TWI642632B - Gynura decolorized metabolite, the composition and the use for decolorization of dye thereof - Google Patents

Abstract

The invention provides a decolorizing metabolite of Hongfengcai liquid, a composition thereof and a dye decolorizing application thereof, wherein the decolorizing metabolite of the red cabbage liquid is preliminarily decolorized by Shewanella sp., and the composition comprises red phoenix. The vegetable liquid decolorizes the metabolite; the composition can generate a electron shuttle and effectively promote the redox reaction, effectively decolorizing and degrading the dye. The compositions of the present invention do not include unnatural chemicals and are environmentally friendly.

Description

Decolorization metabolite of red phoenix liquid, its composition and application of dye decolorization

The invention provides a decolorizing metabolite of Hongfengcao liquid, a composition thereof and a dye decolorizing application thereof, the composition comprising a decolorizing metabolite of Hongfengcao liquid.

The waste water discharged from the process of colored dyes contains 10-15% of dyes, which have a large amount of organic or inorganic compounds, which cause serious harm to the environment. In order to make the wastewater discharge comply with the regulatory requirements, the industry uses bleaching water addition, redox, activated carbon adsorption, or electrolysis to achieve the decolorization effect, but after the decoloring agent, coagulant or both, it cooperates with the wastewater. Will produce agglomerated material stroke sludge deposition caused by secondary pollution.

Most of the dye molecules are compounds having an aromatic structure, and the chromophoric groups are mostly azo groups (-N=N-), and the structure is stable, and the dyes are fixed on the textile fibers by covalent bonding. Some azo dyes or their degraded products have toxicity and potential cancer risk. Therefore, dye wastewater treatment is a public health and environmental protection point that cannot be ignored.

Previous studies have found that chemically synthesized dyes and decolorized metabolites can be used as electron shuttles (ES) to promote electron transfer and promote bio-decolorization and energy recovery in wastewater treatment (Chen et al. 2013 Int J Hydrogen) Energ). However, in the wastewater treatment, additional non-natural chemical synthetic substances such as malachite green, o- and p-aminophenol (2- Aminophenol, 2AP and 4-aminophenol, 4AP), 1-amino-2 naphthol (1A2N), 4-aminonaphthol (4A1N), although effective, can cause secondary pollution and is not consistent with sustainable development. As well as environmental protection application regulations, it is necessary to find ES applications that are ecologically compatible and good-natured to utilize bio-stimulation strategies for wastewater decolorization.

The purpose of this study is to extract the natural product ES from the natural ecology and to decompose and convert the natural ES produced by the natural environment to replace the chemical synthesis that will cause secondary pollution. substance.

The application of Hongfengcai in the past is the application of the extract preparation or the use of its juice as a health food. The Taiwan patent (certificate No. M499072) discloses a food structure containing red phoenix vegetable and fruit powder which promotes the absorption of ferrous ferrous sulphate. Hongfengcai is used as a vegetable and fruit powder additive to promote the absorption of ferrous ferrous sulphate; Taiwan Patent (Publication No. 201515655) discloses a polar extract of Hongfengcai and its preparation method and use, wherein the polar extract of red phoenix can increase iron in plants The effective absorption of the juice reduces the incidence of anemia; however, there is currently no research or public information to disclose the biotransformation of Hongfengcai liquid for other environmentally friendly application techniques.

The object of the present invention is to provide a decolorizing metabolite of Hongfengcai liquid, a composition thereof and a dye decolorizing application thereof, thereby achieving the purpose of decolorizing, degrading and eliminating waste water pollution.

In order to achieve the above object, the present invention provides a method for preparing a decolorizing metabolite of Hongfengcai liquid, which comprises: performing decolorization metabolism after mixing Shewanella sp. and red phoenix juice.

Wherein, the preparation method further comprises: mixing the red phoenix liquid and the bacterium of Shewanella sp., and the decolorization rate thereof is 90%.

Wherein, the added volume ratio of the red phoenix liquid is 1%-10% of the decolorized metabolite of the red phoenix liquid.

Wherein, the preparation method of the red phoenix liquid comprises: removing the supernatant from the red phoenix vegetable and water, and extracting the supernatant liquid, wherein the weight ratio of the red phoenix to the water is 10-20%: 80-90%; the supernatant The filtrate was filtered with a filter paper to obtain a filtrate; and the filtrate was obtained by a filter having a pore size of 0.24 μm.

The present invention further provides a composition for decolorizing a dye, which comprises the aforementioned decolorizing metabolite of Hongfengcao liquid.

Wherein, the composition further comprises a decolorizing metabolic intermediate for increasing the electron shuttle, the decolorizing metabolic intermediate selected from the group consisting of malachite green, o-aminophenol (2AP), p-aminophenol (4AP), 1-amino group. A group consisting of 2-naphthol (1A2N) and 4-aminonaphthol (4A1N). .

The invention further provides a method of decolorizing a dye comprising adding to the dye a composition for dye decolorization as described above.

Among them, the method is used for decolorization of dye wastewater.

Among them, the method is for degrading an azo dye.

Among other things, the method can be applied to a biofuel cell that degrades the dye and produces an electron shuttle.

Fig.1 Experimental procedure of decolorization reaction of microorganisms in Hongfengcai liquid; Fig.2 Cyclic voltammetry of different strains in Hongfengcao water extract before decolorization and after decolorization Figure 3: The cyclic voltammetry of pre-decolorization and decolorization was carried out by different doses of red cabbage solution by decolorization of microbial strain (WLP72); Figure 4: Decolorization of red phoenix metabolites (DM) after discoloration The effect of strain (WLP72, YTL1) on the decolorization and cell growth of reactive black dye No. 5 (RBk5).

Fig. 5 Cyclic voltammetry analysis of different strains of Shewanella sp. in Hongfengcai liquid before decolorization and after decolorization, using commercially available strain BCRC 80907 culture liquid and red phoenix liquid The composition, which measures the cyclic voltammogram, can also produce a redox reaction.

All of the technical and scientific terms described in this specification, unless otherwise defined, are intended to be common to those of ordinary skill in the art. The present invention is exemplified by the following examples, which are intended to be illustrative and not restrictive. Unless otherwise indicated, the materials used in the present invention are readily available commercially, and the following are merely examples of available pipelines.

The preparation method of the red phoenix extract used in the invention comprises the steps of: grinding the red phoenix with water into a mud; centrifuging at 11,000 rpm and 23 ° C for 20 minutes; taking the supernatant and pumping the air (5A filter paper (ADVANTEC)) The pore size was 0.06 mm. The filtrate was obtained; the filtrate was filtered through a filter having a pore size of 0.24 μm, and a red cabbage extract which can be used for the production of an electron shuttle was obtained.

Cyclic voltammetry (CV) is a measure of changing the potential to obtain the direction of the redox current, mainly by applying a cyclic potential, starting from a The measurement of the direction of the reduction current is mainly performed by applying a cyclic potential, applying a potential from a starting potential to an end point potential, and then changing back to the same starting potential at the same rate, which is a cycle, but The method of measurement can be cycled multiple times to achieve a definitive result, and a potential-current map obtained by analysis of a reversible oxidation reaction is plotted. When scanning from a low potential to a high potential, the analyte will generate an anodic peak of the oxidation current, and it can be determined at which potential an oxidation reaction occurs. If the decolorized metabolite liquid has a significant redox peak, it has the property of promoting dye discoloration.

Example 1. WLP72 ( Shewanella haliotis ) has the property of promoting discoloration

4 strains WLP72 ( Shewanella haliotis ), K2 ( Exiguobacterium acetylicum ), YTL1 ( Aeromonas hydrophila ), KB23 ( Aeromonas hydrophila ) were cultured in LB medium, and 1-2% medium volume of Hongfengcao water extract (20g/100ml) was added. Degradation and decolorization; after the decolorization rate reaches 90%, the decolorized bacteria solution is centrifuged (11,000 rpm, 15 minutes, 25 ° C) to separate the bacteria, and the upper layer of clear decolorizing metabolite liquid is taken for cyclic voltammetry. As shown in Fig. 1, the cyclic voltammetry is in the scanning range of 0.4 volts (V) to -0.6 V, the scanning rate is 10 millivolts per second (mV ‧ s ^-1 ), and the number of scanning circles is 3, before the decolorization, The cyclic voltammetry spectrum after decolorization, the results in Figure 2 show that the strain WLP72 (Fig. 2A) has obvious redox peak after decolorization, indicating that the decolorized metabolite can indeed produce an intermediate product that promotes electron transfer, strain K2 (Fig. 2B). YTL1 (Fig. 2C) and KB23 (Fig. 2D) do not have obvious redox peaks after decolorization, which means that the same natural plant extracts and the same decolorization conditions have different decolorization products due to their different metabolic physiological activities. Do you have In promoting the effect of electron transfer, only the strain WLP72 can effectively decolorize, and the cyclic voltammogram has a significant redox peak, indicating that it has the property of promoting dye discoloration. The water extract obtained the best decolorization effect.

Example 2, the effect of different doses of Hongfengcao culture on the decolorization ability of WLP72 to promote dye

The strain WLP72 ( Shewanella haliotis ) was cultured in LB medium, and added with 1-10% medium volume of Hongfengcao water extract (high concentration: 20g/100ml, low concentration: 20g/200ml) for degradation and decolorization; After %, the decolorized bacterial liquid was centrifuged (11,000 rpm, 15 minutes, 25 ° C) to separate the bacteria, and the upper clear decolorizing metabolite liquid was taken for cyclic voltammetry, which was in the scanning range of 0.4. Volt (V) to -0.6V, scanning rate is 10 millivolts per second (mV‧s ^-1 ), scanning circle number is 3, compare the cyclic voltammogram before decolorization, after decolorization, Figure 3 results show that high concentration The redox peak after decolorization of WLP72 metabolic liquid cultured in Hongfengcai water extract was the most obvious, indicating that the effect of strain WLP72 on dye decolorization increased with the increase of the concentration of Hongfengcao water extract in the culture solution. The decolorization effect obtained by adding 5% Hongfengcao water extract to the above culture solution is the best.

Example 3, the effect of decolorized metabolites (DM) on the decolorization ability of strains

In order to further test the effect of decolorized metabolites (DM) on the decolorization of different strains, in this example, the above strains WLP72 and YT11 were taken, and the dye-reactive black dye RBk5 was added for decolorization evaluation to verify that the strain contained Does not contain the effect of red cabbage liquid on decolorization metabolism.

After culturing the bacteria and adding the dye, the cell absorbance (wavelength 600 nm) and the dye absorbance (RBk5 maximum absorption wavelength 600 nm) were measured by a spectrophotometer at a fixed time. The concentration measurement can be estimated in the following manner: The measured value of the original bacteria-containing liquid sample is OD _{500 nm} ^X-Dye = OD _{500 nm} ^Dye + OD _{500 nm} ^X ...(1)

After centrifugation, the upper layer (assuming only the dye) is measured as OD _{500 nm} ^sup = OD _{500 nm} ^Dye ... (2)

After centrifugation, the upper layer liquid (assuming only the dye) is measured as OD _{λ max} ^sup = OD _{λ max} ^Dye ... (3)

The triad can immediately determine the dye absorbance (OD _λmax ^Dye ) and the cell absorbance OD _600nm ^X , and then take appropriate dilution factor to make the dye absorbance in the linear range (OD value about <1.0), the concentration The calibration curve calculates the actual concentration separately.

The growth rate and decolorization rate were calculated per unit cell concentration as follows: growth rate d ln X/dt (1/X)(△(X)/△t)

Decolorization rate SDR=-(1/X)(d[dye]/dt)

X: cell concentration (ODU) X ₀ : initial cell concentration (ODU)

t: time (h)

[dye]: dye concentration (mgL ^-1 )

With the addition of decolorized metabolites (DM), the decolorization rate (SDR) of RBk5 by strains was found to be fast and slow: WLP72-DM (13.85)>WLP72 (11.47)>YT11- DM(9.00)>YT11(5.13), it can be found that the decolorization metabolite (DM) added to the red phoenix can increase the decolorization effect of the strain. The decolorization rate of WLP72-DM is 20.69% higher than that of WLP72, and the decolorization rate of YT11-DM is higher than that of YT11. 75.29%, as shown in Figure 4, and the decolorization rain is related to electron transfer, further inferring that the decolorized metabolites of Hongfengcai have improved electrons. The transfer ability can accelerate the reduction and decolorization, effectively improving the decolorization effect. .

Example 4 Shewanella sp. has a decolorizing effect

According to the foregoing embodiment, WLP72 ( Shewanella haliotis ) has microorganisms capable of efficiently decolorizing the ability to generate an electron shuttle. Hsinchu Food Industry Development Research Institute No. BCRC 80907 ( Shewanella algae ) is the same genus as WLP72. This example tests whether WLP72 has the same efficacy as BCRC 80907.

The strains WLP72 ( Shewanella haliotis ) and BCRC 80907 ( Shewanella algae ) were cultured in LB medium, and the extract of Hongfengcao water (20g/100ml) was added with 1-10% medium volume for degradation and decolorization; after the decolorization rate reached 90%, The decolorized bacterial solution was centrifuged (11,000 rpm, 15 minutes, 25 ° C) to separate the bacteria, and the upper clear decolorizing metabolite liquid was taken for cyclic voltammetry, which was 0.4 volts in the scanning range (V). ) to -0.6V, the scanning rate is 10 millivolts per second (mV‧s ^-1 ), the number of scanning circles is 3, compared with the cyclic voltammogram before decolorization and after decolorization, the results of Figure 5 show that the strains WLP72 and BCRC 80907 There are obvious redox peaks, which represent that the decolorized metabolite can indeed produce an intermediate product that promotes electron transfer, indicating that it has the property of promoting dye discoloration. The decolorization effect obtained by adding 5% Hongfengcao water extract to the above culture solution is the best.

It can be inferred from the above results that the WLP72 separated by the applicant (refer to the Yilan University Chemicals Department Yue Peilin master's thesis, June, 105) and the purchased BCRC 80907 and Hongfengcai liquid have the ability to produce electronic shuttle. It can be effectively decolored, and it can be known that the effect of co-cultivation with red phoenix liquid and the production of a electronic shuttle by the red phoenix liquid is the inherent ability of Shewanella sp. , and the biological material of the present invention belongs to the present invention. Those skilled in the art can obtain and implement the present invention in light of the present specification. That is to say, the potential difference of the decolorizing metabolite can generate a electron shuttle and a redox reaction, and can be used for degrading the dye to achieve the effect of decolorization.

The detailed description of the preferred embodiments of the present invention is intended to be limited to the scope of the invention, and is not intended to limit the scope of the invention. The patent scope of this case.

Claims (10)

The invention relates to a method for preparing a decolorizing metabolite of red phoenix liquid, which comprises: decolorizing metabolism after mixing Shewanella sp. culture solution and red phoenix liquid. The preparation method according to claim 1, wherein the preparation method further comprises: after mixing the Shewanella sp. culture solution and the red phoenix solution, the decolorization rate is 90%. The preparation method according to the first aspect of the patent application, wherein the ratio of the added volume of the red phoenix liquid is 1% to 10% of the culture medium of the Shewanella. According to the preparation method of the first aspect of the patent application, the preparation method of the red phoenix liquid comprises: removing the supernatant after centrifugation of the red phoenix, wherein the weight ratio of the red phoenix to the water is 10-20%. : 80~90%; the supernatant is filtered by a filter paper to obtain a filtrate; and the filtrate is obtained by a filter having a pore size of 0.24 micrometers (μm). A composition for decolorizing a dye, comprising the decolorizing metabolite of Hongfengcao liquid according to claim 1 of the patent application. The composition of claim 5, further comprising a decolorizing metabolic intermediate for increasing the electron shuttle, the decolorizing metabolic intermediate selected from the group consisting of malachite green, o-aminophenol (2AP), A group consisting of aminophenol (4AP), 1-amino 2-naphthol (1A2N), and 4-aminonaphthol (4A1N). A method for decolorizing a dye, comprising adding a composition for dye decolorization as described in claim 5 of the patent to a dye. The method of claim 7 is for the decolorization of dye wastewater. The method of claim 7, wherein the method is for degrading an azo dye. The method of claim 7, wherein the method is applicable to a biofuel cell that degrades the dye and produces an electron shuttle.