First we would create a breeding ground for algae strains to multiply and mothers breeding

population in the nursery. By photo-bioreactors and the technique of photosynthesis would increase the farm in vertical or horizontal tubes to generate large amounts of algae will dry and then they extract the oil content (for biomass varieties) and other groceries packaged for the

subsequent sale in the domestic market. We are at this moment in the preparation phase of the project, preparation of proforma of

materials, supplies, grounds and nurseries, and other photo-bioreactors required to complete the project's financial report.

Also initiated contacts with potential buyers of both products for subsequent regular supply agreements under which we will specifically on the same course of development of

the crop. We hope to be located geographically close sources that emit CO2, which is very much in the development and multiplication of algae production, we estimate for a property of

10,000 m2 (one hectare) about 1900 kgs. algae daily. We are sure and convinced that it will be a great source of renewable energy.

• We intend to produce biomass algae photo-bioreactors for the supply and biodiesel plants. For further processing into products (biodiesel, ethanol, etc.).

• At the moment we are at the stage of presentation and preparation of the specification. • About Biodiesel:

• These products are derived mainly from vegetable raw materials while also being made with animal fat and now we are doing this with bioalgas grown from certified stocks.

• This kind of bio-fuels can come from canola, corn, wheat, and organic debris as the frying oil. Moreover, they are conducting a series of approaches that can develop these fuels rare

organisms like weeds, termites or microalgae, which are those referred to my project. • You have recently learned of the start functioning from several companies that sell, or want to do, biodiesel made from oil obtained from the cultivation of microalgae. These agencies are responsible for more than 50% of photosynthesis on the planet, and also

convert CO2 into biomass green since incorporate it into their own body. The microalgae gather solar energy and accumulate in their fat through photosynthesis, absorb carbon

dioxide and emitting oxygen. • These crops are made in specific equipment, the photobioreactor, who manage the

productivity of these organisms is very high. Cultures were carried out in closed systems and the highly controlled culture conditions (nutrients, temperature, lighting, etc.)..

However, the relationship between humans and the algae is not new. • In Mexico have for many years feeding products made from biomass of these algae called Spirulina and the natives still use the devandita Chad Spirulina for food. On the other hand, species of genera such as Chlorella, Anthrospira, Dunaliella ou Haematococcus are useful in

industry.

• The advantages of algae-based biofuels include: • 1) They represent the cycle of CO2, most of them have better emissions, are

biodegradable and contribute to sustainability, and under stress conditions and with much CO2 in their environment, their production is multiplied.

• 2) They are "friendly" to the environment, no pollution and capture large amounts of CO2, but its greatest advantage is that large quantities are produced daily.

• Normally the density of algae produced crop is 10 to 20 times the normal growing and may

be even greater. • The algae tend to produce a high amount of polyunsaturated fatty acids, which decreases the stability of biodiesel. But polyunsaturated fatty acids have low melting points so in cold

climates is far more advantageous than other types of biofuels. • Production of oil from algae is 200 times greater than in plants. As is also higher biodiesel

production

• It has a high yield and therefore low cost. • The production of biodiesel from algae has the characteristics of reducing emissions of

CO2 and nitrogen compounds from the atmosphere. • Algae can grow on a wide range of conditions for which they are found anywhere on the planet within aquatic plants, artificial substrate such as wood or bottles, in ponds, marshes, swamps, snow, lakes fresh or salt, on rocks, etc.. It is not difficult to find areas to cultivate. In principle, like plants, algae need three basic components for growth: sunlight, CO2 and

water. Finding strains of algae to grow them is not difficult, but it's hard to find strains that permit the production of biodiesel because algae need such a high maintenance and on the

other hand are easily contaminated with other species. Since algae need sunlight cell, CO2 and water to grow, can be cultivated in ponds and lakes.

These types of crops are called "open systems". • The risk of such cropping systems is the high probability of being contaminated by other types of algae, since algae are the largest component in oil are not necessarily the fastest

growing, as some strains of algae contaminants could massively invade the crop. Moreover in this system have little control against environmental conditions such as water

temperature, CO2, light intensity, so that crop growth depends on environmental conditions and generally occurs in the warmer months. In general, for growing in open

systems are sought strains that can grow under conditions in which other agencies would find it difficult to develop as high or low pH, T º specific, specific nutritional requirements,

etc.. It is for this reason that only few species were successfully grown in such systems. The advantage is that open systems are very inexpensive and easy to build because basically

what you do is build ponds or pools on the floor. An alternative system for the growth of algae is by greenhouses (*), also a pool. Although

reducing the acreage solves many issues which are open systems: less likelihood of contamination by unwanted species, can grow a larger number of species, the culture

period as there is greater temperature control and may increase the amount of C02 in the atmosphere, which would also increase the growth rate of algae.

The ponds have systems that allow the algae to keep moving in the middle, so that all receive the same amount of light and nutrients. On the other hand is continuously renewed

amount of C02 and nutrients from the medium. Another type of closed systems of crops are those that incorporate Fotobiorreactores

white light and natural and where conditions are more controlled than in open systems. These systems are very expensive but have high returns in terms of oil production from

algae. Some types of photobioreactors include:

• Plastic tubes or triangular-shaped glass (*): Gases such as C02 and O2 are flowing from the bottom of the hypotenuse and algae culture medium are flowing in the opposite

direction.

• Fotobiorreactores tubular horizontally (*): These are acrylic tubes in which circulates horizontally more algae culture medium to precipitate and are not all receive the same

amount of light and nutrients

• vertical column of bubbles: environmental movement is generated by algae in a vertical column through the flow of gases like carbon dioxide. Lights through light tubes along the

tube, which aims to reduce the cost of cultivation of algae on a large scale and make it simple.

• Fermentation Equipment: Some oil companies gained no photosynthetic growth of algae, but feeding the algae with sugars which then ferment these. One such company that called

Solazyme, a biotech company that is developing techniques to produce fuel for cars and planes from algae.

• It is documented that seaweed used as food and in the fourth century in Japan and in the

sixth century in China. Today, those two countries and the Republic of Korea are the biggest consumers of seaweed as food and their needs are the basis for an industry gathering of six million tonnes of fresh seaweed per year worldwide, with a value of

approximately 3 500 million Euros. • During the past 50 years, demand has grown faster than the ability to meet the needs with natural seaweed stocks (wild). The research on the life cycle of algae has led to the

development of industries engaged in cultivation, which now cover more than 90 percent of the market demand.

• China is the largest producer of edible seaweed, about five million tonnes. Most of this amount is the kombu, produced from Laminaria japonica culture ropes suspended which

occupies hundreds of acres in the ocean. The Republic of Korea grows about 800 000 tons of three different species, 50 per cent of which corresponds to wakame, produced from Undaria pinnatifida whose culture is similar to Laminaria in China. Japanese production

amounts to some 600 000 tonnes, of which 75 percent is the nori, produced from Porphyra species, it is a product of great value (about 11,000 euros per tonne), compared with kombu

(2 000 euros and wakame (5 000 €.). • alginate, agar and carrageenan are thickeners and gelling agents are extracted from seaweed and are the mainstay of the industrial uses of these. The use of seaweed as a

source of these hydrocolloids dates back to 1658 when Japan was discovered in the gelling properties of agar extracted by hot water from a red alga. Pearl moss extracts, another

seaweed, contain carrageenan and were widely used as thickeners in the nineteenth century, while the extracts of brown algae began to occur no longer sold commercially as

thickeners and gelling until the 1930s. Industrial uses of seaweed extracts expanded rapidly after World War II, but sometimes were limited by the availability of raw materials.

• Today is collected about a million tons of fresh seaweed extracts obtained to produce the three aforementioned hydrocolloids.

• They are produced in total 55 000 tonnes of hydrocolloids worth 400 million Euros. • The production of alginate (150 million Euros) is made from extracts of brown algae collected in its entirety, it is too expensive to grow algae to obtain raw materials for

industrial uses. • The production of agar (90 million Euros) is mainly from two types of red seaweed, one

of which has been grown since 1960 or 1970, but in a much larger scale since 1990, and this has allowed the expansion of this industry.

• The production of carrageenan (160 million Euros) was based at first on wild marine algae, particularly the pearl moss, a small alga that grows in cold waters with a limited

resource base. However, since the early 1970s, the industry has grown rapidly because of the availability of other algae that contain carrageenan, successfully grown in temperate countries with low labor costs. Currently, most of the algae used to produce carrageenan are cultivated, although still a small demand for pearl moss and some other wild species

from South America. • In the 1960s, Norway was the first country to produce seaweed meal, made from dried

kelp and dried, used as a feed additive. The drying is usually done in kilns fueled with oil, so oil prices affect the cost. Are collected annually some 50 000 tons of fresh seaweeds, which

are manufactured with 10 000 tonnes of seaweed meal for a value of 3 and a half million Euros.

• The total value of industrial products derived from seaweed is € 400,000,000 (four

hundred million Euros). • The total value of all products of the seaweed industry is estimated at 3.500.000.000 €

(million Euros) • Seaweeds can be classified into three groups based on their color: brown, red and green.

Botanists refer to these large groups feofíceas, Rhodophyceae and Chlorophyceae, respectively. Brown algae are usually large, ranging from the large kelp which often

measured 20 feet long, to the smaller species of 30 to 60 cm, through thick and leathery algae from two to four meters. Red algae tend to be smaller, and usually varies in length from a few centimeters to a meter, but the red algae are not always red: sometimes are

purple and even red-brown, but despite botanists classify it as Rhodophyceae due to other features. Green algae are also small, and the range of variation of size is similar to red algae. • Seaweeds are also called macro-algae, as distinguished from the micro-algae, microscopic in size, often unicellular, and are known mainly by blue-green algae that sometimes grow in

rivers and streams and pollute. • The marine algae found in nature are sometimes called wild algae, to differentiate them

from the algae that are grown.

The Project - Promoter Promoter: Alberto Luzardo Castro – BIOD2

C/Raigosu 24– (33930) Langreo Asturias-Spain

+34 660735324 - +34 984181443 aluzardo@aol.es - www.biod2.luzardomarine.com

The idea was to take into account the demand for vegetable oils to produce biodiesel, and that they were causing other serious problems in the food sector, so I decided to try to

solve part of the problem and developing valid alternatives that were first of steady flow of raw materials, and therefore dismissed the possibility of growing other types of power

plants and devote himself to the production of algae, due to their rapid reproduction and daily harvest.

In this section, we will develop the best marketing strategy as the following diagram:

(SORRY , THIS IN SPANISH)

So far as developed by us (BIOD2) has been the most ambitious project in the production of micro-algae ', using all the CO2 from power stations and steel mills, that would provide all

the CO2 needed for optimal photosynthesis. For example suffices to mention that only the Arcelor-Mittal Steel into the atmosphere

sends 1 million tonnes of CO2 per year, with this source of input to our project would need jacket its annual carbon dioxide.

According to the portal www.biod2.luzardomarine.com , oil production in micro-algae are higher than those for traditional oilseeds. In this sense, estimates indicate that soybean,

rapeseed and oil palm produce 50 m3 km-2 yr-1, 100 to 140 m3 km-2 yr-1, and 610 m3 km-2 yr-1, respectively, while the micro-algae can produce 10 000 to 20 000 m3 km-2 yr-1.

According to our opinion, early studies suggested that the energy produced by the cultivation of micro algae is much higher than that produced by biofuel crops on land, and

that if intensifying research in this technology could produce substantial amounts of biofuels without impacts on land use.

Furthermore, regarding the use of macro-algae for fuel production, an Israeli company has patented a technique which produces a liter of fuel for every 5 kg of a kind of macro-algae

from the Mediterranean Sea. Biogas production

It has been shown to produce methane through anaerobic fermentation, have higher rates than other sources of biomass. In this context, anaerobic digestion for biogas production

and showed good results.

The idea of producing methane gas from algae was proposed in the early '50s. Although progress has been made in research in the production of this fuel, its use has not been

crowded, mainly due to production costs, however, this technique can be used very well for algae grown in polluted areas or with sewage, in order to take advantage of existing

nutrients.

Ethanol is an intermediate product in complete digestion of organic matter and is produced by specific microbial strains in this way the ethanol must be produced under controlled

conditions to avoid contamination problems. USA is currently the largest ethanol producer in the world.

Studies indicate that macro-algae contain a high amount of polysaccharides (about 60% by weight), which are the platform for the production of various chemicals via fermentation.

Thus, ethanol production is the most effective technique for utilization of the carbohydrate fraction in the seaweed.

It tested the production of ethanol through the fermentation of extracts of the macro-alga Laminaria hyperborea, achieving 0.43 g ethanol per gram of substrate. Recently, Japanese scientists from Tokyo University of Marine Science and Technology, presented details of an

ambitious project to produce ethanol on a large scale through the processing of macro-algae sargasso (hondawara) grown in an area of 10 000 km2, is estimated production of 3

billion tonnes.

Ask: At the time the demand for vegetable oils for biodiesel processing plants are at full

production is estimated at about 250,000 tonnes of raw material per month and only when supply exceeds 30% of this need.

All these offerings are divided into:

-oils soybeans, rapeseed, jatropha, palm and recycled is 90% of the total. -algae oils barely reaches 2.5% and the remaining 7.5% is obtained from other plants that

produce oil is of plant or its fruit.

Our process is as we said before 4 stages: -Cultivation, drying, pressing and refining of oil .-

Then he packed and proceeds to keep on deposit until its withdrawal by the buyers - Method of sale provided: EXW –

•

- Firstly we supply a biodiesel plant in And then as the development of the expansion will try the other communities and third phase to export within the European Union production

surpluses.

Sales Strategies: Segment the market (divided into different groups according to specific characteristics,

socio-economic, territorial, personality, ..) Analyze purchase motivations that affect different customers (fashion, economic interest,

convenience, safety, attractiveness ,...) and analyze and explain why, how, when making their decisions to buy a product, and where and how much you buy .

We will seek information about how to make purchasing decisions. (People primeron are reported after contact and, ultimately, purchase and, as the economic weight of the goods with respect to their income, take a more immediate decision, weighing the risk of buying or acting on impulse, always valuing confidence that the product offered, its price and its

image, valuation may be individual or influenced by recommendations or influence.)

Competition: • Only two companies in Spain develop these techniques in algae (one in Alicante, and the

other in Cadiz), both regional and local government support in their research, but a non-productive (they are still in full development of the research).

Conclusion on competition:

Paradoxically, when you start a business without competition is many times more risky to start confronting the existence of competitors. When others have found their way in a

particular sector, which comes after is a more "trite." We may also compare, analyze the advantages and drawbacks of the neighboring business, success strategies, weaknesses, etc. ... and brought to market by offering

something better and more competent. It is important to know the competitors. That is, determine which of them we will face. The

goal is not to destroy the contrary, but steal market share. For each of these aspects, ultimately, should provide a comparison with the business plan, highlighting the case if you are in a strong or weak position with respect to our initiative. We will study whether there are factors that may influence the current market structure,

industry trends, socioeconomic factors, new administrative regulations, demographic trends, etc..

The algae are composed primarily of proteins, carbohydrates, nucleic acids and fatty acids.

The fatty acids are found in membranes, storage products, metabolites, etc.. The percentage of fatty acids varies among species, although there are species whose fatty acids represent 40% of its dry weight. These are fatty acids that are then converted into biodiesel. For the production of these algae are searched containing a high lipid content

and are easily cultivated.

Scenedesmus obliquus 50-56 10-17 12-14 3-6 Scenedesmus quadricauda 47 1.9 - Scenedesmus dimorphus 8-18 21-52 16-40 - Chlamydomonas rheinhardii 48 17 21 - Chlorella vulgaris 51-58 12-17 /14-22 4-5 Chlorella pyrenoidosa 57 26 2 - Spirogyra sp. 6-20 33-64 11-21 Dunaliella salina 57 32 6 Euglena gracilis 39-61 14-18 14-20 Prymnesium parvum 28-45 25-33 /22-38 1-2 Tetraselmis maculata 52 15 3 -

One species of green algae most widely used in the development of biodiesel is

Botryococcus braunii (*). This species produces large amount of hydrocarbons as terpenes, which constitutes about 30 to 40% of its dry weight. The botriococeno is the predominant

hydrocarbon in Botryococcus braunii. It can be used to produce octane, kerosene and diesel. For the production of biodiesel from botriococeno, you must first find a suitable strain of Botryococcus braunii producing a high yield of hydrocarbon. In selecting such

strains, attributes may be lost as disease resistance, competitive disadvantages, and so on. For this reason we need photobioreactors for cultivation of such strains.

Other species of algae that can potentially be used in the production of biodiesel for its high

oil content:

• Scenedesmus dimorphus - This is a favorite for the high performance oils for biodiesel, but one problem is that it produces thick sediments if the crop does not often stirs

• Dunaliella tertiolecta - This strain produces about 37% oils. It is a fast growing strain which means it has a high rate of CO2 absorption.

• Bacilliarophyta (diatom) - A favorite of ASP. The problem is that silicone in water needs, while Chlorophytes need nitrogen to grow

• Chlorofita - Green algae tend to produce starch, rather than lipids. They have very high growth rates at 30 ° C with high intensity of the light water type at 55 mmho / cm.

"The accumulation of lipids in algae occurs during periods of environmental stress,

including growth in media with low nutrient conditions. To induce stress in crops for biodiesel production one of the strategies to reduce the ration of nitrogen compounds, or

induce as variations in temperature, pH, starvation. etc. Some studies suggest that the enzyme acetyl-CoA carboxylase may be involved in the production of fatty acids, so that through genetic manipulation of the gene that codes could increase lipid production by

increasing the activity of the enzyme . Extracting oil from algae is basically the seaweed extract of culture medium (through any suitable separation process) and then use the wet seaweed to extract the oil. There are

three well-known methods of extracting oils from algae

1. Expeller / press: algae after being dried maintain their oil content, they are pressed with an oil press. Sometimes he uses a combination of press and solvent extraction.

2. Method of solvent hexane: This is one of the favorites and solvent extraction is not very

expensive. Once the oil is extracted with cyclohexane press is used to extract the remaining content of the algae. Then by distillation separates oil cyclohexane.

3. Supercritical Fluid Extraction: A method capable of extracting 100% of the oil, but needs a high equipment. CO2 is liquefied to the point of having the properties of a liquid and a gas,

liquefied fluid then this acts as a solvent for oil extraction algal.

Other methods of extracting much less used as the enzymatic extraction, osmotic shock and removal through ultrasound.

The biodiesel production process is based on transesterification of oil (*). The oils are mainly composed of molecules called triglycerides, which are composed of three fatty acid

chains attached to a glycerol molecule. The transesterification consists of replacing the glycerol by a simple alcohol such as methanol or ethanol, so as to produce methyl or ethyl esters of fatty acids. This process helps reduce oil viscosity, which is mainly caused by the presence of glycerol in the molecule. The high viscosity of the oil prevents its direct use in diesel engines, a disadvantage that is overcome by this process. To achieve the required

reaction temperatures between 40 and 60 ° C, and the presence of a catalyst, which can be caustic soda or potash.

Increased production of oils in algae through genetic engineering

As mentioned above, there is at least a known molecular method to increase production of algae oil. The enzyme acetyl-CoA carboxylase is involved in one of the steps in the synthesis of oils in algae. During the American project Aquatic Species Program - Biodiesel from Algae

is cloned which encodes the acetyl-CoA carboxylase gene from a diatom and thus isolate the enzyme. When people were able to successfully clone this gene, researchers in this project achieved a first and successful transformation system in diatoms. Both the gene that encodes acetyl-CoA carboxylase as well as the diatom transformation system were

patented. He got on to express the enzyme in diatoms in the hope of increasing oil levels. However in the experiments were conducted was not a significant change in the level of oil

produced by diatoms, so this method is still in a research process.

Interesting proposing new projects bioalgas techniques, whereby a cement plant would be

the first to capture CO2 from algae. .

A Spanish company Biod2, be capturing CO2 emissions from a cement plant from algae. The company also would plan to use algae for biofuels.

. It's no secret that the cement manufacturing process is both energy intensive and dirty. The global cement production emits only about 5% of emissions of greenhouse gases annually, both as a byproduct of decarbonization of lime (60%) and solid fuel burning furnaces (40%). As demand in countries like India and China continues to grow strongly, global emissions of

cement plants and other industrial sources continue to rise. .

The Spanish company sees a real opportunity in these emissions of greenhouse gases. The cement plant located in northern Spain, via "Biod2" could get the first successful use of carbon dioxide emitted by a major industrial source to produce high-value biomass from

microalgae. .

The Biod2 project would capture carbon dioxide and other emissions from a cement plant to use creating a slime algae rich in nutrients that can be dried and used as fuel.

. Biod2 grow in adjacent facilities fireplaces, is collected, dried using heat also recovered

from the cement plant and then used together with fossil fuels normally used in its cement kilns. The company aims to demonstrate the scalability of the pilot project and show that it

can be used in any industrial chimney. .

Several companies would develop promising technologies in their race to capture successfully divert and reuse indutrial scale emissions of greenhouse gases to produce

biofuels.

Not to mention the millions of euros that would save by emiisiones of CO2 in the form of carbon .-

The average price per tonne of carbon sequestered is between US $ 13-20 (US $ 3.55 to 5.50 per ton of CO2). It is important to note that 3.5 tons of alga production utilizes 1.27 tons of carbon, based on these indicators should be encouraged producers seaweed can also issue

bonds capture carbon, for the environmental services they offer, as a way to increase revenues.

But even required to develop methodologies for determining carbon sequestration by

seaweed depending on the species, geographic location, etc, both natural grasslands, as in the growing areas. In this must involve research centers and the respective state

governments. Both macroalgae and microalgae require CO2 for growth; in this sense, Can algae producers and artisanal fishermen to manage and conserve seagrass meadows issue

bonds carbon capture ?.

One of the advantages of marine algae is their rapid growth and thus will have a greater carbon sequestration, unlike the forests where carbon capture is only performed during the

growth of trees, which are also slow growth.

Among photobioreactors, works of adaptation and annexed to the cement plant chimneys, pipes, tanks and the laboratory would cost approximately 600,000 euros, more rights and

exclusivity Project original idea by some 100,000 euros to the signing of agreement between the parties to develop the project.

The benefits will be based on the amount of CO2 emitted, but keep in mind that for every ton. CO2 absorbed in the process of growing algae, the savings will be 6.91 euros (at

January 23, 2015)

FOTS BELOW AND SUPPORT SCHEMES FOR THE METHOD, EQUIPMENT AND SYSTEMS

USED IN CROP DEVELOPMENT PROJECT

The following is a flowchart of the collection of carbon dioxide (CO2) a greenhouse and aquaculture in the photo-bioreactor to produce algae in ponds (open system)

Aquaculture in a greenhouse to provide large amounts of carbon dioxide (CO2) suitable for recycling

algae in photo-bioreactors for algae produce controlled

PHOTOGRAPHS OF THIS SUPPORT (For reference) 3D map of a Bio-algae plant 10 hectares:

:

"CULTURE OF ALGAE IN LABORATORY"

""GETTING ALGAE HARVEST 1.5 GRS. Per liter"