Biosurfactant Production From Unconventional Resources

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Biosurfactant Production from UnconventionalResources: a Short Overview

ARTICLE JANUARY 2012

CITATIONS

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349

5 AUTHORS, INCLUDING:

Diego Coelho

University of Campinas

13PUBLICATIONS 27CITATIONS

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Edgar Silveira

Universidade Federal de Uberlndia (UFU)


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Roberto Souza

Universidade Federal de Sergipe


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Elias Basile Tambourgi

University of Campinas

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Available from: Diego Coelho

Retrieved on: 06 January 2016
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International Review of Chemical Engineering (I.RE.CH.E.), Vol. 4, N. 2

ISSN 2035-1755 March 2012

Manuscript received and revised February 2011, accepted March 2012 Copyright 2012 Praise Worthy Prize S.r.l. - All rights reserved

175

Biosurfactant Production from Unconventional Resources:

a Short Overview

Diego F. Coelho1, Silvanito A. Barbosa2, Edgar Silveira3,

Roberto R. Souza4, Elias B. Tambourgi5

Abstract The Biosurfactant production has experienced an intensive and fast growing since it

was "discovered". As its own definition, biosurfactants are a class of surface-active molecules that

are produced extracellularly or as part of the cell membrane by microorganisms, i.e., by a

biological pathway. In the last years, several researchers groups' have focused its efforts on

investigation of a medium composition which could maximize the production of these surface

active agents and make it economically feasible. Thus, the purpose of this review is to provide a

comprehensive and updated overview of the use of agro-industrial wastes and renewable

resources as raw material for production of biosurfactants and also to focus on principal

applications of biosurfactants, covering part of its wide range of chemical specialty. Copyright

2012 Praise Worthy Prize S.r.l. - All rights reserved.

Keywords:Biosurfactant, Renewable-Resources, Agroindustrial Wastes

I.

Introduction

The surfactants are SURFace ACTive AgeNTS and

constitute an important class of chemical products widely

used in a great variety of household and industrial

applications and indispensable components of daily life

[1]-[2]. Their production was estimated in 2007 to bearound 10 million tons per year [3]. They are widely

used in the pharmaceutical, cosmetic, petroleum, and

food industries [4]. Surfactants are amphiphilic

compounds that reduce the free energy of the system by

replacing the bulk molecules of higher energy at an

interface, signifying, e.g., which surfactants assist the

solubility of polar compounds in organic solvents.

Surfactants have been used industrially as adhesives,

flocculating, wetting and foaming agents, demulsifiers

and penetrants [5]. They are used for these applications

based on their abilities to lower surface tensions, increase

solubility, detergency power, wetting ability and foaming

capacity [4].The presence of surfactants can lead to anincrease in the concentration of hydrophobic compounds

in the water phase, through the formation of oil water

emulsions and solubilization, where, above the Critical

Micelle Concentration (CMC), biosurfactant molecules

aggregate to form micelles. [6].At concentrations above

the CMC, biosurfactant molecules associate to form

micelles, bilayers and vesicles. The CMC is obtained

where the surface tension remains steady despite the

changes in concentration as represented Fig. 1 [15].

Micelle formation enables biosurfactants to reduce the

surface and interfacial tension and increase the solubility

and bioavailability of hydrophobic organic compounds

[12].

The CMC is commonly used to measure the efficiency

of surfactant. Efficient biosurfactants have a low CMC,

which means that less biosurfactant is required to

decrease the surface tension [15].

The effectiveness of a surfactant is determined by its

ability to lower the surface tension, which is a measure

of the surface free energy per unit area required to bringa molecule from the bulk phase to the surface [7].

Fig. 1. Relationship of Surface tension, interfacial tension,

solubilization and CMC with biosurfactant concentration

Many different types, with several structures (see Fig.

2), of surfactants are already being used in industry, but

it is important to develop even more new compounds to

broaden the spectrum of specific properties and

applications [8].

Most of these surfactants are petroleum based, i.e.,

they are chemically synthesized. That origin makes them

environmentally harmful, decreasing organic degradation
https://www.researchgate.net/publication/229930032_Properties_and_industrial_applications_of_sophorolipids?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/6352701_Microbial_production_and_application_of_sophorolipids_Appl_Microbiol_Biotechnol?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/8201898_Mulligan_CN_Environmental_applications_for_biosurfactants_Environ_Pollut_1332_183-198?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/8201898_Mulligan_CN_Environmental_applications_for_biosurfactants_Environ_Pollut_1332_183-198?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/11016558_Microbial_surfactants_and_their_use_in_field_studies_of_soil_remediation_J_Appl_Microbiol?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/14112145_Microbial_production_of_surfactants_and_their_commercial_potential_MMBR?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/49854063_Review_Environmental_Applications_of_Biosurfactants_Recent_Advances?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/14112145_Microbial_production_of_surfactants_and_their_commercial_potential_MMBR?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/243778501_Surfactants_And_Interfacial_Phenomena?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/243778501_Surfactants_And_Interfacial_Phenomena?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/14112145_Microbial_production_of_surfactants_and_their_commercial_potential_MMBR?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/14112145_Microbial_production_of_surfactants_and_their_commercial_potential_MMBR?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/49854063_Review_Environmental_Applications_of_Biosurfactants_Recent_Advances?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/11016558_Microbial_surfactants_and_their_use_in_field_studies_of_soil_remediation_J_Appl_Microbiol?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/8201898_Mulligan_CN_Environmental_applications_for_biosurfactants_Environ_Pollut_1332_183-198?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/8201898_Mulligan_CN_Environmental_applications_for_biosurfactants_Environ_Pollut_1332_183-198?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/6352701_Microbial_production_and_application_of_sophorolipids_Appl_Microbiol_Biotechnol?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/229930032_Properties_and_industrial_applications_of_sophorolipids?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/227609815_Production_and_application_of_trehalose_lipid_biosurfactants?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==


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D. F. Coelho, S. A. Barbosa, E. Silveira, R. R. Souza, E. B. Tambourgi

Copyright 2012 Praise Worthy Prize S.r.l. - All rights reserved International Review of Chemical Engineering, Vol. 4, N. 2

176

possibly through its toxic effects [6]. However the

leading trend towards using environmental friendly

technologies has enhanced the search for biodegradable

compounds of natural origin [9].

Then, in the past few decades, a heterogeneous group

of surface-active molecules and microbial origin,

referred to as biosurfactants, have gained considerableinterest.

Fig. 2. Chemical Structures of some common biosurfactants

They present several advantages over synthetic

surfactants. The biodegradability of these biological

surfactants is one of major properties, because it prevents

accumulation and toxicity problems in natural

ecosystems, once that the capability of these molecules

to emulsify water and hydrocarbons enhances the

biodegradation of pollutants. Nevertheless, from an

economic standpoint, biosurfactants are not yet

competitive with the synthetics, because the raw material

and production cost are expensive. So far, several

renewable substrates from various sources, especially

from industrial wastes have been intensively studied for

microorganism cultivation and surfactant production at

an experimental scale [10].

The choice of inexpensive raw materials is important

to the overall economy of the process because they

account for a large quote of the final product cost (about

50%) and also reduce, or eliminate, the expenses with

wastes treatment [11].

The purpose of this review is to provide acomprehensive and updated overview of the use of agro-

industrial wastes and renewable resources as raw

material for production of biosurfactants and also to

focus on principal applications of biosurfactants,

covering part of its wide range of chemical specialty.

II. But what actually are Biosurfactants?

As its own definition, biosurfactants are a class of

surface-active molecules that are produced

extracellularly or as part of the cell membrane by

microorganisms, i.e., by a biological pathway.

TABLEI

MAIN TYPES AND EXAMPLES OF SOME BIOSURFACTANTS PRODUCED

BY MICROORGANISMS (ADAPTED FROM [13])

Main Types Examples Microorganisms

Glycolipids

Trehalose lipidRhodococcus

erithropolis

Sophorose lipidTorulopsis magnoliae,

Candida bombicola

Rhamnose lipidPseudomonas

aeruginosa

Mannosylerithritol

lipid

Shizonella

melanogramma,

Pseudozyma

Antarctica

LiposaccharidesEmulsane Acinetobacter

calcoaceticus

AlasanAcinetobacter

radioresistens

Lipopeptides

Surfactin Bacillus subtilis

ViscosinePseudomonas

fluorescens

Polymers

EmulsanAcinetobacter

calcoaceticus

BiodispersanAcinetobacter

calcoaceticus

Liposan Candida lipolytica

Phospholipids

Fatty acids and

neutral lipids

Corynomycolic

acid

Corynebacterium

insidibasseosum

All surfactants have a common structure: a lipophilic

portion, with a hydrocarbon of one or more fatty acids,

bounded to a hydrophilic portion, which can be an ester;

a hydroxyl group; a phosphate; a carbohydrate or a

carboxylate [8]. The fatty acids can be saturated (or not),

hydroxylated or branched.

Unlike chemically synthesized surfactants, which are

classified according to their dissociation pattern in water,

biosurfactants are categorized by their chemical

composition, molecular weight, physic-chemical

properties and mode of action and microbial origin [12].

A large variety of microorganisms are known to

produce biosurfactants, which vary in their chemical
https://www.researchgate.net/publication/11016558_Microbial_surfactants_and_their_use_in_field_studies_of_soil_remediation_J_Appl_Microbiol?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/51637836_Advances_in_utilization_of_renewable_substrates_for_biosurfactant_production_AMB_Express_11-19?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/26472533_Production_of_biosurfactants_using_substrates_from_renewable-resources_Songklanakarin_J_Sci_Technol?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/226313785_Biosurfactant_production_by_microorganisms_on_unconventional_carbon_sources?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/7001671_Adsorption_on_stainless_steel_surfaces_of_biosurfactants_produced_by_gram-negative_and_gram-positive_bacteria_consequence_on_the_bioadhesive_behavior_of_Listeria_monocytogenes_Colloids_Surf_B_Biointer?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/7001671_Adsorption_on_stainless_steel_surfaces_of_biosurfactants_produced_by_gram-negative_and_gram-positive_bacteria_consequence_on_the_bioadhesive_behavior_of_Listeria_monocytogenes_Colloids_Surf_B_Biointer?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/226313785_Biosurfactant_production_by_microorganisms_on_unconventional_carbon_sources?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/26472533_Production_of_biosurfactants_using_substrates_from_renewable-resources_Songklanakarin_J_Sci_Technol?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/51637836_Advances_in_utilization_of_renewable_substrates_for_biosurfactant_production_AMB_Express_11-19?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/11016558_Microbial_surfactants_and_their_use_in_field_studies_of_soil_remediation_J_Appl_Microbiol?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==


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178

III.1. Biosurfactant Production

on Oil-Containing Wastes

World production of oils and fats, founded in all

living cells, are most used in the food industry, is about

2.5-3 million tons and generates great quantities of

wastes, tallow, lard, marine oils or soap stick, and free

fatty acids from the extraction of seed oils. Wastedisposal is a growing problem, which explains the

increasing interest in use of waste in microbial

transformation [24]. Waste or used lubricating oils are a

serious environmental problem. In the environment, the

waste oil can bind to organic matter, mineral particles

and organisms [33]. Mercade et al. [34] reported the

screening and selection of microorganisms capable of

utilizing waste lube oil for producing biosurfactants.

From 18 hydrocarbon-contaminated soil samples, 44

different isolates capable of growth on waste lube oil (as

sole carbon source) were selected. Only 10% of the

strains isolated produced biosurfactants (trehaloseglycolipids, from Rhodococcus sp., and lipopeptides,

from Bacillus sp.). The production of bioemulsifiers by

Candida lipolytica (strains 1055 and 1120) using media

supplemented with 5% Babassu oil and 1% glucose as

carbon source was evaluated by Sarubbo et al. [35].

The bioemulsifiers was produced as secondary

metabolites at the end of the exponential growth phase

and beginning of the stationary growth phase.

Alcanivorax borkumensis utilizes aliphatic

hydrocarbons as its main carbon source for growth and

produces an anionic glucose lipid biosurfactant and thus

potentials of Alcanivorax strains during bioremediation

of hydrocarbon pollution in marine habitats have beenstudied [36]. Frying oil is produced in large quantities for

use both in the food industry and at the domestic scale.

They can act as effective and inexpensive raw materials

for biosurfactant production [20]-[37]. Additionally, any

oil wastes from vegetable oil refineries and the food

industry have a potential to induce microbial growth and

also been used as appropriate substrates for biosurfactant

production. P. aeruginosaPACL strain, isolated from oil-

contaminated soil taken from a lagoon has been grown in

residual waste of soybean oils to produce biosurfactant

by submerged fermentation in stirred tank reactors [38].

Sunflower seed oil and oleic acid can be used for the

production of rhamnolipids by Thermus thermophilusHB8.

The potential production of rhamnolipids has been

demonstrated using Thermophilic eubacterium [39].

Thus, a sound strategy of waste management for the food

and auto industries to reduce is produce biosurfactant

from vegetable oils, used vegetable oil and used lube oil,

reducing the generation of waste.

III.2. Biosurfactant Production

from Agro-Industrial Wastes

Although hydrocarbons have been used as substrates

of choice for the production of biosurfactants and

bioemulsifiers [40]-[41] due the effect of induction of

biosurfactant production, which makes any hydrophobic

substrate accessible to the cell, water-soluble substrates

also have been used [11].

Moreover, for many applications (e.g. food and

cosmetic industry) substrates hydrocarbon based are

unacceptable. Water-soluble substrates are cheaper andare preferred over hydrocarbons since single-phase

fermentations are simpler than biphasic fermentation

[11]. The disposal of wastes is a well-known and

growing problem, and new alternatives for their use are

being studied once the treatment and disposal costs for

these wastes are a vast financial burden to various

industries [42].

Then, the use of alternative low-cost substrates is an

important strategy to facilitate industrial development of

biosurfactant production. To this end, good components

seem to be agroindustrial byproducts or wastes, once

these residues generally contain high levels of

carbohydrates or lipids to support growth and surfactantsynthesis [19].

These agricultural based wastes are influenced by the

agricultural practices and industries and are based in

particular regions or countries. For example, in Brazil,

the production of soap stock (one of the wastes of the oil

neutralization process in soybean oil refining) amounts to

2-3% of the total oil production and is affected by the

fatty acid content of the oil [9]. Brazil is also among the

main producers of vegetable oils, such as soybean oil,

babassu oil and palm oil [43].

Rufino and Co-workers [44] applied a sequential

factorial design to optimize biosurfactant production by

Candida lipolyticausing soybean oil refinery residue as

substrate. This study evaluated the impact of three

cultivation factors, amounts of refinery residue, glutamic

acid and yeast extract. The resultant biosurfactant

showed high emulsifying ability and surface activity,

with stability at wide range of pH (2-12), temperatures

(0-120C) and salinity (2-10% NaCl).

Nitschke et al. [45] evaluated the utilization of oil

wastes for the production of rhamnolipids by

Pseudomonas aeruginosaLBI strain. They used wastes

obtained from soybean, cottonseed, babassu, palm, and

corn oil refinery. The best results were achieved for the

soybean soap stock waste, which generated 11.7 g/l ofrhamnolipids and a production yield of 75%,

highlighting the fact that low cost substrate can be

utilized for rhamnolipid production for application in

high value pharmaceutical and food industry applications

[46]. Using molasses (which is a rich source of available

carbon produced as co-product of sugar industry

generated during sugar manufacturing) and corn-steep

liquor as the primary carbon and nitrogen source, Patel

and Desai [47] produced a rhamnolipid biosurfactant

using P. aeruginosa (Strain GS3). The biosurfactant

production reached a maximum when a combination of

7% (v/v) molasses and 0.5% (v/v) corn-steep liquor

waste used.


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179

TABLEIII

EXAMPLES OF BIOSURFACTANT PRODUCTION USING AGROINDUSTRIAL WASTES (ADAPTED FROM [49])

Substrate Chemical Class Microorganisms Reference

Olive Oil Mill Effluent (OOME) rhamnolipids Pseudomonas sp. [57]

Soybean Oil Refinery Wastes rhamnolipids P. aeruginosa [58]

Molasses lipopeptides B. subtilis [59]

Whey rhamnolipids P. aeruginosa [60]

Waste frying Oil rhamnolipids P. aeruginosa [61]

Waste Lubricating Oil glycolipids Rhodococcus sp. [34]

Deprotenized Whey sophorolipids C. bombicola [62]

Cassava Waste lipopeptides B. subtilis [63]

In their studies, de Gusmo et al. [48] applied a

factorial design to investigate the effects and interactionsproduced by use of vegetable fat waste, yeast extract and

glucose in the production of biosurfactant by Candida

glabrata after 144 h of cultivation. Was achieved a

maximum surface activity with vegetable fat waste at 5%

and yeast extract at 0.2%.

Beyond this study was the first report on the use of a

vegetable fat waste as substrate for biosurfactant

production, the product containing cell-free broth

retained its surface-active properties after incubation at

high temperatures, at a wide range of pH values and salt

concentrations. Structural determination suggests it to be

a mixture of carbohydrates, proteins and lipids and theauthors further concluded its suitability for use in

bioremediation and oil recovery. P. aeruginosa can be

cultivated in Cashew Apple Juice (CAJ) supplemented

with peptone (5.0 g/L) and nutritive broth to obtain

surfactants. Surface tension during the fermentation can

be reduced by 41% when P. aeruginosa is cultivated in

CAJ supplemented with peptone [50] compared to other

amino acid sources. Ohno et al. [51]-[52] reported

production of iturin and surfactin by a strain of B.

subtilis NB 22 using wheat bran and okara (soybean curd

residue produced as by-product of tofu manufacturing

processes). It is composed of water (81.1%), protein

(4.8%), fat (3.6%), starch and sugar (6.4%), fiber (3.3%),and ash (0.8%). Table III shows the main agroindustrial

wastes used as main carbon source and study object of

several researches. Recently, some researchers groups

are directing their studies to genetic engineering [53]-

[54]-[55]-[56], cloning genes and allowing biosurfactant-

producing microorganisms to degrade specific waste and

used it as substrate.

IV. Biosurfactant Production Using

Renewable Substrates

Although the utilization of carbon rich wastes, likeagroindustrial wastes, allow us achieve a double benefit

(reducing the pollutants while produce useful products),

the selection of suitable waste material with the rightproportion of nutrients that permits microorganism

growth and production of target-product is the main

problem associated.

Furthermore, the constituents of the waste could affect

the properties of the final product and, consequently, its

functionality [9].Thus, the development of a promising

and economically feasible process for biosurfactant

production appears to depend upon the use of abundant

and low-cost raw materials and the optimization of the

operational cultivation conditions.

The plant biomass is the main sustainable source of

renewable, providing low cost raw materials, reductionin environmental pollution, and relative ease of

operation. A large body of literature on biosurfactant

production using substrate related to vegetable industries

exists and has a geographical significance in relation to

industries associated and type of biosurfactant produced

[9].

Oliveira et al. [43] used palm oil, a low-cost

agricultural byproduct which is used in as raw material

for soap and food industries, for biosurfactant production

using Pseudomonas alcaligenes (a strain isolated from

crude oil contaminated soil).

They achieved a biosurfactants concentration of 2.3

g/l and E24more than 70% with the hexane, jet fuel andcrude-oil. Abdel-Mawgoud et al. [64] reported the

production of a rhamnolipid by Pseudomonas

aeruginosa isolate Bs20 on soybean oil amended

medium. Perfumo et al. [65] confirmed that the

emulsifying capacity and thermo and halo tolerance

properties presented by the rhamnolipid produced makes

that biosurfactant adequate for use in bioremediation of

hydrocarbon-contaminated sites or in the petroleum.

Monteiro et al. [66] reported the growth and

biosurfactant production using sunflower oil

supplemented mineral medium by the yeast Trichosporon

montevideense, CLOA 72. The glycolipid produced

exhibited good surface and emulsifying activity with
https://www.researchgate.net/publication/51637836_Advances_in_utilization_of_renewable_substrates_for_biosurfactant_production_AMB_Express_11-19?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==https://www.researchgate.net/publication/51637836_Advances_in_utilization_of_renewable_substrates_for_biosurfactant_production_AMB_Express_11-19?el=1_x_8&enrichId=rgreq-38ad4ad6-562e-4af8-838b-b25d51ad5b66&enrichSource=Y292ZXJQYWdlOzIzNTY3NTA0NDtBUzoxMDM5MjQzODgzMzU2MTdAMTQwMTc4ODkxMzU4Nw==


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vegetable oils, toluene, isooctane, cyclohexane, hexane

and hexadecane. Furthermore, the biosurfactant was

thermo and halo tolerant and stable in wide range of pH

values.

Sim et al. [67] studied the utilization of a mixture of

canola oil, soy bean and glucose for rhamnolipid

production by P. aeruginosa UW-1 and reported 10-12fold increase in rhamnolipid production on vegetable oils

in comparison to glucose. Costa et al. [68] observed that

Pseudomonas aeruginosa LBI produces a rhamnolipid

that reduces extensively surface tension and shows good

emulsification while evaluated the use of oil from

Andiroba (Carapa guianensis), Babassu (Orbignya

spp.), Buriti (Mauritia flexuosa), Brazilian Nut

(Bertholletia excelsa), Cupuau (Theobroma

grandiflora) and Passion Fruit (Passiflora alata). The

Brazilian Nut shows the highest rhamnolipid

concentrations (9.9 gl-1), followed by the Passion Fruit

(9.2 g/l) oils. Prieto et al. [69], another Brazilian group,

used a P. aeruginosa LBM10 isolated from a southerncoastal zone in Brazil, to produce a rhamnolipid-type

biosurfactant growing on soybean oil, soybean oil

soapstock, fish oil and glycerol. Production using

soybean oil achieved the maximum yield and the

biosurfactant was stable at a wide range of pH and salt

concentration.

V. Other Substrates Sources

Some researchers groups studies around world carried

out studies with some renewable substrates mainly

confined to a particular geographic region.The glycerol produced as byproduct of biodiesel

industry has accumulated more intensely with recent

surge of this biofuel. The low cost glycerol could be used

as water soluble substrate for biosurfactant production

[9]. The utilization of glycerol as single carbon source by

Pseudomonas aeruginosa in the production of a

rhamnolipid was studied by Nitschke et al. [42], but

yields were less compared to traditional hydrophobic

substrates.

Another production of rhamnolipid was reported by

Rahman et al. [70], achieving 1.77 g/l of biosurfactant by

P. aeruginosa DS10-129. Zhang et al. [71] produced,

also using glycerol as the sole carbon source, 15.4 g/lrhamnolipids using P. aeruginosa growing on a basal

mineral medium, showing clearly the feasibility of

utilizing glycerol as carbon source for growth and

biosurfactant production by microbes. Morita et al. [72]

showed that Pseudozyma antarctica JCM 10317, a

basidiomycete yeast, efficiently produced

mannosylerythritol lipids (MELs) as glycolipid

biosurfactants from glycerol, achieving 16.3 g/l by

intermittent feeding of glycerol.

Lee et al. [73] reported the use of fish oil for

biosurfactant production. They optimized the culture

medium for the Pseudomonas aeruginosaBYK-2 KCTC

18012P for enhanced rhamnolipids production and used

25 g/l fish oil as carbon source. In optimum conditions,

they achieved a yield of 17 g/l of rhamnolipid.

VI. Final Comments

Active at extreme temperatures, pH and salinity; they

are environmentally friendly, biodegradable, less toxicand non-hazardous, these are the main properties which

makes the biosurfactants a potential substitute for the

surfactant produced from petroleum derivatives. Many

researchers continue to investigate the production of

these microbial surface active agents using raw material

widely marketed (industrially synthetized), trying to

reduce the production cost through process

optimizations. However, the combination of an industrial

waste and a cheap substrate is a promising approach to

reduce production cost and their polluting effect at the

same time.

Moreover, industrial by-products and renewable

resources also can be used to produce them with equallyefficiency. Over this review, we can note the large variety

of unconventional substrates that has been employed on

studies of biosurfactant production and the main types of

molecules produced. Precisely the property of generate a

specific structure for each combination of

microorganism; strain and formulation of fermentation

media that make these compounds highly promising,

once that studies can be performed easily to adjust the

molecule for a particular purpose.

These studies clearly indicate the vast potential of the

unconventional substrates for the biosurfactant

production and, most importantly, indicate that theproduction of these molecules needs to utilize a

combination of several techniques (e.g., industrial

strategies, genetic engineering or research to find new

raw materials or microorganisms ) to overcome, from an

economic standpoint, the synthetic surfactants. Current

researches focus on the development of these techniques.

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Authors information1Universidade Estadual de Campinas (UNICAMP), Faculdade de

Engenharia Qumica, Departamento de Engenharia de Sistemas

Qumicos e Informtica.

E-mail:[email protected]

2Instituto Federal de Educao, Cincia e Tecnologia de Sergipe (IFS).

E-mail: [email protected]

3Universidade So Paulo, Faculdade de Cincias Farmacuticas.


4Universidade Federal e Sergipe (UFS), Departamento de Engenharia

Qumica.


5Universidade Estadual de Campinas (UNICAMP), Faculdade de

Engenharia Qumica, Departamento de Engenharia de Sistemas

Qumicos e Informtica.


Diego de Freitas Coelho (Corresponding

author), born in Salvador Brazil, Abril 2th,

1987. Graduated in Industrial Chemistry in 2010

at Universidade Federal de Sergipe, Sergipe,

Brazil. Master degree obtained in 2012 at

Universidade Estadual de Campinas, Campinas,

Brazil. His major field of study is the

purification of biomolecules from renewable raw

materials and agroindustrial wastes. His preview published works

involves the purification of proteolytic enzymes through fractional

precipitation and aqueous two-phase systems. His most recently studies

focuses on processes integration and processes optimization, also to

purify biomolecules. At the moment, he is developing his Ph.D. thesis

in modeling and construction of an annular centrifugal contactor to

purify bromelain from agroindustrial wastes.

Silvanito Alves Barbosa, born in Tobias

Barreto - Sergipe - Brazil, February 12, 1971.

Graduated in Industrial Chemistry in 1997 and

in Chemistry in 2002 at Universidade Federal de

Sergipe, Sergipe, Brazil. Doctor in

Biotechnology in 2011 obtained from the

Universidade Estadual do Cear. His main area

of study is the production of biosurfactants for

use as feedstock in the production of biodetergents in the oil industry.

He is professor at the Instituto Federal de Cincia e Tecnologia de

Sergipe - IFS - acting in the area of Chemical and Petrochemical

Industry and developing works in agreement between Petrobras and the

IFS.


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