Bioprospecting for high PUFA content in Northern marine microalgae. Pia Steinrücken University of Bergen Norway

Transcription

1 Bioprospecting for high PUFA content in Northern marine microalgae Pia Steinrücken University of Bergen Norway 1

2 Focus new microalgae with industrial potential University of Bergen, Norway University of Las Palmas de Gran Canaria University of Antofagasta, Chile 1. Bioprospecting 2. Characterization of selected strains 3. utdoor mass cultivation 2

3 Focus new microalgae with industrial potential University of Bergen, Norway 1. Bioprospecting Polyunsaturated fatty acids H 3

4 Fatty acids SAFA saturated fatty acids H MUFA monounsaturated fatty acids H PUFA polyunsaturated fatty acids H Polar lipids Algae cell P Neutral lipids Triacylglycerol (TAG) Chloroplast Nucleus mitochondria 4

5 Fatty acids SAFA saturated fatty acids H MUFA monounsaturated fatty acids H PUFA polyunsaturated fatty acids H Polar lipids Algae cell P Neutral lipids Triacylglycerol (TAG) Chloroplast Nucleus mitochondria 5

6 Fatty acids SAFA saturated fatty acids H MUFA monounsaturated fatty acids H PUFA polyunsaturated fatty acids H Polar lipids Algae cell P Neutral lipids Triacylglycerol (TAG) Chloroplast TAG Nucleus mitochondria TAG TAG 6

7 PUFAs EPA Eicosapentaenoic acid 20:5 DHA Provide health benefits Cardiovascular health Neural system Immune system Essential in human nutrition Docosahexaenoic acid 22:6 Major source Fish oil EPA Algae EPA DHA DHA EPA DHA Health products EPA DHA aquaculture Food Health products EPA DHA aquaculture Food 7

8 PUFAs EPA Eicosapentaenoic acid 20:5 DHA Provide health benefits Cardiovascular health Neural system Immune system Essential in human nutrition Docosahexaenoic acid 22:6 Major source Fish oil EPA Algae EPA DHA DHA EPA DHA EPA DHA Health products aquaculture Food 8

9 Concept 1. Bioprospecting 9

10 Concept 1. Bioprospecting Sampling Isolation Cultivation Screening Selection 10

11 Sampling 11

12 Sampling 12

13 Sampling Arctic Cruises May 2014 July 2014 Nov2014 August

14 Sampling Bergen 14

15 Sampling Bergen 15

16 Sampling Bergen 16

17 Sampling Bergen 17

18 Isolation 18

19 Isolation 19

20 Isolation 20

21 Cultivation 15 C PAR: 50 µmol photons m 2 s 1 10 C PAR: 50 µmol photons m 2 s 1 Conditions Medium: 80% Sea Water Nutrients (Walne`s) Light : 50 µmol photons m 2 s 1 16:8 light/dark Temp.: 15 C Fjord summer 10 C Arctic, Fjord winter 21

22 Cultivation 15 C PAR: 50 µmol photons m 2 s 1 10 C PAR: 50 µmol photons m 2 s 1 22

23 Cultivation 15 C PAR: 50 µmol photons m 2 s 1 10 C PAR: 50 µmol photons m 2 s 1 23

24 Cultivation 15 C PAR: 50 µmol photons m 2 s 1 25 strains Diatoms Green Algae 149 clonal strains 10 C PAR: 50 µmol photons m 2 s strains Diatoms 24

25 Screening D Day Conditions Medium: 80% Sea Water Nutrients (Walne`s) Light : µmol photons m 2 s 1 continuous Temp.: 15 C Fjord summer 10 C Arctic, Fjord winter 25

26 Screening D Nutrients Nutrients Day Conditions Medium: 80% Sea Water Nutrients (Walne`s) Light : µmol photons m 2 s 1 continuous Temp.: 15 C Fjord summer 10 C Arctic, Fjord winter 26

27 Screening D Nutrients Nutrients 0.2 TAG Day Conditions Medium: 80% Sea Water Nutrients (Walne`s) Light : µmol photons m 2 s 1 continuous Temp.: 15 C Fjord summer 10 C Arctic, Fjord winter 27

28 Screening D Nutrients Nutrients Day Analysis Daily: D 750 Exp & stat: 3x Dry weight 3x Fatty acids End: Molecular Identification partial LSU sequences 28

29 Screening Fatty acid analysis ne step extraction / methylation Internal standard 23:0 FAME Gas chromatography Analysis Daily: D 750 Exp & stat: 3x Dry weight 3x Fatty acids End: Molecular Identification partial LSU sequences 29

30 Screening 15 C 10 Strains Fjord 8 Diatoms 2 Green algae C 10 Strains 8 Arctic 2 Fjord winter Diatoms

31 Screening No ID 10 C 15 C

32 Screening No ID 10 C 15 C

33 Screening No ID 10 C 15 C % Nitzshia laevis % Acrocellulus cornucervis 6 99% Nanofrustulum shiloi 7 99% Phaeodactylum tricornutum % Phaeodactylum tricornutum 10 99% Phaeodactylum tricornutum % Thalassiosira hispida 100% Thalassiosira hispida 99% Attheya septentrionalis % Thalassiosira hispida 100% Thalassiosira hispida 100% Thalassiosira hispida 100% Thalassiosira hispida % Thalassiosira hispida 100% Thalassiosira hispida 33

34 Screening 10 C 15 C No ID 1 88% Micractinium sp % Nitzshia laevis % Acrocellulus cornucervis 6 99% Nanofrustulum shiloi 7 99% Phaeodactylum tricornutum % Phaeodactylum tricornutum 10 99% Phaeodactylum tricornutum % Thalassiosira hispida 100% Thalassiosira hispida 99% Attheya septentrionalis % Thalassiosira hispida 100% Thalassiosira hispida 100% Thalassiosira hispida 100% Thalassiosira hispida % Thalassiosira hispida 100% Thalassiosira hispida 34

35 Screening Biomass 10 C 15 C No ID Growth EPA DHA 1 Industrial benchmark levels: EPA > 18% of total fatty acids DHA > 12% of total fatty acids µ av > 0, Time

36 Screening Biomass 10 C 15 C No ID Growth EPA DHA 1 Industrial benchmark levels: EPA > 18% of total fatty acids DHA > 12% of total fatty acids µ av > 0, Time

37 Screening 10 C 15 C Biomass No ID Growth EPA DHA 1 Industrial benchmark levels: EPA > 18% of total fatty acids DHA > 12% of total fatty acids µ av > 0, Time

38 Screening 10 C 15 C Biomass No ID Growth EPA DHA 1 Industrial benchmark levels: EPA > 18% of total fatty acids DHA > 12% of total fatty acids µ av > 0, Time

39 Screening 10 C 15 C Biomass No ID Growth EPA DHA 1 Industrial benchmark levels: EPA > 18% of total fatty acids DHA > 12% of total fatty acids µ av > 0, Time

40 No Growth EPA DHA ID Screening C 15 C P. tricornutum P. tricornutum P. tricornutum Attheya septentrionalis

41 Selection g l 1 D exp stat Attheya septentrionalis Day total DW total FA % exp 19 % stat 41

42 Selection D stat 0.1 exp Attheya septentrionalis Day m g g 1 D W exp stat 25 % total FA 24 % total FA g l total DW total FA % 19 % exp stat FA 42

43 m g g 1 D W D 750 Selection stat exp P. tricornutum 20 % total FA 7 % total FA exp stat Day exp stat FA 25 % total FA 24 % total FA 0.3 total DW total FA m g g 1 D W g l % 19 % exp stat FA 43

44 Selection D stat 0.1 exp Attheya septentrionalis Day m g g 1 D W exp stat 25 % total FA 24 % total FA g l total DW total FA % 19 % exp stat FA 44

45 Selection D stat 0.1 exp Attheya septentrionalis Day m g g 1 D W exp stat 25 % total FA 24 % total FA 25 % total FA g l total DW total FA 12 % 19 % 26 % exp stat stat FA Increase in EPA content in stationary phase from 3% to 4,6 % to 6,4% of DW 45

46 Perspective 13 Attheya septentrionalis 2. Characterization of selected strains 46

47 Perspective 1. Bioprospecting 2. Characterization of selected strains 47

48 MiraclesBergenTeam Svein Rune Erga Pia Steinrücken Siv Kristin Prestegard Hans Kleivdal Fatty Acids Svein Mjos 48

49 49

50 Selection m g g 1 D W D exp P. tricornutum 20 % total FA exp stat Day m g g 1 D W exp stat FA 25 % total FA 24 % total FA g l total DW total FA % 19 % exp stat FA 50