Industrial use of archaea for integrated Pollution Control-Waste Water, Gases , Soil Bioremediation, and the  Biodiversity Conservation Euring. Bela TOZSER Expert UN&EU Commission, Managing and Engineering Director , Envirosan DC H-7100 Szekszárd, Rizling Str. 11. POB.:199. Hungary E-mail: envsan@t-online.hu Web: envizont.eu

Table of contents Industrial use of archaea Integrated, innovative complex for growth of archae Opportunities of application and perspectives

Archaea Archaea are the smallest independently living single-celled organisms on earth Typical cells range in size from 0.5 to1.0 µm in diameter DNA lies free in the cell cytoplasm A cell membrane comprised of phospholipids surrounds the cell. The cell wall is usually made up of protein and carbohydrate and lipids. Most of the time they undergo asexual reproduction, mostly by dividing in half – some cells divide every 12–20 minutes, others take a lot longer. All organisms require carbon, which provides building blocks for cell materials Archaebacteria are also known as "extremophiles," thanks to their ability to survive in extreme environments such as very hot and very cold climates.

Archaea Colored transmission electron micrograph of the archaea Methanospirillum hungatii undergoing cell division Archaea were originally thought to exist only in harsh environments Were often described as ‚extremophiles’ They are widely distributed and are found alongside bacteria in many environments including soil and water. The best described groups of Archaea are the methanogens that produce methane, and the extremophiles that grow under high salt or extreme temperatures. Pyrodictum, for example, has an optimal growth temperature of 105°C.

Prokaryotic Cells First cell type on earth Cell type of Bacteria and Archaea No membrane bound nucleus Nucleoid = region of DNA concentration Organelles not bound by membranes

The archaebacteria have the following unique combination of traits: Prokaryotic traits: They are about 1 micrometer (μm) in diameter, the size of typical procaryotes. They lack membrane-bound organelles. They have nuclear bodies (nucleoids) rather than true, menbranee bound nuclei. Their ribosomes are 70 S, the size of those found in typical prokaryotes. Eukaryotic traits: Their cell walls completely lack peptidoglycan. Their protein synthesis machinery is sensitive to inhibitors that typically affect only eukaryotes and is resistant to many inhibitors that affect prokaryotes. Some of their proteins, pigments, and biochemical processes closely resemble those found in eukaryotic cells.

Archaebacteria include three groups: The methanogens, strict anaerobes that produce methane (CH4) from carbon dioxide and hydrogen. Extreme halophiles, which require high concentrations of salt for survival. Thermoacidophiles, which normally grow in hot, acidic environments.

Enzymes and proteins from organisms that grow near and above 100 °C Hyperthermophile group includes some methanogenic and sulfate- reducing species Only a few species are saccharolytic. Most of the hyperthermophiles absolutely depend on the reduction of elemental sulfur (S0) to H2S for significant growth, The fermentative pathways appear to depend upon enzymes that contain tungsten

Extremophiles as a source of novel enzymes for industrial application Extremophilic microorganisms are adapted to survive in ecological niches: high temperatures, extremes of pH, high salt concentrations high pressure. These microorganisms produce unique biocatalysts which operateunder extreme conditions Selected extracellular-polymer-degrading enzymes and other enzymes could be used in food, chemical and pharmaceutical industries and in environmental biotechnology. (amylases, pullulanases, cyclodextrin glycosyltransferases, cellulases, xylanases, chitinases, proteinases, esterases, glucose isomerases, alcohol dehydrogenases and DNA-modifying enzymes ) A scanning electron micrograph of a ten-million-year-old Archaea.

Biotechnological applications of archaeal biomasses Biological methanogenesis is applied to the anaerobic treatment of sewage sludge, agricultural, municipal and industrial wastes Methanogens are a group of microorganisms that obtain energy for growth from the reaction leading to methane production. Many bioreactor configurations have been exploited to increase the efficiency of anaerobic digestion, such as the rotating biological contactor, the anaerobic baffle reactor the upflow anaerobic sludge blanket reactor, several large-scale plants are in operation

Archaeal enzymes of biotechnological interest Natural and modified archaeal enzymes present huge possibilities for industrial applications Many archaeal enzymes involved in carbohydrate metabolism - special interest to the industrial biotechnology sector. The starch processing industry can profit from the exploitation of thermostable enzymes. Another promising application of hyperthermophilic archaeal enzymes is in trehalose production. Several other polymer-degrading enzymes isolated from archaea could play important roles in the chemical, pharmaceutical, paper, pulp or waste treatment industries. (xylanases and cellulases) Some archaeal metabolites have potential industrial applications. (proteins, osmotically active substances, exopolysaccharides and special lipids ) Archaeal lipids have been proposed as monomers for bioelectronics

The solution from Envirosan DC. -CliEnvHeP3BiC- Climate-, environment-, and healthprotecting, bioenergetical, biotechnological and biorefinery Complex With the multifunctional climate-, environment-, healthprotecting, bioenergetical, biotechnological and biorefinery system the following functions can be realized: Utilization in the environment protection Bioenergetical utilization Biotechnological – health protectional utilization Environmental utilization Biorefinery utilization

-CliEnvHeP3BiC-

Utilization in the environment protection at the following emissioners: CO2- and other GHG bioconversion, real zero carbon emission and O2 output – carbon quotas can be sold, valuable bioproduct production Cement industry Energy industry Chemical industry Vehicles Food industry Biofuels Lime-burners Biogas plants Pyrolysis plants Zero carbon emission of waste-fields Bioenergetical utilization: With the help of the climate-protecting processes, the system produces energy- and energy sources The increase of the energetical effectiveness of the biogas plant Production of biofuels Hidrothermal gasification Combined processes: trigen energy Bioenergetical and biotechnologigal utilization of thermal waters and the dissolved gases of thermal waters Biofuel production Landfill gas use without any kind of emission Energetical use of soiled gas wells Bio - batteries

 Biotechnological – health protectional utilization Functional food producing Production of C5, C6 sugar with versatile utilization Animal feed, protected protein- and fat production for ruminant animals, fish- and crabfeed, hatcheries, increase of spawn- and caviar production Products with iodin content in case of nuclear radiation Propagation of tribe-yeasts Pharmaceutical groundmaterials, fine-biochemicals, vegetal stem-cells from archeas, medicinal fungus, herbs cells produced in closed/sterile system Balneotherapy, wellness products Beauty-care, skin-care, biocosmetics Biotechnological care of the crop, pest control Complex utilization of carbon-dioxide stations for high-end biotechnological development Microalgaes, archaea for space-traveling and submarines Synthesis-gas-production, biomaterials, biopolymers, bio building blocks, bioresins

Environmental utilization Reduce of the salt content of thermal-, and seawater with use of halofil micro-organisms Intesification of waste water cleaning Nitrogen reduce of the fermentations fluid of the biogas plants, odour reducing/prevention Disposal of harmful wastes Elimination of radioactive pollution Regeneration of polluted soils (heavy metals, biological degradatible pollutions) Propagation of micro-organisms for soil- and pest control for seed-corn treatment Enrichment of soils: biofertilizer Water-, and soil regeneration polluted with red-sludge Water protection, restoration of biodiversity Utilization of non-fermentable organic waste (pyrolysis) Environment rehabilitation Decrease of air pollution Waste management, utilization of organic wastes Air conditioning of closed areas, like spaceships, submarines

Biorefinery utilization biofuel diversification biochemicals biocharbohydtrates bioresins Biochemicals The projects are adoptable for any geographycal locations. The size and complexity of the system is variable, The product-range is broadable. The size, the product-mix and the product volumes are changeable in connection with the market needs. From the container-form system to the bigggest giga-complexes is every form possible. It is posible to connect the system to polluting plants like heating plants, biogas plants, sewage farms, waste-fields etc.

Fully Automatised Dynamical Photocatalitical Bioreactor

Intensification of biomethanisation process of anaerobical digestion- archae contribution up to 30% incresing of process yield High integration of biotechnological, environment and climate protection processes, very high efficiency in pollution control of waste-water : archae and microalgae for increasing of biological treatment efficiency CO2 GHG – atmospheric pollutants and extreme climate phenomena producing gases bioconvarsation to valuable energetical and biotechnological products waste to energy factory of complex with anaerobical intensified digesters , pirolysis reactors for non biodegradable organical wastes, biorefinery capacities for diversification of biofuels contribution to soil bioremediation , biodiversity conservation by high quality biofertilizers

Dynamical photocatalytical bioreactors Capacities are harmonized to pollutant emissions, waste cantities, the complex is caracterized by really zero carbon emission Use to complex valorification of natural resources : CO2- gas fields, thermal, mineral, brine, treated waste watwers, sea water Design of whole complex with multiple use and different size is fully finalized simulated, redy for developing of execution projects Fully authomatized and on-line monitorized processes, complex instrumentation, data processing and IT

Opportunities of application and perspectives Adaptability to specifique needs of China Asia, Europe and Worldwide The implementations represent high social impacts for improving the quality of environment, protection of health, reducing the extreme climate phenomena, production of high amount of renewable energies. The economical efficiency is very high comparable to traditional processes by decresing of reactor capacities, investment and operational costs, valuable bioenergetical and biotechnological products,high amount of tradable CO2 The concrete technico-economical parameters will be determined by feasibility studies- Boundless perspectives by introducing in industrial use of new archae, combinations of process with other microorganisms, development of technical contents The use of this complex ensure the biotechnology and bioenergy based sustainable development

Summary The use of archaea in framework of integrated industrial complexes for pollution control, waste water, gases,soil bioremediation, biofertilization and biodiversityconservation open new perspectives for increase the technical and economical efficiency of climate- environment- health protection, bioenergy production from waste, therefore for sustainable development.

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