Extremophile Research & Biobank CCCryo

Logo CCCryo

The Extremophile Research & Biobank CCCryo Unit is concerned with the adaptation strategies and usefulness of cryophilic (= cold-loving) freshwater micro algae, so-called snow and permafrost algae. The CCCryo the unit's culture collection, which is probably unique in its scope and diversity, provides the basis for work.

Cryophilic algae are exposed in many ways to extreme stress factors in their natural habitats. These include in the first instance cold, increased light and UV radiation, desiccation as well as severe alternations in nutrient and salt content. The goal of this unit is to characterize isolates collected on expeditions with regard to their diverse adaptation strategies to these extreme environmental parameters and to transfer the special metabolites developed by the algae to industrial applications.

For bioproduction on an industrial scale, the unit moreover developed suitable photobioreactors for the sterile mass cultivation of autotrophic organisms.

 

In our work, we assume a comprehensive approach to the research:

  • 1. Basic research: Starting with the collection of algae during expeditions in polar and high alpine regions of the earth, individual strains are insulated and characterised taxonomically, phylogenetically as well as with regard to their culturing demands. With regard to their special adaptation strategies, we analysed the isolates at the RNA, protein as well as primary and secondary metabolite level.
  • 2. Application research: The algae strains are more closely characterised with regard to industrially exploitable metabolites. These include, for instance, antioxidants such as carotenoids, vitamins or multiple unsaturated fatty acids (PUFA), as well as special substances such as natural freeze-protection substances. Then their induction and production is optimised on a laboratory scale.
  • 3. Photobioreactor development (PBR): For production on an industrial scale, universally usable or also especially adapted photobioreactor plants, so-called multiloop and double helix systems, are being developed. Great emphasis is placed on sterile production conditions in order to be able to offer innovative products for the cosmetic and pharmaceutical industry as well as the food and feedstock industry.

This comprehensive approach to research allows us to offer research results from the source up to raw products from a single provider. Independent of the approach described above, the algae strains in our collection at the CCCryo website are available for public and private research companies.

PUFAChain – Development of an Integrating Bio-Production Process for Multiple Unsaturated Fatty Acids (PUFA) from Microalgae

In the scope of this EU project, an international team of nine partners examined the value-added chain from the bio-resource,  the production, harvest and reconditioning process up to and including the final product. The objective is to establish an economically stable production processes for Omega-3 fatty acids (DHA and EPA) that can serve as building blocks for modern oleochemistry.

Production of Algae Biomass as Starting Material for Cosmetic Additives

The Swiss company Mibelle AG Biochemistry develops innovative active ingredient additives for cosmetics. A process was developed together with Mibelle in a long-standing cooperation in order to gain special constituents from snow algae on an industrial scale. In addition, we continuously develop and optimise the production processes.

Photobioreactors

Photobioreactor
© Photo Fraunhofer IZI

Photobioreactor

Photobioreactors were developed to cultivate microalgae under sterile conditions, used in production lines at Fraunhofer IZI as well as by industrial customers since 2011. The reactors, which have a work volume of up to 60 L, facilitate the biotechnological utilisation of a broad range of phototrophic microorganisms, and can be individually adapted to specific production requirements.

Algae Pigments as Natural Dyestuffs for the Food Industry

Carotenoids

Carotenoids

Typical of snow and permafrost algae is the ability to produce secondary carotenoids and other antioxidants such as alpha-tocopherol (vitamin E). In their natural habitats, they react to stress with low nutrients and high light and UV radiation. The various algae strains sometimes show very different pigmentation patterns.

For mass production, the process is typically subdivided into two phases. In the first phase, a high biomass is produced with the optimal provision of nutrients and light. In the second phase, the synthesis of secondary carotenoids is introduced by setting certain stress factors. Usually great quantities of lipids are also built up in which the fat-soluble carotenoids are stored.

Depending on the algae strain, the algae mass contains various amounts of:

  • alpha and beta-carotenoids
  • lutein
  • neoxanthin
  • violaxanthin
  • antheraxanthin
  • zeaxanthin
  • echinenon
  • hydroxyechinenon
  • adinoxanthin
  • canthaxanthin
  • astaxanthin
  • alpha-tocopherol

Interesting for the nutritional supplements and feed stock as well as for the cosmetics industry are predominantly lutein, astaxanthin and alpha-tocopherol (vitamin E).

Mapping of Snow Algae Fields along the Coast of Svalbard

Green snow

Green snow

Red snow

Red snow

In 2013, the sixth expedition to Spitzbergen was carried out. The objective was to obtain a comprehensive overview on the distribution of snow algae fields along the entire coast of Spitzbergen and the nearby islands to the east. It had previously been uncertain why red and green snow, the mass phenomenon of snow algae blossoms, is so very pronounced in some fields and not others. The 2010 expedition had ruled out nutrient availability as the sole contributing factor. With the help of a remote-controlled octocopter system using VIS and IR cameras, as well as chemical soil analyses, it was determined in 2013 that the prevailing geological formations in the various regions of Svalbard and small-scale topography have a decisive influence on the occurrence of snow algae. These extremophile organisms prefer silica-rich grounds (sandstone and gneiss) with low pH values and low carbon content as opposed to chalk-rich regions. However, a special taxonomic group can occur on chalky sub-surfaces. Local climate conditions, such as sufficient precipitation, additionally appear to be important. Such field results provide important clues for the optimal cultivation of these unusual algae.

Ice Structuring Proteins Made of Snow Algae to Improve Quality of Ice Cream and Deep-Frozen Confectionery Products (ISP Food)

This cooperative project with the Fraunhofer IVV in Freising examined the integration of ice structuring proteins (ice structuring proteins, ISP, AFP) made of snow algae in ice cream and other deep-freeze products. ISP have the ability to bind irreversibly on ice surfaces and thus restrict the ice crystal morphology to small crystals. Our goal was to develop a supplement containing ISP for the food industry that prevents the recrystallization process and hence to significantly improve the quality of frozen food products.

Among others things, the work group and its partners have the following resources and devices at their disposal for special kinds of work:

  • Algae culture collection CCCryo with about 400 isolates of cryophilic organisms (algae, cyano-bacteria, fungi and mosses)

    The database of the culture collection CCCryo, as well as ordering information are accessible at the website. The algae are available for public and industrial research institutions.

  • Three PERCIVAL cell culture cabinets (T = -15 °C to +40 °C, light = 0-1.000 µmol m-2 s-1, UV-A/B)
  • One culture room (T = -10 °C to +30 °C, light = 0-200 µmol m-2 s-1) for test cultures
  • One culture room (T = +4 °C to +30 °C, light = 0-400 µmol m-2 s-1) for algae mass cultures
  • One temperature-controlled 12’ double container for algae mass cultures (T = +4 °C to +30 °C), illumination with LED technology up to 800 µmol m-2 s-1
  • In-situ sterilisable glass tube photobioreactors of multi-loop and double-helix design with the airlift principle (1 x 60 L, 2 x 30 L, 3 x 25 L, 6 x 10 L), total volume in the sterile production processes = about 255 L, with an annual capacity of about 100 kg fresh algae mass
  • Cryo-microscope with digital image processing
  • Clifton nanoliter osmometer (Otago osmometers)
  • Freezing unit for controlled cryo-conservation (SYLAB)
  • Cryogenic storage for protective storage and cryo-conservation
  • Gas chromatograph with FID detector (Agilent 7890B)
  • Element analyser (EuroEA CNS)
  • Octocopter UAV system with VIS and IR digital cameras for documenting aerial views

  • Mibelle AG Biochemistry

  • de Vera J-P, Boettger U, Lorek A, Wolter D, Grunow D, Hübers H-W, Spohn T, Noetzel RdlT, Sánchez FJ, Billi D, Baqué M, Rettberg P, Rabbow E, Reitz G, Berger T, Leuko S, Möller R, Bohmeier M, Horneck G, Westall F, Jänchen J, Fritz J, Meyer C, Onofri S, Selbmann L, Zucconi L, Kozyrovska N, Leya T, Foing B, Demets R, Cockell CS, Bryce C, Philips Brown S, Olsson-Francis K, Wagner D, Serrano P, Edwards HGM, Joshi J, Huwe B, Moritz S, Ehrenfreund P, Elsaesser A, Ott S, Meessen J, Feyh N, Szewzyk U, Schulze-Makuch D, Hermelink A, Lasch P. BIOMEX – the Biology and Mars Experiment in space during the EXPOSE-R2 mission on the ISS. GRA. 2014;16.
  • Leya T. Snow Algae: Adaptation strategies to survive on snow and ice. In: Seckbach, J., Oren, A. & Stan-Lotter, H. (eds.): Polyextremophiles, Vol. 27 (2013): S. 401-423, Springer Netherlands. DOI dx.doi.org/10.1007/978-94-007-6488-0_17.
  • Remias D, Wastian H, Lütz C, Leya T. (2013): Insights into the biology and phylogeny of Chloromonas polyptera (Chlorophyta), an alga causing orange snow in Maritime Antarctica. Antarct. Sci. 25 (2013), 5: S. 648-656. DOI dx.doi.org/10.1017/S0954102013000060.
  • de Vera J-P, Boettger U, Noetzel RdlT, Sánchez, FJ, Grunow D, Schmitz N, Lange C, Hübers H-W, Billi D, Baqué M, Rettberg P, Rabbow E, Reitz G, Berger T, Möller R, Bohmeier M, Horneck G, Westall F, Jänchen J, Fritz J, Meyer C, Onofri S, Selbmann L, Zucconi L, Kozyrovska N, Leya T, Foing B, Demets R, Cockell CS, Bryce C, Wagner D, Serrano P, Edwards HGM, Joshi J, Huwe B, Ehrenfreund P, Elsaesser A, Ott S, Meessen J, Feyh N, Szewzyk U, Jaumann R, Spohn T. Supporting Mars exploration: BIOMEX in Low Earth Orbit and further astrobiological studies on the Moon using Raman and PanCam technology. Planetary and Space Science 74 (2012), 1: S. 103-110.
  • Remias D, Aigner S, Leya T, Lütz C, Stuppner H, Schwaiger S. Characterization of an UV- and VIS-absorbing, purpurogallin-derived secondary pigment new to algae and highly abundant in Mesotaenium berggrenii (Zygnematophyceae, Chlorophyta), an extremophyte living on glaciers. FEMS Microbiol. Ecol. 79 (2012), 3: S. 638-648. DOI dx.doi.org/10.1111/j.1574-6941.2011.01245.x.
  • Spijkerman E, Wacker A, Weithoff G, Leya T. Elemental and fatty acid composition of snow algae in Arctic habitats. Frontiers in Microbiology 3 (2012): S. 380. DOI dx.doi.org/10.3389/fmicb.2012.00380.
  • Remias D, Karsten U, Lütz C, Leya T. Physiological and morphological processes in the Alpine snow alga Chloromonas nivalis (Chlorophyceae) during cyst formation. Protoplasma 243 (2010), 1: S. 73-86. DOI dx.doi.org/10.1007/s00709-010-0123-y.
  • Leya T, Rahn A, Lütz C, Remias D. Response of arctic snow and permafrost algae to high light and nitrogen stress by changes in pigment composition and applied aspects for biotechnology. FEMS Microbiol. Ecol. 67 (2008), 3: S. 432-443. DOI dx.doi.org/10.1111/j.1574-6941.2008.00641.x.
  • Reichle C, Schnelle T, Müller T, Leya T, Fuhr G. A new microsystem for automated electrorotation measurements using laser tweezers. Biochim. Biophys. Acta 1459 (2000): S. 218-229.
  • de Nys R, Leya T, Maximilien R, Afsar A, Nair PSR, Steinberg PD. The need for standardised broad scale bioassay testing: a case study using the red alga Laurencia rigida. Biofouling 10 (1996), 1-3: S. 213-224. DOI dx.doi.org/10.1080/08927019609386281.