SPE-179775-MS

Production and Application of Schizophyllan in Microbial Enhanced Heavy

Oil Recovery
S. J. Joshi, Y. M. Al-Wahaibi, S. Al-Bahry, A. Elshafie, A. S. Al-Bemani, A. Al-Hashmi, P. Samuel, M. Sassi,
and H. Al-Farsi, Sultan Qaboos University; M. S. Al-Mandhari, Petroleum Development Oman

Copyright 2016, Society of Petroleum Engineers

This paper was prepared for presentation at the SPE EOR Conference at Oil and Gas West Asia held in Muscat, Oman, 21–23 March 2016.

This paper was selected for presentation by an SPE program committee following review of information contained in an abstract submitted by the author(s). Contents
of the paper have not been reviewed by the Society of Petroleum Engineers and are subject to correction by the author(s). The material does not necessarily reflect
any position of the Society of Petroleum Engineers, its officers, or members. Electronic reproduction, distribution, or storage of any part of this paper without the written
consent of the Society of Petroleum Engineers is prohibited. Permission to reproduce in print is restricted to an abstract of not more than 300 words; illustrations may
not be copied. The abstract must contain conspicuous acknowledgment of SPE copyright.

Abstract
Generally partially hydrolyzed polyacrylamides (HPAM) are used worldwide for polymer-based oil
recovery processes. However there aree some problems associated with HPAM applications such as,
treatment of produced water, oil-water separation and toxicity issues associated with degradation products
of HPAM. Schizophyllan biopolymer is biodegradable and environmental friendly alternative to chemical
polymers used for EOR applications. In present work we studied the schizophyllan production by fungi
Schizophyllum commune ATCC38548, its structural characterization and potential applications in Micro-
bial Enhanced Heavy Oil Recovery (MEOR).
Different minimal production media containing carbohydrate based carbon sources were screened for
better biopolymer production. During the course of experiments we studied microbial growth profile,
biopolymer production, and rheological properties of biopolymer, chemical characterization and appli-
cation of biopolymer in enhancing heavy oil recovery using Berea sandstone cores.
The biopolymer was produced in significant quantities and increased the viscosity of the production
medium after 9 days of incubation. Viscosities were measured using rheometer at different shear rates
(0.1-100 s-1) and temperatures, where it was found to be isothermal over a wide range of temperature with
strong shear thinning behavior. After 14 days of incubation, the viscosity was 21535 – 43 cP at shear rate
from 0.1 – 100 s-1, respectively. Thermogravimetric analysis (TGA) analysis showed that the moisture
content in purified biopolymer was 6.3%; it was quite stable till 100-200 °C and melted at temperatures
higher than 250 °C. It was characterized as schizophyllan by MALDI-TOF-MS and NMR (1H and proton
decoupled 13C NMR) analysis. The repeating tetrasaccharide unit of schizophyllan structure comprised of
homoglucan with ␤-1, 3-linked backbone and single ␤-1, 6-linked laterally glucose side chains at every
third residue. The biopolymer was used for MEOR experiments at 45 °C, using Berea sandstone core
plugs. The biopolymer injection resulted in 28% additional heavy oil recovery over residual oil saturation
(Sor).

Introduction
There are concerns about the shortage of oil production and energy resources along with the incremental
cost of heavy oil production and environmental impact. This raised the attention to look for other feasible
2 SPE-179775-MS

techniques which are environmental friendly, economical and quite efficient methods. Microbial enhanced
oil recovery (MEOR) is considered as one such technique with potential for extending the life of
production wells in a declining oil reservoir (Al-Sulaimani et al., 2011). MEOR technique is based on
utilizing microorganism’s metabolites and bio-products to recover additional or incremental oil from a
reservoir. There are a variety of metabolites produced by microorganisms that can be useful for oil
recovery, such as biosurfactants, biopolymers – polysaccharides, biogases, biosolvent, secreted by
microbes. As compared to conventional enhancing heavy oil recovery operations, which requires expen-
sive chemicals or steam generation equipment, MEOR is flexible and potentially cost-effective, and an
environmentally friendly technique. MEOR mechanisms, similar to the conventional enhanced oil
recovery mechanisms, are (classified) intended to cause alteration of oil/water/rock interfacial properties,
and, changing flow phase and flow behavior (Gray et al, 2008).
Polymer flooding is considered as an established technology for enhanced oil recovery in the oil
industry and the most widely used polymer is polyacrylamide or partially hydrolyzed polyacrylamide
(HPAM). This polymer is effective as a viscosifying agent but it has some limitations like losses due to
adsorption and mechanical degradation, and its poor salinity tolerance (Leonhardt et al, 2014). Starting
from 1980s, some biopolymers were considered and started to be implemented such as xanthan which is
produced by the bacterium Xanthomonas campestris (Leonhardt et al, 2014). Application of biopolymer
in enhancing oil recovery is accomplished by mobility ratio reduction, permeability modification to
improve water flood sweep efficiency and selective plugging of high permeability zones, also known as
thief zones (Shabani-Afrapoli, 2012). One of biopolymer based MEORs’ attraction is that the biopolymer
can be produced using cheap natural available nutrients such as wheat bran and corn cobs, which are
abundantly available as byproducts of the wheat and corn processing industries (Zhong et al., 2013);
activated charcoal; detoxificated rice hull hydrolysate, which is the main byproduct of paddy process (Shu
and Hsu, 2010); and distiller’s dried grains (Sutivisedsak et al., 2013).
Mushrooms comprise a vast and yet largely untapped source of powerful new pharmaceutical products.
In particular, and most importantly for modern medicine, they represent an unlimited source of polysac-
charides with antitumor and immunostimulating properties. Schizophyllan is one of the most famous and
widely used polysaccharide, produced by the fungus Schizophyllum commune, a white-rot ubiquitous
mushroom (Zhong et al., 2013). Some research works were conducted to test the applicability of
schizophyllan in enhanced oil recovery (Leonhardt et al., 2014; Gao, 2015). Schizophyllan is often used
as a model polymer to study the viscosity of neutral linear rods and at a concentration of 1% (w/v) and
it forms a gel at room temperature (Zhong et al., 2013).
The scope of the present study was to investigate the biopolymer production using S. commune
ATCC38548, in different production media under optimum incubation conditions, rheology studies,
physical and chemical structural characterization, and application in MEOR using core flood experiments.

Methods

Microorganism and Culture Conditions

The fungal strain used in this study - S. commune ATCC38548, was procured from American Type
Culture collection (ATCC), USA. The S. commune was grown on yeast malt extract (YME) agar, where
small square (1cm2) of S. commune stock culture was transferred using sterile section blade to the YME
agar plates and incubated at 25°C for 5-8 days. The growth of the fungus was observed everyday by
checking the diameter of the fungus colony. Generally at the end of day 5-8 days the fungus covered the
entire plate (80mm diameter). The S. commune plates were stored in the refrigerator at 4°C, for further
experiments.
SPE-179775-MS 3

Optimization of Biopolymer Producing Media

For the optimization of biopolymer production, six different production media were prepared, their
composition are shown in Table 1. In all six media glucose monohydrate was used as a carbon source with
different combination of organic nitrogen sources.

Table 1—Production Media Composition

Media/Composition (g/l) 1 (Rau, 1999) 2 3 4 (Sutivisedsak et al., 2013) 5 6

Glucose 30 30 30 20 30 30
Yeast Extract 3 3 3 – 3 3
Malt Extract – 3 – 20 – 3
Peptone – – 3 – – –
Urea – – – – 3 –
KH2PO4 1 1 1 – 1 –
MgSO4.7H2O 0.5 0.5 0.5 – 0.5 –

Approximately, 8 mm diameter discs of growing S. commune were cut using sterile plastic tips and
inoculated to each 100 ml production media per 250 ml capacity Erlenmeyer flasks. The flasks were
incubated in incubator shaker at 28°C; 160 rpm. The growth of S. commune was observed visually for the
first 5 days and the flasks were harvested for all analysis starting from day 5 till day 14.
Analytical procedures
In order to determine the mycelial biomass, the samples were filtered under vacuum using fine
crystalline filter paper to ensure that the smallest mycelia will not pass. The filtrated mycelia were kept
to dry in oven at 80 oC, for 2 days to measure the mycelial dry weight. Cell free broth was used for pH,
and viscosity measurements. Initially the viscosity measurements were carried out using cannon-feneske
glass viscometers, and later rheometer (DV3T LV, Brookfield, USA) was used for confirmation of the
best medium and detailed rheology studies.
Biopolymer Extraction
The cell free broth was used for biopolymer extraction. Where 2 volumes of 98% ethanol were added
to the samples for the separation of the biopolymer since it is not soluble in alcohol and it was kept at 4°C
overnight (Figure 1A). Next day, the samples were centrifuged at 14000 rpm for 40 minutes at 4°C and
the pellets were collected. The collected pallets were dissolved in distilled water and kept in the freezer
at ⫺80°C for 2 days, and freeze dried in a lyophilizer (Labconco, USA). Figure 1B shows the freeze-dried
biopolymer pellet. Freeze-dried biopolymer was used for various physical and chemical characterization
studies.

Figure 1—Biopolymer extracted with 98% ethanol (A) and collected as white pellet after freeze-drying (B)
4 SPE-179775-MS

Physical and Chemical characterization

Thermogravimetric Analysis (TGA) TGA analysis was conducted using freeze-dried biopolymer pow-
der produced from media 1. Where briefly 16.148 mg of powder was heated to different temperatures
30°C – 700°C, under nitrogen flow, and the weight loss was measured.
Matrix Assisted Laser Desorption Ionization – Time of Flight-Mass Spectroscopy (MALDI-TOF-MS)
All MALDI-TOF experiments were performed at Central Analytical and Applied Research Unit
(CAARU), Sultan Qaboos University, on UltraFlextreme (Bruker Daltonics, Bremen, Germany) operating
in positive reflectron mode in the m/z range of 50 –2000 Da. Stainless steel MTP 384 target plate was used
for all the molecular weight analysis. The ␣-cyano-4-hydroxycinnamic acid (HCCA) dried droplet
protocol described in Bruker manual was employed for sample preparation and spotting. The MALDI-
TOF spectra were externally calibrated using a commercially available peptide mix (peptide calibration
standard II, Part-No #222570, Bruker Daltonics, Germany). FlexAnalysis Software v3.3 (Bruker Dalton-
ics) was used for visualization and initial data processing.
Nuclear Magnetic Resonance (NMR) spectroscopy The NMR experiments were carried out in Bruker
Avance III HD 700 MHz spectrometer equipped with 5mm TCI H/C/N cryoprobe at CAARU, Sultan
Qaboos University. The proton (1H) NMR experiment was run using zg30 pulse programme operating at
176.08 MHz. Acquisition parameter were as follows: 90° proton pulse width of 8.00 ␮s, relaxation delay
of 1s, 16 scans. The proton decoupled carbon (13C) NMR experiments were carried out using composite
pulse decoupling scheme operating at 176.08 MHz. Acquisition parameter were as follows: 90° proton
pulse width of 8.00 ␮s, relaxation delay of 2s, 512 scans. The Spectra were recorded in CDCl3 at 298K
and processed using TOPSPIN 3.2 software.
Core-plugs and Fluid Properties
Standard Berea core-plugs were used for all core-flooding experiments. The properties of core-plugs
used were: 1.5 in x 3.06 in (Diameter x length); 252-302 mD, gas permeability; 17.67-18.19 ml, pore
volume. The crude oil used was heavy oil provided from one of the oil field. Its properties at 45 and 25
°C, respectively were: 2350-10970 cp, viscosity; 0.941-0.965 g/cc, density; 16.36-14.68 API°; and 1.82
mgKOH/mg TAN.
Core-flood Experiments
The core plugs, after cleaning with chloroform/methanol cleaning mixture, were first saturated at
100% with formation brine using vacuum desiccator for 24h before loading to the core holder.
Initially 1000 psi confining pressure was applied to the core holder. The system lines were flushed
with formation brine to eliminate any air bubbles existing in the system. After that the system was
heated to 45°C and kept for 2 hours to ensure that the temperature is distributed uniformly throughout
the core and any increase of confining pressure was released during heating process. One pore
volume (PV) of brine was flooded first at 0.4cc/min and the differential pressure was measured in
order to calculate the liquid absolute permeability of the core. Oil was flooded at 0.4 cc/min till no
more extra water produced. The produced water volume was considered as the oil initially in core
taking into account the dead volume. For the secondary recovery stage, brine was flooded at
0.4cc/min till no more significant oil volumes produced and the water cut reached 0.9. For the tertiary
stage, 6 PV of biopolymer broth solution was injected at 0.4cc/min. The differential pressure was
recorded and monitored during all the production stages.

Results and Discussion

Worldwide water soluble, high molecular weight, linear anionic polyacrylamide (PAM) and partially
hydrolyzed polyacrylamide (HPAM) are widely used to improve the mobility ratio during oil
production. With the promotion of the polymer oil-displacing technology large quantities of PAM/
SPE-179775-MS 5

HPAM are discharged into the environment with oil extraction wastewater. HPAM present in
production water causes some serious environmental problems such as, it will remain in the produced
water, increasing the difficulty in oil–water separation, the oil content in sewage is greatly increased,
there is a high probability that the wastewater will exceed the local discharge limit and it can also
hardly avoid infiltrating and contaminating groundwater resources. In addition, the costs and
difficulties of produced water treatments increased with the high concentration of the HPAM in the
wastewater (Chen et al., 2003; Zhao et al., 2008; Wen et al., 2010; Bao et al., 2010; Yongrui et al.,
2015). One of the environmentally friendly and effective alternatives is to use biopolymers for EOR
applications.

Biopolymer Production, Extraction and Rheology studies

Six different glucose based media were tested for biopolymer production where the samples were
withdrawn from 5 to 14 days of incubation and analyzed for growth, biopolymer yield and viscosity
measurements. As shown in Figure 2–7, all production media supported good mycelial growth
(observed as small mycelium balls). Other researchers have reported production using flasks with
baffles (Rau, 1999) or adding the glass beads in the flasks (Leathers et al., 2006). In terms of field
application, bioreactors are originally designed with baffles that will enhance the growth and
production of schizophyllan biopolymer. Significant biomass growth (about 10 g/L) was obtained
after 7 days of incubation in media 2, 3, and 4, and after 8 days of incubation in media 1, 5 and 6.
In terms of schizophyllan production, it was found that the biopolymer yield from each media was
comparable with viscosity measurements. Highest schizophyllan production for Media 1, 2, 3, 4, 5,
and 6 were; 5.39 g/l, 8.08 g/l, 4.58 g/l, 9.62 g/l, 2.72 g/l, and, 4.35 g/l after; 12 days, 14 days, 11 days,
14 days, 12 days, and 13 days, of incubation, respectively. It is interesting to note that for all media,
except media 5, a minimum of 9 days of incubation was required to obtain a significant increase of
schizophyllan biopolymer production and increase in viscosity. As shown in Figure 2–7, the viscosity
measurements were carried out for 10 samples from each of the 6 different media from day 5 till day
14, using cannon-feneske glass viscometers. The first significant increase in viscosity was observed
after 9 days of incubation for all the 6 different production media. All of the 6 media resulted in a
good schizophyllan biopolymer production as observed from the significantly increased viscosity
values. Media 1, 2, and 4 showed high viscosities of about 2000 cP after 10 days of incubation.
Whereas, sharp decrease in viscosity was observed in medium 2, from 2000 cP at day 10 to about 25
cP at day 11. Medium 4 showed some increase in viscosity for the first 13 days of incubation
followed by a sharp increase in viscosity at day 14, whereas Medium 5 showed the lowest viscosity
values, where the maximum viscosity obtained was 42cP. It is well-known that the medium pH
affects the production of microbial metabolites, like biopolymers and biosurfactants. In general
bacteria prefer near neutral pH (6-7), and fungi can grow at wide range of pH, including acidic pH
(7). All tested media didn’t show significant changes in pH values during 5-14 days period, and was
observed to be acidic between pH 5 and 6. It was also observed that, from all media a minimum of
about 4 g/l concentration of schizophyllan biopolymer is required to have a sharp increase in
viscosity. This concentration is known as the overlap concentration, a characteristic of all polymers.
6 SPE-179775-MS

Figure 2—The biomass, biopolymer yield, and the viscosity profile of S. commune in production medium 1

Figure 3—The biomass, biopolymer yield, and the viscosity profile of S. commune in production medium 2
SPE-179775-MS 7

Figure 4 —The biomass, biopolymer yield, and the viscosity profile of S. commune in production medium 3

Figure 5—The biomass, biopolymer yield, and the viscosity profile of S. commune in production medium 4
8 SPE-179775-MS

Figure 6 —The biomass, biopolymer yield, and the viscosity profile of S. commune in production medium 5

Figure 7—The biomass, biopolymer yield, and the viscosity profile of S. commune in production medium 6

Based on the above experiments, production Medium 1 showed better results in terms of
biopolymer production and increased viscosity, and it is also cost-effective since it is the cheapest
media in terms of its cost. For all these reasons, Medium 1 was chosen for further experiments.
Highest observed biomass and schiophyllan yield were 15.8 g/L and 5.17 g/L, respectively, after 14
days of incubation. The average viscosity observed after 14 days of incubation was T425 cP, using
cannon-fenske glass viscometer. As the expected viscosity was higher than what was observed using
SPE-179775-MS 9

cannon-fanske viscometer, it was further analyzed using rheometer (Brookfield, USA), at different
shear rates.
Figure 8 shows the daily average viscosity measurements of the collected samples using rheometer
at different shear rates 0.1 – 100 s-1, at 25°C. Maximum viscosity was obtained after 14 days of
incubation. Schiophyllan biopolymer had a sharp shear thinning behavior; viscosity after 14 days of
incubation was reduced from 21535cP to 43cP when shear rate increased from 0.1s-1 to 100s-1,
respectively. This strong elasticity and shear thinning behavior is a characteristic of biopolymers due
to its rigid triple helix structure which makes it favorable for field application where the viscosity of
the injected biopolymer solution at turbulent flow will result in very low viscosity which will reduce
the high back-pressure and will prevent damage to the formation. As a result, the injection pumping
pressure and the viscosity of the solution will increase as it transport away from the wellbore to the
reservoir till it reach the reservoir speed of about 1 ft/day which is laminar flow. The reservoir speed
is approximately equal to 7s-1, where the viscosity of biopolymer solution observed was equal to
635cP. The biopolymer was further tested for rheological properties at different temperatures and
shear rates. The viscosities were measured using rheometer at different shear rates (0.1-100 s-1) and
temperatures (25-60)C), where it was found to be isothermal over a wide range of temperature with
strong shear thinning behavior (Figure 9).

Figure 8 —Viscosity measurements at 25°C for 9th – 14th day samples of S. commune
10 SPE-179775-MS

Figure 9 —The rheology of biopolymer at different shear rates (0.1-100 s-1) and temperatures (25-60)C) using rheometer

Physical and Chemical characterization of Biopolymer

TG Analysis Figure 10 shows the weight loss observed after subjecting powder sample to different
temperatures. At 100°C, 1.02 mg (6.3%) was lost from the sample as water evaporated. A near plateau
was observed and no significant weight loss occurred as the temperature increased from 100°C to 200°C.
Previously reported studies showed that this biopolymer is stable at high temperatures of 135°C due to its
rigid structure type. As the temperature increased to 300°C, a sharp weight loss of about 11 mg (70%) was
observed which indicate that biopolymer molecules melted at this temperature. A second near plateau was
observed as the temperature increased from 300°C to 700°C with little extra weight loss. The total weight
loss at 700°C is equal to 13 mg (80%) which indicates that almost all the biopolymer melted. Rau et al.,
(1999) reported that at temperatures ⬎135°C and at pH ⬎ 12, the triple helix melts to single, randomly
coiled chains, equivalent to the reduction of the average molecular weight by one-third.

Figure 10 —TGA of Schizophyllan (16.148 mg) powder

SPE-179775-MS 11

MALDI-TOF and NMR Analysis MALDI-TOF showed molecular mass/charge of around 14250kD
for our biopolymer (Figure 11). The NMR spectra of the SCG showed anomeric sugar signals at 4.93
ppm due to the ␤-1,3-linked glucose and ␤-1,6-linked glucose. Three proton signals of the glucose
moiety are overlapped (4.5 ppm). One proton signal of the glucose unit appears as a well resolved
doublet resonance at 4.1 ppm. The Characteristic methylene –CH2 signals were present at 3.43 and
3.67 ppm. These signals overlap with the –OH signals of the glucose moiety and the water (3.33
ppm), as shown in Figure 12. The carbon NMR spectrum of our biopolymer is shown in Figure 13.
Where glucose units containing six carbon numbers, along with the peak assignments at 61.38-103.41
ppm were observed. The repeating tetrasaccharide unit of schizophyllan structure comprises of
homoglucan with ␤-1, 3-linked backbone and single ␤-1, 6-linked laterally glucose side chains at
every third residue and this structure is a popular model for rod-like or stiff-worm like polymers
(Kony et al., 2007, Sutivisedsak et al., 2013) It is similar to the reported structure of biopolymer -
Schizophyllan (Yang et al., 2004).

Figure 11—MALDI-TOF-MS spectrum of biopolymer – schizophyllan

12 SPE-179775-MS

Figure 12—The 1H NMR spectrum of biopolymer – schizophyllan

13
Figure 13—The C- NMR spectrum of biopolymer – schizophyllan

Core-flood Experiments
Figure 14 shows the improved oil recovery after injecting biopolymer broth. It is clear that after injecting
6 PV of biopolymer broth without dilutions, 28% extra oil recovery from the oil initially in core resulted
which make the total recovery, after secondary and tertiary flooding, equals to 79.5 % but, as conse-
SPE-179775-MS 13

quences, it resulted in high differential pressure up to 160 psi due to the high viscosity of the bulk
biopolymer broth. Similarly as conventional polymer flooding, biopolymer flooding recovery mechanism
was based on improving the mobility ratio and the sweep efficiency and it cannot result in extra recovery
beyond the residual oil saturation since it has no effect on improving the mobility of the capillary trapped
oil droplets that require surfactant to reduce its interfacial tension. Partially hydrolyzed polyacrylamide
(HPAM) are by far the most used polymer in EOR applications, which is very much affected by the salt
dependency, other factors influencing the viscosity of HPAM solutions are the degree of hydrolysis,
solution temperature, molecular weight and solvent quality (Wever et al., 2011). Biopolymers such as
starch, xanthan (produced by bacteria - Xanthomonas campestris), guar gum have been used in oil
industries for many years (Karmakar and Chakraborty, 2006). Sun et al., (2011) reported exopolysac-
charide production by a genetically engineered Enterobacter cloacae strain having a promising applica-
tion potential for microbial enhanced oil recovery. They reported in-situ MEOR by selective plugging of
core-plugs by polymer produced by genetically engineered bacterium. Sun et al., (2013) reported
additional 3.5%-11.3% oil recovery from sand-pack columns using bacterial exopolysaccharide. We
observed around 28% additional oil recovery using biopolymer – schizophyllan.

Figure 14 —Total oil recovery at 45 °C after water flooding followed by 6 PV of Schizophyllan biopolymer broth

Generally HPAM is used in oil industries for EOR operations, where it is reported to give 10-15%
additional recovery. However there are some problems associated with HPAM applications such as
treatment of produced water. Furthermore, the residual PAM/HPAM in the produced water can slowly
degrade into the toxic acrylamide monomer. It is generally (but not universally) accepted that polyacryl-
amide is safe and non-toxic. However, acrylamide is a potent neurotoxin, which has been studied by
numerous researchers. Many reports indicate that commercial PAM/HPAM can be easily degraded to
acrylamide when it is heated or exposed to ultraviolet irradiation and its intermediate products are
hazardous as their monomer is highly toxic (Chen et al., 2003; Zhao et al., 2008; Wen et al., 2010; Bao
et al., 2010; Jebasingh et al., 2013; Yongrui et al., 2015). The degradation of acrylamide monomer has
cumulative neurotoxicity, and it is identified as a genotoxic carcinogen. Since HPAM can remain in
surface water and groundwater for a long period of time, it may endanger human health. Due to very good
14 SPE-179775-MS

solubility in water, acrylamide can be easily transferred through the aqueous sources and will leach
through soils, but due to its low vapor pressure it is not reported to enter and transported into the
atmosphere. Low levels of acrylamide are reported to be degraded quickly in surface water, but higher
concentrations are difficult to degrade (Chen et al., 2003; Zhao et al., 2008; Wen et al., 2010; Bao et al.,
2010; Jebasingh et al., 2013; Yongrui et al., 2015). Biopolymers such as schizophyllan are reported to be
biodegradable and non-toxic to humans and environment.

Conclusions
We have tested six different glucose based production media for biopolymer production using fungal
strain S. commune ATCC38548, and the following conclusions were made based on the experimental data.
– The S. commune ATCC38548 was able to produce biopolymer in all tested media, where medium
1 was cheaper and supported better growth and higher production.
– Viscosities were measured using rheometer at different shear rates (0.1-100 s-1) and temperatures,
where it was found to be isothermal over a wide range of temperatures (25-60)C) with strong shear
thinning behavior.
– After 14 days of incubation, the viscosity was 21535 – 43 cP at shear rate from 0.1 – 100 s-1.
– The biopolymer was characterized using MALDI-TOF and NMR, as schizophyllan.
– Core flooding tests revealed the potential of schizophyllan in MEOR, where 28% heavy oil (Sor)
was produced from Berea sandstone core-plugs.
It is recommended to conduct further studies on scale-up of biopolymer using cheaper raw materials,
extensive stability studies and effect on enhancing heavy oil recovery.

Acknowledgements
Authors acknowledge the funding from Petroleum Development Oman (CR/SCI/BIOL/11/01) to carry out
the research.

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