FACULTY OF ENGINEERING

HK03 CHEMICAL ENGINEERING PROGRAMME

KC 46603 ENZYME ENGINEERING
SEMESTER 2 2017/2018

GROUP ASSIGNMENT

TITLE: WASTE TREATMENT

SUBMISSION DATE: 11th MAY 2018

LECTURER: DR MARIANI RAJIN

NO. NAME MATRIC NO

1. ANEESCH PREETHA A/P VIDA NAIGAM BK 13110587
2. SAVITRA LAKSMI A/P VEJAYAN BK 14110213
3. SHARMINI NAIR A/P PRATHABAN BK 14110217
4. THURGA DEVI A/P VIJAYAKUMAR BK 14110240
5. YAMUNAH A/P JAIBALAH SINGAM BK 14110248
6. YOGANANTHINY A/P JAGADESON BK 14110249
 TABLE OF CONTENT

Executive Summary 1

1.0 Introduction 2

2.0 Application of Enzymes in Particular Waste Treatment 4

3.0 Advantages and Disadvantages Compared to Conventional Method 10

4.0 Challenges in Enzyme Engineering Application 14

5.0 References 17

2
 Executive Summary

The introduction and implementation of stringent standards for waste discharge

into the environment has urged the need for the development of alternative waste
treatment processes. Enzymes from various microbes have been reported to play a
crucial role in waste treatment applications. Enzymes or biological catalysts can act
specifically and remove recalcitrant pollutants by precipitation and transform it other
products by changing the characteristics of a given waste making it more suitable for
treatment or help in converting waste material to valuable products. Enzymes are
widely used industries for waste treatment. The advantages of enzymes are ability of
the enzymes to work in mild reaction conditions, do not require protection of substrate
functional group and have a long half-life. Furthermore, enzymes were chosen to treat
waste because it is environmental friendly and reduces the amount of pollutants which
results in adverse impacts in human health and environment. There are various types
of waste can be treated by enzymes, however the treatment of solid wastes,
wastewater, soil and hazardous wastes is focused in this assignment. This assignment
also includes the advantages and disadvantages of enzyme application compared to
conventional method and challenges in the application of enzymes in waste treatment.

1
1.0 Introduction
Enzymes are biological catalyst produced by living cells to cause specific biochemical
reactions generally forming the various metabolic processes of the cells and are
indispensable to maintenance and activity of life. Enzymes are highly specific in their
action on substrates and often many different enzymes are required to bring about
sequence of metabolic reactions performed by living cells (Pandey, Singh, Pandey, &
Badruddin, 2017a). Enzymes are usually derived from microorganisms. Each type and
strain of microorganism produces many enzymes which perform different functions
such as hydrolysing, oxidizing or reducing and metabolic in nature. Microbial enzymes
are known to play important role as metabolic or biological catalyst which makes them
to be preferred in various industrial applications.

The end use in various market for industrial enzymes is extremely widespread
with numerous industrial commercial application. Microbes have served and continue
to serve as one of the biggest source for many enzymes. The disadvantages of
industrial processes have become the main reason for enzymes to be preferred in
industries. The disadvantages are processes with low catalytic efficiency, lack of
enantiomeric specificity for chiral synthesis, requirement of high temperature, low pH
and high pressure. Furthermore, the use of various chemical and organic solvents to
improve the catalytic activity of the reaction has increased the amount of waste and
pollutants produced by the companies.

Enzymes seems to be a solution for the problems because they work under
mild reaction conditions. Besides that, enzymes also do not require protection for
substrate functional groups, have a long half-life, high stereo selectivity yielding stereo
and regio-chemically-defined products at an acceleration of 105 to 108-fold and
enzymes also works on synthetic substrates. Moreover, enzymes also can be selected
genetically and modified to improve its key properties such as stability, substrate
specificity and specific activity. Thus, it can be concluded that enzymes have a wide
application in various industries in Malaysia. Enzymes or biological catalyst not only
can be used in the production of products such as yogurt, alcohol and detergent but
it also can be used to treat waste produced from the industries.

2
 Research has been conducted to investigate the possibilities and applications
offered by enzymes in waste treatment. The are three main reasons which lead to the
research. Firstly, the rate of introduction of xenobiotics and recalcitrant organic
pollutants into the environment is on the rise and it is difficult to achieve the required
degree of removal of these pollutants using conventional chemical and biological
processes; therefore, there is a need for the development of alternative treatment
methods that are faster, cheaper, more reliable and simpler to be implement than
current processes. Secondly, there is a growing recognition that enzymes can be used
to target specific pollutants for treatment. Finally, recent biotechnological advances
have allowed the production of cheaper and more readily available enzymes through
better isolation and purification procedures (Nicell, Karam, & Nicell, 2015) .

Waste treatment processes can be characterized as either physico-chemical or

biological processes. Enzymatic treatment falls between these two traditional
categories because it involves chemical processes based on the action of biological
catalysts. There are several advantages of enzymatic treatment; which are application
of biorefractory compounds; operation at high and low contaminant concentrations;
wider range of pH usage, temperature and salinity, reduction in shock loading,
absence of delays associated with the acclimation of biomass, simplicity of controlling
the process and reduction in sludge volume as no biomass is produced (Nicell et al.,
2015).

The potential advantages have allowed the development of enzyme treatment

processes for the treatment of wastewaters, solid wastes, hazardous wastes and soils.
The objective of this assignment is to provide a summary of application of enzyme in
treatment of wastes from different industries and compare the advantages and
disadvantages with conventional method.

3
2.0 Application of Enzymes in Particular Waste Treatment
a) Wastewater Treatment

As mentioned earlier, enzymes are one of the biocatalysts that were produced
by the living cells. These living cells cause specific biochemical reactions and they
formed various metabolic processes of the cells and are indispensable to maintenance
and activity of life. Microbial enzymes have a very important role or function as
metabolic catalysts which caused them to be used in various industrial applications.
Waste treatment can be categorised as solid waste and wastewater treatment(Pandey,
Singh, Pandey, & Badruddin, 2017b).

Wastewater is obtained from the discharge of inadequately treated industrial

and domestic purposes activities. Indirectly, wastewater cause high level of inorganic
pollutants in the ocean or any river discharge. Thus, enzyme engineering plays a vital
role in treating the wastewater by using some active enzymes especially microbial
enzymes as mentioned above. Usually aerobic biological process is one of the most
important or frequent used method in industrial treatment as they have high degree
treatment rather than anaerobic treatment. Anaerobic treatment is done based on the
concept of the resource recovery. Waste treatment processes can be categorised into
two different categories which are chemical or biological processes. So, enzymatic
treatment falls between these two conventional categories as they involve chemical
processes based on the action of biological catalysts.

One of the main industries that play a vital role in applying enzyme engineering
is pulp and paper industry as enzyme technologies can reduce environmental problems
(Kenealy & Jeffries, 2003)The enzymes that can be used for this industry are such as
below:

 Microbial xylanases
- Through eliminating the need for chlorine in the manufacture of
elemental chlorine free (ECF) and totally chlorine free (TCF) printing
and writing paper grades.

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 Lipases

-Control the accumulation of pitch during the production of paper

from pulps with high resin content, such as sulfite and mechanical pulps.
 Xylanases

-Increasing the bleaching speed

 Laccase
-Increase the bonding of fiber to strengthen the paper
 Cellulase

- To increase the brightness of paper

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 b) Hazardous Waste Treatment

Another category that required enzymes application is that management of hazardous

waste. Hazardous waste can be categorised into heavy metals waste and cyanide
waste. Cyanide waste is known as the waste that is obtained from the production of
chemical intermediates, synthetic plastics, rubber or any other leaching process. As
cyanide is a metallic inhibitor and can be one of harmful substances to living organism
especially human, so it should be inhibited from the discharge of effluents with
application of enzymes.

Examples of enzymes that been used for the cyanide waste management
are such as below:

 Cyanidase
- Capable of converting the cyanide to ammonia and formate
- Based on gram negative bacteria
- Have high affinity and high stability towards cyanide
 Cyanide Hydratase
-Known as formamide hydrolyase as it hydrolyses cyanide to formamide
- Through immobilisation enzyme application in industrial effluent

Meanwhile, heavy metals be categorised as hazardous waste too as they

contain arsenic, chromium, lead, and others that can be found from industrial and
mining sectors. They should be avoided from being discharged into any water bodies
as they contain high toxicity rather than other components. The ways to inhibit the
wastes are such as below:

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  Usage of Citrobacter sp

- Done by immobilisation process in polyacrylamide gel where the metals

contain solutions were passed through
- Removal of cadmium and copper
- Around 90% metals removed, and enzymes are being reused

However, these enzymes should be used in correct amount to prevent any

negative effects. So, an enzyme dose of 2.5 to 5 mg dm gave good results, an
increased dose led to the creation of more hydrophilic end groups which negatively
affected the dewatering process.

c) Solid Waste Treatment

Enzyme application can be seen in solid waste treatment. Enzymes in this solid
waste treatment is used to improve the dewatering sludge produced as a by-product
of wastewater treatment. Dewatering is basic to diminish the mass of water contained
in slimes that are to be burned or disposed of into landfills. Enzymes such as
carboyhdrase, lipase and protease are used to improve the dewaterability of
sewerage sludge and to increase the quantity of water release during pressing to
achieve a reduction in sludge volume. Enzyme addition is used to cause the cleavage
of the water binding macromolecules. Phosphatic slimes which contains considerable
amounts of swelling clay-like material and their fine particle size in the solid waste is
difficult to dewater. Peroxidase enzyme is used to improve the dewatering of these
slimes. Peroxidase treatment of the slimes instigated a higher mechanical binding
among slimes particles and that peroxidase altogether advanced the development of
algae and mold with the advantageous impact of aggregation of the particles with

7
high viscosity and gel strength. Higher viscosity and higher gel strength will help in
the post- sedimentation settling.

Horseradish peroxidase (HRP) enzyme is the example of peroxidase type

enzyme. HRP catalysed the oxidation of the toxic aromatic compounds in the solid
waste treatment when it is activated by peroxidase (Nicell, 2001). The reaction
products are polymerized through a non-enzymatic process which produces the water-
insoluble precipitates that can easily remove from water or wastewater by
sedimentation or filtration. HCP is suitable because of its properties which are wide-
ranging pH and temperature.

Besides that, use of cellulose and bacterial enzyme such as lysozyme or

muramidase is used in sludge dewatering. Use of cellulose with penicillin gave poorer
results when compared to the lysozyme enzyme. Lysozyme enzyme apparently able
to floc matrices and causes rapid increase in the dewatering rates.

d) Soil Contamination Treatment

Soil which is the major component of terrestrial ecosystems, which performs

many important functions to the environment. However, soil contamination has
become major problem in the ecosystem. Soil contamination or soil pollution is caused
by the presence of xenobiotic which is the human made chemicals or other alteration
in the natural soil environment. Major causes of this problem are caused by industrial
activity, agricultural chemicals or improper disposal of waste. To overcome this
problem, enzyme application is used.

Some important groups of enzymes which are used in soil contamination are
constituted by oxidative enzymes. They have large role in the formation of humus
material in soil, exchange between plants and soil through the degradation and
synthetic mechanisms. Other than that, they can involve in the reaction between
humus constituents and xenobiotics molecules and form bound residues in water
system and humus materials in soil. One of the application of enzyme in soil
contamination is dehydrogenase.

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 Dehydrogenase is used in treating soil contamination caused by the petroleum
products (Kaczynska et.al,2015). Stimulation of dehydrogenase by compost, both in
contaminated and non-contaminated soils, proves that it may accelerate
microbiological degradation of petroleum-derived contaminants. Dehydrogenase is
often considered as an index of the general microbiological activity of the soil as the
important of dehydrogenase as a pollution indicator is supported by the properties
which is the lack of the ability to accumulate in the extracellular environment.
Dehydrogenase consists in the biological oxidation of organic matter in the soil by
hydrogen transfer from the organic substrate to organic acceptors.

Other than that, to overcome the soil contamination by copper, enzyme

dehydrogenase, urease and acid and alkaline phosphatases is used. Soils
which are contaminated by copper are from industrial and communal waste, sewage
sludge and industrial dust emissions (Wyszkowska et.al,2005). The greater the
amount of mineral and organic colloids in soil, the lesser the negative effects of
contamination with metals. Soil contamination with copper significantly inhibited the
activity of dehydrogenase, urease and acid and alkaline phosphatase. From the paper
of (Wyszkowska et al., 2005), it shows that when compared with phosphatases
enzyme, dehydrogenase and urease appeared to be better indicators of soil
contamination with copper, as their activity was strongly inhibited by copper than the
activity of the phosphatases.

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3.0 Advantages and Disadvantages Compared to Conventional
Method
The conventional method of treating waste are chemical and biological treatment
systems. In particular, the implementation of increasingly stringent standards for the
discharge of wastes into the environment has necessitated the need for the
development of alternative processes for the production of goods and for the
treatment and disposal of wastes compared to the two traditional method mentioned
before.

Chemical transformations involve the application of reagents and reaction

conditions to chemically alter species through a chain of events resulting in the yield
of a desired product. Such processes involve a string of events that are usually well
defined and can be controlled to maximize efficiency. However, such processes often
require the presence of excess quantities of reagents to accomplish the transformation
to the desired extent. In addition, particularly harsh conditions (e.g., high temperature
or extremes of pH) are often required in order to facilitate the chemical
transformations. This can present a problem once the desired transformation has
taken place since the resulting stream may be a low quality mixture that cannot be
disposed into the environment or reused without subsequent treatment. And, finally,
many chemical treatment processes are not highly selective in terms of the types of
pollutants that are transformed during treatment. Consequently, such processes are
usually more economical for the treatment of dilute wastewaters and are often used
as a polishing step prior to waste discharge into the environment (Aitken, 1993).

Another success method has been used for many decades is biological
processes. These processes are designed to take advantage of the biochemical
reactions that are carried out in living cells. Such processes make use of the natural
metabolism of cells to accomplish the transformation or production of chemical
species. The metabolic processes occur as a result of aa sequence of reactions
conducted inside the cell that are catalysed by proteins called enzymes. Biological
processes can be used to carry out processes for which no efficient chemical
transformations have been devised. In addition, biological processes can often be
conducted without the harsh conditions that are necessary during chemical

10
transformations. However, due to the sensitivity of microorganisms to changes in their
environment (e.g. pH, salinity, temperature, and the presence of toxic or inhibitory
compounds), these processes can be difficult to control over long term, and maybe
subject to frequent upsets. Moreover, it also requires supply of macro and
micronutrients for the support of the microorganism growth, and often result in the
formation of large quantities of biomass that ultimately must be discarded into the
environment. The biochemical reactions occur at a rate that is limited by the
metabolism of the microorganism and, thus, are often slower than chemical processes.
Biological systems are commonly used to remove the bulk organic load in wastewaters,
these systems often have difficulty in removing toxic pollutants to consistently low
levels. Therefore, conventional biological processes may not be able to improve water
quality sufficiently to meet wastewater discharge criteria.

In order to overcome the problems associated with chemical and biological

systems, researches focuses on the environmental applications of pure enzymes that
have been isolated from their parent organisms. There are four main reasons to find
the alternative method and the reasons are as following:

 The rate of introduction of recalcitrant organic pollutants into the

environment is on the rise, and it is becoming increasingly difficult to
achieve an acceptable degree of removal of these pollutants using
conventional chemical and biological processes;
 There is a need for the development of alternative treatment methods
that are faster, cheaper, more reliable, and simpler to implement than
current processes;
 There is a growing recognition that enzymes can be used to target
specific pollutants for treatment;
 Recent biotechnological advances are expected to enable the production
of cheaper and more readily available enzymes through genetic
manipulation of microbial and plant cells and through improved
efficiency of isolation and purification procedures

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 Pure enzymes are often preferred over intact organisms containing the
enzymes because pure enzymes can act with greater specificity, their activity can be
better standardized, easier to handle and store and enzyme concentration is not
dependent on bacterial growth rates. This can lead to some important advantages of
enzymatic processes over biological systems such as:

 their application to compounds that resist biodegradation

 their high selectivity allowing for the treatment of targeted pollutants,
their action on, or in the presence of, many substances which are toxic
to microbes
 their operation over relatively wide temperature, pH and salinity ranges
compared to cultures of microorganism
 their operation both at high and low concentrations of contaminants
 their high reaction rate compared to biological processes which enables
the use of smaller systems of lower cost
 the absence of shock loading effects associated with changes in
contaminant concentrations that overwhelm the microorganism’s ability
to adapt
 the absence of delays associated with shutdown/start-up periods that
are normally required to acclimatize biomass to waste streams
 the lower quantity of sludge production since biomass is not generated
 high system stability allowing for simpler process control.

Enzymes also offers a number of advantages over conventional chemical

processes. The advantages are as following:

 high degree of specificity that allows enzymes to remove target

pollutants selectively which precludes undesirables or unnecessary
reactions which would otherwise increase reactant consumption and
correspondingly, increase the cost of treatment
 operation on compounds that are present in trace quantities or that
cannot remove by existing chemical/physical processes
 operation in a catalytic manner resulting in a high reaction velocity and
efficient use of chemical reagents

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  operation under low temperature conditions, thereby reducing energy
requirements for processes normally conducted at elevated
temperatures
 Operation under mild pH conditions, thereby reducing the impact of
corrosion on reaction vessels and avoiding the need for waste
neutralization

While the above advantages are indeed significant, it should be noted that the
majority of chemical and biological processes are not candidates for replacement by
enzymatic processes. This is because conventional methods have a fundamental
advantage over enzymatic systems where they have the ability to simultaneously
transform a broad range of compounds. For instance, many municipal, agricultural,
and industrial wastes consist of a mixture of organic compounds usually classified
under the broad categories of biological oxygen demand (BOD) or chemical oxygen
demand (COD). Once released into receiving water bodies, these collections of organic
compounds result in the depletion of dissolved oxygen in the water column as a result
of natural microbial processes. In many instances, the majority of these compounds
can be efficiently degraded through the combined action of mixed cultures of
microorganism. In contrast, enzymes are biological catalysts whose actions are
tailored exclusively act upon specific chemical species. Thus, enzymatic treatment will
not result in the removal of a broad range of compounds from a waste stream, but
will only accomplish the transformation of an individual compound or class of
compounds. This limits the application of enzymes to accomplish the transformation
of target species that are either problematic due to their toxicity or that have been
identified as the raw materials from which enzymes can produce value-added
products.

Last but not least, the disadvantages are very much lesser compared to the
advantages of enzymatic processes compared to the conventional methods. Thus, it
is believed that enzymatic processes will be a great alternative.

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4.0 Challenges in Enzyme Engineering Application
a) Enzyme Cost

Although there is potential for enzymatic treatments in wastewater treatment systems,

the development of these processes from an engineering perspective is lacking
(A.Nicell, 2006). The major reason is the cost of the enzyme itself. For industrial scale,
many enzymes are needed, and this takes up an enormous amount of cost. The main
reason enzymes are expensive is because of the cost of their isolation, purification
and production. However, Dec and Bollag (1994) have reported a good practical
solution, which is to use plants or plant materials that contain enzymes directly, the
usage of crude enzyme extracts, and the development of cloned cells that can be
stimulated to produce enzymes efficiently in reactor systems.

Like for instance, the use of plant material to decontaminate water polluted
with phenolic compounds through enzymatic reactions. Peroxidases from minced
horseradish, potato and white radish were used to remove 2,4-dichlorophenol up to
concentrations of 850 mg/L (A.Nicell, 2006). Amazingly, the success rates are
incredible.

b) Toxic Reaction Products

The major reason of enzyme usage in waste treatment is to act on specific pollutants
to convert them into products (A.Nicell, 2006). Contaminants being fully mineralised
is much preferable rather than other transformations. However, the ammonia and
format production from cyanide wastes or the conversion of organophosphate
pesticides to easily removable products are also acceptable (A.Nicell, 2006).
Nonetheless, it must be noted that, in some cases, the products of enzymatic reaction
could be more toxic and harmful than the parent compounds. This defeats the whole
purpose of using enzymes.

So, it is desired that the reaction products be less harmful and toxic, or more
biodegradable than the original pollutant. It can also be more amenable to subsequent
treatment. Yet, there is no single enzyme that is expected to catalyse very extensive
transformations of pollutants. Hence, the reaction products are likely to retain some
of the characteristics of the compound (Aitken, 1993). Which is why, characterization

14
of reaction products is needed to assess their impact on downstream processes or on
the environment into which they are released. Sadly, there are only a few studies
conducted to look up on this issue. This is mainly because of the difficulty in identifying
the products of enzymatic reactions and the difficult task of assessing their toxicity.

For instance, there are several studies conducted to evaluate the toxicity of
trace quantities of soluble products that remain after the treatment of phenols with
peroxidases (A.Nicell, 2006). Ghioureliotis and Nicell (1999) showed that there is a
gradual accumulation of dissolved toxic soluble products during the treatment of
phenol. Yet, these products can be removed without any difficulty using activated
carbon as a polishing step. In addition, Ikehata and Nicell (2000) proved that all
solutions of phenols and chlorophenols that were treated with tyrosinase followed by
the addition of chitosan had remarkably lowered the toxicities. These studies are of
great importance and encouraging, however similar studies must be conducted for
other enzymes considered for use in waste treatment applications.

c) Solid Residues Disposal

Waste treatment industry often deals with solid waste which needs to be disposed
safely. As for the enzymatic application in the waste treatment industry there is also
presence of solid wastes. For instance, although there is no any large amount of solid
residues generated like the biological treatment, some solid residues remains at the
end of the reactions such as the formation of polymer precipitates while treating
phenols with peroxidases, spent adsorbents like talc, chitin or activated carbon used
to remove the soluble products of the enzymatic reactions, residues of plant materials
like raw soybean hulls which are used in place of purified enzymes during treatment,
or formation of metal complexes when phosphatase are used to treat heavy metals
(Nicell).

To treat or solve the solid residues disposal issues during the enzymatic
reactions some solutions have been applied which are as follows. The polymer or
adsorbents can be incinerated to recover energy if the dangerous combustion by-
products emissions can be hindered. However, the residues of plant materials can be
used as compost and soil conditioners, if the pollutants do not leach from them at
considerable rates. As for the issues regarding the heavy metals complexes, it is more

15
difficult to handle due to the unavoidable leaching problem from landfills. But there
are attempts done to recover the metals for industrial reuse might solve this issue.
However, until today none of these problems regarding solid residues disposal has
been approached sufficiently.

d) Restricted Applicability of Enzyme Technology in Waste Treatment

Waste treatment is an unavoidable field study in waste management. Hence, to

manage the waste, treatment is very important. However, there is no any treatment
available which is universally applicable which causes the waste management field to
be highly complex and there are limited number of reliable processes to be used for a
specific application. Hence, it important to develop new process technologies that can
meet the increasing specific needs in the industry. Although the enzyme technology
has been specific and specialised, it plays an important role in the waste treatment
industry.

The specialised market of the enzymatic treatment has limited the application
of enzymes that are already developed commercially for other purposes (Aitken,
1993). Research and development on enzyme generation for waste treatment industry
are restricted only for large-scale uses such as enzymes that can improvise the
digestibility or dewaterability of wastewater treatment sludges (Aitken, 1993).
Moreover, enzymes with broad substrate specificities can be suitable for the waste
treatment markets due to the pollutants diversity found in wastes. This kind of broad-
specificity enzymes can change a range of compounds in a given class. Some examples
of enzymes include the organophosphate pesticide hydrolases and phenol-oxidizing
enzymes.

Regardless of the relatively restricted enzyme technology application in the

waste treatment industry, there are enzymatic processes available that are beneficial.
For most enzymes that are declared as candidates for waste treatment applications,
little work has been done beyond demonstrating target compound transformation
(Aitken, 1993). Other than that, there are many technical problems which needed to
be explored before judging the enzymatic treatment processes to be technically and
economically feasible since this is only the first step toward development of a viable
commercial process.

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5.0 References
Aitken, M. D. (1993). Waste treatment applications of enzymes: opportunities and
obstacles. The Chemical Engineering Journal, 52(2), 49–58.
https://doi.org/10.1016/0300-9467(93)80057-U

Dec, J. and Bollag, J.-M., “Use of Plant Material for the Decontamination of Water
Polluted with Phenols”, Biotechnology and Bioengineering, Vol. 44, 1994, pp.
1132- 1139.

Ghioureliotis, M. and Nicell, J.A., “Assessment of Soluble Products of Peroxidase

Catalyzed Polymerization of Aqueous Phenol”, Enyzme and Microbial
Technology, Vol. 25, 1999, pp. 185-193.
Ikehata, K. and Nicell, J.A., “Assessment of the Products of Tyrosinase-
Catalyzed Oxidation of Phenols”, Process Biotechnology Vol. 16, No. 4, 2000,
pp. 533-540.
Jim A. Nicell, Ph.D., P.Eng. ENVIRONMENTAL APPLICATIONS OF ENZYMES
Department of Civil Engineering and Applied Mechanics, McGill University, 817
Sherbrooke Street West, Montreal, Quebec, Canada, H3A 2K6. (n.d.).

Kaczyńska, G., Borowik, A., & Wyszkowska, J. (2015). Soil Dehydrogenases as an

Indicator of Contamination of the Environment with Petroleum Products. Water,
Air, and Soil Pollution, 226(11). https://doi.org/10.1007/s11270-015-2642-9

Kenealy, W. R., & Jeffries, T. W. (2003). Enzyme Processes for Pulp and Paper: A
Review of Recent Developments Enzyme Applications in Pulp and Paper. Wood
Deterioration and Preservation Advances in Our Changing World, 210–239.
https://doi.org/10.1021/bk-2003-0845.ch012

Nicell, J. A. (2001). Environmental applications of enzymes. Interdisciplinary

Environmental Review, 3(1), 14. https://doi.org/10.1504/IER.2001.053866

Nicell, J. A., Karam, J., & Nicell, J. A. (2015). Review Potential Applications of

Enzymes in Waste Treatment, (September).

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Pandey, K., Singh, B., Pandey, A. K., & Badruddin, I. J. (2017b). Application of
Microbial Enzymes in Industrial Waste Water Treatment. International Journal of
Current Microbiology and Applied Sciences, 6(8), 1243–1254.

Wyszkowska, J., Kucharski, J., & Lajszner, W. (2005). Enzymatic Activities in Different
Soils Contaminated with Copper, 14(5), 659–664.

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