Cellulose: structure, morphology, and
crystalline forms
CHEM-E2140
Eero Kontturi
29th October 2015
Outline
(1) Molecular structure of cellulose
- molecular vs. supramolecular structure
(2) Crystalline forms (allomorphs): - cellulose I
- cellulose II
- cellulose III
- cellulose IV
- consequences of the crystalline forms
(3) Morphology of native cellulose: microfibril
- models for microfibrillar cross sections
- crystalline-amorphous structure
- microfibril aggregation / coalescence
Molecular structure of cellulose
Non-reducing
end group
Reducing
end group
Reducing
OH
HO
HO
O
O
OH
end group
OH
O
HO
OH
Anhydroglucose
HO
OH
OH
HO
O
H
OH
n /2
OH
OH
Cellobiose
OH
Reducing end group is in equilibrium
with a cyclic hemiacetal
HO
OH
O
O
OH
Molecular vs. supramolecular structure
Supramolecular structure
Molecular structure
OH
Anhydroglucose
OH
O
HO
O
O
HO
OH
O
n /2
OH
Cellobiose
- (14)--D-glucopyranose
- high native DP (~5000-15000)
Structure revealed:
Freudenberg
Liebigs Ann. Chem. 1928, 461, 130.
Haworth Nature 1930, 126, 438.
- individual cellulose chains linked
together by intermolecular bonding
Structure revealed (cellulose I ja I):
Nishiyama et al.
J. Am. Chem. Soc. 2002, 124, 9074.
J. Am. Chem. Soc. 2003, 125, 14300.
Crystalline forms of cellulose
Cellulose allomorphs
NATIVE
FORM
Regeneration or
alkali treatment
Cellulose I
Cellulose I
Cellulose II
IRREVERSIBLE
Liquid NH3 or
diamine treatment
REVERSIBLE
Cellulose IIIII
Cellulose IIII
Heat treatment
Cellulose IVI
New evidence suggests that cellulose IV
is a slightly disordered form of cellulose I.
Methods for measuring the crystalline
forms and crystallinity of cellulose
X-ray diffraction
Solid state Nuclear Magnetic Resonance (NMR) spectroscopy
specifically: cross-polarization magic angle spinning (CP-MAS) 13C NMR
Most applied methods, generally regarded as the most reliable
Electron diffraction
Neutron scattering
IR spectroscopy
Raman spectroscopy
Native cellulose:
Cellulose I
Cellulose I
Cellulose I crystal
Cellulose chains form sheets which are connected with each other
Radial cross section of a cellulose I crystallite:
66 model (not confirmed!)
36 cellulose chains
Within the sheets:
hydrogen bonds
Between the sheets:
van der Waals bonds
Cross sectional image taken from: Gross and Chu J. Phys. Chem. B 2010, 114, 13333.
Cellulose I: main hydrogen bonding
patterns
Main hydrogen bonds:
Sheet in cellulose I
35 intramolecular bond
lends rigidity to the cellulose
chain
1
2
26 intramolecular bond
3
6
36 intermolecular bond
keeps the sheets together
NOTE: Cellulose chains run
parallel in cellulose I crystal
Distinction: cellulose I and cellulose I
Two forms of native crystalline cellulose exist: I and I.
Atalla and Vanderhart Science 1984, 223, 283.
Crystallographic details in 1
resolution (cellulose I ja I):
Nishiyama et al.
J. Am. Chem. Soc. 2002, 124, 9074.
J. Am. Chem. Soc. 2003, 125, 14300.
I: one chain triclinic
I: two chain monoclinic
- dominant in, e.g.,
bacterial cellulose
and algae
- dominant in higher plants
(e.g. wood, cotton)
Distinction: cellulose I and cellulose I
Baker et al. J. Struct. Biol. 1997, 119, 129.
Distinction: cellulose I and cellulose I
Cellulose I is the predominant form in higher plants (wood, cotton
etc.) and tunicate (cellulose in tunicate animals)
Cellulose I is the predominant form in algae and in cellulose emitted
by microbes (bacterial cellulose)
NOTE: Cellulose I and cellulose I ALWAYS coexists with each other in
nature, usually within the same microfibril.
Regenerated cellulose:
Cellulose II
Cellulose II
NATIVE
FORM
Cellulose I
Regeneration or
alkali treatment
Cellulose II
Cellulose I
Man-made form of cellulose
Preparation by: - dissolving the cellulose / regeneration
- swelling in concentrated alkali (e.g. > 10% NaOH)
Silk-like texture of cellulose II materials means that they
are widely applied in textile industry.
Distinction between cellulose I and II
(a) cellulose II
Cellulose
(b) cellulose IIII
Cellulose
1
2
3
6
2
6
Hydrogen bonding patterns are different.
Distinction between cellulose I and II
Cellulose chain has a direction:
OH
Anhydroglucose
HO
HO
HO
O
O
OH
Non-reducing end
OH
Reducing
end group
OH
HO
OH
OH
HO
O
H
OH
n /2
OH
OH
Reducing end
Cellobiose
Cellulose I
Cellulose II
Parallel
Anti-parallel
Distinction between cellulose I and II
Cellulose II
Preparation by: - dissolving the cellulose / regeneration
- swelling in concentrated alkali
How is it possible for the cellulose chains to transform from
parallel to anti-parallel without dissolution?
Cellulose I
Cellulose II
swelling
Parallel
Anti-parallel
Distinction between cellulose I and II
Cellulose crystals in parallel microfibrils run in opposite direction
NaOH swells the crystals
Anti-parallel arrangement is thermodynamically more favourable than
parallel arrangement
HYPOTHESIS:
Cellulose chains in parallel microfibrils intermingle and form new
antiparallel crystals
Cellulose I
Cellulose II
swelling
Parallel
Anti-parallel
Evidence for antiparallel cellulose II
Single cellulose I microfibril
CELLULOSE I
Two cellulose I microfibrils
Swelling in NaOH (aq)
(i.e. mercerization)
CELLULOSE II
Single cellulose II microfibril
Kim et al. Biomacromolecules 2006, 7, 274.
Evidence for antiparallel cellulose II
Dissolved end-functionalized cellulose:
Exposure to gold surface/
regeneration
Sulphur binds with gold.
Diffraction pattern
of cellulose I
Au
Forced parallel orientation
Yokota et al. Adv. Mater. 2007, 19, 3368.
Cellulose III
Conversion into cellulose III
NATIVE
FORM
Regeneration or
alkali treatment
Cellulose I
Cellulose II
Cellulose I
IRREVERSIBLE
Liquid NH3 or
diamine treatment
REVERSIBLE
Cellulose IIII
Cellulose IIIII
Parallel chains
Anti-parallel chains
Hess and Trogus Ber. 1935, B68, 19861988.
Conversion: cellulose I cellulose III
Cellulose I
Cellulose IIII
Ammonia-cellulose I complex
NH3 (l)
NH3 removal
(e.g. evaporation)
Wada et al. Macromolecules 2006, 39, 2947.
Distinction: cellulose I and cellulose III
Cellulose I
Hydrogen bonds only between
cellulose molecules within the
sheets
van der Waals bonds between the
sheets
Cellulose IIII
Hydrogen bonding also between
the sheets
Wada et al. Macromolecules 2006, 39, 2947.
Distinction: cellulose I and cellulose III
Cellulose I
Cellulose IIII
Chundawat et al. J. Am. Chem. Soc. 2011, 133, 11163.
Reversibility of cellulose III
Cellulose III can be converted back to its starting material
Cellulose I
Cellulose II
Cellulose I
Hydrothermal treatment
at ~160C
or
Thermal treatment at >200C
Cellulose IIII
Cellulose IIIII
Parallel chains
Anti-parallel chains
Chanzy et al. Carbohydr. Res. 1987, 160, 1.
Wada et al. Macromolecules 2001, 34, 3271.
Conversion: cellulose III cellulose I
Black: cellulose IIII
Grey: cellulose I
Wada et al. Macromolecules 2001, 34, 3271.
Cellulose IV
Conversion: cellulose III cellulose IV
NATIVE
FORM
Regeneration or
alkali treatment
Cellulose I
Cellulose II
Cellulose I
Liquid NH3 or
diamine treatment
Cellulose IIII
Cellulose IIIII
Heat treatment in
glycerol at 260C
Cellulose IVI
Cellulose IVII
Hutino and Sakurada Naturwissenschaften 1940, 28, 577.
Conversion: cellulose III cellulose IV
recent evidence
Recent evidence from FT-IR spectroscopy, solid state NMR spectroscopy,
X-ray diffraction and diffraction simulations:
Wada et al. Biomacromolecules 2004, 5, 1385.
Newman Cellulose 2008, 15, 769.
Credible proof that cellulose IVI is not a genuine allomorph
Cellulose IVI is seen as a distorted form of cellulose I
Consequences of the different
crystalline forms
Stiffness properties
Form
Cellulose I
Cellulose II
Cellulose IIII
Cellulose IIIII
Elastic modulus
138 GPa
88 GPa
87 GPa
58 GPa
The values are estimates for pure crystalline forms.
Native cellulose I is decidedly stiffer and stronger than the man-made forms
Nishino et al. J. Polym. Sci. B 1995, 33, 1647.
Enzymatic hydrolysis: cellulose I vs. II
Hydrolysis of
cellulose II is much
faster and proceeds
to a greater extent
than that of
cellulose I.
NOTE: Cellulose II hydrate is a form of cellulose II with water molecules inside
its crystalline lattice.
Wada et al. Polym. Degrad. Stabil. 2010, 95, 543.
Enzymatic hydrolysis: cellulose I vs. II
vs. III
Glucose yield from enzymatic hydrolysis of different cellulose allomorphs
C-I: cellulose I
C-I*: ammonia-cellulose complex
C-II: cellulose II
C-III: cellulose III
AC: amorphous cellulose
Chundawat et al. J. Am. Chem. Soc. 2011, 133, 11163.
Morphology of cellulose:
Native cellulose microfibril
Microfibrils in the plant cell wall
Individual fibre
10 m
Cellulose microfibril
Diameter: 2-20 nm
Cross sectional models of crystalline
cellulose microfibrils
66 (36) chain models
35 nm
3.8 nm
Ding and Himmel J. Agric. Food Chem. 2006, 54, 597.
Sugiyama et al. Macromolecules 1991, 24, 4168.
Based on Atomic Force Microscopy
much debated model.
Based on diffraction studies
More accepted model
Cross sectional models of crystalline
cellulose microfibrils
24 chain model
Based on diffraction, NMR, and FTIR spectroscopy data
Debate is still open
Fernandes et al. PNAS 2011, 108, E1195
Cross sectional models of crystalline
cellulose microfibrils
Terminological note:
36 and 24 chain models refer to the smallest cellulose microfibrils,
such as those present in wood cells
Often these smallest microfibrils are referred to as elementary fibrils
In many species, the microfibrils are larger but they are multiplicates of
the elementary fibrils
Often the microfibrils (or elementary fibrils) aggregate, forming larger
microfibrillar bundles
Crystallinity and width of microfibrils
Collected data from analyses of some native cellulose sources
Source
Degree of
crystallinity
Microfibril
width*
Microfibril
width**
Algal cellulose
>80 %
10 nm
10-35 nm
Bacterial cellulose
65-79%
5 nm
4-7 nm
Cotton linters
56-65%
5 nm
7-9 nm
Ramie
44-47%
5 nm
3-12 nm
Hemp
60%
3-5 nm
3-18 nm
Flax
56%
4-5 nm
3-18 nm
Dissolving pulp
43-56%
4-5 nm
10-30 nm
*) Deduced from X-ray diffraction (reflection from 110 lattice plane)
**) Deduced from transmission electron microscopy images
Zugenmaier, In: Crystalline cellulose and cellulose derivatives;
Springer: 2008; p. 208.
Crystalline-amorphous transitions in a
cellulose microfibril
Longitudinal order: fringed-fibrillar
model of native cellulose microfibrils
Crystallographic data presents evidence
that cellulose within microfibrils is not
totally crystalline.
Proposition:
cellulose runs through alternating
crystalline and amorphous regions.
Fringed-fibrillar model of native cellulose
microfibrils
Not just one model
Hearle 1958
Dolmetsch 1968
Hess and Kiessig 1953
... to name but a few.
Fringed-fibrillar model evidence?
Level-off degree of polymerization (LODP): cellulose depolymerizes rapidly
down to a certain level after which the degradation is minimal.
HCl HYDROLYSIS
5OC
18OC
LODP
80OC
40OC
Proposition: amorphous domains are hydrolyzed leaving the
crystalline domains intact.
Battista Ind. Eng. Chem. 1950, 42, 502.
Fringed-fibrillar model evidence?
DP of ramie fibres after
HCl hydrolysis
Small angle x-ray scattering
pattern of untreated ramie
LODP
Crystallite length (i.e. length of crystalline domains)
by SAXS agrees with the level-off
degree of polymerization (LODP).
Nishiyama et al. Biomacromolecules 2003, 4, 1013.
Fringed-fibrillar model evidence?
DP of ramie fibres after
HCl hydrolysis
LODP
However,
- the yield loss upon controlled
acid hydrolysis is very small
- SAXS pattern implies a very
short disordered region
Proposition: disordered regions
are merely 4-5 anhydroglucose
units in length.
Nishiyama et al. Biomacromolecules 2003, 4, 1013.
Fringed-fibrillar model so what?
The disordered segments can be selectively targeted with acid hydrolysis.
H+
H+
H+
H+
H+
RESULT:
cellulose nanocrystals
LODP
LODP depends on the native
source of cellulose.
Rnby Discuss. Faraday Soc. 1951, 11, 158.
Fringed-fibrillar model so what?
Alternating crystalline-amorphous regions explain well
the macroscopic mechanical properties of cellulosic materials.
The length and width of the crystalline domains
depend on the native source of the material.
Elastic properties of isolated cellulose
nanofibrils depend on their native source.
Page J. Pulp Paper Sci. 1983, 3, TR15.
Fringed-fibrillar model implications?
When you see data of the degree of crystallinity of cellulose, its physical
meaning is unclear
If the degree of crystallinity is, e.g., 64%, it does not mean that 64% of the
cellulose is crystalline and 36% is amorphous
Probably much of the material responsible for the amorphous response
resides on the microfibril surface
Cellulose I and cellulose II degrees of crystallinity should never be
compared with each other
Systematic sets of data can be compared with each other if the crystalline
forms, the analytical method, and the raw materials are similar
Aggregation or coalescence of
cellulose microfibrils inside the cell wall
Appearance of a cellulose microfibril
Aggregates: 12-20 nm
Individual microfibrils: ~3.5 nm
200 nm
AFM image of a surface of
bleached birch kraft pulp;
sample untreated.
TEM image of longitudinal
cross-section of chlorite
delignified pine cell wall;
freeze-dried and stained.
Heyn J. Ultrastructure Res. 1969, 26, 52.
Appearance of a cellulose microfibril
Aggregates: 12-20 nm
TEM image of radial
cross-section of wood cell wall.
Zimmermann et al.
J. Struct. Biol. 2006, 156, 363.
Individual microfibrils: ~3.5 nm
TEM image of longitudinal
cross-section of chlorite
delignified pine cell wall;
freeze-dried and stained.
Heyn J. Ultrastructure Res. 1969, 26, 52.
Implications of microfibril aggregation
Felling/
drying
Why is this important?
Reduced surface area
Aggregation
Reduced accessibility
Schematic cross sections of microfibrils
Fewer reaction sites
Difficulties to extract
cellulose nanofibrils
Suchy et al.
Biomacromolecules 2010, 11, 515.
Summary
Cellulose exists in several crystalline allomorphs:
- cellulose I and I (native forms)
- cellulose II (prepared regeneration or alkaline treatment)
- cellulose IIII and IIIII (prepared by liquid ammonia treatment)
Cellulose allomorphs are physically different and they differ in reactivity
Structure of cellulose microfibril is an unsettled issue (36 and 24 chain
models)
Cellulose in microfibrils is not totally crystalline: the order is frequently
interrupted by longitudinal disorder (fringed-fibrillar model)
Microfibrils have an inherent tendency to coalesce with each other
Disorder and coalescence affect the macroscopic physical properties of
cellulosic materials
