BIOMASS RECALCITRANE

LIGNIN

A Twenty-first Century Challenge Isroi (http://isroi.com)

Cell Wall Model: Cellulose, Hemicellulose and Lignin

Cell Wall Model: Cellulose, Hemicellulose and Lignin

Cell Wall Model: Cellulose, Hemicellulose and Lignin

Pseudocolor SEM micrograph of lignin redeposited on cellulose (10,000X).

www.nrel.gov/.../technology.cfm/tech=18

Lignin:

Natures second abundant organic substances Product of phenylpropanoid pathway Lignin monolignol/lignin-forming pathway steps evolved during transition (evolution) of plants from the water habitat to the land habitat.

LIGNIN: Moleculer Basis and role in plant adaptation

Plant evolution

Diagram showing changes in cell wall composition during the course of evolution.

Sarkar P et al. J. Exp. Bot. 2009;60:3615-3635

The Author [2009]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Sarkar P et al. J. Exp. Bot. 2009;60:3615-3635

LIGNIN: Moleculer Basis and role in plant adaptation

Figure 3. Phylogenetic tree showing the distribution of lignin monomer composition across major lineages. *, Sunits are only found in cell cultures of Ginkgo, not in wood (Novo Uzal et al., 2009). **, Lignin-like structures are reported in some mosses and green algae, but the presence of real lignin in these nonvascular species remains questionable; red algae have been barely studied (Weng et al., 2008b; Martone et al., 2009).

Vanholme R. et.al. Plant Physiol. 2010:153:895-905

Copyright 2010. 2010 American Society of Plant Biologists. All rights reserved.

Red Seaweed

Like many land plants, this red seaweed produces lignin, a primary component of wood. (Credit: Kathy Ann Miller)

Selaginela
http://www.science20.com/news_releases/biofuels_research_makes_a_plant_evolution_discovery

Selaginela

http://esciencenews.com/
Both lignin and cellulose are found in the rigid cell walls of the xylem cells (those that conduct water) in the primitive plant, Selaginella. (Credit: Zina Deretsky, National Science Foundation; Selaginella cross section SEM by Jing-Ke Weng, Clint Chapple, Purdue University; Lignin structure from Wout Bergjan, John Ralph, Marie Baucher (Annual Review of Plant Biology, Vol. 54:519-546, June 2003); Cellulose structure from http://www.chusa.jussieu.fr/disc/bio_cell/)

Fossil Ginkgo leaves from the Jurassic of England. Some 160 million years ago.

Modern Ginkgo leaves.

Corn Stover

Wood

http://siteresources.worldbank.org

Softwood

Hardwood

Larger, longer cells Water transported by cells

Contains large-diameter vessels Water transported by these vessels

Lignin: Major monolignols of lignins.

H-unit P-hydroxyphenyl

G-unit guaiacyl

S-unit syringyl

Hatfield R., Vermerris W. Plant Physiol. 2001:126:1351-1357

Lignin: Major monolignols of lignins.

(A) Monolignol esters 30-32 found in grasses.

(D) Several dominant substructures present in native lignins.

Lignin Structure:

Freudenbrerg (1965)

Lignin Structure:

Brunow et al. 1998

Lignin Structure:

Lignin Biosynthesis Pathway

Lignin Pathway Evolution

Humphreys, J. M., C. C. S. Chapple. 2002. Rewriting the lignin roadmap. Plant Bio. 5, 224229.

Lignin Pathway Evolution

Humphreys, J. M., C. C. S. Chapple. 2002. Rewriting the lignin roadmap. Plant Bio. 5, 224229.

Vascular Plant Diversification and Lignification

Vascular Plant Diversification and Lignification

Pinus taeda

Medicago sativa

Vascular Plant Diversification and Lignification

Loblolly pine (Pinus taeda)

Tamarack (Larix laricina)

Ebony (Diospyros ebenum)

Douglas fir

compression

Black cottonwood

tension

normal normal

Monilignol Biosynthesis

Monilignol Biosynthesis

7.3.2

Monilignol Biosynthesis

7.3.3

Monilignol Biosynthesis

7.3.4

Monilignol Biosynthesis

7.3.5

Monilignol Biosynthesis

7.3.6

Monilignol Biosynthesis

7.3.7

Monilignol Biosynthesis

7.3.8

Monilignol Biosynthesis

http://aob.oxfordjournals.org/content/91/6/673/F3.expansion

Inherent Shortcomings in lignin analyses: a critical juncture and the urgent need

Lignin Isolation Procedure: Strong acid (sulphite pulping proceses) Strong alkaline (kraft pulping) Milled wood lignin Bjrkman lignin

Lignin Structure: Lignin subunit and lignin structural analyses by NMR spectroscopy

Figure 2. Representation of a lignin polymer from poplar, as predicted from NMR-based lignin analysis (adapted from Stewart et al., 2009).
Vanholme R. et.al. Plant Physiol. 2010:153:895-905

Copyright 2010. 2010 American Society of Plant Biologists. All rights reserved.

Quantification of lignin amounts, lignin degradation protocols, and synthetic dehydropolymerizates

Klason Lignin Various non-lignin componens, such as tannins, was determined Acetyl bromide Thioglycolic acid

Modulation of monolignol pathway and peroxidase enzymatic step

Lignin, a key component of plant cell walls, is normally synthesized from three simple monolignols (top). The hydroxyl group (OH), shown in red, must remain unmodified for these precursors to link up. Liu's team created a novel enzyme (green "ribbon" structure) that can methylate this specific hydroxyl group. This enzyme may therefore lead to ways to interfere with lignin biosynthesis in plants to make them easier to break down for biofuels.

Chang-Jun Liu

Native Lignin Macromolecular Configuration

1965

Freudenbrerg (1965)

Lignin Structure: 1970s-1980s

Lignin Structure:

Adler (1977)

Lignin Structure:

Partial structure of a hypothetical lignin molecule from European beech (Fagus sylvatica ). The phenylpropanoid units that make up lignin are not linked in a simple, repeating way. The lignin of beech contains units derived from coniferyl alcohol, sinapyl alcohol, and para-coumaryl alcohol in the approximate ratio 100:70:7 and is typical of angiosperm lignin. Gymnosperm lignin contains relatively fewer sinapyl alcohol units. (After Nimz 1974.)

Lignin Structure: 1990s and 2000s

Brunow et al. 1998

Lignin Structure: Lignin subunit and lignin structural analyses by NMR spectroscopy

Figure 2. Representation of a lignin polymer from poplar, as predicted from NMR-based lignin analysis (adapted from Stewart et al., 2009).
Vanholme R. et.al. Plant Physiol. 2010:153:895-905

Copyright 2010. 2010 American Society of Plant Biologists. All rights reserved.

Future Outlook

Future Outlook
1. Monolignol transport to the cell/wall 2. Lignin initiation sites 3. Lignin Primary Sequences/New Chemistries 4. Re-oxidation of the growing lignin chains 5. Monolignol radical and lignin primary chain interactions, proposed template polymerization, and lignin association 6. Transcriptional control of individual cell wall formation processes, biomechanics, and biodegradation of plant cell walls.

Biomass Recalcitrance

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Alkali nitrobenzene oxidation/thioacidolysis/permanganate oxidation degradation procedures