Environmental Biotechnology

Seven-sessions organized by

Bruce E. Rittmann
Professor and Director Center for Environmental Biotechnology Biodesign Institute at Arizona State University Tempe, Arizona 85287-5701 www.biodesign.asu.edu

Much of the material is taken from Environmental Biotechnology: Principles and Applications by B. E. Rittmann and P. L. McCarty, McGraw-Hill Book Co., New York (2001) Center for Environmental Biotechnology Vision Document, at www.biodesign.asu.edu

EB1 -- Introduction to Environmental Biotechnology

Environmental Biotechnologies Provide Valuable Services to Society

Treating industrial and municipal wastewaters to protect water resources, ecosystems, and human health Restoring sites contaminated with hazardous materials Reclaiming impaired water resources Capturing renewable resources, particularly energy Producing environmentally benign products

Science and Technology Foundation for Environmental Biotechnology

1 Metabolic Basis--The bacterias food is our pollutant 2 Microbial Ecology and Its Control--Create the
conditions to select for the right microorganisms

3 Biomass Retention--Develop systems to take

advantage of natural aggregation as flocs and biofilms

1 Metabolic Basis
The principle: a pollutant to us is some microorganisms substrate. Substrate means a material involved in generating energy to grow and sustain the microorganisms. It is like food or fuel. Substrate, fuel, or food involves sending electrons from an electron donor to an electron acceptor. On the one hand, virtually every pollutant is an electrondonor or an electron-acceptor for some group of microorganisms. On the other hand, a substrate can be a true fuel, or a source of energy that we can capture.

Treatment Examples
Pollutant/Role
Biodegradable organic matter (BOD)/donor Ammonium/donor Nitrate/acceptor TCE/acceptor

What We Add
Acceptor: e.g., O2 Acceptor: O2 Donor: organic compound or H2 Donor: H2 or organic H2 source

Energy-Capture Examples
Pollutant/Role
Biodegradable organic matter (BOD)/donor Biodegradable organic matter (BOD)/donor Biodegradable organic matter (BOD)/donor

Energy Outlet
Methane (CH4) Biohydrogen (H2) Electricity (i)

2 Microbial Ecology and Its Control

We deal with large, open systems. Microorganisms continually enter most processes. We have only partial control of the type and concentration of pollutants (fuels) that are input. Pure culture is not a relevant concept in practice. We deal with mixed cultures that often change. Therefore, the game is microbial ecology and steering it towards the types of microorganisms that do the job we want done.

Microbial Ecology as Science

As a scientific discipline, microbial ecology tries to answer these questions: Who is there? (Community structure) What could they do? (Community potential) What are they doing? (Community function) What are their interactions with each other and their environment? (Community interactions)

The Players in Microbial Ecology

The main players are from the Bacteria and Archaea domains, which comprise the protista or prokaryotes, which are single-celled organisms roughly 1 m is size We are just beginning to quantify and appreciate the phylogenetic diversity among those two Domains. Prof. Hausner will tell you about this aspect in EB3 and 4.

Bacteria have many different shapes (coccus, rod/bacillus, spirillum, and filaments/chains of cells of different shapes). But, all of them are small,in the order of 1 m for an individual cell.

QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture.

QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture.

Tools to Control the Microbial Ecology

Adding the proper other substrate in the right amount Modest adjustments to pH, temperature, and nutrients Efficient retention of biomass in general and the most critical microbial types in particular

3 Biomass Retention
In general, biomass is retained because it aggregates into suspended flocs or attached biofilms. Aggregation is a natural process that involves the production of extracellular polymeric substances (EPS), which are complex polymers involve protein, carbohydrate, and nucleic acid.

Biomass Retention
Flocs typically are a few 100 m is size. They are slightly heavier than water and can be removed from the water stream by settling and retained in the system. They also can be retained by filtration. Biofilms can be up to a few 100 m thick and are retained by being attached to a large amount of surface area in the process.

Dramatic photomicrographs of a floc and a biofilm on a membrane

30 m

Biomass Retention
The many different types of processes used in environmental biotechnology reflect, first and foremost, how to retain the microorganisms. The retention approach must be consistent with the means to supply the other substrate, as well as constraints imposed by economics, space, and operating skill.

EB2 Microorganisms and Metabolism

Chemical Composition of Prokaryotic Cells (R&M pg14)

Biomass Total Water Dry Matter Dry Organic Mater Carbon Oxygen Hydrogen Nitrogen Inorganic Matter P2O5 (as P) K2 O Na2O MgO CaO SO3 (2.2g) 0.7g 1.0g 0.9g 1.0g 1.5g 400g 300g 100g

90g
40.5 to 49.5g 19.8 to 25.2g 4.5 to 6.3g 8 to 13g

TSS

VSS

COD (BODL) at fd

10g
5.0g

Xin
Fixed, Mineral SS

Prokaryotic and Eukaryotic Cell Structures

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The eukaryote cell is much larger and differentiated.

Differentiating Cell Types

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The metabolic type of a microorganism is mainly determined by it electron donor and acceptor

QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture.

Basic electron and energy flows for microorganisms -- overview

fso + feo = 1 Energy Production feo Electron Donor (S, Substrate) Acceptor, SO fe = feo + fm

Reaction End Products

fm fso Active Biomass (Xa) Acceptor, SO Cell Residual, Inert Biomass (Xi)

C,N Cell Synthesis Growth

fs = fso - fm

The donor is oxidized, with the electrons (fe) transferred to the acceptor. This yields energy captured as ATP.
fso + feo = 1 Energy Production feo Electron Donor (S, Substrate) Acceptor, SO

fe = feo + fm

Reaction End Products

fm fso Active Biomass (Xa) Acceptor, SO Cell Residual, Inert Biomass (Xi)

C,N Cell Synthesis Growth

fs = fso - fm

The energy and more electrons from the donor (fs) are invested to synthesize new active biomass.

fso + feo = 1 Energy Production feo Electron Donor (S, Substrate)

Acceptor, SO

fe = feo + fm

Reaction End Products

fm fso Active Biomass (Xa) Acceptor, SO Cell Residual, Inert Biomass (Xi)

C,N Cell Synthesis Growth

fs = fso - fm

The synthesis of new biomass consumes elemental nutrients, like C, N, and P.

fso + feo = 1 Energy Production feo Electron Donor (S, Substrate)

Acceptor, SO

fe = feo + fm

Reaction End Products

fm fso Active Biomass (Xa) Acceptor, SO Cell Residual, Inert Biomass (Xi)

C,N P Cell Synthesis Growth

fs = fso - fm

Active biomass slowly decays (sort of like dying). Decay consumes more acceptor, as the biomass gains maintenance energy by oxidizing itself.
fso + feo = 1 Energy Production feo Electron Donor (S, Substrate) Acceptor, SO

fe = feo + fm

Reaction End Products

fm fso Active Biomass (Xa)

Decay

Acceptor, SO Cell Residual, Inert Biomass (Xi)

C,N Cell Synthesis Growth

fs = fso - fm

Decay also generates residual, inert biomass (sort of like dead cell bodies), which accumulates suspended solids that are not metabolically active.
fso + feo = 1 Energy Production feo Electron Donor (S, Substrate) Acceptor, SO

fe = feo + fm

Reaction End Products

fm fso Active Biomass (Xa) Acceptor, SO

Decay

C,N Cell Synthesis Growth

fs = fso - fm

Cell Residual, Inert Biomass (Xi)

Solids Retention Time (SRT)

SRT is the master variable for most environmental biotechnologies. It has units of time (e.g., days) and is the average time that active biomass is in the system. SRT = (total active biomass)/(net rate of activebiomass production) SRT = 1/(specific growth rate) = 1/ We will define SRT more quantitatively in EB5.

The SRT cannot be too short: The active biomass washes out, and no substrate is removed.
100 S [SRT min]lim 90 SRTmin 80 3.00 70 Xa, Xi, Xv (mg/L) 2.50 60 S (mg/L) Xv Xi Xa 4.00

3.50

50

2.00

40

Xio 1.50

30 1.00 20 0.50 10 So

0 0 2 4 6 8 10 SRT (d) 12 14 16 18 20

Smin 22

0.00

A long SRT increases substrate removal, but also enriches the total biomass in inert biomass. For a long SRT, much of the biomass can be inert, which removes no substrate.
100 S [SRT min]lim 90 SRTmin 80 3.00 70 Xa, Xi, Xv (mg/L) 2.50 60 S (mg/L) Xv Xi Xa 4.00 3.50

50

2.00

40

Xio 1.50

30 1.00 20 0.50 10 So

0 0 2 4 6 8 10 SRT (d) 12 14 16 18 20

Smin 22

0.00

The metabolic type of a microorganism is mainly determined by it electron donor and acceptor

QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture.