ANTIMICROBIAL AND STABILITY STUDY OF BIOSURFACTANT … · 2019-01-03 · ANTIMICROBIAL AND...

Available online at www.jpsscientificpublications.com

Volume – 4; Issue - 1; Year – 2018; Page: 1347 – 1358

DOI: 10.22192/iajmr.2018.4.1.3

Indo – Asian Journal of Multidisciplinary Research (IAJMR)

ISSN: 2454-1370

© 2018 Published by JPS Scientific Publications Ltd. All Rights Reserved

ANTIMICROBIAL AND STABILITY STUDY OF BIOSURFACTANT

PRODUCED FROM Pseudomonas aeruginosa PBS29

P. Poonguzhali*1, S. Rajan

1 and R. Parthasarathi

2

1Department of Microbiology, M.R. Government Arts College, Mannargudi - 614 001, Tamil Nadu,

India. 2Department of Microbiology, Faculty of Agriculture, Annamalai University, Chidambaram - 608 002,

Tamil Nadu, India.

Abstract

The biosurfactant production using the test strain Pseudomonas aeruginosa PBS29 performed

using acid precipitation followed by solvent extraction. The effectiveness of various solvents such as

Diethyl ether, Chloroform/Methanol (2:1 ratio), Dichloromethane, Ethyl acetate and Acetone were

evaluated and confirmed that Chloroform/Methanol as ideal solvent comparatively. The CMC value of

the biosurfactant was found to be 60 mg/ml. The extracted biosurfactant showed its stability over a wide

range of temperature, pH, sodium chloride concentration, Foaming activity and effect of metal ions such

as lead and zinc. Alkaline pH showed a higher stability than the acidic pH. The effect of temperature on

the biosurfactant found to stable till 100 ºC. Emulsification activity has not been affected till 20 %

Sodium chloride concentration.

Key words: Biosurfactant, CMC, Stability, Surface tension, Emulsification index and Antimicrobial

activity.

1. Introduction

Biosurfactant has gained its importance

in the recent years due to impact in various

fields. The lower toxicity, eco-friendly,

solubility and easy degradability nature has

contributed the biosurfactant to occupied a

unique position for its extending application in

various fields such as petrochemical and oil

industries, detergents, laundry formulations,

paper and textile industries, household cleaning

products, food, medical and pharmaceutical,

cosmetics, agricultural products like herbicides,

pesticides and also in bioremediation. The better

foaming properties, stability at extremes of pH,

salinity and temperature and ecological

compatibility makes the biosurfactant to

*Corresponding author: P. Poonguzhali

Received: 30.12.2017; Revised: 07.01.2018;

Accepted: 18.01.2018.

E.mail: [email protected]

overcome the use of chemical surfactants (Patil

et al., 2014). Biosurfactants exhibit a diverse

chemical structure, including glycolipids,

lipopeptides, phospholipids, fatty acids, or

neutral lipids, among others (Geys et al., 2014).

Rhamnolipid belongs to the glycolipid

type of biosurfactant possessing both

hydrophobic and hydrophilic portion

constituting of the fatty acid of variable length

linked with the carboxyl end of the rhamnose

molecule. Based on the number of rhamnose

molecules (one or two) linked, it has been

categories into mono and di-rhamnolipid

(Adetunji et al., 2017). The antimicrobial

activity of the biosurfactant is being significant

for its usage in the field of medicine and

agriculture. There exists certain evidence

insisting on the non-specific immunity induced

by the rhamnolipid against pathogens (Sachdev

and Cameotra, 2013). The present study is

attempted to characterize the biosurfactant

produced by Pseudomonas aeruginosa PBS29

http://www.jpsscientificpublications.com/

Poonguzhali/Indo – Asian Journal of Multidisciplinary Research (IAJMR), 4(1): 1347 – 1358 1348


and to evaluate its stability. This investigation

also reveals the antimicrobial activity against

bacterial pathogens.

2. Materials and methods

Culture conditions and Biosurfactant

production

The bacterial strain Pseudomonas

aeruginosa PBS29 (GenBank Accession

Number MG273769) was used for the study.

The strain was selected based on its potential

biosurfactant activity on the preliminary

investigation (Poonguzhali et al., 2017), with the

reduced surfaced tension below 35 mN/m as

described by Joshi et al. (2008). The Mineral salt

medium was used for the biosurfactant

production. Fermentation conditions established

by inoculating 2 % (v/v) inoculum (overnight

grown broth culture of Pseudomonas aeruginosa

PBS29 corresponding to 107

CFU/ml),

supplemented with 1 % hydrocarbon, at pH 7

and was incubated at 37 °C for about 7 days in a

rotary shaker with 120 rpm.

Biosurfactant extraction

The biosurfactant was extracted from the

supernatant by acid precipitation and solvent

extraction method followed by rotary

evaporation to pool the brownish content of the

biosurfactant as described by Aparna et al.

(2012) with modification. The culture broth was

filtered (Millipore filter with 0.2 μm pore size)

and centrifuged at 10000 g for 20 min to obtain

the cell free supernatant for further process. The

pH of the supernatant was reduced to 2 by the

addition of 6 M HCl and kept at 4 °C overnight for precipitation. It was neutralized by the

addition of sodium bicarbonate and further

extracted using the equal volume of solvent in a

separating funnel for phase separation. The

organic phase was allowed for rotary

evaporation. The resulting product was freeze-

dried and stored. To determine the efficacy of

various solvents such as Diethyl ether,

chloroform/methanol (2:1 ratio),

dichloromethane, ethyl acetate and acetone were

employed for accessing their extraction through

their emulsification activity.

Characterization of biosurfactant

The extracted biosurfactant was

characterized by estimating the chemical

constituent of the biosurfactant. The protein

concentration of the biosurfactant was estimated

according to Lowry et al. (1951). The total

carbohydrate was also determined by phenol-

sulfuric acid method (Hanson et al., 1981) and

the fatty acid (Lipid) content was estimated

adopting the method of Sadasivam and

Manickam (1991).

Determination of Critical Micelle

Concentration

The extracted biosurfactant was serially

diluted with distilled water to achieve various

concentrations ranging from 0 to 120 mg/ml.

The surface tension of the dilution was measured

using Du-Nuoy-ring method till a constant

surface tension value was attained (Desai and

Banat, 1997). The Critical Micelle Concentration

was obtained by plotting the surface tension

versus the concentration of the biosurfactant and

expressed in terms of g/L. SDS was used as the

positive control and distilled water as negative

control.

Stability study of biosurfactant

The cell free supernatant was employed

to determine the stability of the biosurfactant.

The thermal stability was evaluated by heating

the supernatant at 0 °C, 25 °C, 50 °C, 75 °C, 100

°C and125 °C for about an hour and then cooled

to room temperature. Similarly, pH of the

supernatant was adjusted to 2, 4, 6, 8, 10 and 12

respectively. The sodium chloride concentration

was adjusted to 0 %, 5 %, 10 %, 15 %, 20 % and

25 % by artificial addition to the supernatant.

The Surface tension and Emulsification index

(E24 %) was accessed for all the above

mentioned conditions. The effect of (1 %) metal

ions such as lead and zinc (w/v) on the growth of

the production strains was also accessed on

Mineral salt agar medium. All the experiments

were carried out in triplicate.

Foaming activity

Foam stability of the biosurfactant was

examined at the different time interval. Based on

the foam height, shape, size of the bubble

constituting the foam and time taken for

dissolution of bubble and foam property were

assessed.

Antimicrobial activity

The antimicrobial activity of the

biosurfactant was performed by disc diffusion

method according to Das et al. (2008). The

pathogens obtained from Government Hospital,



Cuddalore were inoculated on Tryptic Soy Broth

and incubated at 37 °C for about 2 to 4 hrs.

Lawn culture of each pathogen was made on the

Muller Hinton agar plates. Sterile disc of 6 mm

size was placed at equidistance on each plate.

The Minimal Inhibitory Concentration (MIC) of

the biosurfactant against the clinical pathogens

was accessed by inoculating 10 μl of the

biosurfactant at different concentration ranging

from 2 μg/ml, 4 μg/ml, 8 μg/ml and 16 μg/ml

(diluted with distilled water to obtain different

concentration) on to the sterile disc and further

incubated at 37 °C for 24 hrs. The zone of

inhibition around the disc was measured to

determine the MIC of each pathogen.

3. Result and Discussion

In order to investigate the stability and

antimicrobial activity of the biosurfactant, the

potent strain Pseudomonas aeruginosa PBS29

were employed. Similarly, several studies

insisted on Pseudomonas species as the potent

biosurfactant production (Silva et al., 2014; El-

Sheshtawy and Doheim, 2014; Raza et al., 2014;

Singh and Tripathi, 2013; Huang and Liu, 2013).

There was also report of other organisms like

Serratia rubidaea SNAU02 (Nalini and

Parthasarathi, 2017), Klebsiella sp. (Jain et al.,

2013), Bacillus subtilis (Mnif et al., 2013; Fahim

et al., 2013), Bacillus methylotrophicus USTBa

(Chandankere et al., 2013).

The efficacy of biosurfactant extraction

employing different solvents such as diethyl

ether, chloroform/methanol (2:1 ratio),

dichloromethane, ethyl acetate and acetone

evaluated in the present investigation revealed

maximum extraction achieved using the solvent

chloroform/methanol (2:1 ratio), which was

identified from its emulsification index of about

62 % (Figure - 1). This result was in accordance

with Pathaka and Nakhate (2015) who reported

the chloroform/methanol (2:1 ratio) as the best

solvent compared with ethyl acetate (1:1 ratio).

While the study of Trummler et al. (2003)

suggested ethyl acetate as the ideal solvent for

extraction of biosurfactant. However, several

studies (Sahoo et al., 2011; Chander et al., 2012;

Aparna et al., 2012; Arora et al., 2015) indicated

the maximum extraction and recovery of

rhamnolipid/other types of biosurfactant using

chloroform/methanol.

Characterization of biosurfactant

The Characterization of the extracted

biosurfactant revealed the estimated amount of

246.28 µg/0.1 ml of carbohydrate, 59.34 µg/0.1

ml of proteins and 274.1 µg/0.1 ml of lipid

respectively. The total glycolipid concentration

was determined as 1832.4 µg/0.1 ml. While the

reference strain, Pseudomonas aeruginosa

MTCC 2453 expressed 1809.1 µg/0.1 ml of total

glycolipid concentration. The rhamnose test was

performed by Orcinol method (Chandrasekaran

and Bemiller, 1980) for quantifying rhamnolipid

which revealed the yellowish orange color

formation as an indication of rhamnolipid as

suggested in the study of Kalyani et al. (2014).

Critical micelle concentration was

obtained at a point where there was no further

reduction in the surface tension. Regardless,

CMC was determined at the increasing dilution

of biosurfactant or at its decreasing

concentration. However, the study of Patowary

et al. (2017) explained the CMC value of

biosurfactant produced by Pseudomonas

aeruginosa 56 mg/L and insisted that there was

no further decrease in surface tension even after

increasing the concentration of the biosurfactant

beyond 56 mg/L.

Even within the presence of a small

concentration of biosurfactants, a CMC can be

attained, from that variation within the surface

tension was ascertained (Sobrinho et al., 2008).

In the present study, CMC value obtained was

60 mg/L, (Figure - 2) which was much lower

than the Sodium dodecyl sulfate (SDS), which

further proved the better biosurfactant activity of

the current study. The CMC potentially of the

reference strain was also compared with the test

strain and was found to be satisfied.

Stability of Biosurfactant

The produced biosurfactant was

subjected to the different range of temperature,

pH, NaCl concentration to access their stability.

However, the biosurfactant in this study showed

a high stability over the wide temperature range

of 0 °C, 25 °C, 50 °C, 75 °C 100 °C and 121 °C

(including the autoclavable temperature). A

similar result was observed in the study of

Khopade et al. (2012) who reported the stable

biosurfactant under extreme temperatures.



Moreover, the report of Borodoli and

Konwar et al. (2012) insisted the stability of the

biosurfactant producing Pseudomonas

aeruginosa strains even at the temperature of

100 °C for the different time interval ranging

from 5 to 60 min with respect to surface tension.

The decrease in viscosity due to salinity

increased the NaCl concentration which affects

the quality of emulsion, thereby reducing the

emulsification capacity (Sarubbo et al., 2006).

Heating of the biosurfactant to 100 °C has not

been producing any considerable difference on

the biosurfactant performance and was found to

be stable.

The biosurfactant activity was also

evaluated over a pH range of 2 to 12. The

activity of biosurfactant confirmed that it was

not affected by the change in pH. But the acidic

pH showed a reduction in emulsification index

though not much variation when compared with

the synthetic surfactant (SDS). The Stable

emulsion in the presence of salt has also been

suggested from the biosurfactant producing

strain Bacillus licheniformis JF-2 (Ilori et al.,

2005). Ghojavand et al. (2008) reported that the

biosurfactant activity of Bacillus subtilis PTCC

1696 has not been varied even at higher NaCl

concentration (20 % w/v) with the surface

tension of 34 mN/m and the similar findings

were also observed in the present study. In a yet

another study, Khopade et al. (2012) observed

slight change at an increased concentration of up

to 8 % (w/v) NaCl. Most of the earlier studies

also reported high stability of biosurfactant at

higher salinity (Al-Sulaimani et al., 2011;

Darvishi et al., 2011; Al-Wahaibia et al., 2014).

While assessing the Stability, the

extracted biosurfactant showed its stability over

foaming activity at various time intervals (Table

- 1). A stable growth of the biosurfactant

producing strain has also been exhibited at 1 %

(w/v) metal ions such as lead and zinc when

supplemented (filter sterilized) in the Mineral

salt agar medium. Furthermore, the stability of

the biosurfactant at the wide range of pH,

temperature and salinity (Figure - 3) make it

appropriate for the extreme circumstances that

could be encountered in the various field during

application as indicated by Darvishi et al.

(2011). Foaming property of the biosurfactant in

the current investigation (Table - 2) was found to

be good with stable foam till 72 hrs.

Antimicrobial activity of Biosurfactant

The considerable use of biosurfactants

comprised their function as anti-adhesive agents

towards pathogens, making them valuable for

treating many diseases medically and as

therapeutic agents (Singh and Cameotra 2004).

Rhamnolipids are recognized to be active against

the Gram-negative bacteria P. aeruginosa,

Enterobacter aerogenes, Serratia marcescens

and Klebsiella pneumonia, in addition, to be

employed towards Gram positive Micrococcus

sp., Streptococcus sp., Staphylococcus sp. and

Bacillus species (Benincasa et al., 2004). There

was varied biosurfactant production from

microbial source employing different growth

conditions and their structures were responsible

to endorse variation in their action potentials

(Das et al., 2014). In the present investigation,

the biosurfactant produced was tested against the

genera Bacillus, Staphylococcus, Salmonella,

Shigella and compared with the reference strain

(Table - 3 and 4). The MIC of Staphylococcus

obtained was 4 μg/ml, while for other test

pathogens, the MIC obtained was 8 μg/ml.

Though the biosurfactant produced exhibited its

effectiveness towards all the test organisms, yet

Staphylococcus found to have a more inhibitory

effect (Figure - 4) among others.

The change in the antimicrobial zone of

inhibition against the Gram positive bacterium

tested was more prominent than that the Gram

negative organism. The growth of Bacillus

subtilis NCTC 10400 was inhibited by

rhamnolipids and sophorolipids (Rienzo et al.,

2016). Rhamnolipid from soyabean oil waste

source has an antimicrobial activity towards

bacterial pathogens such as Bacillus cereus, and

Staphylococcus aureus in the study of Rodrigues

et al. (2006). Thus, the antimicrobial property

provided evidence for their role in the presence

of its potentiality in biotechnological and

biopharmaceutical applications owing to their

biological nature.



Table - 1: Stability study of the biosurfactant from the selected strain

Isolates Pseudomonas aeruginosa PBS29

Emulsification activity (E24%) 71.36 ± 0.3

CMC value (mg/l) 60

Rhamnose test +

Foam stability Very good

Em

uls

ific

ati

on

In

dex

* (

E2

4%

) &

Su

rfa

ce T

en

sio

n**

(mN

/m)

pH

2 29.85 ± 0.5* 64.73 ± 0.6

**

4 42.47 ± 0.3 52.11 ± 0.3

6 71.16 ± 0.6 33.4 ± 0.5

8 70.33± 0.3 33.95 ± 0.4

10 69.28 ± 0.4 35.82 ± 0.2

12 68.79 ± 0.5 35.31 ± 0.7

Temperature

(°C)

0 68.34 ± 0.3 38.56 ± 0.5

25 71.56 ± 0.6 34.9 ± 0.3

50 72.13 ± 0.3 33.22 ± 0.4

75 71.64 ± 0.1 34.63 ± 0.2

100 70.57 ± 0.2 35.81 ± 0.1

125 68.84 ± 0.2 37.34 ± 0.4

Sodium

chloride

concentration

(%)

0 69.60 ± 0.5 36.85 ± 0.6

5 71.31 ± 0.4 33.17 ± 0.7

10 71.18 ± 0.3 33.44 ± 0.5

15 70.33 ± 0.7 33.61 ± 0.3

20 70.05 ± 0.4 34.18 ± 0.3

25 24.58 ± 0.2 67.81 ± 0.2 Values indicate Mean ± Standard deviation

Table - 2: Foam stability of the Biosurfactant

Foam Properties 24 hrs 48 hrs 72 hrs

Foam stability +++ +++ ++

Foam height (cm) 2.1 1.8 1.3

Foam Size Tiny bubbles Tiny bubbles Medium bubbles

Inference Excellent Stability Excellent Stability Good Stability +++ Highly stable; ++ Moderately stable.

Table - 3: Minimal Inhibitory Concentration of the biosurfactant exhibiting its Antibacterial

activity against pathogens

Bacterial

pathogens

Zone of Inhibition (mm)

MIC of Biosurfactant (µg/ml) using Pseudomonas

aeruginosa PBS29

2 4 8 16

Bacillus sp. Nil Nil 11.33 ± 0.6 28.33 ± 0.6

Staphylococcus sp. Nil 12.33 ± 0.6 13.67 ± 0.6 21.67 ± 0.6

Salmonella sp. Nil Nil 10.33 ± 0.6 14.33 ± 0.6

Shigella sp. Nil Nil 11.67 ± 1.2 27.67 ± 1.2 Values indicate Mean ± Standard deviation



Table - 4: Antibacterial activity of the Reference strain against pathogens

Bacterial pathogens Zone of Inhibition (mm)

MIC of Biosurfactant (µg/ml) using Reference strain

2 4 8 16

Bacillus sp. Nil Nil 9.67 ± 0.6 24.00 ± 1.0

Staphylococcus sp. Nil 11.33 ± 1.5 12.33 ± 0.6 20.67 ± 0.6

Salmonella sp. Nil Nil 8.33 ± 0.6 12.67 ± 1.2

Shigella sp. Nil Nil 10.67 ± 0.6 25.33 ± 0.6

Values indicate Mean ± Standard deviation

Figure – 1: Comparative extraction of biosurfactant employing different solvents

Figure - 2: (a) CMC of the biosurfactant from Pseudomonas aeruginosa PBS29

(b) CMC value obtained for SDS using different concentration



PBS29

Figure – 3: Stability study of the biosurfactant produced from Pseudomonas aeruginosa

PBS29

(a)

(c)

(b)



Figure - 4: Antimicrobial activity of the biosurfactant against bacterial pathogens employing (a)

Pseudomonas aeruginosa PBS29 and (b) Reference strain

4. Conclusion

The biosurfactant produced from

Pseudomonas aeruginosa PBS29 demonstrated

the ability of chloroform/methanol in the

biosurfactant extraction and was found to be

ideal among other solvents employed. The test

strain Pseudomonas aeruginosa PBS29 also

exhibited stable growth of the with 1 % metal

ion (Lead and Zinc). The present study exhibited

the stability of the extracted biosurfactant at a

wide range of pH, temperature, and NaCl

concentration. Low CMC value than the

synthetic surfactant was also significant. The

antimicrobial property of the biosurfactant was

one of the noteworthy instrumentation for the

therapeutic usage. In conclusion, assessing the

(a)

(c)

(b)



combination of these properties has been always

a valuable tool for the application of the specific

biosurfactant in various sectors including

Industrial, Medical and Pharmaceutical,

Biotechnological, Food and Agricultural

purpose.

Acknowledgment

We extend our sincere thanks to the

Department of Microbiology, Faculty of

Agriculture, Annamalai University, Annamalai

Nagar, Chidambaram for providing the

laboratory facility. We also register our earnest

gratitude to the Department of Microbiology,

M.R. Government Art College, Mannargudi for

their valuable support throughout the work.

Conflict of Interest

The authors declare that there are no

conflicts of interest.

5. References

1) Adetunji C, Oloke JO, Pradeep M, Jolly

RS, Anil KS. (2017). Characterization

and optimization of a rhamnolipid from

Pseudomonas aeruginosa C1501 with

novel biosurfactant activities.

Sustainable Chemistry and Pharmacy, 6:

26 - 36.

2) Al-Sulaimani H, Al-Wahaibi Y, Al-

Bahry SN, Elshafie A, Al-Bemani A,

Joshi S, Zaragari S. (2011). Optimization

and partial characterization of

biosurfactant produced by Bacillus

species and their potential for enhanced

oil recovery. SPEJ, 16(1): 672 - 682.

3) Al-Wahaibia Y, Joshib S, Al-Bahryb S,

Elshafieb A, Al-Bemania A, Shibulal B.

(2014). Biosurfactant production by

Bacillus subtilis B30 and its application

inenhancing oil recover. Colloids and

Surfaces B. Biointerfaces, 114: 324 –

333.

4) Aparna A, Srinikethan G, Smitha H.

(2012). Production and characterization

of biosurfactant produced by a novel

Pseudomonas sp. 2B. Colloids Surface,

95: 23 - 29.

5) Arora SK, Sony J, Sharma A, Taneja M.


of biosurfactant from Pseudomonas spp.

International Journal of Current

Microbiology and Applied Sciences, 4(1):

245 - 253.

6) Bharali P, Konwar BK. (2011).

Production and physicochemical

characterization of a biosurfactant

produced by Pseudomonas aeruginosa

OBP1 isolated from petroleum sludge.

Applied Biochemistry and Biotechnology,

164: 1444 – 1460.

7) Chandankere R, Yao J, Choic MMF,

Masakorala K, Chan Y. (2013). An

efficient biosurfactant-producing and

crude-oil emulsifying bacterium Bacillus

methylotrophicus USTBa isolated from

petroleum reservoir. Biochemical

Engineering Journal, 74: 46 - 53.

8) Chandrasekaran EV, Bemiller JN.

(1980). Constituent analyses of

glycosaminoglycans in Methods in

Carbohydrate Chemistry. Whistler, R.L.,

Ed. Academic Press, New York. 8: 89 -

96.

9) Darvishi P, Ayatollahi S, Mowla D,

Niazi A. (2011). Biosurfactant

production under extreme environmental

conditions by an efficient microbial

consortium, ERCPPI-2. Colloids Surface,

84(2): 292 - 300.

10) Das P, Mukherjee S, Sen R. (2008).

Antimicrobial potential of a lipopeptide

biosurfactant derived from a marine

Bacillus circulans. Journal of Applied

Microbiology, 104: 1675 –1684.

11) Das P, Yang X, Ma LZ. (2014). Analysis

of biosurfactants from industrially viable

Pseudomonas strain isolated from crude

oil suggests how rhamnolipids congeners

affect emulsification property and

antimicrobial activity. Frontiers in

Microbiology, 5: 696 - 704.

12) Desai JD, Banat IM. (1997). Microbial

production of surfactants and their

commercial potential. Microbiology and

Molecular Biology Reviews, 61: 47 – 64.

13) El-Sheshtawy HS, Doheim MM. (2014).

Selection of Pseudomonas aeruginosa

for biosurfactant production and studies

of its antimicrobial activity. Egypt

Journal of Petroleum Products, 23: 2356

– 2364.



14) Fahima S, Dimitrova K, Vauchela P,

Gancela F, Delaplace G, Jacquesa P,

Nikov I. (2013). Oxygen transfer in three

phase inverse fluidized bed bioreactor

during biosurfactant production by

Bacillus subtilis. Biochemical

Engineering Journal, 76: 70 - 76.

15) Geys R, Soetaert W, Van Bogaert I.

(2014). Biotechnological opportunities in

biosurfactant production. Current

Opinions in Biotechnology, 30: 66 – 72.

16) Ghojavand H, Vahabzadeh F, Roayaei E,

Shahraki AK. (2008). Production and

properties of a biosurfactant obtained

from a member of the Bacillus subtilis

group (PTCC 1696). Journal of Colloid

and Interface Science, 324: 172 - 176.

17) Gudina E, Teixeira J, Rodrigues L.

(2010). Isolation and functional

characterization of a biosurfactant

produced by Lactobacillus paracasei.

Colloids Surface, 76(1): 298 - 304.

18) Hanson RS, Phillips JA, Gherhardt P.

(1981). Manual of Methods for General

Bacteriology. American Society for

Microbiology, Washington, DC. p. 328.

19) Huang W, Liu Z. (2013). Biosorption of

Cd(II)/Pb(II) from aqueous solution by

biosurfactant-producing bacteria:

Isotherm kinetic characteristic and

mechanism studies. Colloids and

Surfaces. Biointerfaces, 105: 113 – 119.

20) Ilori M.O, Amobi CJ, Odocha AC.

(2005). Factors affecting biosurfactant

production by oil degrading Aeromonas

spp. isolated from a tropical

environment. Chemosphere, 61: 985 –

992.

21) Jain RM, Mody K, Joshi N, Mishra A,

Jha B. (2013). Effect of unconventional

carbon sources on biosurfactant

production and its application in

bioremediation. International Journal of

Biological Macromolecules, 62: 52 - 58.

22) Joshi S, Bharucha C, Jha S, Yadav S,

Nerurkar A, Desai AJ. (2008).

Biosurfactant production using molasses

and whey under thermophilic conditions.

Bioresource Technology, 99(1): 195 -

199.

23) Kalyani ALT, Sireesha GN, Sankar GG

and Prabhakar T. (2014). Isolation of

Biosurfactant producing Actinomycetes

from terrestrial and Marine soils.

International Journal of Pharmaceutical

Science and Research, 5(9): 4015 - 4022.

24) Khopade A, Biao R, Liu X, Mahadik K,

Zhang L, Kokare C. (2012). Production

and stability studies of the biosurfactant

isolated from marine Nocardiopsis sp.

B4. Desalination, 285: 198 - 204.

25) Lowry OH, Rosenbrough NJ, Farr AL

and Randall RJ. (1951). Protein

measurement with the Folin phenol

reagent. Journal of Biological Chemistry,

193: 265.

26) Mendes AN, Filgueiras LA, Pinto JC and

Nele M. (2015). Physico-chemical

properties of Rhamnolipid Biosurfactant

from Pseudomonas aeruginosa PA1 to

Applications in Microemulsions. Journal

of Biomaterials and Nanobiotechnology,

6: 64 - 79.

27) Mnif I, Elleuch M, Chaabouni SE, Ghribi

D. (2013). Bacillus subtilis SPB1

biosurfactant: Production optimization

and insecticidal activity against the carob

moth Ectomyelois ceratoniae. Crop

Protection, 50: 66 - 72.

28) Pathaka AN, Nakhate PH. (2015).

Optimization of Rhamnolipid: A New

Age Biosurfactant from Pseudomonas

aeruginosa MTCC 1688 and its

Application in Oil Recovery, Heavy and

Toxic Metals Recovery. Journal of

Bioprocess Biotechnology, 5: 229 - 241.

29) Patil S, Pendse A, Aruna K. (2014).

Studies on optimization of biosurfactant

production by Pseudomonas aeruginosa

F23 isolated from oil contaminated soil

sample. International Journal of Current

Biotechnology, 2(4): 20 - 30.

30) Patowary K, Patowary R, Kalita MC,

Deka S. (2017). Characterization of

Biosurfactant Produced during

Degradation of Hydrocarbons Using

Crude Oil As Sole Source of Carbon.

Frontiers in Microbiology, 279(8): 1 - 14.

31) Poonguzhali P, Rajan S, Parthasarathi R.

(2017). Screening and characterization of

Biosurfactant producing bacteria from



Hydrocarbon and Pesticides

contaminated soils from Cuddalore

district. Life Science Archives, 3(6): 1207

– 1217.

32) Raza ZA, Rehman A, Hussain MT,

Masood R, Haq Au, Saddique MT, Javid

A, Ahmad N (2014) Production of

rhamnolipid surfactant and its application

in bioscouring of cotton fabric.

Carbohydrate Research, 7(8): 110 – 122.

33) Rienzo MAD, Stevenson P, Marchant R,

Banat IM. (2016). Antibacterial

properties of biosurfactants against

selected Gram positive and negative

bacteria. FEMS Microbiology Letters,

363: 1 - 8.

34) Rodrigues L, Banat IM, Teixeira J,

Oliveira RR. (2006). Biosurfactants:

potential applications in medicine.

Journal of Antimicrobial Chemotherapy,

57: 609 - 618.

35) Sachdev DP, Cameotra SS. (2013).

Biosurfactants in agriculture. Applied

Microbiology and Biotechnology, 97(3):

1005 - 1016.

36) Sahoo S, Datta S, Biswas D. (2011).

Optimization of Culture Conditions for

Biosurfactant Production from

Pseudomonas aeruginosa OCD1.

Journal of Advanced Scientific Research,

2(3): 32 - 36.

37) Sarubbo LA, Luna JM, Campos-Takaki

GM. (2006). Production and stability

studies of the bioemulsifier obtained

from a new strain of Candida glabrata

UCP 1002. Electronic Journal of

Biotechnology, 9(4): 400 - 406.

38) Silva EJ, Silva NMPR, Rufino RD, Luna

JM, Silva RO, Sarubbo LA. (2014).

Characterization of a biosurfactant

produced by Pseudomonas cepacia

CCT6659 in the presence of industrial

wastes and its application in the

biodegradation of hydrophobic

compounds in soil. Colloids and Surfaces

B: Biointerfaces, 117: 36 – 41.

39) Singh DN, Tripathi AK. (2013). Coal

induced production of a rhamnolipid

biosurfactant by Pseudomonas stutzeri,

isolated from the formation water of

Jharia coalbed. Bioresource Technology,

128: 215 - 221.

40) Singh P, Cameotra S. (2004). Potential

applications of microbial surfactants in

biomedical sciences. Trends in

Biotechnology, 22: 142 - 146.

41) Sobrinho HBS, Rufino RD and Luna JM.

(2008). Utilization of two agro industrial

by-products for the production of a

surfactant by Candida sphaerica

UCP0995. Process Biochemistry, 43: 912

– 917.

42) Suresh Chander CR, Lohitnath T,

Mukesh Kumar DJ, Kalaichelvan PT.


of biosurfactant from Bacillus subtilis

MTCC441 and its evaluation to use as

bioemulsifier for food biopreservative.

Advances in Applied Science Research,

3(3): 1827-1831.

43) Trummler K, Effenberger F, Syldatk C.

(2003). An integrated

microbial/enzymatic process for

production of rhamnolipids and L-(+)-

rhamnose from rapeseed oil with

Pseudomonas sp DSM 2874. European

Journal of Lipid Science and

Technology, 105(10): 563 – 571.



Access this Article in Online

Quick Response

Code

Website www.jpsscientificpublications.com

DOI Number DOI: 10.22192/iajmr.2018.4.1.3

How to Cite this Article:

P. Poonguzhali, S. Rajan and R. Parthasarathi. 2018. Antimicrobial and stability study

of Biosurfactant produced from Pseudomonas aeruginosa PBS29. Indo - Asian Journal of

Multidisciplinary Research, 4(1): 1347 – 1358.

DOI: 10.22192/iajmr.2018.4.1.3

http://www.jpsscientificpublications.com/

Date post:	21-May-2020
Category:	Documents
Upload:	others
View:	13 times
Download:	0 times

ANTIMICROBIAL AND STABILITY STUDY OF BIOSURFACTANT … · 2019-01-03 · ANTIMICROBIAL AND...

Documents