Technical Field:

The present invention relates to the fields of ore enrichment, flotation, enrichment of carbonated ores/calcite-magnesite, biotechnology, microbiology, bioflotation. Flotation enrichment of carbonate minerals with the mining companies that can use that performs the sodium/potassium oleate flotation chemicals, including chemicals that are produced by the chemical, and particularly engaged in the production of flotation to increase the efficiency, and can replace the chemical structure characterized by biological surfaceactive reagents is aimed at improving the use of the Collector.

State of the Art:

Flotation is a mineral separation (enrichment) process that takes place in an environment called the pulp, a mixture of water and minerals. It is a physicochemical process based on the fact that solid surfaces repel water naturally or with the help of chemicals. Flotation, cleaning of wastewater today, etc. it is quite widely used in many non-mining areas. In the field of mining-ore preparation; It provides the most effective separation of 0.2 mm and below and can be applied to large tonnages; therefore, it is one of the most widely used methods in the industry, inoculation of the vaccine from culture by 2%.

Flotation is not an isolated development, but, on the contraiy, in the second half of the 19. Century it is one of the great innovations that brings a horizon-opening change to mining and ore preparation and ensures the increase of mineral production. Because there was a great acceleration in the industrial revolution, and this acceleration also led to a large increase in the consumption of minerals and metals, this period has been an exciting period in the mineral industry.

The development of flotation as an industrial process can be studied in 3 periods. As a first period, between 1800 -1900, it can be given that the first attempts to decontaminate and agglomerate the precious mineral in the industry were made. During this period, oil was not yet used and oil consumption was too much, so it could not be applied on a large scale as an extremely costly process. From 1900 to 1920, many studies were conducted on the flotation of lead-zine minerals in Broken Hill, Australia (1901-1915) and the flotation of copper mines in Huge mines in Western America (1911-1925) due to the need to economically concentrate fine sulfide minerals. During this period, which can be described as the second period of developments in flotation, flotation has become an industrial technology and worldwide electricity distribution has become possible due to the copper obtained by flotation. After 1960, 2 major inventions were realized. X-ray and radioisotope Wave analysis systems developed, due to this system, information about the process can be quickly retrieved, and correct process control is provided to a high volume of air flowing in the opposite direction of the pulp and bubble columns, the separation of the pre-flotation machines with small air bubbles pulp cells have been introduced, such as new high-energy airborne. In this way, a much higher flotation rate was achieved with high energy cells compared to columns.

Accordingly, the previous research on the process can be grouped into two main hea

1. Improvement of system control by adapting technological developments (online analysis, etc. developments) to the processing.

2. Increasing process efficiency by developing different and more effective flotation devices. In particular, the decrease in the efficiency of the flotation process in the extremely thin (slim) dimension has led researchers to the topic of an effective flotation method in the extremely thin dimension.

The flotation process has previously been defined as the transportation of fine-sized grains to the foam zone by adhering to the air bubbles supplied to the medium while suspended in the aqueous medium. However, many of the minerals, on the other hand, like water, while only a few (mica, talc, coal, etc.) it has a naturally water-repellent surface. In order to float the water with foam flotation of minerals, several chemicals called “collectors” are used that cover the surface and make it water repellent. In flotation, except for collectors that make mineral surfaces water-repellent; to ensure selectivity, different chemicals such as suppressors, foams, pH adjusters are also used to create a balanced foam zone.

Considering the operation of the process described above, two significant disadvantages can be mentioned. The first is that too much water is consumed, and the other is that the reagents used are usually very expensive and environmentally toxic chemicals that only certain companies produce.

Especially after the Second World War, as a result of intensive developments in technology, increased production, industrialization, the natural balance of the world began to deteriorate. As the effects of this began to be seen, many attempts to protect the environment were started and their implementation with increasingly strict laws was also provided for by various regulations. This has had its effect in the mining sector as well as in all sectors. As a result of this effect, there are strict requirements for the regulation of waste dams, etc. In addition to introducing many applications, process practitioners have also focused on research on more environmentally friendly alternatives. Advances in biotechnology have also made biological processes more environmentally friendly alternatives that can replace many traditional processes.

In flotation, this effect has manifested itself in the form of the search for reagents that are more environmentally friendly, more compatible with the environment. Bioflotation, on the other hand, is one of the topics being investigated as an alternative to traditional flotation, although it has not yet been into industrial application. Bioflotation can be briefly defined as the flotation process in which microorganisms or their metabolites are used instead of the traditional chemicals used in flotation. Studies conducted on the subject have shown that apart from environmental benefits, bioflotation applications may also be more economically advantageous than traditional methods. In the literature, the yield ratios of chemicals such as sodium oleate, dodecylphosphate, Dodecyl-N- carboxyethyl-N-hydroxyethyl-imidazoline, which are the main reagents used as collectors (collectors) in flotation, are usually obvious. As an example of the possible economic advantage of bioflotation, flotation costs can be reduced by using bioreactors as an alternative to these chemicals.

As a result, the advantages that bioflotation can provide compared to flotation with conventional reagents:

1. The use of environmentally friendly reagents.

2. Possible lower costs for waste storage and reclamation after the process (direct discharge to the environment may be possible).

3. Some bioreactives provide higher efficiency than their traditional alternatives.

4. There may be lower costs for some bioreactives compared to their traditional alternatives.

In order for biotechnology to be integrated into flotation, mention should be made of the discovery that some microorganisms and/or their metabolites interact selectively with mineral surfaces. The advantages of these microorganisms or their metabolites because they do not cause environmental pollution, are environmentally and human-friendly have been evaluated and their use in flotation and flocculation methods using interaction with the mineral surface has been raised. It is included in the literature that this microorganism or its metabolites can be used as bioreactors, especially in flotation, such as foaming agents, depressants (suppressants), and collectors. According to these data, it is possible to collect studies on the use of microorganisms and reagents of microbial origin in flotation in three main groups depending on the purpose of use:

As a first group, studies on the availability of some microbial metabolites that are known to reduce the surface tension of water as a foamer can be mentioned. For example, in a study published by Hassan et al., they investigated the availability of a metabolite produced by the bacterium Bacillus subtilis, which uses gas oil as a source of carbon and energy, as a foamer, and showed that this bioreactive is more effective than MIBC in terms of foam height and stability.

As a second group, studies on microorganisms or metabolites that make the mineral surface more hydrophilic as a result of their interaction with the mineral surface can be considered. It has long been known that bacteria of the type Thiobacillus ferrooxidans dissolve pyrite. In parallel, Misra and Chen used such bacteria as a flotation inhibitor and demonstrated its effectiveness (Misra and Chen, 1996). Zheng et al., on the other hand, have shown that the microorganism Mycobacterium phlei is a good dolomite suppressor in the separation of apatite and dolomite (Zheng et al., 1998). In a study published by Ytice et al, they used a microorganism of the type Acidothiobacillus ferrooxidans and found that the copper concentrate tenor increased by 22% with the collapse of pyrite (Ytice et al., 2006). In a study conducted by Vasanthakumar et al., the selectivity of sphalerite selectively from galena with Bacillus subtilis adapted to sphalerite was shown (Vasanthakumar et al., 2017). The third group of microbial reagents is a study in which microorganisms or their metabolites are used as collectors. In a study published by Misra et al., the bacterium Mycobacterium phlei, which is known to exhibit hydrophobic properties, was used as a collector in the flotation of hematite.

In this study, it has been observed that there is an excellent relationship between decantation yield and adhesion of bacteria to hematite. Misra et al. studied the separation of very fine coal grains from pyrite by selective flocculation and colon flotation with the bacterium Mycobacterium phlei. In a study conducted by De Mesquita et al., the potential of a non-pathogenic hydrophobic bacterium, Rhodococcus opacus, as a flotation reagent for the hematite quartz system was investigated.

In the studies, it was revealed that there are significant changes in the mineral surface with microorganism-mineral interactions, zeta potential, and contact angle measurements. However, this change was observed in quartz at a very small pH in the range of (pH: 1.3-3.5). Therefore, at pH:5, the microorganism is only selectively adsorbed to the hematite surface and separation from quartz can be possible with a highly effective hematite flotation. In a study conducted by Chockalingam et al., it was shown that pyrite can be separated from quartz and calcite by flocculation or flotation method after its interaction with Bacillus polymyxa or bacterial metabolites. In a publication by Merma et al., the flotation of apatite and quartz minerals with a bacterium of the type Rhodococcus opacus, a Gram (+) bacterium, was studied, and it was shown that the apatite yield reached 90%, while the quartz yield remained at 14% under optimal conditions. As a result of this study, it was determined that the microorganism used is a very promising biotoiler and biopurifier for phosphate flotation on an industrial scale. In a hematite enrichment study conducted by Lopez et al. in 2015, in which Rhodococcus rubber was used as a collector, they showed that the bacterium in question has a serious potential for metal sulfide flotation in the future. In another study, in which Rhodococcus opacus was used as a collector, the malachite mineral was floated with fairly high selectivity and efficiency.

If the stages of the process are examined one by one, the situation is as follows:

1. After preparing the water-ore mixture in a certain solid ratio, it is necessary to adjust the pH for the preparation of the medium at the initial stage. pH adjustment is usually performed for two purposes. The first of these is the following: The effectiveness of ionizing reagents depends on the degree of ionization, and it, in turn, usually depends on pH, so the ambient pH should correspond to the fact that the reagent is in an ionic state. For example, amines are effective at acid pH, while anionic reagents are more effective at basic pH. The second goal is also related to selectivity. For example, if adsorption of the collector to the mineral surface occurs by electrostatic attraction, the zeta potentials of minerals are regulated by pH and selective adsorption of the collector to any desired mineral is achieved. However, adjusting the pH will have a cost such as acid/base consumption. In addition to the cost of reagents, this arrangement will also increase the repair and maintenance costs by causing problems such as wear on the equipment if it is operated at excessive pHS. Also, if necessary, the pulp temperature should be regulated. In particular, flotations of industrial raw materials cam be more efficient at certain temperatures. In addition, the properties of the water used are also an important factor for flotation efficiency, especially in industrial raw material flotation. Because it negatively affects the yield, it is undesirable to have a high hardness of the water in the flotation of industrial raw materials, and therefore NaOH is preferred instead of CaO, which is cheaper for an alkaline environment. This, in turn, is an element that increases costs. Today, because water resources are rapidly decreasing, water use has become quite important. Water consumption is also high in flotation. Therefore, the use of excess water in flotation is a disadvantage of the application (Bentli, 2006). At this stage, the addition of the necessary suppressive and collecting reagents is carried out. Here, on the other hand, many different problems and issues are stated. These problems and issues can be listed as follows: The reagents used are usually expensive products that are produced only by certain companies. In this aspect, the price of the reagent used and the amount consumed per ton of ore are very important parameters that directly affect operating costs. Another important issue is the interaction of the reagent with the environment. The use of environmentally harmful reagents imposes quite a lot of side costs on the enterprise (measures to be taken in the transportation and storage, additional measures in the field of wastewater and waste dams, etc.). Flotation is a method with a lot of energy use. In this situation, the cost is also calculated for enterprises. To minimize energy usage, the flotation process should take place quickly and the desired efficiency should be obtained in a short time. The time given for the reagents used to perform the desired task, that means, the time given for the conditioning process, is important. Longer conditioning requirements for a fixed capacity, the required tank volume increases, and this is required for both the initial investment and operating costs with the propeller can lead to significant increases in energy and maintenance costs. An increase in the flotation rate will reduce the flotation cell volumes for a constant capacity. This will again provide an advantage in both initial investment and operating costs. Taking the concentrate. Reagents used during flotation can easily cause environmental pollution. In the flotation plant, wastes are usually stored in a waste dam after they are removed to a certain solid ratio in tikiners. That is why biopolymers, which are one hundred percent soluble in nature and can disappear with minimal harm, are becoming important in all areas of the industry. Bioplastics, biodetergents such as biosurfactants have also been used in the industry. As the flotation process requires a lot of water consumption, as much water is returned to the system as possible. There is an application numbered WO2018209416A1 related to “Ore flotation method using a bioreactive obtained from Gram-positive bacteria”. In the study, metabolites obtained from Rhodococcus opacus and Rhodococcus erythrhopolis microorganisms were used. The work mainly mentions the separation of hematite, an oxide mineral, from quartz.

There is an application numbered CN102284372A, related to the “flotation method of carbonated minerals”. The collectors mentioned in the specification are oleic acid, tall oil, and dodecylamine, and it is understood that flotation is completely related to the use of chemicals.

In decontamination studies, the angle of contact between the mineral surface and the air bubble is an important indicator of mineral hydrophobicity. Therefore, the change in the angle of contact in the absence and presence of the reagent also determines the effect of the reagent on the hydrophobicity of the mineral. Higher contact angles mean higher (intermediate) dec voltages and better adhesion between particles and bubbles. The higher the contact angles, the higher the (intermediate) dec voltages and the better the adhesion between particles and bubbles. Briefly, the flotation medium should contain substances that activate surface tension. Another of these substances should also reduce the water-air surface tension so that it foams, and its amount should be sufficient to create a stable/stationary foam.

Reviewing these situations, the reagents used as standard must perform all these operations. Flotation in this case makes the reagents expensive, and many of them are also imported. The rapid effect of biosurfactants on surface tension and their easy availability make biosurfactants more advantageous than standard reagents.

As a result, a new technology is needed that can overcome the disadvantages mentioned above.

Description of the Invention:

The presented invention is a method of improving the flotation efficiency in carbonated minerals that can overcome the above-mentioned disadvantages by the use of “surfactin”, a bioreactive whose effectiveness has been proven, which provides an advantage in the cases described below. In the classical calcite/carbonate flotation, oleate is used as a collector. The challenges of classical calcite flotation are listed below.

It is studied at high pHs. This means that it increases the consumption of reagents, as well as accumulating excess alkaline waste that is more incompatible with nature in waste dams. In addition, it is also a factor that increases costs in terms of repair and maintenance of equipment at the plant.

Bioflotation has yielded high yields at lower PHS. In this case, it will alleviate the disadvantages caused by working at high pHs. The water temperature is important in carbonate flotation. This requires extra precautions so that the pulp temperature can remain at certain values. As a result, it increases operating costs. It will also significantly increase the initial investment costs by requiring the establishment of a completely closed facility according to the climatic conditions of the region where the enterprise is located, or it will cause the operation of the enterprise to stop during certain seasons.

In bioflotation, where ’’Surfactin" was used as a collector, high efficiency was obtained at a wider ambient temperature (18-20 ^c C). This will allow it to reduce operating costs for pulp temperature and provide a more comfortable working environment.

Oleate is chemically adsorbed to the carbonate surface and requires conditioning times of 15 minutes and above on a laboratory scale. This, in turn, leads to the fact that the total volume of the conditioning tank for a fixed hourly capacity is large; it increases the initial input. In addition, in activities such as mixing, larger volumes mean more energy consumption.

In the case where ’’Surfactin" was used as a collector, fairly good efficiency values were obtained with 4 minutes of conditioning. This, in turn, reduces both the initial input and operating costs for a fixed hourly feed.

The consumption of oleate is quite high in carbonate flotation.

The consumption per ton of ore for ’’Surfactin" is much less, it will be able to reduce the cost increase of bioflotation caused by the difference in unit prices of reagents.

In carbonate flotation, even under the most controlled and appropriate conditions, the yield remains at 60-70%, especially on an industrial scale. In the ore studied, only 5000 g/t collector, 23 °C temperature, and 55% efficiency at pH 10.6 were obtained.

With the use of ’’Surfactin" as a collector, 80% efficiency was achieved in much more acceptable conditions and using a collector amount of 360 g /T. This, in turn, reduces the reagent costs per unit of concentrate much lower.

The biggest obstacles preventing bioprocesses from moving to a large scale, especially in mining for all sectors, where they are not implemented on an industrial scale, can be listed as the production time and cost of the biorective. The metabolite used is a biopolymer that is currently being produced industrially for different sectors and is being used on an industrial scale. This, in turn, eliminates the problem of production time, which is a disadvantage for the proposed method to be adapted to the industry.

When all the listed advantages are evaluated, the surfactant, although the production cost is higher than oleate, is seen to be financially competitive with oleate on an industrial scale due to the reduction in operating costs it offers along with the advantages it will bring. Description of the Figures:

The invention will be described concerning the accompanying figures so that the features of the invention will be more clearly understood and appreciated. But the purpose of this is not to limit the invention to these certain regulations. On the contrary, it is intended to cover all alternatives, changes, and equivalences that may be included in the scope of the invention as defined by the accompanying claims. It is emphasized that the details shown are shown only to describe the preferred embodiments of the present invention. As a result, the shaping of methods; is presented to provide the most convenient and easily understandable definition of the rules and conceptual features of the invention. In these drawings;

Figure 1. The subject of the invention is the view of the process steps of the production method.

Figure 2. The subject of the invention is the view of the process steps of the production method

Figure 3. The subject of the invention is the view of the process steps of the production method.

Figure 4. The subject of the invention is the view of the process steps of the production method.

Figure 5. The subject of the invention is the FTIR spectrum view of a bioreactive; a) the FTIR spectrum view of a standard surfactant; b) the FTIR spectrum view of a manufactured surfactant.

Figure 6. The subject of the invention is the NMR spectrum view of a bioreactive; a) the NMR spectrum view of a standard surfactant; b) the NMR spectrum view of a manufactured surfactant.

Figure 7. This is the XRD analysis of the studied calcite sample.

Figure 8. It is a minor element analysis of a calcite sample.

Figure 9. The parameters and levels were evaluated in the study.

Figure 10. The design matrix and the results of the experiment.

Figure 11. The results of the analysis of variance for the obtained yield (%) are the correlation values of the obtained model.

Figure 12. Predicted/Actual graph of the harmony of the estimated data obtained from the model equation with the experimental results.

Figure 13. The main effects of variables on yield (%).

Figure 14. In terms of efficiency, the estimated data obtained from the model equation and verification (verification) are the test results.

The figures to help understand the present invention are numbered as indicated in the attached image and are given below along with their names.

Description of References:

10. Passivation

20. Release into Incubation

30. Inoculation.

40. Ham Biyoreaktifin Hazirlanmasi

50. Obtaining a Supernatant 60. Lowering the pH to 2

70. Centrifugation

80. Addition of Ethyl Acetate

90. Vortexation

100. Drying

110. Performing NMR Analysis

120. Performing FTIR Analysis

130. Grinding of Carbonate/Calcite Mineral

140. Performing XRD Analysis

150. Performing XRF Analysis

160. Addition of Bioreactive to Flotation Pulp

170. Receipt of the Floating Product

Description of the Invention:

The invention nutrient agar (G/L meat extract, 5 g peptone, 1 g, Sodium Chloride 5 g, 2 g yeast extract, 15 g agar) with stock cultures of bacteria Bacillus subtilis (pasajlanma a) (10), 24-hour, 35 °C, 150 rpm in nutrient agar enviroment (G/L meat extract 1, peptone 5 g, Sodium Chloride 5 g, yeast extract 2 g for the release of incubation (20), sterilized mineral salt enviroment (G/L; ammonium chloride 15 g, potassium dihydrogen phosphate 4.3 g, dipotassium hydrogen phosphate 3.4 g, potassium chloride 1.1 g, Sodium Chloride 1.1 g, yeast extract 0.5 g, Calcium Chloride 0.24 g, zinc sulfate synthesis 0.29 g, manganese sulfate monohydrate 0.17 g, magnesium sulfate synthesis 0.5 g, glucose 20 g of culture vaccine 2% to be inoculated with (30), and 35 °C at 150 rpm release for at least 72 hours incubation (30) raw biyoreaktif the preparation of the result (40), after incubation of microorganisms growing in a mineral salt environment at +4 °C and supernatant was removed from the enviroment by centrifugation at 10,000 rpm for 15 minutes at least obtained (50), be reduced to pH 2 with 2 N HCL supematantin (60), were transferred to centrifuge tubes supematantin pH reduced to 2 and at +4 °C for 10 minutes and centrifuged at 10,000 rpm (70), and pellets discharged on supematantin the addition of ethyl acetate (80), pellets vortekslenme till it dissolves thoroughly up to (90) can be centrifuged at 10,000 rpm for at least 1 minute (70), separate a tube of dried supematantin taken into centrifuge concentrator (100), Fourier transform infrared spectroscopy analysis of the obtained alternately biyosurfektan (110), followed by nuclear magnetic resonance analysis obtained biyosurfektan (120), calcite flotation experiments of the samples that will be used in closed-circuit grinding with a milling 0.150 mm (130), calcite of a sample of X-ray diffraction analysis (140), x-ray fluorescence spectrometry, element analysis of the samples by minor calcite (150), Addition of bioreactive agent as collector to carbonate/calcite flotation pulp (160) and flotation of sample by air-taking floating product (170) consists of process steps (Figure 1 - Figure 14).

Detailed Description Of The Invention:

The subject of the invention is carbonate, especially calcite flotation. In the general carbonate flotation, Na/K oleate is used as a collector. A sample of calcite with a purity of about 97% was taken before, and after it was brought to a size of -0.2 mm, flotation experiments were carried out with a series of related and statistical experimental design methods and a series of surfactant biosurfactants. Comparison of the proposed method, which is the subject of the invention, and the classical method is given separately for each stage of the process according to the results of laboratory studies:

1. According to laboratory studies, a 55% yield was obtained at pH 11 in oleate flotation, while high yields were obtained at pH 9 with surfactant (no need for pH regulation using additional chemicals). In traditional flotation, the yield increase was achieved between 23-25°C at ambient temperature, but much better yields were obtained at room temperature in the new proposed system. Due to the temperature and pH advantage provided by the surfactant, the equipment used will be used for a longer period, and the extra operating cost will also be eliminated.

2. The consumption of oleate reached 4000-5000 g per ton of ore, but 50-55% yield was not achieved. With the biotopper of 360 g/t surfactant, which can be called about 1/10 of this value, the yield has increased to 80% and above. Considering the unit costs of reagents, oleate will probably be cheaper, but considering the quantities used, oleate will lose this advantage significantly. Another important advantage for the collector is that the surfactant can be produced with a reactor and system to be installed at the mining facility and used directly in the flotation.

3. Minute conditioning with surfactant used as a biotoiler was sufficient to achieve an 80% yield. However, the yield value remained below 50% when the oleate was given a conditioning time of less than 15 minutes. This, in turn, means a reduction of about 20-25% in the conditioning tank for constant capacity. The easy and fast adsorption of biosurfactants and the high flotation efficiency obtained will reduce the energy usage in terms of both duration and total volume in conditioning. Therefore, a shorter conditioning period will have a positive impact on both initial investment and operating costs.

4. Experiments determining the speed constants for comparing flotation rates have not been conducted. However, it has been noted as an experimental observation that the swimming time of surfactin and calcite is significantly lower than that of oleate. Although numerical data cannot be provided, it is thought that higher capacities can be met with smaller cell volumes. If the situation is evaluated for carbonate flotation plants that are fixed, there may be a serious increase in their installed capacity.

5. No differences are expected regarding the intake of concentrate.

6. As stated in the first clause, the pH of the waste will be at levels closer to neutral. In addition, biosurfactants are more preferred reagents during flotation than oleate as a reagent. In this way, it is on the list of materials that can be used without creating environmental pollution (Silva et al., 2018). Therefore, in cases of storing waste and even mixing the water in waste dams into streams or groundwater in cases such as a possible flood disaster, bioreactive will provide a more trouble-free environment both in terms of pH and collector.

In a study published by Sarvamangala et al., the effect of the Bacillus subtilis microorganism and its metabolite on the flotation of high purity alumina, silica, calcite, and hematite was investigated. Comparative evaluation of the data on calcite flotation, which is the part of this research related to the proposed patent, with the proposed method article is given below:

1. In the study conducted by Sarvamangala et al., both the microorganism and its metabolite showed a suppressive effect on calcite. As a collector, oleate, a classic carbonate collector, was used in the study.

In the proposed patent study method, the surfactant bioreactive replaces oleate and is used as a carbonate collector.

2. In the study conducted by Sarvamangala et al., the efficiency of 95% obtained with calcite collector decreased to 74% with 1-hour conditioning with microorganism cells and to 50% with 1-hour conditioning with metabolite taken without cells. With these results, it can be said that both bioreactives exert a suppressive effect on calcite. However, the decrease in yield seems to be insufficient yet to be able to say that bioreactors can be used effectively on an industrial scale. However, for example, when the effect of the same reagents on the hematite mineral was studied in the same study, it was observed that microorganisms reduced the yield from 95% to 5%. These data also suggest that bioreactive is a potential good hematite suppressor. In the proposed patent study method, the surfactant is used as a collector. Therefore, it is quite different from the calcite bioflotation in the mentioned research. In addition, the yield value obtained using a biotoiler in the proposed method is much higher than the yield value obtained with oleate, which is currently being used on an industrial scale. This makes the proposed method attractive from the point of view of the industry.

3. In the study given above, 1-hour conditioning times are mentioned. This period will be able to push the viability of the bioreactive as an element that increases costs on an industrial scale.

In the proposed patent study method, the conditioning period is less than half the time required by the alternative oleate. This makes the surfactant bioreactive more advantageous in terms of operating costs.

4. In the previous study, the bacterium “Bacillus subtilis” and its extracellular protein were used as bioreactively metabolites.

In the method of the proposed patent study, surfactin biosurfactant, a different metabolite of the same bacterium (Bacillus subtilis), was used again.

A bioflotation study in which a bioreactive was used as a collector in magnesite and calcite flotation in the literature was published by Botero et al. in 2007. In this study, the Rhodococcus opacus bacterium itself was used as a collector and flotation experiments were carried out in a Hallimond tube and with a 0.8 g sample. According to the data obtained, after 30 minutes of conditioning, a 92% yield at pH 5 in magnesite and a 55% yield at pH 7 in calcite were achieved (Botero et al., 2007). The comparison of this study and the proposed method in patent study is given in the following articles.

1. In the method applied by Botero et al., the appearance of the appropriate pH of 5 in magnesite and 7 in calcite can be a serious problem, especially on an industrial scale. Because the natural pH values of carbonated minerals are usually 8 and above. The acid added to the medium is: according to its reaction, the dissolution of the carbonate mineral results in the outflow of water and carbon dioxide. During this reaction, the pH also increases. This means that during the flotation process, it is necessary to constantly add acid to the medium from the first stage until it foams. Acid addition, on the industrial scale, reagent consumption, such as the difficulty of process control in the enterprise (additional) details that will bring cost.

In the method proposed in the patent study, quite good results were obtained in magnesite near natural and calcite near natural pH.

In the publication made by Botero et al., the bioreactive conditioning time was kept for 30 minutes. An increase in this period means an increase in both initial investment and operating costs on an industrial scale. Considering that the study is a microflotation experiment with a 0.8 g sample, even moving the scale to a laboratory scale between 1- 5 1 will increase these times. It is quite likely that these times will increase much more on an industrial scale, where control is much more difficult. Increases in initial investment and operating costs due to the increase in the required time will make it difficult to move the method to an industrial scale.

In the method proposed in the patent study, quite good yields can be obtained with 4- minute conditioning, and these data are not micro flotation, but laboratory-scale mechanical flotation data.

The last study in the literature related to the bioflotation of carbonates was published by Abdel-Khalek et al. in 2009 (Abdel-Khalek et al., 2009). The proposed comparison of this study and the patent study is presented in the following clause :

1. In the study conducted by Abdel-Khalek et al., it is related to the flotation of phosphate with oleate and the determination of an appropriate biostimulant to be used in the suppression of magnesite in this process. In the study, more than one microorganism was examined and the bacterium Corynebacterium diphtheriae was proposed as a biostimulant of magnesite. This microorganism has a pathogenic nature. This situation conflicts with the main purpose of bioprocesses, which are at the center of research, especially for their environmental advantages.

The microorganism and its metabolite used in the method proposed in the patent study are non-pathogenic. 2. The pH values mentioned in the study conducted by Abdel-Khalek et al. are quite low for carbonated ores. Studying these pH values reveals results parallel to the results of the study published by Botero et al. in 2007.

In the method proposed in the patent study, the pH value is very advantageous in terms of ease of operation due to its proximity to the natural pH of the pulp.

3. In this study, the purpose of using bioreactive is to suppress magnesite.

In the method proposed in the patent study, the surfactant biosurfactant, a metabolite of microorganisms, works as a highly effective carbonate collector.

In general, the whole process consists of 5 main steps. These:

1. Raw bioreactive preparation

2. Bioreactive extraction and characterization

3. Mineral screening stage

4. Sample preparation

5. Flotation process

Raw Bioreactive Preparation: stock cultures of Bacillus subtilis bacteria used in the study passivation (10) of nutrient agar (G/L meat extract, 5 g peptone, 1 g, Sodium Chloride 5 g of yeast extract (2 g), agar (15 g), activation, and rebel culture for the preparation of nutrient broth (G/L meat extract 1, peptone 5 g, Sodium Chloride 5 g, yeast extract 2 g) medium was used. Mineral salt medium for the production of biosurfactants (g/L; ammonium chloride 15 g, potassium dihydrogen phosphate 4.3 g, dipotassium hydrogen phosphate 3.4 g, potassium chloride 1.1 g, sodium chloride 1.1 g, yeast extract 0.5 g, calcium chloride 0.24 g, zinc sulfate heptahydrate 0.29 g, manganese sulfate monohydrate 0.17 g, magnesium sulfate heptahydrate 0.5 g, glucose 20 g) were used. The vaccine culture obtained by transferring from the bacterial culture contained in the nutrient agar medium to the sterile liquid medium was incubated at 35 °C and 150 rpm in the nutrient broth medium for 24 hours. After incubation, the vaccine was inoculated 2% from the culture into the prepared sterilized mineral salt medium and left to incubate for 72 hours at 35 °C and 150 rpm.

Bioreactive Extraction: The microorganism developed in a mineral salt environment after incubation was centrifuged at +4 °C for 15 minutes at 10,000 rpm and removed from the environment and supernatant was obtained. With 2 N HC1, the pH of the supernatant was reduced to 2. The supernatant with a pH 2 was then transferred to centrifuge tubes and centrifuged at +4 °C at 10,000 rpm for 10 minutes. After centrifugation, the supernatant was discarded and ethyl acetate was added to the pellet. The pellet is vortexed until thoroughly dissolved. After the vortex was centrifuged at 10,000 rpm for 1 minute. The resulting supernatant was taken into a separate centrifuge tube and dried in a concentrator.

Bioreactive Characterization: The FT-IR spectra of the bioreactive produced by experimental study with surfactant used as a reference were recorded using the PerkinElmer Spectrum Two FTIR spectrometer, in the range of 4000-400 cm-1, using the KBr pellet technique. The FTIR spectrum was measured on KBr pellets prepared by vacuum pressing of 1 mg of dried powder sample and 100 mg of KBR (spectrometer quality). The FTIR analysis results of the reference sample and the experimental sample are indicated in Figure 5a and Figure 5b, respectively. The results obtained show that the FTIR profile of the biosurfactant obtained by the experimental study coincides with the FTIR profile of the surfactant used as a reference.

The proton (1H) NMR spectrum of samples of the bioreactive produced by experimental study with the surfactant used as a reference in the deuterated chloroform solvent was recorded in the GEOL ECZ 500R spectrometer at room temperature. Their operating frequency is 500.13 MHz for the 1H core. Tetramethylsilane (TMS) was used as the internal standard for all NMR analyses and was observed at 0 ppm. The results of the 1H NMR analysis of the reference sample and the experimental sample are indicated in Figure 6a and Figure 6b, respectively. The results obtained show that the NMR profile of the biosurfactant obtained by the experimental study coincides with the NMR profile of the surfactant used as a reference.

Mineral screening: It is shown in the literature that the surfactant obtained from the Bacillus subtilus bacterium reduces the surface tension of water from 72 mN/m to 27 mN/m at a concentration of 20 pM. This data shows that the stated reagent will be a good foamer in flotation. But later, when the explicit chemical structure of the metabolite was examined, it was found that one side was hydrophilic and one side was hydrophobic. When these structures were also examined, it was concluded that they have the potential to be used as collectors in oxide-carbonate mineral flotation. On top of this, different oxidized-carbonated minerals (silicate, calcite, and magnesite) were subjected to flotation by using the obtained biological surfactant as a collector. As a result of these experiments, it was found that the bioreactive in question cannot float silicate, but it can float calcite and magnesite, which are carbonate minerals. The next stage of the study is based on the flotation of calcite samples.

Sample preparation: A natural CaCO3 sample containing trace amounts of impurities was used in the experiments. The X-ray diffraction analyses of the studied calcite sample were performed on a Panalytic EMPYREAN XRD device. The XRD pattern is shown in Figure 7. The chemical analysis results of the ore used are given in Figure 8. As can be seen from the table, the sample contains about 97.27% CaCO3 with a CAO content of 54.471%. Other ratios such as 0.113% SiO2 and 0.048% Fe203 also indicate the purity of the sample. The minor element analysis of the calcite sample used in the study was carried out using the X-ray fluorescence spectrometer (Panalytical ZETIUM). The calcite sample to be used in flotation/flotation experiments was ground to -0.150 mm by closed-circuit grinding. (110)

Flotation Process: In flotation experiments, the existing Denver brand mechanical flotation cell was used in ESOGU Ore Enrichment Laboratories. After the surfactant used as a bioreactive was produced at the ESOGU Biotechnology Laboratory, research was conducted on the purpose for which this bioreactive can be used in flotation and in which minerals. These researches are mainly focused on the pHs at which the surfactant works and its open molecular structure. According to the obtained data, it was concluded that it has a foaming property and can be used in oxide-carbonate mineral flotation. In addition, flotation experiments were carried out in the laboratory, in which -0.15 mm high purity quartz, calcite, and magnesite samples and surfactant bioreactive were used as collectors. Flotation experiments have shown that while the bioreactive does not affect quartz, it flotates both carbonate minerals with fairly high yields. It has also been observed that the surfactant does not require the use of a foamer, it can form a fairly strong and stable foam zone. After that, the effect of some parameters on yield in calcite bioflotation was studied by using statistical experimental design methods. To obtain a reference, classical flotation tests of calcite with oleate have also been carried out, in particular, the effect of pH, temperature, and conditioning time on yield has been tried to determine by simple experiments.

Reagents used:

- It is known that the hardness of the water in carbonate flotation reduces the flotation efficiency. Therefore, alkaline values higher than the natural pH (8-8.5) were obtained with NaOH. The pH was reduced with a 30% solution of H2SO4 in experiments to look at the reagent response at neutral pHs.

- As a collector, the prepared surfactant was used. In addition, some experiments have been conducted with sodium oleate to determine how temperature, conditioning time, and pH affect yield in classical carbonate flotation.

- Since both reagents used have a foaming property, no foaming agent was added.

In the experiments, the effect of three different (numerical) parameters (factors) was studied by using statistical experimental design methods. The statistical design method chosen is the “Central Composite Design (CCD)”, which is a “Response Surface Methodology (RSM)”. In the creation of the design, the a value used to determine the axial points were selected as 2. Accordingly, 3 levels were determined for each parameter; a total of 5 levels were determined together with the -a and +a axial points. Accordingly, a total of 20 experiments were programmed. The 3 parameters whose effect will be studied in the experiments, together with their levels, are given in Figure 9. The parameters studied in the experiments were selected as pH, solid ratio, and collector amount. The Design Expert 10.0 package program was used to create a design matrix, statistical analysis of the results, and create mathematical models. The results of the design table and the response variable “Efficiency (%)” are given in Figure 10. The results were evaluated according to the ratio of floating calcite to feed. All parameters were subjected to variance analysis for the response variable. The range of the analysis was ranged for a 95% confidence interval, and in this confidence interval, the ANOVA table and the model were created, the effect of which was reduced by removing meaningless terms from the model. When creating this table, the hierarchy was preserved. The results of the analysis of variance obtained according to the flotation yield, the reduced ANOVA table created is given in Figure 11 along with the value of R2, which indicates the compatibility of the model with the actual values. The graph “Estimated/Actual" data versus estimated data”, which shows the compatibility of the estimated efficiency values related to the resulting model equation and the experimental results, is given in Figure 12. When looking at the ANOVA table (Figure 11), it is seen that the harmony (R2 is over 93%) is quite high dec the experimental data and the values estimated according to the model equation for the response variable. This is also seen in the graph given in Figure 12. In Figure 13, the main effects of the variables whose effects were examined on the response variable (Yield; %) are shown graphically. As can be seen from the figure, the range of the ratio did not have much effect on the yield in the studied range, but the other two parameters caused a parabolic effect. Finally, the pR2 value of about 75% indicates that the prediction power of the model is also high (Figure 11).

After creating the model equations, optimization of the conditions for the goal of maximizing the response variable “flotation efficiency” with the help of the corresponding modules of the package program used, and estimation of the response variable under optimal conditions was carried out. At the final stage, a verification study (verification experiments) was also carried out to verify the response variable in the conditions proposed as optimal conditions (Figure 14). in the range of 5% confidence interval, the best yield for flotation yield was calculated as 75.92% at a solid rate of 39%, pH 9.3 and amount of 290 g/t collector at a solid rate of 39%. In these conditions, the experiment was repeated and the flotation yield rate was found to be 77.2%. The fact that this value remains in the range of 95% confidence interval and is especially close to the estimated value of 75.92% shows the repeatability of the experiments and the predictive power of the model; in summary, the success of the created model.

Previously, it was noted that the metabolites of microorganisms reduce the surface tension due to their structure (amphipathic). This situation has shown the potential of using bioreactives as surfactants for different purposes in flotation processes. The innovative part introduced into the literature in the proposed patent study is that surfactant, a metabolite of microorganisms can be used as a collector in the flotation of carbonate minerals such as calcite and magnesite. The advantages it provides, especially in terms of yield and flotation conditions, make this proposed bioreactive a very serious competitor to oleate on an industrial scale.

The advantages of the surfactant bioreactive, which makes it a serious competitor to oleate, the classic carbonate collector, according to the flotation stages, are summarized below.

- Pulp preparation: It requires more moderate conditions in terms of pH and temperature.

- Collector addition: Its consumption per ton of ore is very small compared to oleate (from 1/5 to 1/10).

Conditioning: A minimum of 15 minutes of treatment with oleate is required, while 4 minutes of treatment with surfactin is sufficient.

- Flotation time: Although no experimental studies have been carried out yet on the flotation kinetics, it has been observed that it is very fast compared to oleate flotation.

Yield: The yield obtained by bioreactive in the studied calcite sample is much higher than that of the classical reagent.