MicroMBT: Discovery and training of microbial biocatalysts for biomass conversion using Moving Bed Technology (MBT)

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: Discovery and training of microbial biocatalysts for biomass conversion using Moving Bed Technology (MBT) Marine Biotechnology ERA-NET Stakeholder meeting 13-14

1 : Discovery and training of microbial biocatalysts for biomass conversion using Moving Bed Technology (MBT) Marine Biotechnology ERA-NET Stakeholder meeting October 2016, Brussels by Peik haugen Funded by the Marine Biotechnology ERA-NET 3-year duration An action under the European Commission's Seventh Framework Programme

2 CONSORTIUM Peik Haugen (Coordinator) Concetta De Santi Nils-Peder Willassen Ragnhild D Whitaker Bjørn-Steinar Sæther Fredrik Almqvist Henrik Antti Elin Moe Ricardo Louro Miguel Teixeira José Pedro Malanho da Silva

3 BACKGROUND (The need) White fish (Wild catch) From:

4 BACKGROUND (The need) Available cheap biomasses (avg. 0.2 /kg) 940,000 tons in total (716,000 used) 2011 (Norway alone) Approx. half [530,000 tons]: Ensilage (270,000) Fishmeal (260,000) 160,000 tons Oil/consumption 180,0000 tons Waste/Dumped (cod)

5 BACKGROUND (The need) The global fish protein market Animal Feed Flavour, foam, gelling etc Functional ingredients Cosmetics, cosmeceuticals Nutrients nutrition Pet food Marine proteins Human Bioactive peptides Bioprocessing Enzymes Blod pressure, immuno stimulants etc. Substrate, fermentation (Quorn) Reproduced from: RUBIN - Rapport nr Internasjonal markeds og industrianalyse for marine ingredienser. 2011

6 BACKGROUND (The need) Biotep: A national test plant for bioprocessing Marine biomass Processing Hydrolysates Enzymatic hydrolysis Separation of the aqueous phase from solids and fats 0.1 /kg 3 /kg

7 OUR SOLUTION (The idea) Hydrolysates BIOREACTOR IMMOBILIZED SYNTHETIC BACTERIAL COMMUNITIES PRODUCT 1 PRODUCT 2 PRODUCT 3 PRODUCT 4 PRODUCT 5 PRODUCT 6 PRODUCT 7... PUFA MMPs Siderophores Wax-esters Astaxanthin Other pigments K3 K1 K1 micro The main objective: To develop a sustainable biorefinery process by using synthetic microbial communities and Moving Bed Technology to transform low cost marine rest raw materials to high value compounds

WORK PLAN BUILD RUN CHARACTERIZE SCALE-UP MEDIA FORMULATION & SET UP OF LAB-SCALE BIOREACTORS OPERATION & OPTIMIZATION OF LAB-SCALE BIOREACTORS PRODUCTS

8 WORK PLAN BUILD RUN CHARACTERIZE SCALE-UP MEDIA FORMULATION & SET UP OF LAB-SCALE BIOREACTORS OPERATION & OPTIMIZATION OF LAB-SCALE BIOREACTORS PRODUCTS IDENTIFICATION AND CHARACTERIZATION PRODUCT LEADS (METABOLITES, STRUVITE, ENZYMES, ) SET-UP OF PILOT-SCALE MBT REACTOR UNIT NEW TECHNOLOGIES, TOOLS & PRODUCTS

9 SCIENCE & TECHNOLOGY (How will we do it) Arctic/sub-Arctic microorganisms In-house collection of approx marine bacterial isolates Taxonomically classified based on the 16S rrna gene Selected 100 isolates: Complete genomes Functional screening of 9 activities DNase Cellulase Amylase PUFA Xylanase Secondary metabolite potential predicted Protease Chitinase Gelatinase Esterase Siderophore production

SCIENCE & TECHNOLOGY (How will we do it) 2,5 2,0 Shewanella sp.

0,5 0,0 0 20 40 60 80 100 120 140 160 180 Salmon 2,5 2,0 Time

KH04J14 Marine Broth ASW 3% ph 8_6g/L COD SKIN ASW 3% ph 8_6g/L

SANDEELS ASW 3% ph8_6g/l SPRAT ASW 3% ph8_6g/l HERRING ASW 3%

20 40 60 80 100 120 140 160 180 Time (hours) Marine Broth ASW

10 SCIENCE & TECHNOLOGY (How will we do it) 2,5 2,0 Shewanella sp. SP043 Hydrolysates from marine biomass OD600nm 1,5 1,0 Sprat 0,5 0, Salmon 2,5 2,0 Time (hours) Polaribacter sp. KH04J14 Marine Broth ASW 3% ph 8_6g/L COD SKIN ASW 3% ph 8_6g/L SALMON (P) ASW 3% ph 8_6g/L SALMON (A) ASW 3% ph 8_ 6g/L SANDEELS ASW 3% ph8_6g/l SPRAT ASW 3% ph8_6g/l HERRING ASW 3% ph8_6g/l SHRIMPS Cod OD600nm 1,5 1,0 Shrimp Sandeels 0,5 0, Time (hours) Marine Broth ASW 3% ph 8_6g/L COD SKIN ASW 3% ph 8_6g/L SALMON (P) ASW 3% ph 8_6g/L SALMON (A) ASW 3% ph 8_ 6g/L SANDEELS ASW 3% ph8_6g/l SPRAT ASW 3% ph8_6g/l HERRING ASW 3% ph8_6g/l SHRIMPS Marine Broth ASW 3% ph 8_6g/L COD SKIN ASW 3% ph 8_6g/L SALMON (P) ASW 3% ph 8_6g/L SALMON (A) ASW 3% ph 8_ 6g/L SANDEELS ASW 3% ph8_6g/l SPRAT ASW 3% ph8_6g/l HERRING ASW 3% ph8_6g/l SHRIMPS Herring

11 SCIENCE & TECHNOLOGY (How will we do it) Synthetic microbial communities Kaldnes K1/ 24 wells polystyrene plate Pall Micro24 reactor w/k1 micro beads A B C A: fresh beads B: Used beads C: Stained fresh (left) or used (right) beads

12 SUMMARY We are developing a new method to convert cheap marine rest materials to a range of valuable products Hydrolysate We have a library of Arctic bacteria with potential to produce valuable compounds Marine hydrolysates are used to formulate growth media for marine Arctic bacteria We are currently identifying communities of bacteria that can form robust and stable biofilms in a moving bed biofilm reactor Spent media are analyzed for products at UmU/the Swedish Metabolomics Center BIOREACTOR LIPIDS/Oil Omega-3 Wax esters ENZYMES -ases SEC METABOLITES Siderophores Pigments

13 Thank you for listening!

Biorefining of marine biomass

Biorefining of marine biomass ERA-net Marine Biotechnology, Brussels 13th October 2016 Ole Jørgen Marvik, PhD Special Advisor Biobased Industries The main Government agency for innovation and industry