Knots in trees An exceptionally rich source of bioactive polyphenols

Transcription

1 Knots in trees An exceptionally rich source of bioactive polyphenols Bjarne Holmbom Åbo Akademi IAWS Annual Plenary Meeting St. Petersburg, 16 June 2009 B. Holmbom ÅA 1

2 Knot As it happened in B. Holmbom ÅA 2

3 1998 A crooked spruce tree was cut down A knot was sawn out

4 Analysis: The spruce knot contained 10% of lignans! The start for extensive research on knots B. Holmbom ÅA 4

5 Analysis 10% lignans! More spruce knots Knots from other tree species Technical sep. of knots 1998 Applications Holmbom et al., 2007 B. Holmbom ÅA 5

6 Willför et al., 2003 a Knot in stem Lignans, mg/g Outer branch Tree 3, LK cm Upper w. Tree 6, LK Compr. w cm

7 Much higher concentrations of lignans in the north Piispanen et al., 2008 B. Holmbom ÅA 7

8 Lignans in Picea abies % 6-29% 0.1 % 0.0 % 70-85% of the lignans MeO O O HO HO Hydroxymatairesinol (HMR) Willför et al., 2003 a OH OMe

9 Analysis 10% lignans! More spruce knots Knots from other tree species Technical sep. of knots 60+ species 1998 Applications B. Holmbom ÅA 9

10 250 Picea hydrophilic knot extractives, mg/g LK: Living knot DK: Dead knot Other comp. Other lignans HMR LK DK LK DK LK DK LK DK P. abies P. glauca P. mariana P. sitchensis B. Holmbom ÅA 10

11 Larix hydrophilic knot extractives, mg/g Other comp. Other lignans Secoisolaricir LK DK LK DK LK DK L. decidua L. lariciana L. sibirica B. Holmbom ÅA 11

12 Pinus hydrophilic knot extractives, mg/g Other comp. Other lignans Lariciresinol LK DK LKa LKb LK DK LK DK LK DK P. banksiana P. contorta P. resinosa P. sibirica P. sylvestris B. Holmbom ÅA 12

13 Lignans in Pinus sylvestris % Heartw. Sapw. <0.05 % 33 knots 7 trees In pine knots also Resin acids 9 30% Stilbenes 1 7% MeO HO 80-95% of the lignans O OH O Willför et al., 2003 b OH OMe (-)-Nortrachelogenin (NTG)

14 mg/g d.w Hydrophilic extractives in hardwood knots Betula pendula Populus grandidentata Populus tremula Populus tremuloides Eucalyptus globulus Acacia crassicarpa Acacia mangium Fagus sylvatica Pietarinen et al., 2005 a Pietarinen et al., 2005 b B. Holmbom ÅA 14

15 mg/ g d.w Populus grandidentata Flavonoids in Populus species Others Populus tremula Dihydrokaempferol + glycoside Populus tremuloides Kaempferol + glycoside Naringenin + glycoside OH HO O OH OH O Dihydrokaempferol OH HO O OH O Naringenin Pietarinen et al., 2005 a

16 mg/g d.w Flavonoids in Acacia mangium and A. crassicarpa Oligomeric flavonoids Biflavonoids Keto-teracacidin Teracacidin Isomelacacidin Melacacidin 0 Acacia mangium Acacia crassicarpa Pietarinen et al., 2005 b B. Holmbom ÅA 16

17 Analysis 10% lignans! More spruce knots Knots from other tree species Technical sep. of knots ChipSep Production of knots 1998 Applications B. Holmbom ÅA 17

18 Stemwood Chipping Screening Pulping Over-thick chips TMP CTMP Kraft pulp ChipSep Knots Normal wood B. Holmbom ÅA 18

19 The ChipSep Process FI Pat Appl. Aug by C. Eckerman and B. Holmbom Pulping Over-thick chips Rechipped Chip Dryer Normal wood Sep. tank Warm air Knotwood B. Holmbom ÅA 19

20 Analysis 10% lignans! More spruce knots Knots from other tree species Technical sep. of knots 1998 Applications HMR as dietary supplement Antioxidants and biocides B. Holmbom ÅA 20

21 The spruce lignan Hydroxymatairesinol (HMR) Strong antioxidant Inhibits breast cancer growth (Univ. of Turku) Hormos Medical Corp., Turku, product development Clinical test on humans: no adverse effects New dietary ingredient clearance from US FDA 2004 LicenseHormos Linnea S.A. in 2005 MeO Production of knots (Separation Research) HO HO OH O O OMe B. Holmbom ÅA 21

22 Farm production of knots

23 Chips in Continuous ChipSep separator Knot product Normal wood out B. Holmbom ÅA 23

24 B. Holmbom ÅA 24

25 On the market in Recent research has shown that plant lignans have a positive influence on the development of breast, prostate and colon cancer which rely specifically on oestrogens in order to progress. Lignans also help to maintain good cardiovascular health and to moderate other oestrogendependent health problems such as menopause symptoms and osteoporosis. B. Holmbom ÅA 25

26 Knots: the richest source of polyphenols in nature MeO HO HO O O MeO HO OH O O MeO HO OH O OMe OH OMe OH OMe OH HMR Nortrachelogenin Lariciresinol MeO HO OH OH OH OMe MeO HO OH O O OMe In many species also flavonoids and stilbenes Secoisolariciresinol Matairesinol B. Holmbom ÅA 26

27 Next polyphenol products? HMR also in functional foods Natural cosmetics Technical antioxidants Natural biocides B. Holmbom ÅA 27

28 Analysis 10% lignans! More spruce knots Knots from other tree species 6-29% lignans 60+ species 5-15% polyphenols Technical sep. of knots ChipSep Production of knots 1998 Applications HMR as dietary supplement Antioxidants and biocides A new platform for production of bioactive compounds from trees B. Holmbom ÅA 28

29 Key publications I Pohjamo, S.P., Hemming, J.E., Willför, S.M., Reunanen, M.H.T. and Holmbom, B.R. (2003), Phenolic extractives in Salix caprea wood and knots, Phytochemistry 63:2, Willför, S., Hemming, J., Reunanen, M., Eckerman, C. and Holmbom, B. (2003 a), Lignans and lipophilic extractives in Norway spruce knots and stemwood, Holzforschung 57:1, Willför, S., Hemming, J., Reunanen, M. and Holmbom, B. (2003 b), Phenolic and lipophilic extractives in Scots pine knots and stemwood, Holzforschung 57:4, Willför, S.M., Ahotupa, M.O., Hemming, J.E., Reunanen, M.H.T., Eklund, P.C., Sjöholm, R.E., Eckerman, C.S.E., Pohjamo, S.P. and Holmbom, B.R. (2003), Antioxidant activity of knotwood extractives and phenolic compounds of selected tree species, J. Agr. Food Chem. 51:26, Holmbom, B., Eckerman, C., Eklund, P., Hemming, J., Nisula, L., Reunanen, M., Sjöholm, R., Sundberg, A., Sundberg, K. and Willför S. (2004), Knots in Trees A New Rich Source of Lignans, Phytochem. Rev. 2(3): Pietarinen, S., Willför, S. and Holmbom, B. (2004) Wood resin in Acacia mangium and Acacia crassicarpa wood and knots, Appita J. 57:2, Lindberg, L.E., Willför, S.M. and Holmbom, B.R. (2004) Antibacterial effects of knotwood extracts on paper mill bacteria, J. Ind. Microbiol. Biotechnol. 31, Eklund, P.C., Willför, S., Smeds, A., Holmbom, B.R. and Sjöholm, R. (2004) A new lariresinol-type butyrolactone lignan derived from hydroxymatairesinol and its identification in spruce wood, J. Nat. Prod. 67:6, Willför, S., Reunanen, M., Eklund, P., Sjöholm, R., Kronberg, L., Fardim, P., Pietarinen, S. and Holmbom, B. (2004), Oligolignans in Norway spruce and Scots pine knots and Norway spruce stemwood, Holzforschung 58: Willför, S., Nisula, L., Hemming, J., Reunanen, M. and Holmbom, B. (2004), Bioactive phenolic substances in industrially important tree species. Part 1: Knots and stemwood of spruce species, Holzforschung, 58: Willför, S., Nisula, L., Hemming, J., Reunanen, M. and Holmbom, B. (2004), Bioactive phenolic substances in industrially important tree species. Part 2. Knots and stemwood of different fir species. Holzforschung 58: B. Holmbom ÅA 29

30 Key publications II Pietarinen, S., Willför, S., Sjöholm, R. and Holmbom, B. (2005), Bioactive substances in important tree species. Part 3. Knots and stemwood of Acacia crassicarpa and Acacia mangium. Holzforschung, 59: Willför, S., Elund, P., Sjöholm, R., Reunanen, M., Sillanpää, R., Von Schoultz, S., Hemming, J., Nisula, L. and Holmbom, B. (2005), Bioactive substances in important tree species. Part 4. Identification of two new 7- hydroxy divanillyl butyrolactol lignans in some spruce, fir and pine species. Holzforschung, 59: Pietarinen, S., Hemming, J., Willför, S., Vikström, F. and Holmbom, B. (2005, Wood resin in Bigtooth and Quaking Aspen wood and knots. J. Wood Chem. Technol. 25(1-2): Markus, H., Mäki-Arvela, P., Kumar, N., Kul kova, N.V., Eklund, P., Sjöholm, R., Holmbom, B., Salmi, T. and Murzin, D.Yu. (2005), Hydrogenolysis of hydroxymatairesinol over carbon-supported palladium catalysts, Catal. Lett. 103(1-2): Willför, S.M., Sundberg, A.C., Rehn, P.W., Holmbom, B.R and Saranpää, P.T. (2005), Distribution of lignans in knots and adjacent stemwood of Picea abies, Holz-Roh Werkst. 63: Markus, H., Mäki-Arvela, P., Kumar, N., Heikkilä, T., Lehto, V.-P., Sjöholm, R., Holmbom, B., Salmi, T. and Murzin, D.Yu. (2006), Reactions of hydroxymatairesinol over supported palladium catalysts, J. Catalysis 238(2): Pietarinen, S.P., Willför, S.M., Ahotupa, M.O., Hemming, J. and Holmbom B.R. (2006), Knotwood and bark extracts : strong antioxidants from waste materials. J. Wood Sci. 52: Willför, S.M., Smeds, A.I. and Holmbom, B.R. (2006), Chromatographic analysis of lignans. J. Chromatogr. A 1122: Pietarinen, S.P, Willför, S.M., Vikström, F.A. and Holmbom, B.R. (2006): Aspen knots, a rich source of flavonoids. J. Wood Chem. Technol., 26(3): Smeds, A.I., Eklund, P.C., Sjöholm, R.E., Willför, S.M., Nishibe, S., Deyama, T. and Holmbom, B. (2007) Quantification of a broad spectrum of lignans in cereals, oilseeds and nuts. J. Agric. Food Chem. 55: Neacsu, M., Eklund, P.C., Sjöholm, R.E., Pietarinen, S.P., Ahotupa, M.O., Holmbom, B.R. and Willför, S.M. (2007): Antioxidant flavonoids from knotwood of Jack pine and European aspen. Holz-Roh Werkst 65: 1-6. Willför, S., Hafizoğlu, H., Tümen, I., Yazici, H., Arfan, M., Ali, M. and Holmbom, B. (2007): Extractives of Turkish and Pakistani tree species. Holz-Roh Werkst. 65:3, B. Holmbom ÅA 30

31 Key publications III Neacsu, M., Micol, V., Pérez-Fons, L., Mallavia, R., Holmbom, B. and Willför, S. (2007) A novel antioxidant phenyl disaccharide from Populus tremula knotwood. Molecules 12(2): Välimaa, A-L., Honkalampi-Hämäläinen, U., Pietarinen, S., Willför, S., Holmbom, B. and von Wright, A. (2006): Antimicrobial and cytotoxic knotwood extracts and related pure compounds and their effects on foodassociated micro-organisms. Int. J. Food Microbiol. 115(2): Piispanen, R., Willför, S., Saranpää, P., Holmbom, B. (2008) Variations of lignans in Norway spruce (Picea abies [L.] Karst.) knotwood: within-stem variation and the effect of fertilisation at two experimental sites in Finland. Trees 22, Book chapter Holmbom, B., Willfoer, S., Hemming J., Pietarinen, S., Nisula, S., Eklund, P. and Sjoeholm, R. (2007) Knots in trees a rich source of bioactive polyphenols. In: Materials, Chemicals and Energy from Forest Biomass (Ed. D.S. Argyropoulos), ACS Symposium Series 954, ACS, 2007, pp Patents C. Eckerman and B. Holmbom: Förfarande för återvinning av kvistnötsmaterial från överstor flis / Method for recovery of knotwood material from over-size chips. Finn. Pat No ( ), US Pat. No 6,739,533 ( ) B. Holmbom, C. Eckerman, J. Hemming, M. Reunanen, K. Sundberg and S. Willför: A method for isolating phenolic substances or juvabiones from wood comprising knotwood. PCT Int. Appl. 2002, WO A1, 12 December 2002, 31 pp. S. Willför, C. Eckerman, J. Hemming, B. Holmbom, S. Pietarinen and A. Sundberg: Novel use of knotwood extracts, Finn. Pat. Appl. Nr , Nov. 12, 2003 B. Holmbom ÅA 31

32 Major financial support Academy of Finland (Finnish Centre of Excellence Programme) Tekes (Finnish Funding Agency for Technology and Innovations) Keksintösäätiö (Foundation for Finnish Inventions) Hormos Medical UPM-Kymmene B. Holmbom ÅA 32

33 Team Work ÅA, Wood and Paper Chemistry Rainer Ekman ( ), Markku Reunanen, Christer Eckerman, Jarl Hemming, Andrey Pranovich, Lari Vähäsalo, Stefan Willför, Kenneth Sundberg, Suvi Pietarinen, Linda Nisula, Thomas Holmbom, Reija Harlamow ÅA, Organic Chemistry Rainer Sjöholm, Patrik Eklund, Annika Smeds University of Turku Risto Santti, Sari Mäkelä, Niina Saarinen, Markku Ahotupa Personnel at Hormos Medical and UPM-Kymmene B. Holmbom ÅA 33