Our
technology platform allows for an efficient isolation of unialgal cultures,
even from highly complex cyanobacterial blooms. Isolated strains are being
classified according to their taxonomy. At this stage about one third is pre-selected
for further analyses. Our proprietary protocols for strain preservation enable
a reliable preservation of almost all known cyanobacterial species.
Screening
Pre-selected
strains enter a screening procedure, employing proprietary PCR methods for the
detection of biosynthetic genes for natural products as well as HPLC/MS analyses
of cell and media extracts after cultivation under certain conditions. Again
about one in three strains entering this phase is selected for upscale
cultivation to yield biomass for bioactivity assays.
Cultivation
Positively
screened strains are scaled up to culture volumes of up to 1000 liters. For
selected strains, cultivation conditions are optimized with respect to natural
product formation and yield. When required the biosynthetic genes are
induced by specific growth conditions to further enhance production of known
target compounds.
Purification / Fractionation
Following
harvest and freeze-drying of up-scaled production strains the target compounds
are purified in a multi-step process. An elaborated extraction and
fractionation procedure in combination with our automated CPC, LC/MS and liquid
handling technology yields libraries of pure substances or pre-purified extract
fractions suitable for microplate based automated assays.
Analysis / Dereplication
All
isolated compounds or fractions are analyzed employing state-of-the-art
equipment, including HPLC coupled with high-resolution multi-stage mass
spectrometric detection of the compounds. The identification of known
cyanobacterial natural products is facilitated using commercially available databases
as well as Cyano Biotechs proprietary in-house database of cyanobacterial
natural products. Structures can be elucidated using a combination of tandem-HRMS and one- and two-dimensional NMR experiments.
Bioactivity Assays
Our
MTP libraries of pure compounds as well as fractions are screened in various
assays for bioactivity with a focus on antibacterial, antifungal and cytotoxic
activity. However, the company is also testing their libraries for antioxidative activity, kinase and phosphatase inhibition as well as for activity
against parasites causing malaria, sleeping sickness or leishmaniasis.
