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Microbial lipases are an important group of biotechnologically valuable enzymes, because of the versatility of their applied properties and ease of mass production. Lipases of microbial origin are widely diversified in their enzymatic properties and substrate specificity, which make them very attractive for industrial applications. This review describes the applications of microbial lipases in detergents. Enzymes can reduce the environmental load of detergent products as the chemicals used in conventional detergents are reduced; they are biodegradable, non-toxic and leave no harmful residues. Besides lipases, other enzymes are widely used in household cleaning products, in laundering, medical, agriculture, etc. This article also reviews the use of enzymes, especially lipases as detergents and different types of lipase containing detergents available in the market.
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African Journal of Biotechnology Vol. 9(31), pp. 4836-4844, 2 August, 2010
Available online at http://www.academicjournals.org/AJB
DOI: 10.5897/AJBx09.026
ISSN 16845315 © 2010 Academic Journals
Review
Enzymes used in detergents: Lipases
Fariha Hasan, Aamer Ali Shah*, Sundus Javed and Abdul Hameed
Department of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan.
Accepted 2 April, 2010
Microbial lipases are an important group of biotechnologically valuable enzymes, because of the
versatility of their applied properties and ease of mass production. Lipases of microbial origin are
widely diversified in their enzymatic properties and substrate specificity, which make them very
attractive for industrial applications. This review describes the applications of microbial lipases in
detergents. Enzymes can reduce the environmental load of detergent products as the chemicals used in
conventional detergents are reduced; they are biodegradable, non-toxic and leave no harmful residues.
Besides lipases, other enzymes are widely used in household cleaning products, in laundering,
medical, agriculture, etc. This article also reviews the use of enzymes, especially lipases as detergents
and different types of lipase containing detergents available in the market.
Key words: Detergents, enzymes, lipases.
INTRODUCTION
Lipases constitute the most important group of biocatalysts
for biotechnological applications (Benjamin and Pandey,
1998). Lipases have been isolated from many species of
plants, animals, bacteria, fungi and yeast. The enzymes
from microorganisms are used in various industries such
as dairy, food, detergents, textile, pharmaceutical, cosmetic
and biodiesel industries, and in synthesis of fine chemicals,
agrochemicals and new polymeric materials (Saxena et
al., 1999; Jaeger and Eggert, 2002). Research on microbial
lipases, has increased due to their great commercial
potential (Silva et al., 2005).
Lipases are added to detergents such as household
and industrial laundry (Kumar et al., 1998) and in
household dishwashers, where their function is in the
removal of fatty residues and cleaning clogged drains
(Vulfson, 1994). The cleaning power of lipase (or other
enzyme containing) detergents increases markedly. The
enzymes such as proteases, amylases, cellulases and
lipases are added to the detergents to improve their
efficiency (Ito et al., 1998).
Laundry detergents are becoming more and more
popular because of their increasing use in washing
machine, where it impart softness, resiliency to fabrics,
antistaticness, dispersible in water and mild to eyes and
*Corresponding author. E-mail: alishah_75@yahoo.com. Tel:
+92-51-90643065. Fax: +92-51-9219888.
skins. There are many different brands or types of laundry
detergents, and mostly they claim some special qualities
(Bajpai and Tyagi, 2008). The detergents industry,
regulated by the European Commission (EC) 648/2004,
Detergent Directive, is considered as the largest enzyme
consuming sector. Concerns over persistence of detergent
chemicals in the environment and its possible conta-
mination of ground water, other fresh sources and their
subsequent health related issues has raised speculation
over biodegradability. This has acted as an impetus for
greater consumption of enzymes as Europe is witnessing
increasing preference for greener detergents.
The need for greater control over microorganisms and
enzymes for industrial purposes has led to a greater
focus on Genetic Engineering and Recombinant DNA
technology. This technology, allows genetic modification
of microorganisms to produce the desired enzyme under
specific conditions. This helps either to produce a particular
type of enzyme or enhance the quantity of enzyme
produced from the single recombinant microorganism.
Understanding the structure of enzymes and modifying
them to extract benefits is categorized as Protein
Engineering. Studies are being conducted on ways to
improve or modify protein structure and its function thus
finally the enzyme. Biotechnology is therefore, being in-
creasingly viewed as a possible solution against traditional
chemicals processes. The production of enzymes from
natural sources and their environment friendly charac-
teristics has led the industry to believe that enzymes are
Hasan et al. 4837
Table 1. Four classes of enzymes are generally used in detergents.
Proteases
Most widely used enzymes in the detergent industry remove protein stains such as grass,
blood, egg and human sweat which have a tendency to adhere strongly to textile fibers.
Amylases
Used to remove residues of starch-based foods like potatoes, spaghetti, custards, gravies
and chocolate.
Lipases
Decompose fatty material. Lipase is capable of removing fatty stains such as fats, butter,
salad oil, sauces and the tough stains on collars and cuffs.
Cellulases
Modify the structure of cellulose fiber on cotton and cotton blends. When it is added to a
detergent, it results in; color brightening, softening and soil removal.
indeed a sustainable alter-native to chemicals in industrial
processes. The factors that aid their increased application is
the availability of wide enzyme types, environmental
friendly behaviour, minimised energy consumption, easily
controlled processes, possible modification of enzyme
characteristics, environ-mentally friendly by-products,
minimal greenhouse gas emissions, decreased use of
non-renewable sources and proliferation of chemicals in
environment. The multi faceted benefits availed through
using enzymes is bound to convert into a greener world
(Kumar, 2009).
ENZYMES USED IN DETERGENTS
Biotechnology based cleaning agents such as enzyme-
based agents, are widely used in industries. The biotech-
nology based cleaning agents (ETBPP, 1998) are cheaper
and less harmful to the environment. They have specific
cleaning action and can also be used at lower tempe-
ratures. They produce effluents with lower COD and non-
corrosive nature. Enzyme-based cleaners are becoming
increasingly popular in the food industry as compared to
caustic or acid cleaning regimes (D'Souza and Mawson,
2005).
The enzyme based detergents have better cleaning
properties as compared to synthetic detergents. They are
active at low washing temperatures and environment
friendly also (Kumar et al., 1998). The enzymes in the
detergents do not lose their activity after removing stain.
The enzyme containing detergents also improve the
fabric quality and keeping color bright.
The enzyme based detergents are used in small quantity
as compared synthetic chemicals. They can work at very
low temperature, environment friendly and completely
biodegradable.
Four classes of enzymes are generally used in detergents
as given in Table 1 (Gormsen et al., 1991).
Removal of proteins by non-enzymatic detergents can
result in permanent stains due to oxidation and denaturing
caused by bleaching and drying. Proteases hydrolyze
proteins and break them down into more soluble
polypeptides or free amino acids. As a result of the
combined effect of surfactants and enzymes, such hard
to remove stains can be removed from fibers.
An alkaline extracellular enzyme obtained from alkaliphilic
Bacillus strains is used in detergents. Other enzymes like
proteases, alpha amylases and cellulases are produced
on large scale and have been used in detergents (Ito et
al., 1998). The domestic dishwashing detergents contain
different chemicals like chelating agents, surfactant,
polycarboxylates (PCA), phosphonates, active chlorine
compounds etc. These substances have high environ-
mental risks, particularly via sewage biosolids, poor
biodegradability, increased organic load to sewage works,
toxicity (CEEP-phosphates in dishwasher detergents, 2007).
Sodium hydroxide or caustic soda (NaOH) is the most
widely used alkaline detergent in the food and beverage
industry due to its low price and high cleaning efficiency
for fatty-type and protein soils. Nitric acid and phosphoric
acid are most commonly used by the food and beverage
industries which lead to the increased level of TDS and
sodium discharge in effluent. To avoid any damage to soil
and vegetation, it is important to reduce the use of
traditional chemicals in food and beverage industries
(Palmowski et al., 2005).
Enzymes are used in detergents to increase the cleaning
ability of detergents. Enzymes can be used instead of
chlorine bleach for removing stains on cloth. The enzyme
protease was produced from alkaliphilic Bacillus clausii
KSM-K16 and strain KP-43 and Bacillus sp. strain KSM-
KP43 and have been incorporated into laundry
detergents. Subtilisin-like serine proteases belonging to
family A of subtilase super family has been used in
laundry and dishwashing detergents (Saeki et al., 2007).
A number of alkaliphilic Bacillus produce alkaline cellulase
(carboxymethylcellulase) that is used as an additive for
improving the cleaning effect of detergents. Enzymatic
properties of some cellulases fulfilled the essential
requirements for enzymes to be used practically in
laundry detergents (Ito, 1997).
Among other enzymes used in some detergents are
Guardzyme (a peroxidase) which inhibits dye transfer
and Carezyme removing the fuzz that builds up on cotton
clothes. Enzymes used in detergents must be effective at
low levels, compatible with various detergent components
and to be active at wide range of temperatures (Kumar et
al., 1998).
A number of detergent formulations such as anionic or
nonionic surfactants and the powdered lipase are used
4838 Afr. J. Biotechnol.
Table 2. Compositions of an enzyme detergent.
Constituent
Composition (%)
Sodium tripolyphosphate (water softener, loosens dirt)
38.0
Sodium alkane sulphonate (surfactant) (cationic/ anionic/ non-inonic/ amphoteric)
25.0
Sodium perborate tetrahydrate, sodium percarbonate (oxidising agent, oxygen bleach)
25.0
Soap (sodium alkane carboxylates)
3.0
Sodium sulphate (filler, water softener)
2.5
Sodium carboxymethyl cellulose/sodium polyacrylate/ polyethylene glycol (dirt-suspending
agent/ (Antiredeposition agents)
1.6
Sodium metasilicate (binder, loosens dirt)
1.0
Bacillus protease (3% active) (any other enzyme)
0.8
Fluorescent brighteners
0.3
Foam-controlling agents
Trace
Perfume
Trace
Water
100%
Source: Jain, 2008.
for the removal of fatty soils from fabrics. These include
sodium nitrilotriacetate, sodium tripolyphosphate, sodium
silicate, sodium citrate, and potassium diphosphate.
Optional ingredients include a detergent builder such as,
foam boosters, alkalies (e.g. sodium carbonate); optical
brighteners (e.g. stilbene derivatives); stabilizers (e.g.
triethanolamine); fabric softeners (e.g. quaternary
ammonium salts) together with bleaching agents and
systems (such as sodium perborate and ethylene
diaminetetra acetate). Different fragrances, dyes, foam
depressors and corrosion agents are also used
additionally (Thom et al., 1990). Use of liquid laundry
detergents has been increasing over the years (Bajpai
and Tyagi, 2007). For more than forty years, enzymes in
encapsulated form have been used in detergent products,
such as laundry formulations. They have increasing
importance as they are biodegradable and function at low
temperatures and offer environmental benefits (Basketter
et al., 2008). A very small amount of the enzymes is used
in detergent preparations and it must withstand the other
components of the detergents, such as surfactants,
oxidants, optical brighteners and also stable at pH values
between 8 and 10.5. (Table 2) (Jain, 2008).
Alkaline yeast lipases are preferred because they can
work at lower temperatures as compared to bacterial and
fungal lipases (Ahmed et al., 2007; Amino et al., 1998;
Saxena et al., 1998). Cold active lipase detergent formu-
lation is used for cold washing which reduces the energy
consumption and wear and tear of textile fibers (Feller
and Gerday, 2003). Enzymes can reduce the environ-
mental load of detergent products because they save
energy by enabling a lower wash temperature to be used
(Vakhlu and Kour, 2006). Addition of cold active lipase in
detergent become biodegradable, leaves no harmful
residues, have no negative impact on sewage treatment
processes and do not have risk for aquatic life (Joseph et
al., 2007).
Various factors affect the performance of enzymes in
detergents; its composition, type of stains to be removed,
washing temperature, washing procedure and water
hardness. Some of the additives of the detergents include;
builders (Sequestering builders- polyphosphates, citrate,
precipitating builders- sodium carbonate, sodium silicate),
zeolite, alkaline agents- sodium carbonate, sodium
silicates, corrosion inhibitor- sodium silicate, processing
aids- sodium sulfate, water, alcohol, xylene sulphonate,
colorants, suda control agents, Opacifiers, bleaching
agents-sodium percarbonate, sodium perborate, hydrogen
peroxide (Bajpai and Tyagi, 2007).
Enzymes in detergent formulations are stable at high
pH and temperature and remove protein and lipid stains.
Enzymatic detergent requires low workable temperature,
low mechanical energy and is less toxic and non-
corrosive (Sekhon and Sangha, 2004). Thermostable
enzymes are usually stable in solvents and detergents;
therefore, they also have considerable potential for many
biotechnological and industrial applications (Haki and
Rakshit, 2003).
The first industrial extremozymes used in textile deter-
gents, were the alkaline cellulases produced by bacteria
isolated from an east African soda lake, which was the
first industrial extremozymes for use in textile detergents,
and were commercialized by Genencor (Antranikian et
al., 2005). Enzymes used in household detergents have
stability at extreme temperatures and pH. The enzymes
must fulfill different requirements such as activity and
stability, substrate specificity and enantioselectivity, for
their application in industry (Podar and Reysenbach,
2006).
The cold active lipases will have a larger share of
enzymes of industrial importance in the coming years
with the increasing interest in psychrophiles (Joseph et
al., 2007). These enzymes offer a great industrial and
biotechnological potential due to their capability to catalyze
reactions at low temperature (Gomes and Steiner, 2004).
LIPASES IN DETERGENTS
There has been a tremendous increase in the significance
of the biotechnological applications of lipases since the
last two decades (Jaeger and Eggert, 2002; Jaeger and
Reetz, 2000; Jaeger et al., 2001), due to the versatile
catalytic behavior of lipases (Bosley, 1997). Lipases form
an integral part of the industries ranging from food
(Jaeger et al., 1994), pharmaceuticals (Chang et al.,
1999) and detergents (Venegas, 1993) to oleo-chemicals
(Houde et al., 2004), tea industries (Saxena et al., 1999)
agriculture, cosmetics, leather and in several bioreme-
diation processes. Because of the vast applications,
newer microbes are to be screened for production of
lipases having desirable properties. Molecular analysis of
the lipase gene helps to understand the structure and
activity relationship of the enzyme, which will help the
researchers to construct new lipases for biotechnological
applications (Tripathi, 2003) according to the demand
and need of industry.
Detergents are common commercial products used in
dish washing, bleaching compositions (Nakamura and
Nasu, 1990), in dry cleaning solvents (Abo, 1990), leather
cleaners (Kobayashi, 1989), cleaning of contact lens
(Bhatia, 1990), clogged drain clearing in domestic/
industrial effluent or food processing treatment plants
(Bailey and Ollis, 1986), in degradation of organic wastes
from exhaust pipes, toilet bowls, etc. (Moriguchi et al.,
1990). Lipases and cellulases in certain detergents
remove dirt/ cattle manure from domestic animals (Abo et
al., 1990). Lipases are also used in degreasing and water
reconditioning in combination with oxido-reductases that
require lesser amount of surfactants and work at low
temperatures (Novak et al., 1990).
The world market for enzymes is expected to reach $7
billion in 2013, with average annual increase of 6.3%,
mostly in the pharmaceutical and biocatalyst enzymes.
Biotechnology will continue to be of importance, helping
to sustain demand for research and biotechnology
enzymes (World Enzymes, 2009). Whereas, according to
another report by Global Industry Analysts, Inc., world
market for industrial enzymes is expected to exceed $2.9
billion by 2012. Another report by San Jose, California
(PRWEB) (2008), states that United States is going to be
the fastest growing market for industrial enzymes with a
CAGR of 5% in 2001 - 2010 while Europe was reported
as the largest regional market having an estimated share
of 30.93% in 2008. Sale of industrial enzymes in Asia-
Pacific was estimated at US$327 million in 2008. Lipases
(for their use in the detergent and cosmetics markets),
were expected to have a CAGR of 9.13% in 2001 - 2010
(http://www.prweb.com/releases/industrial_enzymes/carb
ohydrases_proteases/prweb1569084.htm).
It is estimated that every year, about 1000 tons of lipases
Hasan et al. 4839
are added to approximately 13 billion tons of detergents
(http://www.aukbc.org/beta/bioproj2/introduction.html).
Lipases at present account for less than 5% of the
market that is expected to increase because of wide
range of different applications (Vakhlu and Kour, 2006).
China's demand for detergent has grown in the past
decade and both production and demand will continue to
grow in the next five years (Detergent Market Research,
2006).
Rohm in 1913 patented the use of pancreatic enzymes
for laundry cleaning and marketed as a presoak detergent
under the brand name Burnus. It was sold for about 50
years in the European markets. Rohm developed the first
detergent, containing enzymes as well as the method for
washing protein stained cloth in it. Protein degrading
enzyme (trypsin) was extracted from pancreatic glands
and developed a detergent formulation known as Pan-
creatin. Pancreatin preparation also contains enzymes
having protease, lipase and amylase activity
(http://home.intekom.com/pharm/alcon/polyzym.html). In
1959, a Swiss chemist Dr. Jaag developed a product
called Bio-40 that instead of trypsin contained a protease
obtained from bacterial. In 1965 - 1970, use and sale of
detergent enzymes grew very fast, and slowed down as
the workers started to develop allergies. In 1975,
encapsulation of the granules of enzyme started (Jain,
2008).
BACTERIAL LIPASES IN DETERGENTS
Lipases for detergents are specifically selected on the
basis of low substrate specificity, stability under alkaline
(pH 10 - 11, 30 - 60°C) conditions, in the presence of
surfactants (linear alkyl benzene sulfonates) and enzymes
(proteases), constituents of many detergent formulations.
Continuous screening and protein engineering leads to
the discovery of lipases with the desired properties
(Wang et al., 1995; Kazlauskas and Bornscheuer, 1998;
Cardenas et al., 2001). Detergent enzymes should be
cost-effective and safer to use. Bacterial lipases added to
household detergents reduces or replaces synthetic
detergents, which have considerable environmental
problems (Jaeger et al., 1994).
Normally fat stains are not easy to remove at low
temperatures using conventional detergents, therefore
lipases are required which are active at lower temperatures
and can be used in detergent formulations. The use of
lipase active at low temperature in detergent formulation,
reduces the energy consumption as well as the wear and
tear of textile fibers (Feller and Gerday, 2003, Chaplin,
2004) and maintain the texture and quality of fabrics
(Bjorkling et al., 1991). Lipases from Candida (Nishioka
et al., 1990) and Chromobacterium (Minoguchi and
Muneyuki, 1989) are also used in detergents.
Lipases which are stable and work at alkaline pH (8 to
11), suitable wash conditions for enzymated-detergent
4840 Afr. J. Biotechnol.
powders and liquids have good potential for use in
detergent industry (Jaeger et al., 1994; Gerhartz, 1990;
Hasan et al., 2006). Lipase produced by Acinetobacter
radioresistens was found to be optimally active at pH 10
and showed stability in the range of pH 6 - 10; therefore,
it has a greater potential to be used in the detergent
industry (Chen et al., 1998). Hasan et al. (2007) reported
100% stability of lipase produced by Bacillus sp. FH5, at
pH 10. This enzyme showed promising results when
used in combination with different commercially available
conventional detergents (Javed, 2007). A. radioresistens
produced lipase having optimum activity at pH 10 and
stability in the range of pH 6 - 10, and showed a great
potential to be used in the detergent industry (Chen et al.,
1998).
Extracelluar lipase produced by Micrococcus sp. ML-1
was commonly used in combination with various commonly
used detergents, to enhance the removal of oily stains
from various types of fabrics. Soybean oil-dye stain,
followed by dye stain in mustard oil, groundnut oil and
coconut oil were removed quite easily. The detergents;
Surf, Nirma and Wheel, differed in the oil-dye removes
potential stains from cotton, terrrycot and polyester
(Neeru and Gupta, 2001). The ability of detergents to
wash, remove fatty food stains and sebum from fabrics, is
enhanced by addition of lipases (Andree et al., 1980;
Hemachander and Puvanakrishnan, 2000).
SYSCO makes cleaning products for foodservice industry.
Sysco® Action Suds with enzymes removes tough stains
and makes the fabric white and gives brightness. This
enzyme product has low-alkalinity and works with optical
brighteners, soil re-deposition agents and color-safe
bleach to give longer life to fabric. Sysco® enzyme grease
and waste digester is a natural, multi-strain bacterial
enzyme formulation, effective in both hard and soft water,
unclogs drain lines and digests any accumulated grease
and waste (Cleaning and sanitation Catalog, 2004).
Olin Corporation (Cheshire, UK) invented a detergent
composition comprising the microbial lipase SD2, dode-
cylbenzene sulfonate and gelatin (Holmes, 1991). Novo-
zymes has also marketed lipases (Lipex® and Lipolase
®), used in detergent industry (Novozymes report 2006).
US Agricultural Research Service scientists obtained an
enzyme from a bacterium isolated from shipworm glands
that digests proteins in common laundry stains (Hardin,
1992).
Commercial detergent formulations with high-tempe-
rature optima have been produced from P. mendocina
(Lumafast) and Pseudomonas glumae are used (Jaeger
et al., 1994). In 1995, Lumafast and Lipomax, lipases
from P. mendocina and Pseudomonas alcaligenes,
respectively, were produced by Genencor International,
AU-KBC Research Center, Life Sciences, Anna
University, Chennai, India (http://www.au-kbc.org/beta/
bioproj2/uses.htm). Gerritse et al. (1998) reported an
alkaline lipase from P. alcaligenes M-1, capable of
removing fatty stains when used in a washing machine.
European Patent Office (EPO) Solvay Enzyme Products,
Inc. 1992-01-29/1990-07-25, is a nonionic and/or anionic
detergent formulation that contains lipase from Pseudo-
monas plantarii (Bycroft and Byng, 1992).
Composition of a U.S. Patent 4,707,291, a detergent,
comprise a mixture of an anionic and a nonionic detergent-
active compound in combination with a lipase similar to
the enzyme produced by Pseudomonas fluorescens IAM
1057, P. gladioli or Chromobacterium viscosum (Detergent
formulations containing alkaline lipase, 1992). Lion has
released an improved version of its powder type laundry
detergent "Top" with two different acting enzyme cleansing
agents (http://www.accessmylibrary.com/comsite5/bin/
pdinventory.pl?pdlanding=1andreferid=2930andpurchase
_type=ITManditem_id=0286-23617629).
FUNGAL LIPASES IN DETERGENTS
During 1980-1990, many different enzymes containing
detergents were developed for softening of clothes. In
1988, Novo Nordisk developed a lipase produced by the
fungus Humicola. Commercial detergent formulations
with high-temperature optima have been produced from
Humicola lanuginosa (Lipolase) (Jaeger et al., 1994). As
the quantity of this enzyme was less than required for its
commercial application, therefore, the yield of enzyme
was increased by cloning the gene coding for this lipase
in Aspergillus oryzae. This fungus produced increased
quantity of the enzyme that can be used commercially in
detergents(http://www.efbweb.org/topics/genetic/menu4_
3.htm). The first commercial lipase, Lipolase, was intro-
duced by Novo Nordisk In 1994. This enzyme was
produced from Trichoderma lanuginosus and was
expressed in A. oryzae (Hoq et al., 1985). They also
produced a lipase containing detergent ‘LipoPrime®’.
Laundering with lipase containing detergents is
generally carried out in alkaline conditions; therefore,
alkaliphilic lipases are preferred for example, lipase
obtained from the A. oryzae (Gerhartz, 1990; Umehara et
al., 1990). A presoak formulation was developed using an
alkaline lipase produced by Trichosporon asahii MSR 54,
used for removing oil stains at ambient temperature
(Kumar et al., 2009).
ABS fungal lipase is used in a many sanitary and waste
treatment liquid formulations. This product exhibits
activities in a wide range of pH. Enzymatic methods are
preferred to chemical methods in many sanitary applications
such as grease trap applications; other applications include
detergents, pre-spotters and industrial cleaning compounds
(http://www.americanbiosystems.com/products/abs_fung
al_lipaseL.php).
OTHER LIPASE CONTAINING DETERGENTS
SEBrite-L detergent containing lipase helps in removing
cosmetics, sebum and fatty food stains from garments,
salad oil, animal fat and butter, suitable to use at lower
washing temperatures and neutral pH. Enzyme-containing
detergents are also used in disinfecting and cleaning
hard surfaces in food manufacturing plants and buildings,
contaminated by bacteria and fungi. SEBalase-M and
SEBrite-M detergents with protease and lipase enzymes,
respectively, are used to effectively remove proteinaceous
debris and fat lubricating oils from medical instruments
(http://www.specialtyenzymes.com/detergents.shtml).
Lipases that deal with greasy soil on cloths leave
residual "sick" odors on the cloth. Some detergent
compositions are produced which have fragrances and
have the ability to counter the bad odors due to lipase in
detergent products (Yet2.com).
Enzyme containing laundry detergents marketed by
Procter and Gamble (P&G) includes; tide, ivory snow,
dreft, cheer, and era, perform well. The other benefits to
consumers are better performance, the abilities to
remove fatty stains at low wash temperatures, improve
whiteness, soften fabrics, brighten their colors and
perform in the presence of chemicals (bleach, builder,
surfactant, etc.). P&G searches for microorganisms/
genes which can fulfill consumer need, e.g. an alkaline
cold wash enzyme by a limited number of cycles of
directed evolution is a major challenge in using muta-
genically improved strain of microorganisms/a gene.
Thousands of mutants have to be tested to find the best
strain (http://www.scienceinthebox.com/en_UK/glossary/
Enzymes2_en.html).
Immobilized lipase (Patent # 6,265,191, issued
07/24/2001) is used on surfaces for easy removal of oil
and to change the wettability of fabric surfaces. Lipase in
detergents forms a fabric-lipase complex on the surface
of cloth and removes oil stain and makes them resist
denaturation in the presence of surfactants and de-
activation by heat. This complex resists the removal of
enzyme from the surface of fabric during washing and it
remains active stains after drying of fabric at higher
temperatures, during storage and wear of fabric. Lipase
impedes the oil re-deposition during laundering of fabric.
Oil hydrolysis by-products are removed during washing of
fabric at a basic pH or in the presence of a surfactant
(http://www.kstate.edu/tech.transfer/macc/LaundryEnzym
es.htm).
Two laundry products (a liquid and a solid enzyme) by
Gateway ProClean Inc. USA, breaks down soil into
simpler forms which can then be easily removed. Lipase
in these products remove fat-containing stains of frying
fats, salad oils, butter, fat-based sauces, soups, human
sebum or certain cosmetics (http://www.gatewayproclean.
com/GatewayProCleanInc.USA).
During manufacturing beverages such as wine, fruit
juice, beer and milk are commonly filtered through ultra-
and nano-filters. These filters are fouled with protein,
cellulose, starches, fats, and pectin residues and are
cleaned and disinfected with caustic and chlorine. These
Hasan et al. 4841
can be effectively cleaned using enzyme-containing
detergents like SEB-Prolase P or SEBalase-PB (protease),
CelluSEB-T (carbohydrase), SEBamyl-B (alpha-amylase),
SEBrite-L (lipase), and ExtractSEB-R (pectinase) (http://
www.specialtyenzymes.com/detergents.shtml).
Novozyme launched a low-temperature detergent
enzyme suitable that can be used in the detergents
usually used for hand-washing in developing countries
like China and India. Another product, Polarzyme, can
clean clothes in cold water, and also it has attractive price
(Press Release, 2005).
Enzymes and detergents are used to clean medical
instruments since the last two decades. Multiple enzyme/
detergent instrument cleaners can also remove the
microbial biofilms (Enzymatic instrument cleaners in
health care) (http://www.highbeam.com/doc/1G1-
83061938.html).
Another common application of enzymatic detergents is
cleaning of gastrointestinal endoscopes before disinfection.
Misuses of enzyme detergents include failure to dilute the
detergent, over dilution, use of expired detergent, in-
adequate exposure time, and failure to rinse the detergent
properly from the instrument (http://www.highbeam.
com/doc/1G1-83061938.html).
ISI enzymatic detergents contain the enzymes that can
remove the stubborn organic materials. A pre-cleaning
enzymatic foam developed by ISI, Hemaway enzymatic
foam, is ideal for use in the immediate cleaning of
surgical instruments. Another product by ISI, Hemaway
Tri-Max contain three enzymes for enhanced cleaning
and removing tough organic materials from surgical
instruments (Hutchisson and LeBlanc, 2005). A unique
enzyme formulation, Endozime® AW Plus, can rapidly
remove fat, blood, starch, carbohydrates and protein from
scopes and surgical instruments, cleans the most difficult
instruments (that is, orthopaedic, laproscopic) and is
safer to use on the most delicate (that is, ophthalmic,
microsurgical) instruments. It is a mixture of enzymes
(protease, amylase, lipase and carbohydrase), buffers
and nonionic detergents (http://www.ruhof.com/Catalog
Products.asp?nProductsID=79).
Protease and lipase in the lens cleaning system
considerably enhances the removal of proteinaceous and
lipid material that accumulates on the contact lens
(http://www.enzymetechnicalassoc.org/benefits_paper.pd
f). POLYZYM
®
enzymatic cleaning tablets loosen organic
deposits on the lens surface and remove it easily. Clear-
Lens (TM) Lipo is also a lipase preparation available for
cleaning contact lenses (http://www.novonordisk.com/
Reports/investors/annualreports/ar1997/eb/
prodintros.htm).
Besides increasing detergency, lipase component in
detergents also prevents scaling. The cost of production
of these enzymes is very high. This limitation may be
overcomed by employing molecular techniques, thus
enhancing the production of these enzymes at high levels
and in purified form (Houde et al., 2004).
4842 Afr. J. Biotechnol.
Table 3. Commercial enzymes used in detergent formulations and other applications.
Trade Names
Manufacturer
Alkaline proteases/ subtilisins
Alclase
Novo Nordisk , Denmark
Savinase
Novo Nordisk, Denmark
Esperase
Novo Nordisk, Denmark
Liquanase
Novo Nordisk, Denmark
Everlase
Novo Nordisk, Denmark
Ovozyme
Novo Nordisk, Denmark
Polarzyme
Novo Nordisk, Denmark
Maxacal
Gist-brocades, Delft, The Netherlands
Maxatase
Gist-brocades, Delft, The Netherlands
Opticlean
Solvay Enzymes GmbH, Hannover, Germany
Optimase
Solvay Germany
Protosol
Advanced Biochemicals Ltd., Thane, India
Alkaline protease “Wuxi”
Wuxi Synder Bioproducers Ltd., China
Proleather
Amano Pharmaceuticals Ltd., Nagoya, Japan
Protease P
Amano, Japan
Durazym
Novo Nordisk, Denmark
Maxapem
Solvay, Germany
Purafect
Genencor International Inc., Rochester, USA
Amylases
BAN
Novo Nordisk, Denmark
Termamyl
Novo Nordisk, Denmark
Stainzyme
Novo Nordisk, Denmark
Duramyl
Novo Nordisk, Denmark
Maxamyl
Gist-brocades, Delft, The Netherlands
Solvay amylase
Solvay, Germany
Lipases
Lipolase
Novo Nordisk, Denmark
Lumafast
Genencor, USA
Lipofast
Advanced Biochemicals, India
Cellulases
Celluzyme
Novo Nordisk, Denmark
Endolase
Novo Nordisk, Denmark
Mannanase
Mannaway
Novo Nordisk, Denmark
(Novozyme report, 2006; Kumar et al., 1998).
Extensive and continued research is going on to
develop lipases, which will work under various suitable
conditions as fat removers. There is a lot to be done to
extend and increase the use of enzymes in detergents
globally. Unfortunately, the enzyme detergents are not
used much in developing countries which are compa-
ratively more hot and dusty, which requires frequent
washing of clothes. Enzymes such as glucose oxidase,
lipoxygenase and glycerol oxidase can generate hydrogen
peroxide in situ. Peroxidases may help enhance the
bleaching efficacy of peroxide (Chaplin, 2004).
There is still a need of extensive and continuous
screening for new microorganisms that have the ability to
produce lipolytic enzymes with enhanced activities at low
temperature, have stability at high temperatures, active at
alkaline conditions, higher stabilities in the presence of
other additives usually present in a detergent. The most
important thing among all is to create awareness in the
developing world about the ‘pros of enzyme detergents
and ‘cons’ of the chemical detergents they are
continuously exposed to.
Developments in enzymology and recombinant DNA
technologies, led to the current era of bioengineered
enzymes. Many industries of the world are involved in the
manufacture and trade of enzymes used in detergents.
Presently, three major giants, Novo Nordisk (Novozymes
A/S, North America), Genencor International Inc.
(California, USA) and DSM N.V. Enzymes (Netherlands),
are leading production of industrial enzymes. Novo
Nordisk (Novozymes A/S) markets a range of enzymes
for various industrial purposes (Table 3) (Kumar et al.,
1998), mostly derived from microorganisms, such as
various strains of alkalophilic Bacillus sp. or genetic
and/or protein engineered organisms (Kumar et al.,
1998). They also produce enzymes, other than lipases,
which are used in detergent industry. Many of these
enzymes are produced by fermentation of genetically
modified microorganisms (Table 3).
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... Laundry detergents are becoming more and more popular because of their increasing use in washing machine, where it is soft, resilient to fabrics, antistatic, dispersible in water and mild to skins [4]. There are concerns over persistence of detergent chemicals in the environment and its possible contamination of ground water. ...
... Their subsequent health related issues have raised speculation over biodegradability. This has brought about an increasing preference for greener detergents and inadvertently, the need for greater consumption of enzymes as a detergent component [4]. ...
... Enzymes can reduce the environmental load of detergent products as the chemicals used in conventional detergents are reduced; they are biodegradable, non-toxic and leave no harmful residues [4]. More focus has been drawn to keratinases produced from bacteria because of their actions on both insoluble keratin substrates as well as a wide range of other protein substrates [5]. ...
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... other enzymes; (iv) broad substrate specificity; (v) solubility in water, etc. [30,166]. They are required as detergent additives to remove the tough fatty residues, e.g., of oils, fats, butter, the stains on collars and cuffs, and to clean clogged drainage pipes [97,163,167]. Their addition also has other benefits like rendering the detergent eco-friendlier as it takes less time to remove stains so the process is less energy consuming, producing effluents with lesser COD and imparting softness, anti-staticness to fabrics as well as maintaining fabric quality [167]. Many bacterial lipases have been isolated and tested according to the above-mentioned criteria and are being used commercially as detergent additives. ...
... They are required as detergent additives to remove the tough fatty residues, e.g., of oils, fats, butter, the stains on collars and cuffs, and to clean clogged drainage pipes [97,163,167]. Their addition also has other benefits like rendering the detergent eco-friendlier as it takes less time to remove stains so the process is less energy consuming, producing effluents with lesser COD and imparting softness, anti-staticness to fabrics as well as maintaining fabric quality [167]. Many bacterial lipases have been isolated and tested according to the above-mentioned criteria and are being used commercially as detergent additives. ...
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... Using the enzymes in immobilized form allows their easy retention and repeated use in continuous reaction systems, which results in a decreased overall cost of the process. Enzyme immobilization, which is the attachment of enzyme molecules to solid supports [7], has been proposed for better reusability by improving enzyme stabilization, as it has been reported to enhance the enzymes' thermal and shear stability [8], as well as simplifying the separation [7]. Furthermore, immobilization inside a porous support could protect enzyme molecules from exposure to harsh media. ...
... Using the enzymes in immobilized form allows their easy retention and repeated use in continuous reaction systems, which results in a decreased overall cost of the process. Enzyme immobilization, which is the attachment of enzyme molecules to solid supports [7], has been proposed for better reusability by improving enzyme stabilization, as it has been reported to enhance the enzymes' thermal and shear stability [8], as well as simplifying the separation [7]. Furthermore, immobilization inside a porous support could protect enzyme molecules from exposure to harsh media. ...
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Immobilization has been proposed as a way to simplify the separation and repeated reuse of enzymes, which is essential for their feasible application at industrial scales. However, in their immobilized form, enzyme activity is fully utilized, due primarily to the additional diffusion limitations. Here, the immobilization of lipase on zeolite and its use in catalyzing oil hydrolysis is studied. Adsorption isotherms were investigated, and the data identified the model that best describes the process, which is the Sips model. The adsorption capacity of zeolite was determined as 62.6 mg/g, which is relatively high due to the high porosity of the support. The rate of enzymatic hydrolysis of olive oil, using the immobilized lipase, was determined at a pH of 7 and a temperature of 40 °C and was compared to that when using free enzymes. The results determined the parameters for a diffusion-reaction model. The effects of both the surface reaction and diffusion were found to be significant, with a slightly higher effect from surface reactions.
... Microbial enzymes such as lipase, protease, cellulase, amylase, mannanase and peroxidase are added to detergents to catalyse chemical bond breakdown under high temperature (60°C) and highly alkaline (pH 9-11) conditions. These enzymes help remove protein stains, insoluble starch in dishwashing, oils and fats, and they also help detergents work better (Hasan et al., 2010;Keshwani, Malhotra & Kharkwal, 2015). ...
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Microorganisms' biocatalytic capacity has been used to make bread, wine, vinegar, and other familiar items for generations, although the biochemical foundation of their constituents is unknown. Microbial enzymes have gained appeal for their widespread application in industries and medicine due to their stability, catalytic activity, and ease of production procedures and optimization. Enzymes' usage in a variety of sectors (e.g., food, agriculture, chemicals, and medications) is fast rising due to its shorter processing time, low energy input, cost efficiency, nontoxic, and environmentally benign features. Toxic chemical substances in industrial and residential wastes (nitriles, amines, phenolic compounds and so on) can also be degraded or transformed by microbial enzymes.
... Other studies have shown that these fungal lipolytic enzymes can reduce an environmental load of detergent products as they gain energy that enables the use of a lower wash temperature (Singh & Mukhopadhyay, 2012;Vakhlu & Kour, 2006). In addition, lipases from Aspergillus oryzae, Humicola lanuginosa, and R. oryzaeare used in detergents, allowing better washing performances and energy saving (Hasan et al., 2010). Das et al. (2016) tested the ability of A. tamarii lipase to clean the peanut oil stain used for deep frying. ...
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Many researchers have found fungi as a reliable source of lipase due to the versatility of their properties, ease of mass production, thermal stability, pH stability, broad substrate specificity, retained activity in organic solvents, and their low cost extraction procedure. This review paper presents an overview about the main aspects of fungal lipases screened from several types of strains, as well as their use as biocatalysts. Additionally, some biochemical properties will be reported. As commonly known, lipases can be produced from animals, plants and microorganisms. Compared to other lipases, those obtained from fungi have been found to be more productive, a fact that encouraged the massive production of most fungal lipases due to their considerable commercial importance during the past few years. This paper is concerned about some of the major characteristics that made fungal lipases desirable products in the industrial fields. Due to the enantioselective properties of fungal lipases and their ability to remain active under extreme temperature, pH and organic solvents, enzymes are capable to synthesize esters as well as to catalyze a variety of chemical reactions that include esterification, transesterification, acidolysis and aminolysis in aqueous and non‐aqueous media. Furthermore, lipases are considered to have a commercial importance for biotechnological application fields, which makes them increasingly popular in food, detergent, cosmetic, organic synthesis, and pharmaceutical domains. The biotechnological potential of lipases has made the latter a coveted choice in industries for the present and future as biocatalysts. In addition, a classification of these fungal enzymes is also highlighted in this review. Moreover, the impact of an immobilization strategy of these fungal strains to achieve higher yields and to improve their production is discussed. Finally, fungal enzymes have played a crucial role from ancient times to today in different fields using several types of biological systems, which gives them a great interest for the production of these enzymes in large amounts with low cost and easy viability to enlarge their use in many industries. Likewise, some future perspectives on lipase production will also be discussed by focusing on special cases on lipase engineering. This article is protected by copyright. All rights reserved.
... Aplikasi komersial utama dari enzim lipase hidrolitik yaitu kegunaanya sebagai deterjen [39] karena kemampuannya dalam menghidrolisis lemak, sehingga enzim lipase secara luas digunakan untuk industri laundry dan deterjen rumah tangga. Enzim lipase deterjen secara khusus dipilih karena spesifisitas substrat yang rendah, sehingga mampu menghidrolisis lemak dari berbagai komposisi, memiliki kemampuan untuk bertahan pada kondisi pencucian yang ekstrim yakni pada pH 10-11 dan pada suhu 30-60°C serta memiliki kemampuan untuk bertahan dari kerusakan karena surfaktan dan enzim-enzim lain yang merupakan komponen penting dari deterjen enzim [40]. ...
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Enzim lipase adalah hidrolase asil yang berfungsi dalam pencernaan dan pengolahan lemak. Enzim lipase memiliki akivitas yang dapat menghidrolisis berbagai lemak dan minyak, setiap satu unit per mL (U/mL) dari aktivitas enzim lipase dapat membebaskan 1 µmol asam lemak bebas per menit. Aktivitas enzim lipase pada kondisi optimum yang diperoleh dari pengukuran aktivitas enzimatik pada modifikasi suhu serta pH. Enzim lipase dapat ditemukan pada tumbuhan, hewan, dan mikroorganisme. Kelapa (Cocos nucifera L.) merupakan salah satu tanaman penghasil enzim lipase yang dapat menghasilkan Virgin Coconut Oil (VCO). VCO dapat diubah menjadi diacylglicerol (DAG) dengan hidrolisis ataupun trans-esterifikasi memakai metode kimiawi atau enzimatik. DAG adalah jenis minyak sehat yang dapat mengurangi kandungan Low Density Lipoprotein (LDL), trigliserida (TG) serta menjadi penghambat plasminogen. Kata kunci : Lipase, Diacyglycerol (DAG), Virgin Coconut Oil (VCO)
... Lipases are used in several biotechnological applications, and some of them are carried out in conditions that could be withstood by the crustacean extracts studied here. Some examples are milk fat lipolysis (Fraga et al., 2018), oil wastes (Wang et al., 2007) and wastewater (Affes et al., 2017) treatments and detergent industry (Hasan et al., 2010). Lastly, Ot amylase activity showed stability at 10, 30 and 50 • C, although high activity is observed solely during the first 30 min of the assay. ...
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Marine natural compounds are required by industries to improve and reduce costs of many biotechnological processes. In this work, specific activity of midgut gland digestive enzymes was measured in seven species of decapods. Alkaline and acid peptidase, lipase and amylase activities were registered in Pleoticus muelleri (Pm), Artemesia longinaris (Al), Paralomis granulosa (Pg), Lithodes santolla (Ls), Munida gregaria (Mg), Neohelice granulata (Ng), and Ovalipes trimaculatus (Ot). The highest activities for all the analyzed digestive enzymes were obtained in Pg, Ls, Mg and Ot, which were used for optimum temperature and pH measurements and to test their stability at different temperatures and pH. In general, optimal enzyme temperatures were between 30 and 50 °C in the four crustaceans. Peptidases and lipases from these species showed in general alkaline optimal pH except for those from Ot which presented high activity at the whole pH range (2–12). Amylase from Ot presented its optimal at pH 6–8 and 30–50 °C. Thermal stability of peptidases was elevated at 30 °C for the four species tested (Pg; Ls; Mg and Ot), lipases were stable mainly at 30–50 °C and amylase was stable at 10 °C, 30 °C and 50 °C during the first 30 min of the assay. In general, stability of the four enzymes from Pg was greater at alkaline pH than at acid. The other species showed stability at all pH for the four enzymes. The range of temperature and pH of the tested enzymes activity are coincident with those used in various biotechnological processes and therefore indicates the potential of these enzymes to be used for industrial procedures. Further, most of the species studied are converted into fishery residues, either by processing or by-catch, so their reuse would also contribute to the sustainability of the activity.
... The detergent industry is the largest market for enzyme production in the world [40,41]. All types of detergents, ranging from those produced for household laundering to reagents used for cleaning of contact lenses, use proteases as a standard ingredient [41]. ...
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