in the background art) was 0.344, 0.856, 1.256, 1.016. The total carotenoid content calculated by the linear regression equation of the SPSS 19.0 statistical software through T test analysis has no significant difference (P) from the total carotenoid content measured by the traditional method>0.05)。
4. The application comprises the following steps: collecting 3 groups of E.pelagi bacteria liquid, and measuring optical density values of 0.586 and 0 at wavelength of 600nm in ultraviolet spectrophotometer087, 1.103, into the bacterial concentration (× 10) obtained in the equation of step 16pieces/mL) is 488, 45 and 948, and the total carotenoid content (mu g/mL) of the bacteria liquid at the concentration is 0.433, 0.034 and 0.847 by substituting the pieces/mL) into the equation in the step 2. Culturing the bacterial liquid of the 3 groups of bacteria of E.pelagi overnight at a constant temperature of 25 deg.C, measuring the optical density values of 0.608, 0.102 and 1.125 at a wavelength of 600nm in an ultraviolet spectrophotometer, and substituting into the equation of step 1 to obtain the bacterial concentration (x 10)6piece/mL) is 508, 58 and 968, and the total carotenoid content (mu g/mL) of the bacteria liquid at the concentration is 0.451, 0.046 and 0.865 by substituting the piece/mL) into the equation in the step 2. The method proves that the bacterial activity is maintained, and the bacterial concentration and the carotenoid content are increased to a certain extent after overnight culture.
Example 2: the method is adopted to determine the total carotenoid content of the E.pelagi bacterial liquid at the temperature of 30 DEG C
Time: 5/10/2021, site: shantou university
The operation steps are as follows:
e.pelagi bacterial liquid concentration rapid determination: because the same kind of bacteria is used, the linear relation between the optical density value of the bacteria liquid and the concentration of the bacteria liquid in the embodiment 1 is directly used, and the linear regression equation between the optical density value and the concentration of the bacteria liquid is obtained as follows: Y889.25X-32.754 (R)2＝0.9808)。
2. Establishing a linear regression equation of the concentration of the bacterial liquid and the total carotenoid content of the bacterial liquid:
diluting the E.pelagi bacterial liquid in 6 concentration gradients according to a certain proportion, accurately transferring 500 mu L of the diluted bacterial liquid, and measuring the optical density values of the E.pelagi bacterial liquid by an ultraviolet spectrophotometer to be 0.837, 0.880, 1.229, 1.418, 1.470 and 1.506 under the condition that the wavelength is 600 nm. Obtaining the concentration (multiplied by 10) of the bacterial liquid according to the linear regression method of the optical density value and the concentration of the bacterial liquid6pieces/mL) 712, 750, 1060, 1228, 1274, 1306.
Determining the total carotenoid content (μ g/mL) in the bacteria solution with different concentration gradients in step 1 as 0.681, 0.730, 1.019, 1.077, 1.279 and 1.324 by conventional method (as described in background), and using it as y in step 1And (3) taking the concentration of the obtained bacterial liquid as x, drawing a standard curve, and establishing a linear regression equation for determining the total carotenoid content (mu g/mL) of the E, pelagi bacterial liquid: y is 0.001x +0.007 (R)2＝0.9712)。
3. And (3) effect inspection: randomly taking a certain amount of bacteria liquid with different concentrations, measuring the optical density values of 0.887, 1.263 and 1.444 of the bacteria liquid by an ultraviolet spectrophotometer under the condition that the wavelength is 600nm, and substituting the measured values into the equation in the step 1 to obtain the concentration of the bacteria liquid (multiplied by 10)6pieces/mL) are 756, 1090, 1251. Substituting the obtained concentration into the equation in step 2 to obtain the total carotenoid content (μ g/mL) of the bacteria liquid at the concentration of 0.763, 1.097 and 1.258. Whereas the total carotenoid content (μ g/mL) as determined by conventional methods (as described in the background) is 0.825, 0.928, 1.251. The total carotenoid content calculated by the linear regression equation of the SPSS 19.0 statistical software through T test analysis has no significant difference (P) from the total carotenoid content measured by the traditional method>0.05)。
4. The application comprises the following steps: collecting 3 groups of bacterial liquid of E.pelagi bacteria, measuring optical density values of 0.632, 1.398, and 1.123 at wavelength of 600nm in ultraviolet spectrophotometer, and substituting into the bacterial concentration (x 10) obtained in the equation of step 16piece/mL) of 529, 1210 and 966, and substituting the piece/mL of the formula in the step 2 to obtain the total carotenoid content (mu g/mL) of the bacteria liquid at the concentration of 0.536, 1.217 and 0.973. Culturing the bacterial liquid of the 3 groups of bacteria of the e.pelagi bacteria at a constant temperature of 30 ℃ overnight, measuring the optical density values of 0.662, 1.458 and 1.189 under the wavelength of 600nm of an ultraviolet spectrophotometer, and substituting the optical density values into the bacterial concentration (multiplied by 10) obtained in the equation of the step 16piece/mL) is 556, 1264 and 1025, and the total carotenoid content (mu g/mL) of the bacterial liquid at the concentration is 0.563, 1.271 and 1.032 by substituting the piece/mL) into the equation in the step 2. The method proves that the bacterial activity is maintained, and the bacterial concentration and the carotenoid content are increased to a certain extent after overnight culture.
In the above 2 embodiments, the total carotenoid content of bacterial liquids with different concentrations at the same temperature can be directly calculated by using a linear regression equation. Compared with the traditional method, the method has the advantages of rapidness, simplicity, convenience, environmental protection, low cost, capability of keeping the biological activity of the carotenoid and the like.
In addition, the idea of calculating the total carotenoid content of other bacterial liquids by a linear regression equation is also covered by the idea of the invention.
The above disclosure is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the scope of the present invention, therefore, the present invention is not limited by the appended claims.
Claims (7)
1. A method for rapidly determining the content of total carotenoids in a bacterial liquid is characterized by comprising the following steps:
A. preparing standard bacterial liquid with different concentrations, measuring the optical density value of the standard bacterial liquid as X, drawing a standard curve by taking the concentration of the standard bacterial liquid as Y, and establishing a linear regression equation for rapidly measuring the concentration of the bacterial liquid: y ═ AX + B.
B. And (2) measuring the total carotenoid content of the standard bacterial liquid by adopting a traditional method as y, drawing a standard curve by taking the concentration of the standard bacterial liquid as x, and establishing a linear regression equation for rapidly measuring the total carotenoid content of the bacterial liquid: and y is ax + b.
C. And (4) obtaining an optical density value of the unknown bacterial liquid, substituting the optical density value into the linear regression equation established in the step A to obtain the concentration of the unknown bacterial liquid, and substituting the linear regression equation established in the step B to obtain the total carotenoid content of the unknown bacterial liquid.
2. The method for rapidly determining the total carotenoid content in a bacterial liquid according to claim 1, wherein the bacteria are bacteria capable of producing carotenoids.
3. The method for rapidly determining the total carotenoid content in a bacterial liquid according to claim 1, wherein the bacteria comprise E.
4. The method for rapidly determining the total carotenoid content in a bacterial liquid according to claim 1, wherein the optical density value is obtained by an ultraviolet spectrophotometer.
5. The method for rapidly determining the total carotenoid content in the bacterial liquid according to claim 1, wherein in the step A, the concentration of the bacterial liquid is obtained by using a plate colony counting method, and the bacterial liquid is prepared into the standard bacterial liquid.
6. The method for rapidly determining the total carotenoid content in the bacterial liquid according to claim 1, further comprising the following steps:
D. checking: and D, obtaining the total carotenoid content of the unknown bacterial liquid by adopting a traditional method, wherein the total carotenoid content of the unknown bacterial liquid obtained in the step C has no significant difference compared with the total carotenoid content of the unknown bacterial liquid obtained in the step C.
7. The use of the method according to claim 1 for rapidly determining the total carotenoid content in a bacterial suspension, wherein the total carotenoid content of any bacterial suspension is rapidly obtained.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111057601.2A CN113916820B (en) | 2021-09-09 | 2021-09-09 | Method for rapidly determining total carotenoid content in bacterial liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111057601.2A CN113916820B (en) | 2021-09-09 | 2021-09-09 | Method for rapidly determining total carotenoid content in bacterial liquid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113916820A true CN113916820A (en) | 2022-01-11 |
| CN113916820B CN113916820B (en) | 2024-01-09 |
Family
ID=79234864
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111057601.2A Active CN113916820B (en) | 2021-09-09 | 2021-09-09 | Method for rapidly determining total carotenoid content in bacterial liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113916820B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101042348A (en) * | 2006-03-24 | 2007-09-26 | 中国科学院长春光学精密机械与物理研究所 | Device for nondestructively detecting carotenoid concentration in human body |
| CN102137677A (en) * | 2008-03-19 | 2011-07-27 | 弗洛瑞安·施威格特 | Method for the extraction and detection of liposoluble ingredients contained in biological materials |
| CN102680420A (en) * | 2012-05-30 | 2012-09-19 | 国药集团威奇达药业有限公司 | Method for rapidly determining biotins in miniaturized manner |
| US20130052636A1 (en) * | 2011-08-22 | 2013-02-28 | Spectral Platforms, Inc. | Rapid detection of metabolic activity |
| US20130137119A1 (en) * | 2011-11-30 | 2013-05-30 | The United States Of America, As Represented By The Secretary, Department Of Health And Human | Detection of bacterial contamination in a sample |
| CN104964957A (en) * | 2015-07-02 | 2015-10-07 | 华南理工大学 | Method for rapidly and nondestructively detecting astaxanthin in C.zofingiensis cells |
| CN108913746A (en) * | 2018-07-19 | 2018-11-30 | 威海利达生物科技有限公司 | By improving red phaffia rhodozyma biomass synthesizing astaxanthin and method for measuring |
| CN112239777A (en) * | 2019-07-16 | 2021-01-19 | 汕头大学 | Qualitative and quantitative detection method for carotenoid producing bacteria |
| CN112980917A (en) * | 2019-12-16 | 2021-06-18 | 天津大学 | Method for quickly quantifying escherichia coli in water |
-
2021
- 2021-09-09 CN CN202111057601.2A patent/CN113916820B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101042348A (en) * | 2006-03-24 | 2007-09-26 | 中国科学院长春光学精密机械与物理研究所 | Device for nondestructively detecting carotenoid concentration in human body |
| CN102137677A (en) * | 2008-03-19 | 2011-07-27 | 弗洛瑞安·施威格特 | Method for the extraction and detection of liposoluble ingredients contained in biological materials |
| US20130052636A1 (en) * | 2011-08-22 | 2013-02-28 | Spectral Platforms, Inc. | Rapid detection of metabolic activity |
| CN103733050A (en) * | 2011-08-22 | 2014-04-16 | 光谱平台有限公司 | Rapid detection of metabolic activity |
| US20130137119A1 (en) * | 2011-11-30 | 2013-05-30 | The United States Of America, As Represented By The Secretary, Department Of Health And Human | Detection of bacterial contamination in a sample |
| CN102680420A (en) * | 2012-05-30 | 2012-09-19 | 国药集团威奇达药业有限公司 | Method for rapidly determining biotins in miniaturized manner |
| CN104964957A (en) * | 2015-07-02 | 2015-10-07 | 华南理工大学 | Method for rapidly and nondestructively detecting astaxanthin in C.zofingiensis cells |
| CN108913746A (en) * | 2018-07-19 | 2018-11-30 | 威海利达生物科技有限公司 | By improving red phaffia rhodozyma biomass synthesizing astaxanthin and method for measuring |
| CN112239777A (en) * | 2019-07-16 | 2021-01-19 | 汕头大学 | Qualitative and quantitative detection method for carotenoid producing bacteria |
| CN112980917A (en) * | 2019-12-16 | 2021-06-18 | 天津大学 | Method for quickly quantifying escherichia coli in water |
Non-Patent Citations (5)
| Title |
|---|
| FU-SHIU KUO 等: "Effects of light sources on gorwth and carotenoid content of photosynthetic bacteria Rhodopseudomonas palustris", BIORESOURCE TECHNOLOGY, vol. 113, pages 315 - 318 * |
| L.G.EROKHINA 等: "The content and composition of phycobilisome pigments in cells of ancient viable cyanobacteria from Arctic permafrost", MICROBIOLOGY, vol. 67, no. 6, pages 682 - 687 * |
| 董自艳 等: "紫外-可见分光光度法快速确定细菌菌液的浓度", 中国药品标准, vol. 15, no. 2, pages 120 - 121 * |
| 董自艳: "紫外-可见分光光度法快速确定细菌菌液的浓度", 重要药品标准, vol. 15, no. 2, pages 120 - 121 * |
| 郝常明 等: "光合细菌类胡萝卜素发酵的研究", 天津轻工业学院学报, no. 4, pages 1 - 7 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113916820B (en) | 2024-01-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20150052639A1 (en) | Culture method for Antrodia cinnamomea | |
| Ryu et al. | Statistical optimization of microalgae Pavlova lutheri cultivation conditions and its fermentation conditions by yeast, Candida rugopelliculosa | |
| CN104530102B (en) | It is a kind of to detect the fluorescence copper complex of sulphion and its application in organism | |
| EP3134478B1 (en) | Improvements in the synthesis of phycocyanins | |
| Chapman et al. | A high‐throughput and machine learning resistance monitoring system to determine the point of resistance for Escherichia coli with tetracycline: Combining UV‐visible spectrophotometry with principal component analysis | |
| CN109680022A (en) | The preparation method of chlorella polysaccharide | |
| Shi et al. | A qualitative and quantitative high-throughput assay for screening of gluconate high-yield strains by Aspergillus niger | |
| Chakraborty et al. | Isolation and characterization of pigment producing marine actinobacteria from mangrove soil and applications of bio-pigments | |
| CN113621527B (en) | Novel smoke tube bacterium SZ-4 and application thereof | |
| Sharma et al. | Combined use of Fourier transform infrared and Raman spectroscopy to study planktonic and biofilm cells of Cronobacter sakazakii | |
| Arslan et al. | Determination of bioactive properties of protein and pigments obtained from Spirulina platensis | |
| CN113916820A (en) | Method for rapidly determining content of total carotenoids in bacterial liquid | |
| Zhou et al. | Isolation and identification of pigment-producing filamentous fungus DBFL05 and its pigment characteristics and chemical structure | |
| CN109211805B (en) | Verification analysis method for traceability of mussel carotenoid extract | |
| Yan et al. | Yellow pigment of Metarhizium anisopliae and its application to the dyeing of fabrics | |
| CN103217398A (en) | Classification and identification upon 13 pathogenic bacteria by using Fourier transform infrared spectroscopic technology | |
| CN108918456A (en) | A kind of Astaxanthin In Haematococcus Pluvialis synthesis process research method based on infrared spectroscopy micro-imaging technique | |
| Durrani et al. | Biological characterization and protein profiles of two model bacteria by SDS-PAGE and FT-IR | |
| Choi et al. | Water-soluble red pigment production by Paecilomyces sinclairii and biological characterization | |
| CN110887921A (en) | Method for efficiently and rapidly analyzing characteristic volatile components of eucommia leaves and fermentation product thereof | |
| CN101936960B (en) | Analytical method of components of fatty acid contained in listeria cells | |
| CN106479913B (en) | Gordonia bronchialis and application thereof | |
| CN101864380A (en) | Blue pigment producing bacteria and method for preparing crude preparation by using the same | |
| NL2024514B1 (en) | Enterococcus faecalis with biological antibacterial activity | |
| CN103411915A (en) | Infrared spectroscopy identification method of traditional Chinese medicine preparation-Jinshuibao capsule |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |