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International Journal of Pharmacy Research and Technology

2013, Volume 3, Issue 2, 05-11

ISSN 2250 – 0944 (Online)
ISSN 2250 – 1150 (Print)
Research Article

‘Gellasan’: a Novel Polyionic Matrix for Lipase Immobilization

Solanki S N1*, Pokharkar V B2, Solanki N R1

Department of Biotechnology, Vidyabharati Trust College of Pharmacy, Umrakh, Ta: Bardoli, Dist: Surat,
Department of Biotechnology, Poona College of Pharmacy, Bharati Vidyapeeth University, Erandwane, Pune, Maharashtra.
* Corresponding Author E-mail: Mob No: +919016454096
Received: 20/05/2013, Revised: 05/06/2013 Accepted: 18/06/2013
The aim of the present study was to investigate the application of ‘Gellasan’ - chitosan crosslinked gellan gum as a
novel matrix for lipase immobilization. The biocatalyst used was Porcine Pancreas Lipase (triacylglycerol esterase) (EC Gellan being heteroanionic and chitosan being cationic, the interaction between them resulted in the formation of
ionic crosslinked Gellasan beads. The hydrogel beads were prepared and optimized using a 32 factorial design by ionotropic
gelation technique. The beads were characterized by IR studies, Scanning Electron Microscopy. The lipase activity was
assayed using para-nitrophenylpalmitate as substrate with the highest activity of 261.1U/g of support. The average particle
size was found to be in the range of 192-226 µm. The beads showed a pH dependent swelling with a significantly higher
swelling index at pH 7.4 in phosphate buffer than in 0.1 N HCl. The percent release of the enzyme for the freeze-dried beads
was found to be 2.5-19% while of air-dried beads were 1.5-16% in 0.05 M Tris HCl buffer pH 7.4 within 8 hrs. Thus, the
two polymers gellan and chitosan were compatible for lipase immobilization and may become a potential delivery system to
control the release of drugs in future.

Keywords: Chitosan; Factorial design; Gellan Gum; Gellasan; Ionotropic gelation; Lipase.

INTRODUCTION properties[6]. Gellan gum in either substituted or

Owing to the rapidly expanding and wide spread unsubstituted form produces gels even at low
applications of enzymes in the various spheres like concentrations when hot solutions of the gum are cooled.
biotechnological, pharmaceutical and biomedical [1]. Its aqueous solutions undergo thermoreversible gelation by
Enzymes have become a powerful catalytic tool in the cooling in presence of metallic salts which play a critical
hands of researchers. But the use of enzymes has been role in the gel formation since gellan has a charged group in
restricted to their unstable nature and requirement of every repeating unit . The substituted form produces soft,
stringent temperature and pH conditions. The technique of elastic gels while the unsubstituted form produces hard,
‘Enzyme Immobilization’ offers several advantages viz. brittle gels [7]. Low acyl gellan gels at temperature between
improvement of enzyme stability, easy separation of the 30 & 500C. The high acyl gellan has poor aggregation
catalyst from the product stream, repeated use of the properties hence, it gels around 700C [8]. In pharmaceutical
biocatalyst and the possibility of continuous processes [2]. field, it is used as gelling agent, disintegrant, adhesive
Thus the technology has given an outlet to the various mucosal delivery, transdermal delivery, film coating,
applications in immunosensors, immunodiagnostic assays controlled release tablets, microencapsulation, ocular drug
in biomedicine for analytical systems like biosensors, and delivery [9-11]. It has also been used as agar substitute in
finally has paved a way clinically for enzyme therapy [3]. microbiological media and plant tissue culture [12].
Gums seem to be the ideal matrix for entrapment Similarly, gellan electrophoresis gels have also been used in
because the crosslinks of the gel provide the essential 3-D the isolation of DNA[13].
mesh and network to hold the cells or enzyme in place. This Chitosan [poly (β-(1→4)-2amino-2dedoxy-d-glucose)]
property makes gums very popular for immobilizing is a natural cationic polysaccharide derived from chitin
biologically active molecules. Gums have extensively been which is a copolymer of glucosamine and N-acetyl
used for the immobilization of living cells. Similarly, glucosamine units combined together. It is naturally present
certain enzymes have also been entrapped in the matrices of in abundance in the outer shells of the crustaceans[14].
gums[4]. Chitosan has numerous applications in pharmaceutical
However, most of the entrapment techniques have the systems and medicine. It has been used as a film coating
limitations that they entrap the protein with low efficiency material[15], excipient in the direct compression tablets, as a
and they frequently require to use the volatile organic tablet disintegrant for the improvement of drug
solvents which may alter the enzyme structure [5]. dissolution[16] and as microspheres [17], to enhance the
Gellan gum has also been extensively investigated for transnasal absorption of peptides and other polar drugs like
the same purpose. Gellan is an extracellular polysaccharide insulin, calcitonin and morphine metabolites[18] and as a
produced by Pseudomonas elodea and is composed of carrier in drug targeting[17] due to its biocompatibility, non-
tetrasaccharide repeat units: 1,3- β-D-glucan, 1,4- β-D- toxicity, ability to absorb liquids & ability to form
glucoronic acid, 1,4- β-D-glucose and 1,4- -α-L-rhamnose protective films and coatings it has been used as a wound
in the molar ratio of 2:1:1. Gellan gum is non-toxic and has dressing[19]. According to reports, the release of chitosan
received FDA approval in 1992 for food use. It has been microparticles could be controlled by crosslinking the
tremendously used in food industry due to its gel forming matrix with glutaraldehyde and other crosslinkers but these

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agents have undesirable effects. Glutaraldehyde can cause method was used for preparation of placebo gellasan beads
irritation to the mucosal membranes due to its toxicity[20]. (without entrapped lipase).
Moreover, Gupta and Ravikumar (et al. 2000) reported that
chitosan has antacid and antiulcer characteristics which Freeze-Drying of Gellasan beads.
prevents and weakens the drug interaction in the The protocol of freeze drying involved storage of fresh
stomach[21]. Thus, to overcome the limitation of chemical beads at –40C for 1h and then at –400C in deep freeze for 3
crosslinking, ionic crosslinking interaction has been used. hrs. The beads were then kept on a precooled shelf (-250C
Previously, it has been reported that chitosan is a good to -300C) of a Freeze Dryer (Heto Lyo Pro 3000). Primary
candidate for the immobilization of lipase. Similarly, drying was begun at –300C to –400C by application of
chitosan exhibits higher entrapment efficiency, higher vaccum (0.07 hPa) followed by secondary drying (40C).
loading and higher activity[22]. Hence, lipase was chosen as The drying process was completed in 20 hrs. The freeze
a model enzyme in this study. drying cycle was optimized by determining the moisture
Lipases have received much attention in a wide range content (1-2%) of the beads.
of applications[23-25]. Lipases have been immobilized by
different methods like covalent binding, adsorption, IR Study
entrapment, crosslinking, on various supports[26, 27]. The IR spectrum of gellan gum powder, chitosan
Porcine pancreas lipase (PPL) has been immobilized by powder and lipase powder along with the blank and loaded
entrapment on polyacrylamide with the highest activity of beads was recorded using JASCO FT/IR 5300 fourier
2.187 U/g of solid. The immobilization stabilized the transform infrared spectrophotometer to study the binding
enzyme against heat and urea treatment[28]. PPL was also sites available on gellan gum and chitosan molecules. The
immobilized on celite for the synthesis of butyl butyrate in possible interaction between them and lipase was studied.
a non-aqueous medium, the activity of the immobilized Comparison of this spectrum was done with that of blank
lipase was less than the activity of free lipase[29]. Since, gellasan beads and lipase loaded gellasan beads.
lipases have immense advantages to produce several
chemical products and with the sole intention to tap their Enzyme Activity Assay.
therapeutic potential, we were motivated to study lipase The hydrolytic activity of lipase was measured using
activity in aqueous and organic media. para-nitrophenylpalmitate (p-NPP) as substrate by
Pencreac’h and Baratti method 30. In brief, 20 mg beads
MATERIALS AND METHODS were added to a reaction mixture containing 3ml p-NPP
Materials solution (0.04 M in 2-propanol), 1ml 0.05M Tris HCl
Porcine Pancreas lipase (PPL, Type II, crude) (EC buffer (pH 7.4 at 370C) and 3ml DI water. A negative and para-nitrophenylpalmitate (p-NPP) & BSA control (placebo beads) and a positive control (10 mg free
were purchased from Sigma Chemical Co. (St. Louis, MO). lipase) were also studied simultaneously and the reaction
Gellan Gum (Kelcogel F- Low acyl) was kindly donated by mixture was incubated for 30 mins in an incubator shaker
C.P Kelco Pvt. Ltd, Mumbai, India. Chitosan sample (95% (Brunswick, Germany) at 50 rpm. The activity was
deacetylated) was a gift from A.E. Connock Pvt. Ltd, calculated at 400 OD. One enzyme unit was defined as the
England. Tris base were purchased from Himedia, Mumbai. amount of protein liberating 1µmol of para-nitrophenol per
Folin Reagent, Acetic acid, sodium potassium tartarate minute in these conditions. The entrapment, release and
were all products of Merck, Mumbai. activity studies were carried out in triplicate.

Preparation of Gellasan beads. Percent Yield and Entrapment Efficiency

The beads were prepared by ionotropic gelation Percent Yield, a ratio of actual weight and expected
technique using a 32 full factorial design. The two variables weight was calculated for dried beads. The capacity of
used were: gellan gum (1, 1.5, 2 %w/v) and chitosan (0.3, gellan and chitosan to immobilize the enzyme is called
0.6, 1 %w/v). Various batches were prepared by using all Immobilization Efficiency (IE). It was estimated as the ratio
possible combinations of different levels and variables. between the activity or the protein content of the complex
Appropriate quantity of gellan gum was dispersed into of gellasan and enzyme ( viz. the beads) and the sum of the
preheated deionized water at 800C by gentle stirring. The activities of the beads, free enzyme in excess in the chitosan
temperature was then raised to 900C to achieve complete solution and the free enzyme in the washing solutions.
hydration of the polymer. Similarly, hydration of chitosan The percentage loading/ immobilization efficiency of
was achieved by dispersing appropriate quantity of chitosan the gellasan beads was determined by using the following
into acetic acid (2%v/v) and kept overnight. The hydrated equation [1].
and heated solution of gellan gum was cooled to 450C and E % =
to it a dispersion of 0.5g lipase powder in 1ml 0.05M Tris Act Beads
× 100
HCl buffer, pH 7.4 was added. This mixture was stirred and ∑ Act Beads + Act. Chitosan iltrate + Act Washing
extruded dropwise using a 10ml plastic syringe of bore size Where, E is the immobilization efficiency, and Activity of
18G into the chitosan solution cooled at 50C, kept on a beads is the activity of the complex of gellan- chitosan and
controlled speed stirrer (Eurostar Power control-visc, IKA, the enzyme. The protein content of the enzyme was
Labortechnic, Staufen, Germany). The beads were cured for analyzed by Modified Lowry Method using BSA as a
30 mins in the same solution after which they were standard.
separated by filtration. Washing was completed twice using
DI (deionized) water. Air-dried and freeze- dried beads Swelling study
were used for further studies. The filtrate and the washings Water sorption capacity of gellasan beads was
were collected for the lipase loss measurements. A similar determined by swelling the gel beads in appropriate buffer
solution. For this, 0.1g of the beads from all the batches

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were placed in 50 ml, HCl (0.1N, pH 1.5) and Tris-HCl band at 1415 cm-1 and 3526 cm-1 indicating the presence of
buffer (0.05M, pH 7.2) and allowed to swell at room hydroxylic group and primary amine functional group
temperature for 1 hr. The swollen beads were separated respectively. Similarly, the IR Spectra of gellasan beads
from the media and blotted on the filter paper and then showed a broad suppressed peak of amine of chitosan and
weighed immediately. This process was then repeated for carboxylic group of gellan and appearance of amide band at
next 5 hrs at the interval of 1 hr and kept for 8 hrs. The 3566 cm-1 suggesting the possible interaction between the
percentage of swelling of the beads were calculated by the carboxylic group on the glucoronic acid residue of gellan
formula [1]: and the free amino group of chitosan resulting in a
polyelectrolyte complex [31].
Esw = [(We - Wo) / Wo] × 100
Where, Esw = Swelling of beads at equilibrium
We = Weight of Gellan beads at equilibrium swelling
Wo = Initial weight of beads

Release Study
The gellasan beads were suspended in a vial
containing 10 ml Tris-HCl buffer (0.05M, pH 7.2) at 370C
in an incubator shaker. 0.1 ml samples were withdrawn and
replaced each time with Tris-HCl buffer to maintain the
sink conditions throughout the experiment. The samples
and the blank and freeze-dried beads were run
simultaneously and the protein content was estimated by
Modified Lowry Method.

Scanning Electron Microscopy (SEM)

The surface and morphology of the placebo, loaded
and freeze-dried beads were studied using SEM (JEOL
JSM 840 Model, Japan). Beads were coated to 200A0
thickness with gold-palladium layer by sputter coating and Evaluation Parameters for Gellasan Beads
operated at an acceleration voltage of 10 kV. Various parameters viz. percent yield, IE, activity and
T50 of gellasan beads prepared using factorial design were
Fig. 1 IR Spectra of (A) Gellan gum powder (B) Chitosan evaluated. Results were further analyzed by multiple linear
powder (C) Lipase Loaded regression (MLR) using MINITABR (ver14). The data were
fitted in the equation:
IR Studies Y=β0 + β 1X1 + β 2X2 + β 11 X12 + β 22 X22 + β 12 X1X2
It is clearly evident from the IR Spectra (Fig.1) that,
there were no interactions of the enzyme lipase with gellan The coefficients β0, β1, β2, β12 were computed using
and chitosan thereby, retaining the intact structure of the MINITABR which is a computer programme for multiple
enzyme since the loaded beads showed the presence of regression analysis. In this a stepwise regression analysis
bands at 1749 cm-1, 1581 cm-1 and 1308 cm-1which was done and those variables, which do not contribute
represent the Amide I, II and III bands respectively . IR significant to the effect, are eliminated.
Spectrum of the gellan gum powder showed the presence of MLR was used to identify statistically significant
bands at about 2928 cm-1 and 1018 cm-1 due to the term. β0 is the arithmetic mean response. β1 and β2 are the
carboxylic (-OH) and C-O functional group respectively coefficients of factor X1 and X2. The results of multiple
while the IR Spectrum of chitosan powder showed a sharp regression analysis are summarized in Table 2.

Table 1 Loading Efficiency of Factorial Batches

Batch Yield Average Particle Activity Specific Activity Loading T50
No. (%) Size (µm) (U/mg) (U/mg) Efficiency (%) (hrs)
S-1 72.76 192.61 0.2611 0.6541 77.04 8
S-2 61.43 195.99 0.2201 0.5516 69.70 6
S-3 55.85 198.11 0.136o 0.3408 55.77 13
S-4 66.82 197.35 0.2725 0.6829 53.99 10
S-5 62.8 210.26 0.1700 0.4261 76.04 15
S-6 64.97 212.72 0.1576 0.3949 72.94 13
S-7 77.83 216.84 0.0226 0.0566 61.15 12
S-8 77.76 219.21 0.0114 0.0285 51.80 7
S-9 43.2 226.39 0.0069 0.0173 44.65 12

The average particle size of beads for various batches was and Figs. 4 and 5, the activity and the T50 were governed
obtained in the range of 190-250 µm. Percentage Yield by both the variables. The percent yield decreases as the
(Fig.2) and IE (Fig. 3) of various batches were in the range concentration of both polymers increase, this can be
of 43-77% and 45-76% respectively. As shown in Table 1 explained; as the polymer concentration increases, the

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viscosity of the dispersion also increases,

ncreases, thereby leading to
the difficulties in processing and decrease in yield of beads.

Table 2 Regression Analysis of Different Evaluation

Parameters for Gellan-Chitosan
Chitosan Beads (n=3)
Yield Activity Immobilization
Coefficient T50
(%) (IU) efficiency (%)
Β0 42.54 -0.05457 21.35 -47.67
β1 21.26 0.796 72.02 101.78
β2 43.31 -0.52 28.47 -43.34
β12 -27.86 0.15 7.36 -7.02
β11 -0.233 -0.36 -30.55
30.55 -32
β22 -20.72 0.12 -37.29
37.29 38.88 Fig. 2 Surface Graph Showing the Effect of Variables on
R2 0.633 0.931 0.512 0.751 the Percentage yield
Significance 0.001 0.001 0.001 0.001
Significance terms p< 0.005
n= number of samples tested for each experiment.

A curvilinear plot (Fig.3)) is obtained for the effect of

polymer concentration on IE which shows that as the
polymer concentration increases upto 0 level (Table 1), IE
increases therafter decreases substantially. This could be
attributed to the fact that, the stoichiometric binding of
gellan and chitosan occurs at 1.5% w/v of gellan and 0.6%
w/v of chitosan hence, above this or below this
concentration, binding sites available to form a cross-linked
Fig. 3 Surface Graph Showing the Effect of Variables on
matrix may be insufficient to hold the enzyme thus
the Immobilization Efficiency
resulting into leaking and lesser IE. Several researchers
have also reported a low enzyme loading in these
entrapment techniques[32]. The entrapped enzyme tends to
leach out before the gel has time to settle thereby
decreasing the IE. Thus, taking benefit of the fact that the
mixture of gellan and enzyme solution becomes
increasingly viscous on cooling, the effect of the enzyme
entrapment was increased to 75% by maintaining the
chitosan solution at 50C 31. However, the mixture of gellan
and enzyme was maintained at room temperature to
facilitate the drop formation.
It could be pointed out from Fig.4 that, highest activity
(291 U/mg of support) was found at gellan and chitosan Fig. 4 Surface Graph Showing the Effect of Variables on
ions of 1.5%w/v and 0.6% w/v respectively. At the Activity.
low concentrations, less crosslinking of the matrix leads to
leaking of enzyme from support. Similar results were
obtained at high concentrations due to, low loading
efficiency and formation of a more compact matrix, m the
diffusional limitations are caused since, the substrate cannot
penetrate inside the matrix [22,31].
Similarly, the results of T50 (Fig. 5)
5 are in accordance
with IE. Since, the batch with maximum loading gives
maximum release due to better cross linking. It exhibits a
surface release phenomenon as the binding sites on the
matrix are free.

Swelling of Beads
Swelling behaviour of the two crosslinked polymer Fig. 5 Surface Graph Showing the Effect of Variables on
beads is illustrated in Fig. 6.. The swelling index of the the T50.
beads was higher in phosphate buffer and the beads showed
shrinkage at acidic pH 33. Similarly, the equilibrium The effect of the polymer concentration on swelling
swelling was obtained after 4-55 hours in acidic pH while in was observed and it was seen that as the polymer
alkaline pH it was attained within 2-33 hours. concentration increases, less cross linking occurs above and
Results showed
howed that the order of swelling was 2%> below the stoichiometric concentration. At the
1.5%> 1%. However, the processing of solution containing stoichiometric concentration, all the sites on gellan bind
2% gellan was difficult and hence, not considered for with chitosan thereby, resulting into a completely cross
further studies. linked matrix. Similar observation was reported by Magnin
et al. (2003). Hence, the 0.3% and 1% chitosan batches

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showed lesser swelling due to the more cross linking. index increases. Hence, from percent yield, IE, activity, T50
Therefore, as the gellanllan and chitosan concentrations and swelling studies, we conclude that Batch S-5 S is the
increase, the cross linking density decreases and swelling optimized batch.



Swelling index
1500 1000



0 2 4 6 8
0 2 4 6 8
T i m e (H r s)
Time (Hrs)

1% G- O.3% C 1% G- O.6% C 1% G- 1% C 1.5% G- O.3% C 1.5% G- O.6% C

1.5% G- 1% C 2% G- O.3% C 2% G- O.6% C 2% G- 1% C

Fig. 6 Swelling Study of Gellasan beads in different pH buffers. A: pH -7.2 and B: pH -1.5

Gellasan beads showed a steady increase upto 8 hrs. There

was no significant difference in lipase release profile of the
dried and fresh beads for any of the gellan-chitosan
batches. But the freeze-dried
dried beads showed a higher
percentage of release after 30 mins (burst effect) due to the
enzyme lost at the surface during freeze-drying.
freeze This was in
accordance with the studies conducted by Neau et al 2002.
It was also found that the activity of the beads (195 U/g of
support) decreased after freeze drying probably because of
the exposure of enzyme at low temperature variation.

Scanning Electron Microscopy (SEM)

Fig. 8 (a) (b) and
nd (c) are the external structures of the
loaded freeze-dried, air-dried
dried and blank beads respectively,
Fig. 7 Release Study of Gellasan Beads. obtained with and without lipase by (SEM). Fig 8 (d) shows
the internal structure of sphere without lipase, showing the
Release Studies porous network composed of fibres. fibres Fig.8(e) shows the
Since, swelling studies indicated the possibility of use presence of fibres as well as globular structures. The
of beads at the intestinal pH, we were further motivated to globular structures indicate the presence of intact enzyme
evaluate the potential of the loaded beads in therapy and lipase and fibres are indicative of the complexation between
therefore its release kinetics. Reports on the entrapment of gellan and chitosan. Fig 8(f) (f) shows surface view of the
the enzyme in gels iss not very efficient since, the entrapped loaded beads having a compact structure.
enzyme leaches out in course of time [31]. This is due to the CONCLUSION
large pore size of the matrix. We have approached this The complexation between the two hydrogels; Gellan
problem with an alternate point of view. It is well known and Chitosan resulted into Gellasan - a suitable matrix, for
that lipase is a therapeutic enzyme, a digestive. So, by successful immobilization of lipase. The immobilization
altering the pore size of the matrix, by varying the efficiencies and percent yield of the
he beads were satisfactory
concentrationn of the cross linker, we can allow the enzyme and the process is amenable to easy scale-up.
scale The activity
to leach out (‘release’) in the free form, from the matrix of lipase was found to improve after immobilization.
into the release medium. By this, we could well achieve a Maximum swelling index was obtained at the intestinal pH
controlled release system which will release the enzyme for suggesting the in-vivo use of the beads in future. The
a sufficiently longer period of time. Also, as enzymes being statistical
stical study indicated that the parameters, gellan and
proteins, are susceptible to the attack of the pH conditions chitosan concentrations selected, had a significant effect on
in-vivo.. The immobilization of enzymes into the matrix will the enzyme activity, loading efficiency, percentage release
not only render pH stability to it, but will also serve as a and the yield of the beads. However, a predominant role on
‘carrier’ for the enzyme delivery. With this objective, we enzyme activity afterr immobilization was played by
conducted the release study. polymer concentration. Activity increases with increase in
The release of lipase from air dried and freeze-dried
freeze gellan concentration upto level 0, but beyond that it
beads in-vitro was conducted for 8 hours. It can be seen decreases drastically due to the diffusional limitations. S-5
(Fig. 7) that batch S-55 showed the best release profile with batch was found to be optimized, due to the highest highes
the maximum release. This is in accordance
accorda with the swelling index, immobilization efficiency, T50, percentage
swelling studies. Since, maximum swelling is attributed to yield and optimum activity. Release studies of the gellasan
maximum release as reported by Badwan et al. 1999 [34].

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beads depict a controlled release pattern due to high order release pattern upto 8 hours. But, the freeze-dried
crosslinking. It was also seen that S-5,
5, exhibits a slow first beads showed higher release than the air-dried
air beads.

(a) (b) (c)

(d) (e) (f)

Fig. 8 Scanning electron microscopy of gellan-chitosan
gellan beads. (a) (b)and (c) are the external structures of the blank beads,
dried and loaded beads respectively, obtained with and without lipase by Scanning Electron Microscopy (SEM).
Figure (d) shows the internal structure of sphere without lipase, showing the porous network composed of fibres.
fib Similarly,
seen in Figure (e) but the presence of globular structures sphere contains lipase. These are lipase loaded beads. The presence
of these globular structures indicate the complexation between gellan, chitosan and the enzyme lipase. Figure (f) shows
surface view of the loaded beads having a compact structure.

IR studies indicated the successful binding between Technology, (VCH Verlagsgesellschaft, Weinheim),
the carboxylic (-OH)
OH) of glucuronic acid residue of gellan 349- 402 (1987).
gum and the primary amine residue of chitosan. The loaded
loa 3. Nelson DL, Cox MM & Lehninger, Principles of
beads showed the intact structure of lipase with no covalent biochemistry, 4th Ed. Ch 6 Enzymes. (W.H. Freeman,
binding of lipase with the polymers. Similarly, SEM New York), 190-195 (2004).
confirmed the results, by the presence of fibres indicating 4. Bharadwaj
dwaj TR, Kanwal M & Lal R, Natural gums and
the complexation between gellan and chitosan. While modified natural gums as sustained release carriers,
globular structures indicatee presence of the enzyme lipase. Drug Dev Ind Pharm, 26(10) 1025-1038
1025 (2000).
Accordingly, Gellan gum, a novel microbial 5. Betigeri SS & Neau SH, Immobilization of lipase using
polysaccharide and Chitosan a heteropolysaccharide proved hydrophillic polymers in the form of hydrogel beads,
to be excellent matrices from the point of view of enzyme Biomat, 23, 3627-3636
3636 (2002).
immobilization. Also, the entrapment in these matrices 6. Grasdalen H & Smidsrod O, Gelation of gellan gum,
promises to be simple, non-toxic,
toxic, versatile, biodegradable Carbohydr Polym, 7, 371 – 393 (1987).
and as well as a non-immunogenic
immunogenic method which will gain 7. Kasapis S & Miles N, Structural aspects and phase
a therapeutic dimension in the coming decade.
decade behaviour in deacylated and high acyl gellan systems,
Carbohydr Polym, 38, 145-154
154 (1999).
Acknowledgements 8. Chandrasekaran R & Millane RP, Crystal structure of
The authors thank A.E. Connock Pvt. Ltd, England gellan, Carbohydrate Res 17, 15 – 51 (1988).
and C.P Kelco Pvt. Ltd, Mumbai, India.
ia. for providing gift 9. Santucci E, Alhaique F, Carafa M & Coviello T, Gellan
samples of Chitosan and Gellan Gum respectively. They for the formulation of sustained delivery beads, J
also extend a deep gratitude to Dr. A.A.Prabhune, NCL Control Rel, 42, 157-164
164 (1996).
(National Chemical Laboratory, Pune) for her kind 10. Li J, Kamath K & Dwovedi C, Gellan film as an
suggestions. Financial assistance for this work was implant for insulin delivery, J Biomat Appl, 15, 321 –
provided by AICTE (All India Council for Technical 343 (2001).
Education, India). 11. Pandit JK & Balsubramniam J, In vitro and in vivo
evaluation of the Gelrite gellan gum-based
gum ocular
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