MetaBiopOR - Bioprospecting of oil reservoir metagenomes for novel thermostable biocatalysts of industrial relevance

The project MetaBiopOR aimed at utilizing the exclusive access of the project consortium of SINTEF, NTNU and Statoil to an existing collection of unique DNA samples (metagenomic DNA) and derived DNA sequence data from microbial communities isolated from oil samples from three different oil reservoir wells on the Norwegian Continental Shelf. The main aim of the project was to perform bioprospecting for novel thermostable enzymes of potentially high industrial value by functional screening of libraries of metagenomic DNA fragment libraries and by mining the available DNA sequence information for relevant genes encoding such enzymes. The development and implementation of new, efficient methods for such approaches were also a part of the project. The search for novel thermostable lipases with industrial applications for example in biofuel production, and novel thermostable carbonic anhydrases with potential for biocatalyst-assisted CO2 capture processes were our main focus, and novel enzymes of both classes were identified in the project. The novel enzymes were biotechnologically produced and characterized with respect to beneficial properties like their thermostability. In addition, the metagenomes were also screened successfully for members of other enzyme classes, among others thermostable proteases and carbohydrate active enzymes to demonstrate the broad potential that lies in oil reservoir metagenomes. A number of new cellulases were discovered and are now candidates for further characterization and evaluation in potential follow-up projects towards their potential future application for example in industrial biorefinery processes.

The National Strategy for Marine Bioprospecting 2010 has highlighted bioprospecting of Norwegian oil reservoirs as relevant for marine bioprospecting. In response to that, the proposed project aims at utilizing our exclusive access to an existing collecti on of unique metagenomic DNA samples and DNA sequence data derived from the HEIDRUN oil reservoir on the Norwegian Continental Shelf. On that basis, we will develop and implement innovative and optimized generic methods to efficiently handle and functiona lly access oil reservoir derived metagenomic DNA, applicable in subsequent searches for novel thermostable biocatalysts of relevance for the Norwegian industry. In the proposed project, we will apply both functional screening of comprehensive metagenomic DNA libraries and DNA sequence data mining of the HEIDRUN metagenomic DNA sequence data pool to identify novel members of specifically two classes of enzymes with a high potential for an implementation in future industrial bioprocesses: (i) novel thermost able lipases with a broad industrial application spectrum e.g. in biofuel production processes, and (ii) novel thermostable carbonic anhydrases with potential applications in biocatalyst-assisted CO2 capture and storage processes. From initial sequence da ta mining experiments, we already have strong evidence for the presence of respective biocatalytic functions encoded in the metagenomic DNA from the HEIDRUN oil field, and we are now planning to move forward and extract these functions for an in depth cha racterization. Novel biocatalysts identified by our approach will be recombinantly produced and subsequently characterized with respect to their biochemical and biophysical properties (e.g. thermo- and solvent stability, substrate spectrum, mode of functi on). This analysis will be the basis for the subsequent evaluation of their potential in future industrial bioprocesses.