Biodegradation of Silly Putty

Biodegradation of
Silly Putty
BPE AAT
SHWETHA JAYAKUMAR
What is Silly Putty?
● A Sillicone polymer that has unusual physical properties. It bounces, but it breaks
when given a sharp blow, and it can also flow like a liquid.
● It contains a viscoelastic liquid silicone, a type of non-Newtonian fluid, which makes
it act as a viscous liquid over a long time period but as an elastic solid over a short
time period.
PROBLEM:
● Abrasive-flow-based polishing was first introduced by McCarty in
1960s as a method to deburr and polish metal products
● This method employs a medium made up of fine abrasive particles
mixed with silly putty.
● This technique is used in the aerospace and automotive industries to
finish complex shapes and extremely small orifices
● So how do you discard the Silly Putty that has already been used in
finishing or polishing?
LET’S TRY BIODEGRADATION!!

● In soil, the PDMS polymer can hydrolyze to small, water
soluble siloxanols with the ultimate product being the
monomeric dimethylsilanediol (DMSD).
● This hydrolysis is probably abiotic, because it can take
months to years in wet soil, but only days as the soil dries.
● The hydrolysis product, DMSD, can microbially degrade to
CO2 and inorganic silicate, the latter of which should merge
with the silicate already present in the soil. [1]
Degradation of PDMS to DMSD [2]
Londo sandy clay loam having 12% moisture, degraded
200 centistoke (~s)'~C-labeled PDMS, slowly over six
months to yield about 3% of applied l4C as
low-molecular-weight, watersoluble products. When
the soil was allowed to dry in one week from 12 to 3%
moisture, the degradation rate was much more rapid,
and after several days at 3% moisture about half of the
applied 14C was water desorbable. HPLC-GPC of
tetrahydrofuran (THF) soil extracts showed that PDMS
had been degraded to low-molecular-weight molecule.
Biodegradation of DMSD [3]
A: Four Soil Samples were taken:
1. Sample 1: Sandy soil from Santa Barbara, California

2. Sample 2: Mixture - Forest + Residential Garden Soil + Composted Cow
Manure from Cobleskill, New York
3. Sample 3: Residential Garden Soil from Guilderland, New York
4. Sample 4: Sludge Disposal area from Glendale, Ohio
B: Before weighing moisture contents of the soil were measured and adjusted
1. Santa Barbara Soil: 14%

2. Cobleskill Soil Mixture: 20%
3. Glendale Soil: 40%
The moisture content of Guilderland Soil was not measured. But the moisture
content was very high when collected.
C: Setup of Gledhill Flasks:
● An appropriate amount of water was removed from the Guilderland

soil, by blowing air over the sample.
● Sterilized controls of both the Cobleskill and the Santa Barbara soils
were prepared by autoclaving the soils for 1 h on each of three
successive days, adding the soil to the Gledhill flasks, and the next
day autoclaving the soil-containing flask for an additional hour.
● In addition, sodium azide (2 ml of a 2.5% aqueous solution) was also
added to the autoclaved controls.
● After soil preparation, 50g of respective soil, 2.5 ml of 2,000ppm
solution of [14C]dimethylsilanediol, and 100l of ammonium chloride
(50 g/liter) were mixed together in a 500ml modified Gledhill flask.
D. Screen for a primary carbon source:
● In order to isolate soil microorganisms capable of biodegrading dimethylsilanediol,

the activity observed in the soil samples needed to be transferred to liquid culture.
● Before this could be done, a primary carbon source that could support the growth of
the soil microorganisms in liquid culture and that allowed the continued conversion of
dimethylsilanediol had to be identified.
● Numerous carbon sources were screened for their ability to enhance the
biodegradation of [14C]dimethylsilanediol in soil. These carbon sources were added to
separate samples of either Guilderland or Santa Barbara soil which were spiked with
[14C]dimethylsilanediol, and the production of CO2 was monitored.
● Many of the carbon sources did not enhance the rate of 14CO2 production. However, for
each soil type, at least two carbon sources increased the production of 14CO2 greater
than 90% above the mean. For the Guilderland soil, these carbon sources were acetone
and 2-propanol, and for the Santa Barbara soil, the carbon sources that enhanced
activity were 2,3- butanediol, 2-propanol, and dimethylsulfone.
E. Transfer of Activity to Liquid Culture:
● The contents of the vials from the most active samples from the carbon source
screening (Guilderland soil, acetone and 2-propanol; Santa Barbara soil, 2,3-butanediol,
dimethylsulfone, and 2-propanol) were taken and soil particles were removed by
shaking with glass beads.
● The supernatant was added to a medium containing 500 or 1000ppm of
corresponding carbon source and 100ppm of [14C]dimethylsilanediol.
● Medium was put into a 250ml Erlenmeyer presterilized plastic flasks with screw caps.
● Air was supplied to the cultures when the flasks were opened to remove traps, sample
the culture, or add a carbon source. A plastic, presterilized test tube standing upright
inside the flask and containing 3 ml of 2 M KOH was used as the CO2 trap.
● All liquid culture flasks were incubated at room temperature and were continuously
shaken (except during sampling) on a bench-top laboratory shaker.
F. Isolation of active microorganisms:
● The most active liquid cultures were spread on agar plates containing the same minimal
medium used in liquid cultures and nonlabeled dimethylsilanediol (100 ppm).
● For cultures receiving dimethylsulfone, the agar was made with 500 ppm of dimethylsulfone.
● Biocides (1,000 ppm) were sometimes added to the agar medium of the plates for the
Guilderland microorganisms in order to separate fungus and bacteria in a mixed culture.
● A few days after inoculation, the plates were examined for growth. Single colonies were
picked and replated. Picking and replating were continued until the colonies on the plates
were homogeneous.
● These cultures were restarted in liquid culture to ensure purity and to ensure that activity was
not lost during the purification.
● This cycle was repeated three times before the microorganisms were considered isolated,
pure, and active.
G. Identification and characterization of isolated organisms:
Once the isolated microorganisms were pure, they were plated on BBL R2A medium slants
(without biocides) and allowed to grow at room temperature. After adequate growth had
occurred, the slants were sent to American Type Culture Collection (ATCC) for identification
and characterisation.
The organism from the Santa Barbara soil was a bacterium, which was identified by ATCC as
an Arthrobacter sp..
The fungal culture isolated from the Guilderland soil was identified by ATCC as Fusarium
oxysporum Schlechtendahl.
REFERENCES
[1] Article: Degradation of Silicone Polymers in Nature, Environmental Information -
Update, Health Environment & Regulatory Affairs (HERA), Dow Corning.
[2] R.G. Lehmann, S. Varaprath and C.L. Frye. 1994. Degradation of Silicone Polymers
in Soil. Environmental Toxicology and Chemistry, Val. 13, No. 7, pp. 1061-1064.
[3] Sabourin, C. L., J. C. Carpenter, T. K. Leib, and J. L. Spivack. 1996. Biodegradation of

dimethylsilanediol in soils. Applied and Environmental Microbiology, 62: 43524360
YO U !!
TH A N K

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Biodegradation of

LET’S TRY BIODEGRADATION!!

1. Sample 1: Sandy soil from Santa Barbara, California

1. Santa Barbara Soil: 14%

● An appropriate amount of water was removed from the Guilderland

● In order to isolate soil microorganisms capable of biodegrading dimethylsilanediol,

[3] Sabourin, C. L., J. C. Carpenter, T. K. Leib, and J. L. Spivack. 1996. Biodegradation of

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