Strains Cellulophaga sp. KP1 and Polaribacter sp. VR6 as a source of biotechnologically interesting enzymes

Encimi morskih bakterij so v industrijski biokatalizi zanimivi tako zaradi zmožnosti obdelave polisaharidov morskih alg za farmacevtsko in živilsko industrijo, kot tudi zaradi svojih procesnih lastnosti, npr. aktivnosti pri visoki slanosti, visokem tlaku, nizkih temperaturah itd. Tukaj predstavljamo seva Cellulophaga sp. KP1 in Polaribacter sp. VR6, pridobljena iz obalne vode Koprskega zaliva. Sodita v deblo Bacteroidetes, številčno pomembne razgrajevalce polisaharidov tako mikro kot makroskopskih alg, ki jih pogosto najdemo na morskih delcih, algah in okoliški vodi. Preučili smo ju tako iz biokemijskega kot bioinformacijskega stališča s sekvenciranjem celotnega genoma. Sekvenciranje celotnega genoma je nov način iskanja zanimivih encimov, pri čemer lahko v enem genomu najdemo veliko encimov z enakim substratom, a drugačnimi mehanizmi delovanja, kar je za biotehnologijo še posebno zanimivo. Iskali smo encime CAZY (ang. Carbohydrate Active enZYmes), saj so ti za biotehnologijo najzanimivejši. Glede na te encime smo primerjali naša seva z ostalimi predstavniki debla Bacteroidetes. Izbrali smo dva encima iz genoma seva Cellulophaga sp. KP1 (agarazo in karagenazo), ju rekombinantno izrazili ter jima preverili aktivnost.Enzymes of marine bacteria are of great interest for treatment of marine algal polysaccharides in pharmaceutical and food-related industries not only because of their substrate polysaccharide but also because of the extreme processing properties (low temperature, high pressure, high salinity …). Here we present marine strains Cellulophaga sp. KP1 and Polaribacter sp. VR6, isolated from coastal waters of Gulf of Koper. The isolated strains are of the phylum Bacteroidetes, an important group of algal polysaccharide degraders, commonly found on marine particles, algae and floating in water. Our study covered biochemical and bioinformatical analysis (with whole-genome sequencing) of the above strains. Sequencing of entire genomes is a new approach for finding interesting enzymes. With this approach it is possible to find multiple enzymes with similar function but different mechanism of action, which means different conditions of degradation of certain a polysaccharide. This is especially promising for biotechnological applications. We searched for CAZY enzymes (Carbohydrate Active enZYmes) and compared numbers of certain enzyme families with similar bacteria from the same phylum. We recombinantly expressed two interesting enzymes from strain Cellulophaga sp. KP1 (agarase and carrageenase) and confirmed their activity