Amino acid modifiers in

guayule rubber compounds

Colleen McMahan
Research Chemist
Lead Scientist, Domestic Natural Rubber
United States Department of Agriculture
Agricultural Research Service
Western Regional Research Laboratory
Albany, CA USA
USDA-ARS Objective
To develop commercially-viable natural rubber-producing crops suitable for
cultivation in the temperate climate of the United States.

cis 1,4-polyisoprene

guayule guayule natural rubber

Keys to sustainability
Tools of genetics, biotechnology ++ utilization of rubber and co-products
Opportunity for American and European farmers
Natural rubber the 4th most valuable agricultural import in the USA (2011)
Markets for alternative natural rubber are driving demand
US guayule growing region estimated at over 1,000,000 acres

79

faostat.fao.org
 Is guayule natural rubber a replacement for
Hevea natural rubber?
1. Guayule rubber has lower protein content and
is free from Type I latex allergens (Siler et al. Plasticity Retention Index: Guayule/Hevea blends

70
1996, Hamilton and Cornish 2010). 60
50

2. Guayule rubber is less thermally stable 40

PRI
30

(Schloman et al 1996a, Keller et al. 1981). 20

10
0
3. Guayule rubber cures more slowly (Schloman 0 20 40 60 80 100

et al. 1996b, Cornish et al. 2008). % Guayule

4. Guayule rubber has very low gel content Green tensile properties

5 HEVEA
(McMahan et al. 2007). 4
PI
SBR

Stress (MPa)
GUAYULE
5. Guayule rubber induces SIC more slowly 3
2

(Shimomura et al. 1982). 1

0
0 500 1000 1500 2000
Strain (% )
The difference between Hevea and guayule rubber is not the rubber,
its the non-rubber constituents: proteins, amino acids, etc.
Onizawa, Curable Rubber Compositions with Amino Acids (1981)
amino acids as vulcanization accelerators, in some cases obviating the need for activators (ZnO).
Othman et al. (1993)
Addition of certain types of amino acids increased compound modulus.
Abad et al. (2002)
improved tensile strength retention attributed to in radiation-cured NR compounds to antioxidant properties of cysteine, alanine and
asparagine.
Schloman et al. (2005)
surfactant and washing treatment of Hevea latex reduced the extent of vulcanization in correlation to removal of allergenic proteins.
Tuampoemsab and Sakdapipanich (2007)
improved PRI for deproteinized natural rubber after addition of a Hevea latex extract attributed the result to proteins as natural
antioxidants in NR proteins
Amnuaypornsri et al. (2008)
Superior properties of NR are not directly caused by proteins and phospholipids but their interactions with rubber chain ends to form the
network structure.
Strain-induced crystallization (SIC) of Hevea rubber (Toki 2014).
Removal of proteins reduce material strength and SIC.
Hevea structure model: Amnuaypornsri et al. (2008)
Guayule natural rubber
establishing technological fit for use
Recently, significant new industrial investment has taken place for guayule production
in the southwestern U.S. due to price instability for petroleum-based and imported
rubber and to demand for biobased products.
Sustainable reintroduction of guayule rubber in the industrial supply chain would
benefit from a clearer understanding of the similarities and differences between rubbers
from the two plant sources.
The availability of low protein guayule NR enables an alternative model to compare
the impact of non-rubber constituent biological compounds interaction with a polymer
of natural rubber stereochemistry.
Here. low-protein guayule natural rubber (GNR) was blended with a series of amino
acids varying in chemical structure to study the impact of specific interactions (e.g.
hydrogen bonds, disulfide bonds, and hydrophobic interactions) that may be formed in
the blends.
Materials
L-Proline 98.5+%
SAFC Supply Solutions, MO, USA

L-Glutamine FCC grade PhytoTechology

Laboratories, KS, USA
Guayule natural rubber latex
L-Cysteine HCl 99.94% NutraBio, NJ, USA
Yulex Corporation, USA

Hevea natural rubber latex

Glycine 98% Centex LATZ
ACROS Organics, NJ, USA Centrotrade, USA

trans-4-Hydroxy-L-Proline Fresh Hevea latex

99% Instituto Agronmico de Campinas
Sigma-Aldrich Co., MO, USA So Paulo, Brazil
 Amino acids in blends
trans-hydroxy-
Amino acid Glycine L-Glutamine L- Proline L-Cysteine
L-proline
Formula C2H5NO2 C5H10N2O3 C5H9NO2 C3H7NO2S
C5H9NO3

Structure

Molecular weight, Da
75.07 146.15 115.13 121.16 131.13

Hydrophobicity
0.501 0.251 0.711 0.68 0.527
index

Hydrophobicity index from Black and Mouldt (Analytical Biochemistry, 1991) based on arginine =0 and
phenylalanine =1; higher number means more hydrophobic.
Latex blends preparation

Blends were RT stirred 48-72 hours then air-dried prior to use.

GNR
L-Gln
2% L-Gln +
GNR
Test specimen drying
Compound formulation
Ingredient Tradename/Supplier phr (dry)
Guayule latex Yulex Corporation 100
Potassium hydroxide solution (10%) Fisher Scientific 0.5
Sulfated methyl oleate salt surfactant Darvan SMO (1) 1
Sodium alkyl sulfate/sodium lauryl sulfate surfactant Darvan WAQ (1) 1
Sulfur dispersion Bostex 378 (2) 1
Zinc oxide dispersion Zinc oxide dispersion (1) 1
Dithiocarbamate accelerator Setsit 5 (1) 0-1
Dithiocarbamate accelerator Setsit 104 (1) 0-1
Antioxidant Agerite superlite (1) 0-1
107.5
1= RT Vanderbilt, Norwalk, CT 2= Akron Dispersions, Copley, OH
 Characterization methods
Characteristic Method Parameters Instrumentation
Dynamic viscosity ASTM D6204B+C * (dynamic Advanced Polymer Analyzer
(polymer + blends) complex viscosity) Alpha Technologies, Akron, OH
USA
Cure parameters (compounds) Cure parameters
Plasticity Retention Index ASTM D3194-04 Wallace Plastimeter
PRI Wallace Instruments,
Redhill, Surrey
UK
% Gel ASTM D3616-95 % gel (insoluble) -
Green strength of raw films Instron Tensometer
ASTM D412-06a Tensile strength Instron, Norwood MA
Mechanical properties of Die D USA
compounds
Protein stability SDS-PAGE Gel image -
 + Gly:Pro:OH-Pro 2.0
+ Gly:Pro:OH-Pro 1.0
GNRL Control
+ OH-Pro 2.0
% change in * @ 100C, 1.0 Hz

+ OH-Pro 1.0
+ OH-Pro 0.2
GNRL Control
+ Cys 2.0
+ Cys 1.0
+ Cys 0.2

lowest value + 2.0% cysteine = 1781 Pa-sec

GNRL Control
+ Pro 2.0
Results: Dynamic Viscosity

Controls average * = 2830 Pa-sec

+ Pro 1.0
+ Pro 0.2
GNRL Control
+ Gln 2.0
+ Gln 1.0
+ Gln 0.2
GNRL Control
+ Gly 2.0
+ Gly 1.0
+ Gly 0.2
GNRL Control

30

20

10

-10

-20

-30

-40

-50
 G
N
RL

100
200
300
400
500
600
700

0
Co
n
+ trol
G
ly
+ 0.2
G
ly

cysteine = 35.8
G + 1.
0
N G
RL ly 2
Co .0
n
+ trol
G
ln
+ 0.2
G
ln
G + 1.
0
N Gln
RL 2
Co .0
n
+ trol
Pr
o
+ 0. 2
Pr
o
G + 1.
0
N P ro
RL 2
.
Co 0
n
+ tro
Results: Plasticity Retention Index

Cy l
s
Controls average PRI = 9.6; highest value for +2.0%

+ 0.2
Cy
s
% change in PRI

+ 1.0
G Cy
N
RL s 2
.0
+ Co
O ntr
H o
+ - Pro l
O
H 0. 2
+ - Pro
O
H 1
+ GN - P . 0
G r
ly RL o 2
:
+ Pro Co . 0
G
ly :O H ntr
+ :Pro -P ol
G r
ly :O H o 0
:P .
ro -P r 2
:O o
H 1. 0
-P
ro
2.
0
Results: Green strength
HydroxyProline/Guayule Blends 100% Gua Proline/Guayule Blends 100% Gua

Green Strength + 2.0% OH-Pro Green Strength + 2.0% Pro

0.4 0.5

0.4
0.3

Stress MPa
Stress MPa

0.3
0.2
0.2

0.1 0.1

0
0.0
0 400 800 1200 1600
0 400 800 1200 1600
Strain % Strain %

Glycine: Proline: HydroxyProline 100% Gua

100% Gua Cysteine/Guayule Blends
Guayule Blends + 2.0% Cys
+ 2.0% AA Green Strength
Green Strength 0.4
0.4

0.3 0.3

Stress MPa
Stress MPa

0.2 0.2

0.1 0.1

0.0 0
0 400 800 1200 1600 0 200 400 600 800 1000 1200
Strain % Strain %
 + Gly:Pro:OH-Pro 2.0
100% Hevea

+ Gly:Pro:OH-Pro 1.0
+ Gly:Pro:OH-Pro 0.2
GNRL Control
Controls average gel = 7.9% (sd 1.6); +2.0% cysteine = 12.3%

+ OH-Pro 2.0
+ OH-Pro 1.0
+ OH-Pro 0.2
100% Guayule

GNRL Control
+ Cys 2.0
+ Cys 1.0
+ Cys 0.2
GNRL Control
+ Pro 2.0
+ Pro 1.0
+ Pro 0.2
GNRL Control
+ Gln 2.0
+ Gln 1.0
+ Gln 0.2
Results: % gel

GNRL Control
+ Gly 2.0
+ Gly 1.0
+ Gly 0.2
GNRL Control

150

100

50

-50

-100
Results:
Cure curves for cysteine/guayule blends @ 100C
100% Gua
Cysteine and Guayule Compound Blends + 0.2% C ys
Cure Curve w/o Acc + AO + 1.0% C ys
+ 2.0% C ys
5

4
Strain [S']

2
0 10 20 30 40
Time [min]
Results:
Cure curves glutamine/guayule blends @ 100C
100% Gua
Glutamine/Guayule Compound Blends + 0.2% Gln
Cure Curve w/o Acc + AO + 1.0% Gln

6 + 2.0% Gln

5
Strain [S']

1
0 10 20 30 40
Time [min]
Results: Cure curves @ 1.67 Hz, 0.5 arc, ~160C
100% Gua 100% Gua
Glycine/Guayule Compound Blends +0.2% Gly Glutamine/Guayule Compound Blends +0.2% Gln
Cure Curve +1.0% Gly Cure Curve +1.0% Gln
+2.0% Gly +2.0% Gln
5.0 5.0

4.0 4.0

Strain [S']
Strain [S']

3.0 3.0

2.0 2.0
0 10 20 30 40 0 10 20 30 40

Time [min] Time [min]

100% Gua
HydroxyProline/Guayule 100% Gua Glycine:Proline:Hydroxy-Proline/
+ 0.2% OH-P ro +0.2% Gly:P ro :OH-P ro
Compounded Blends Guayule Compound Blends
+1.0% OH-P ro
Cure Curve +1.0% Gly:P ro :OH-P ro
+2.0% OH-P ro Cure Curve
5.0 5.0 +2.0% Gly:P ro :OH-P ro

4.0 4.0
Strain [S']

Strain [S']
3.0 3.0

2.0
2.0
0 10 20 30 40
0 10 20 30 40
Time [min] Time [min]
 Results: Heat stability of Hevea latex proteins
Fresh Hevea latex Centex Hevea latex
total proteins total proteins

10 l

l
Co r
r

r
ro

C
C

C
ro
ke
ke

ke
nt

nt
ar
ar

ar
0

0
0

0
Co

12

14

16

12

14
10

16
M
M

M
Eaton and Grantham (1915) reported adding heat-coagulable proteins in their original form had only
slightly improved slow curing crepe, but after putrefaction the same proteins had a marked accelerating
action on vulcanization.
Addition of amino acids to guayule latex.
Glutamine and Guayule Blends
ASTM D6204 B -Viscoisty * at 1.0 Hz
3500

acted as a plasticizer reducing bulk 3000

2500

* (Pa.s)
viscosity up to 40% (glutamine) 2000
1500
1000
500
0

acted as an antioxidant, in all cases (best 0.0 0.2 1.0

% Amino Acid
2.0

for cysteine) Glycine and Guayule Blends

Plasticity Retention Index

25

impacted cure depending on the 20

15

PRI
composition and R-group structure 10
5
0

Without accelerator 0.0 0.2 1.0

% Amino Acid
2.0

acceleration (cysteine) Cysteine and Guayule Blends

scorch inhibition (glutamine) 40

35
Plasticity Retention Index

With added accelerator 30

25

PRI
20

.reduced cure state (most) 15

10

moderated reversion
5
0
0.0 0.2 1.0 2.0

(glutamine) % Amino Acid

Summary: Addition of amino acids to guayule latex
L-Glycine
trans-hydroxy-L- L-Proline
Amino acid Glycine L-Glutamine L-Proline L-Cysteine
proline trans-hydroxy-L-
proline
Formula C2H5NO2 C5H10N2O3 C5H9NO2 C3H7NO2S C5H9NO3 -

Structure -

5-12%, systematic
Gel Content - - Up from 4-11% - -
increase

Bulk Lower Reduced, esp. at

Lower at 2% Lower at 2% Lower at 1% Lower at 2%
Viscosity systematically 1%
PRI Improved Improved ++ Improved Improved ++ Improved Improved
Green
- - Slight increase - Slight increase Slight increase
Strength
State of cure slight
Reversion Yes bump, NO Yes Yes Yes Yes
reversion
Conclusions
Amino acids can be considered biobased modifiers in latex and bale rubber
compounds for use as plasticizers, antioxidants, and as vulcanization aids.
L-glutamine, in particular, served as a plasticizer, antioxidant, and anti-
reversion agent in guayule compounds.
There are over 500 known amino acids; additional studies are required to
fully assess their technological, economic, and life cycle potential.

Continued studies can also provide further insight as to the mechanisms of

non-rubber constituents interaction with polymers in Hevea (and other)
natural rubber compositions.
 Dankeschn!
This work was supported by the United
States Department of Agriculture,
Agricultural Research Service through a
collaboration with the University of
Nevada-Reno.

Materials were generously donated by:

Yulex Corporation
Instituto Agronmico de Campinas
RT Vanderbilt Company Prof. David Shintani
Centrotrade Ms. Dhondup Lhamo
Akron Dispersions