Potential Uses of Immobilized Bacteria, Fungi, Algae, and Their

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Chapter 15

Potential Uses of Immobilized Bacteria, Fungi, Algae, and Their Aggregates for Treatment of Organic and Inorganic Pollutants in Wastewater Manab Das and Alok Adholeya* The Energy and Resources Institute, I H C, Darbari Seth Block, Lodhi Road, New Delhi 110003 *E-mail: [email protected].

Bioremediation of wastewater using microorganisms and their aggregates is recognized to be an efficient green treatment (biological origin) mean with a relatively low cost compared to conventional physical and chemical treatment processes. Microorganisms such as bacteria, fungi and often algae are used for removal of targeted pollutants from wastewater. Microorganism can be used in following two ways (1) Direct mixing of free microorganisms with waste water, and there is no separation between microorganisms and treated water (2) Microorganism immobilized in bedding materials or encapsulated within a matrix, and there is a distinct separation between microorganisms and the treated water. However, use of immobilized or encapsulated cells is considered more effective than free cells as it leads to higher biomass loading, easier operation of solid-liquid separation, higher biodegradation rates, better operation stability, greater protection from toxic substances, increased plasmid stability of immobilized cells. Lignocellulosic biomasses, ceramics, polymers from both natural and synthetic origin are commonly used as bedding materials or for entrapment of microorganisms within it. These immobilized cells show immense potential to clean up a wide range of pollutants including phenolic compounds, hydrocarbons, propionitrile, organic and inorganic dyes, N,N-dimethylformamide, pyridine, benzene, toluene and © 2015 American Chemical Society In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

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xylene (BTX), heavy metals and also unwanted amount of nutrients such as nitrogen and phosphorus from wastewater streams. Integrated process of assimilation, adsorption and biodegradation is the sole responsible mechanism behind wastewater remediation by immobilized microorganisms. However, much more multifaceted investigations are still required in this field to develop more systematically integrated technologies and increase treatment efficiencies.

Introduction Global populations are expected to exceed 9 billion by 2050 having projected 6.4 billion contributions from urban areas alone and about 1.4 billion from slums. This ever increasing population growth along with urbanization, rapid industrialization, expanding and intensive food production methods are all putting pressure on water resources and on the other hand, inadequate infrastructure and management system increasing the unregulated or illegal discharge of contaminated water within and beyond national borders. Total fresh water on our planet is very limited, only 2.5% of all the water on earth is fresh water. Moreover, only 1% of this fresh water is available for withdrawal and human use. Despite of this fact, about 70-90% of the available fresh water is used in food production and a much of this water is returned to the system having additional nutrients and contaminants load. In addition, joint contributions from human and industrial waste in downstream make this scenario more critical. These wastewaters not only contaminates fresh water resources but also cause disturbances in coastal ecosystem, threatening food security, access to safe drinking and bathing water and providing a major health and environmental management challenge (1). Thus, it is essential to do proper treatment of wastewater prior to disposal into streams, lakes, seas and land surfaces. Most common methods of physical and chemical treatment of wastewater includes advanced oxidation, electro-coagulation and flocculation but they have their own limitations such as high cost, generation of secondary pollutants and also complex operation. On the contrary, biological treatments are associated with low cost and simple in operation. Hence, it has been employed extensively during past few decades for treatment of organic pollutants in wastewater. Suspended or activated sludge process and immobilized microorganism system are the two widely used methods in biological treatment of waste water. The activated sludge processes have several drawbacks including low biomass concentration, easy wash out and hence various novel technologies like acoustic cavitation, ultraviolet irradiation and magnetic field have been developed to increase efficiency of activated sludge processes. But application of these modern technologies becomes limited due to complex construction and high operation costs. On the other hand, higher biomass loading, easier operation of solid-liquid separation, higher biodegradation rates, better operation stability, greater protection from toxic substances, increased plasmid stability make immobilized microorganism systems most popular (over 320 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

activated sludge process) among water researchers worldwide (2, 3). This chapter will review application of immobilized microorganisms for treatment of wastewater more precisely industrial wastewater contaminated with both organic and inorganic pollutants.

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Immobilized Microorganism System An immobilized cell is defined as a cell that by natural or artificial means is prevented from moving independently of its neighbours to all parts of the aqueous phase of the system under study (4). Different techniques (Figure 1) that are extensively used in immobilization of microbial cell includes adsorption, covalent coupling, cross linking of microorganisms, encapsulation into a polymer gel, and entrapment in a matrix (2, 3).

Figure 1. Methods of immobilization of microbial cells.

Adsorption The simplest immobilization method is based on physical interaction between microorganisms and carrier surface. It is a reversible process and can lead to the peeling of adsorbed microorganisms during the operation. 321 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

Covalent Coupling In covalent coupling, microorganisms are immobilized with bonding reaction of reactive groups (e.g. –NH2 or –COOH) at the surface of biological cell, for instance protein. The coupling leads to increase in stability of microorganism but the bioactivity of microorganism decreases rapidly during post-operation process.

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Cross Linking of Microorganisms It is used to link bio-macromolecules each other with covalent bonds by using multifunctional reagents, such as glutaraldehyde, bisdiazobenzidine and hexamethylene diisocyanate. The method is simple but very difficult to control. Encapsulation into a Polymer Gel Many synthetic polymers such as polyacrylamide, polyvinyl alcohol; natural polymers like collagen, agar, agarose, cellulose, alginate, carrageenan etc. are often used to encapsulate microorganisms. Diffusion limitation is one of the inevitable drawbacks associated with encapsulation method. Entrapment in a Matrix Entrapment of the microorganisms in porous polymer carrier is often used to capture the microorganisms from suspended solution and then obtain the immobilized microorganisms. Due to porous structure of polymers, the pollutants and various metabolites produced easily diffuse through the matrix. Factors Responsible for Immobilization of Microorganism Immobilization of microorganism can be further categorized as “passive” (using the natural tendency of microorganism to attach to surfaces-natural or synthetic-and grow on them) and “active” (flocculant agents, chemical attachment, and gel encapsulation (5). Different types of active and passive immobilization method and their characteristics are presented in Table 1. Passive Immobilization Fimbria (pili), capsules (glycocalyx), various holdfast structures, stalks, cell wall component and slimes are variety of structures are responsible for natural attachment of microorganism to a surface. Besides, several other forces are also responsible for passive immobilization: electrostatic interactions, covalent bond formation, hydrophobic interactions (6). Ionic and hydrogen bonding are the most common electrostatic interactions involved in the initial stage of adsorption. Although being individually weak as compared to covalent bonds, they are nevertheless capable of producing relatively firm binding, if number of bonds is sufficiently large (4). 322 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

323

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Table 1. Different Type of Immobilization Methods and Their Characteristics (3, 6) Diffusion restriction

Immobilization Methods

Biomass loading

Toxicity problems

Ease of application

Mechanical stability

Carrier binding Physical adsorption

No

Low

Low

Reversible immobilization

Covalent bonding

No

Low

severe toxicity

High

Simple

Covalent cross linking

No

Moderate

severe toxicity

High

Simple

Micro encapsulation

Relatively low

High

severe toxicity

Low

Complex

Membrane separation/ entrapment

Fouling can cause severe diffusion restrictions

High

No

High

Complex

Entrapment within polymers

Relatively high: varies with the polymer material kind and construction

High

Moderate

Varies greatly with the kind of polymer

Simple

Simple

Examples of carrier/ matrix support Wood, sawdust, polygorskite, montmorilonite, hydromica, porous porcelain and porous glass treated with polycations

Entrapment in a matrix

In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

Alginates, k-carrageenan, agar, agarose, polyacrylamide, polyvinyl alcohol, polyethylene glycol, polycabamoyl sulphonate, microporous membrane filter

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In general, covalent bonds are formed between organic groups located on the bedding materials and microbial ligands (large quantities of different reactive groups present at the outer surface of microbial cells and cell walls) (7). Hydrophobic interactions between the microorganisms and the bedding material are very important as attachment of microorganism and bedding materials takes place in aqueous environment. The water molecule (separator between microorganism and bedding material’s surface) gets removed making possible interaction of hydrophobic group from both microorganism and bedding material. It has been reported that hydrophobic surfaces are more prone to colonization of dental and soil bacteria than are their hydrophilic counterparts (8). Sometime partial covalent bonding between the microbial cells and the bedding material play important role in immobilization. Often, in presence of aqueous solution hydroxides are formed instead of metal oxides incorporated in the bedding material and suitable amino or carboxyl groups on the cell surface can replace those hydroxyl groups (6, 9).

Active Immobilization Methods of active immobilization of microorganisms involve microencapsulation, membrane separation, covalent bonding and crosslinking, and entrapment within polymers. It is a kind of artificial immobilization of microorganism using inherent or induced surface properties of carriers/bedding materials and also of targeted microorganisms.

Carriers Used in Immobilization Both natural and synthetic carriers are used for immobilization of viable microbial cells (Table 2). However, the selection of carriers depends on various factors; it should have good mechanical strength, light weight, flexibility in overall shapes, nontoxic, nonbiodegradable in test conditions as well as cost effective. A comparison between natural and synthetic polymers is presented in Table 3. In general, there are two types of carrier materials used in immobilization of microbial cells :organic and inorganic.

Organic Carriers Mostly, organic carriers have a higher absorptivity compared to inorganic materials. Presences of larger varieties of reaction groups such as carboxyl, amino, hydroxyl on organic carriers are responsible for additional adsorption capacity. Often entrapment of microorganisms is carried out using organic polymers that are more abundant than inorganic materials. Organic polymers can be grouped in two sub categories: natural and synthetic polymer and both of them should be hydrophilic in nature, so that substrates can diffuse into the polymer beads during treatment of wastewater stream (14). 324 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

Table 2. Examples of Different Natural and Synthetic Carriers Used to Immobilize Microorganisms for Use in Biodegradation (2, 10–13) Carrier material

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Agar

Targeted compounds/ reaction

Microorganisms

Inorganic cyanides

Pseudomonas putida

Acrylamide

Pseudomonas sp., Xanthomonas sp.

3-Chloroaniline

Pseudomonas acidovorans CA28

4-Chloro-2-nitrophenol

Mixed culture

Dichloroacetic acid

Xanthobacter autotrophicus

Inorganic cyanides

Pseudomonas putida

p-Cresol

Pesudomonas sp.

Pentachlorophenol

Flavobacterium sp.

Phenol

Candida sp., Pseudomonas sp., Fusarium flocciferum,

Pyridine

Pimelobacter sp.

n-Valeric acid

Alcaligenes denitrificans

Sodium cyanide

Pseudomonas putida

Tertiary treatment of municipal wastewater

Chlorella sorokinian Azospirillum brasilens

Tributylin

Cholorella sp.

Triethyl amine

Arthobactor protoprotophormiae

Diatomaceous earth pellets

Glyphosate

Mixed culture

Diatomaceous earth biocarrier

p-Nitrophenol

Pseudomonas sp.

4-Chlorophenol

Alcaligenes sp. A 7-0, A 7-1, A 7-2

Cyanuric acid

Pseudomonas sp NRRL B-12228

Hydrocarbons

Candida parapsilosis

Granular clay, slag of lava

PAHs

Mixed culture

k-carrageenan

Inorganic cyanides

Pseudomonas putida

Sodium dodecyl sulfate

Pseudomonas C12B

Volatile fatty acids

Alcaligenes denitrificans

Alginate

Granular clay

Polyacrylamide

Continued on next page.

325 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

Table 2. (Continued). Examples of Different Natural and Synthetic Carriers Used to Immobilize Microorganisms for Use in Biodegradation (2, 10–13) Carrier material

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Polyurethane

Targeted compounds/ reaction

Microorganisms

Aliphatic and polycyclic aromatic hydrocarbons

Prototheca zopfii

Chlorophenol

Mixed culture, Rhodococcus sp.

Pentachlorophenol

Flavobacterium sp.

Dechlorination of spent sulphite bleach effluents

Streptomycetes sp.

Table 3. A Comparison between Natural and Synthetic Polymers Used for Immobilization of Microbial Cells (2, 3, 6) Characteristics

Polymers

Desired criteria

Natural

Synthetic

Solubility

Low

High

Low/not

Biodegradability

Low

Medium

Low

Stability

High

Low

High/Medium

Diffusivity

High

High

Medium/Low

Growth

Possible

Good

Good/Moderate

Immobilization procedure

Simple

Simple

Laborious

Toxicity

Non-toxic

Non-toxic

Depends largely on chemical composition and methods of preparation

Cost

Low

Low

Moderate/High

Carrageenan, Ca-alginate, agar, cellulose

Polyvinyl alcohol (PVA), polyethylene glycol (PEG), polycarbomoyl sulphonate (PCS), polyacrylamide (PAM)

Examples

Alginate, chitosan, agar, collagen, agarose and carrageenan are common examples of frequently used natural organic carrier materials for immobilization of microorganisms. The source of these polysaccharides is mainly algae and is prepared by gelation of soluble polymers either by cooling and/or in presence of a solution that contains different ions (6). 326 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

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Synthetic polymers can be prepared by gelation of monomers using a wide range of chemical or photochemical reactions. These polymers are not easily biodegradable and have much better mechanical performance compared with natural polymers. However, diffusivity is considered to be lower in synthetic polymers. Synthetic gel such as polyacrylamide (PAM), polyvinyl alcohol (PVA), polyethylene glycol (PEG), polycarbonyl sulphonate (PCS) and synthetic plastics like polyacrylonitrile (PAN), polyethylene (PE), polypropylene (PP), polyurethane (PU), and polyvinyl chloride (PVC) have been explored significantly as carrier materials (3).

Inorganic Carriers Inorganic carries are generally selected to immobilize microorganisms by electrostatic attachment between the cells and the carrier materials. Inorganic carrier materials are not only cost effective but also can resist to microbial degradation and have good thermostability performance. Porous glass, ceramics, clay, activated charcoal, anthracite, zeolite are most frequently used inorganic carriers (3). It has also been found that incorporation of specific metals into the carrier material allows better attachment of microbial cells to carrier materials. For example, incorporation of Fe (II) within a bedding material can increase the binding of yeast cells almost by 50% (9).

Mechanism of Pollutant Removal from Wastewater The main pollutant removal mechanism by microorganisms (e.g. bacteria, fungi, algae) include assimilation, biosorption and biodegradation. Ingestion of unwanted nutrients such as nitrogen, phosphorus, carbon from wastewater by microorganisms and or microbial aggregates and use of the same for its own growth is termed as assimilation. The complex surface structures of (e.g. branched, filamentous, spherical, oval, mushroom sheet) and presence of different functional groups (e.g. bacterial surfaces contains carboxyl, phosphoryl, hydroxyl, and amino functional groups) help microorganisms/microbial aggregates to adsorb heavy metals, organic and inorganic materials. This phenomenon is defined as biosorption and can occur through various processes such as complexation, chelation, co-ordination, ion exchange, flocculation and/or precipitation, reduction. Biodegradation is defined as the decomposition/chemical disbanding of organic materials by microorganisms/microbial aggregates. Often, biodegradation is associated with a complex series of biochemical reactions and targeted compound can be degraded aerobically and/or anaerobically. However, often assimilation, biosorption and biodegradation occur simultaneously during the removal of pollutant (15). 327 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

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Application of Immobilized Algae, Bacteria, and Fungi for Wastewater Treatment Immobilization of algae/microalgae for use in variety of biotechnological application started over 40 years ago. To keep living cells within a matrix metabolically active as long as possible and with limited mobility during its functioning are considered as main objectives of immobilization of algal cells. In most cases, entrapped algae get benefited from immobilization. Apart from avoiding grazing by aggressive zooplankton, reduction in competition for nutrients with other microbial species, several improvements in the metabolism, function, and behavior are observed in immobilized algae (5). A few examples of immobilized algae/microalgae used for removal of diverse kind of pollutants from wastewater are presented in Table 4.

Table 4. Examples of Immobilized Algae Used for Removal of Different Kind of Pollutants from Wastewater (5, 11, 16, 17) Immobilizing matrix

Alginate

Targeted pollutants

Algae species

Nitrogen

Chlorella vulgaris, Chlamydomonas reinhardtii

Phosphorus

Dunaliella salina, Nanochloris sp.

Nitrogen and phosphorus

Anabaena doliolum, Scenedesmus obliquus, S. intermedius

Tributylin (TBT)

Cholorella sp.

Phenanthrene, fluoranthene and pyrene

Selenastrum capricornutum

Cadmium

C. vulgaris, Tetraselmis chuii

Cesium

Chlorella salina

Cobalt Copper

Chlorella vulgaris, Tetraselmis chuii

Gold

Chlorella homosphaera

Lead

Chlorella vulgaris, Chlamydomonas reinhardtii

Manganese

Chlorella salina

Nitrogen and phosphorus

Scenedesmus quadricauda, C. Kessler, Spirulina maxima

Cadmium

Chlorella vulgaris, Scenedesmus acutus

Nitrogen and phosphorus

Phormidium laminosum

Carrageenan

Cellulose fibbers

Continued on next page.

328 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

Table 4. (Continued). Examples of Immobilized Algae Used for Removal of Different Kind of Pollutants from Wastewater (5, 11, 16, 17) Immobilizing matrix Loofa sponge

Targeted pollutants

Algae species

Lead

Chlorella sorokiniana

Nickel

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Copper Polyacrylamide

Lead

Chlorella regularis

Zinc Uranium Mixed hydrocarbons

Polyurethane

Polyurethane foam

Prototheca zopfii

n-alkanes Cobalt

Scenedesmus obliquus

Cadmium

Chlorella vulgaris, Scenedesmus acutus

Zinc Polyvinyl alcohol Silica gel

Copper

Sargassum baccularia

Cadmium

Spirulina platensis

Mercury

Chlorella vulgaris

Use of immobilized bacteria is also an important aspect in the field of biological wastewater treatment. There are now plenty of evidences (Table 5) that bacterial cells immobilized onto or into carrier matrix possess severe beneficial properties over suspended free cells, particularly high viability and ability to withstand environmental stresses and increased catalytic activity (31).

Table 5. Example of Immobilized Bacteria Used for Removal of Different Kind of Pollutants from Wastewater Bacteria

Type of pollutants

Immobilizing matrix

Reference

Arthobacter protophormiae

Triethylamine

Alginate

(12)

Mixed culture

Coumaphos, chlorferon, diethylthiophosphate

Alginate

(18)

Pseudomonas putida

Benzene, Toluene, o-xylene

Agave fiber/ polymer

(19)

Continued on next page.

329 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

Table 5. (Continued). Example of Immobilized Bacteria Used for Removal of Different Kind of Pollutants from Wastewater

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Bacteria

Type of pollutants

Immobilizing matrix

Reference

Paracoccus sp. Strain KT 5

Pyridine

Bamboo based activated carbon

(20)

Ochrobacterium sp. DGVK1

Dimethylformamide

PVA-alginate

(21)

Klebsiella oxytoca

Propionitrile

Alginate, cellulose triacetate

(22)

Bacilus cereus

Phenol

Alginate

(23)

Pseudomonas sp.

Azo dyes

Sol-gel silica

(24)

Microbe B350

Cu (II)

Polyurethane

(25)

Bacillus strain CR-7

Pb (II), Al (III), Cr (VI), Cu (II), Fe (III), Zn (II), Ni (II), Cd (II), Co (II), Mn (II)

Alginate

(26)

Cupriavidus sp., Sphingobacterium sp., Alcaligenes sp.

Zn, Cd

Alginate, pectate, synthetically cross linked polymer

(27)

Arsenic oxidizing bacteria (AOB)

As (III)

Polyvinyl alcohol

(28)

Bacillus sp., Pseudomonas sp., Serratia sp.

Hg, Cr, Ni

Alginate, polyacrylamide

(29)

Pseudomonas putida YNS1

Cu, Cd, Phenol

Alginate-silica

(30)

Fungi can produce a large variety of extracellular proteins, organic acids and other metabolites and also capable of adapting severe environmental stresses. All these characteristics recognize fungi as potential microorganism for treatment of industrial wastewater. However, in terms of application ease and effectiveness, a free fungal cell shows severe drawbacks as fungus mycelium are too exposed to environmental stresses. Consequently, a good alternative could be immobilization of fungal biomass on suitable support with the aim to maintain its viability and improve its activity (32). Some examples on use of immobilized fungi for treatment of different kind of pollutants are presented in Table 6. 330 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

Table 6. Examples of Immobilized Fungi Used for Removal of Different Kind of Pollutants from Wastewater

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Fungi

Type of pollutants

Immobilizing matrix

Reference

Yarrowia lipolytica

Oil, COD

Ca - alginate

(33)

Geotrichum candidum

Phenol and COD

Ca - alginate

(34)

Phanerochaete chrysosporium

Phenolic compounds

Wood chips op Italian poplar

(35)

Trametes pubescens, Pleurotus ostreatus

Industrial and model dyes

Polyurethane foam

(36)

Cunninghamella elegans

Synthetic dye

Ca - alginate

(37)

Phanerochaete chrysosporium

Remazol brilliant blue R

Loofa sponge

(38)

Aspergillus niger

Cu (II), Cd (II)

Poly vinyl alcohol

(39)

Trichoderma viride

Cr (VI), Ni (II), Zn (II)

Ca - alginate

(40)

Trochoderma harzianum

U

Ca - alginate

(41)

Polyporus squamosus

Cr, Mn, Fe, Ni, Cu, Pb

Ca - alginate

(42)

Phanerochaete chrysosporium

Pb (II)

Loofa sponge

(43)

Rhizopus cohini

Cr (VI)

Saw dust, polyurethane, alginate

(44)

Suitability of Immobilized Microbial Cells for Use in Bioreactors User’s flexibility to design and manufacture immobilized microbial matrix of different size and shapes with desired cell density make it possible to use immobilized microorganisms as a discrete phase in the bioreactor and also to decouple their hydrodynamic behavior from other existing phases. Thus, a wide range of bioreactor configurations including stirred tank, packed bed, fluidized bed, gas agitated and membrane bioreactor can be used with immobilized microorganisms (Table 7). 331 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

Table 7. Example of Immobilized Microbial Cell Bioreactors

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Design of bioreactor

Type of pollutants

Immobilizing matrix

Microorganism

Reference

Fluidized bed

Phenol

Alginate

Pseudomonas putida

(45)

Membrane

Cr (VI)

Agar-agar

Pseudomonas sp.

(46)

Fluidized bed

Azo dyes

polyvinylalcohol

Aeromonas hydrophila, Comamonas testosterone, Acinetobacter baumannii

(47)

Packed column

Cr (VI)

Sawdust, polyurethane and alginate

Rhizopus cohnii

(44)

Fluidized-bed bioreactor

Aliphatic and PAHs

sawdust, polyvinyl alcohol, polyacrylamide

Rhodococcus sp.

(31)

Stirred tank reactor

Anthraquinone dye drimarene blue K2RL

Scotch-Brite

Aspergillus niger SA1

(48)

Continuous flow column reactor

Propionitrile

Alginate, cellulose tri acetate

Klebsiella oxytoca

(22)

Airlift bioreactor

Dimethylsulfoxide

polyvinylalcohol

Pseudomonas sp. W1

(49)

Continuous packed bed column

Cr (VI), Ni (II) and Zn (II)

Ca - alginate

Trichoderma viride

(40)

Continuous packed bed reactor

N,N-dimethylformamide

Polyvinyl alcoholalginate blend

Ochrobactrum sp. DGVK1

(21)

Batch and fluidized bed column reactor

Cr, Ni, Cu, Cd

Alginate

Yeast, Pseudomonas aeruginosa, Bacillus subtilis, Escherichia coli

(50)

Other advantages of use of immobilized microorganism in bioreactor includes high biomass loading, continuous reactor operation without risk of cell washout, protection of cells from toxic substrates, reaction selectivity, increased biodegradation rate and enhanced operation stability (3, 51). 332 In Water Challenges and Solutions on a Global Scale; Loganathan, et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2015.

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Conclusion Today wastewater is a worldwide concern as most wastewaters are hazardous to living being and environment and must be treated properly before it ends into streams, lakes, seas and land. Biological agents like bacteria, fungi and algae play an important role in wastewater treatment and found very effective in degradation and or attenuation of a wide range of pollutants in wastewater. In this context, immobilization of microscopic biological agents offers several advantages over freely suspended cells in biological treatment system. Easy separation of cells from reaction media, reuse of active cells, minimum wastages of cells, higher volumetric reaction rates because of higher local cell concentration are most important features of biological wastewater treatment with immobilized cells. Additionally, immobilization of viable cells increases the stability of a microbial system, allowing its application under extreme environmental conditions, and development of continuous bioprocesses, avoiding release of contaminants during biodegradation process. However, still we are in very nascent stage of large scale application of immobilized microorganisms for wastewater treatment and more interdisciplinary effort should be given to develop efficient and cost-effective mean for waste water treatment through immobilized cells.

Acknowledgments Authors are very grateful to Dr. R.K. Pachauri, Director General, TERI, for providing the infrastructure and environment for preparation of the manuscript. We apologies to all those researchers whose findings we overlooked or could not include because of accessibility limitations.

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