P C dProper Care and Feeding of theBio-community

Presented by:Richard B SmithRichard B. Smith

Monitoring & MaintenanceE i i IEngineering, Inc.

Bi hBiography

Graduated with a BCE from Georgia Tech Graduated with a BCE from Georgia Tech. Experienced in design and construction of

t d t t t t t f ilitiwater and wastewater treatment facilities. Began designing and operating soil and

d t t t t t i 1990groundwater treatment systems in 1990. Started using biological processes in soil and

d t i 1997groundwater in 1997. Licensed by DOE to use the PHOSter

Process in 2009.

I t d tiIntroduction

Biologically based processes are become Biologically-based processes are become more widely accepted.Still b “bl k b ” Still can be seen as a “black box”.

The better we understand what is necessary f i bl b t i l l ti th b ttfor a viable bacterial population the better we apply the various technologies available.

Obj tiObjectives

Review of the elements necessary to build Review of the elements necessary to build and sustain a population of archaea/bacteria.L k t l l f f th l t Look at molecular forms of these elements required to make them bio-available.R i th t k l d f th Review the current knowledge of the breakdown pathways of various important hydrocarbon/metal contaminantshydrocarbon/metal contaminants.

P tProcaryotes

These are single cell These are single cell organisms with no defined nucleusdefined nucleus.

DNA is circular.N h No chromosomes.

No organelles. 3 types of rRNA/50

types of protein.

B t i A hBacteria vs. Archaea

Bacteria: A hBacteria: Ester bonds in

membrane.

Archaea:o Ether linkages in cell

membrane.membrane. Walls made of

peptidoglycan.o No peptidoglycan in cell

walls.o Three RNA polymerases

One RNA polymerase. o Three RNA polymerases.o Extremophiles. (not

exclusively)B h d i fo Better at the reduction of more complex carbon compounds.p

B t i /E k /A hBacteria/Eukarya/Archaea

L 1 WLesson 1: Wrap-up

Summarize important points Summarize important points. Allow time for questions.

H dHydrogen

Most prevalent element Most prevalent element. Cells use H+ gradients to drive ATP

( d i t i h h t ) th i l t(adenosine triphosphate) synthesis, solute transport and flagella rotation.H i i di ti f l ti t ti f pH is indicative of relative concentration of

H+.

C bCarbon

About 50% of the dry weight of all life is About 50% of the dry weight of all life is carbon.Fi d f b di id i i b Fixed from carbon dioxide, inorganic carbon or hydrocarbon contaminants.

NitNitrogen Atmospheric Atmospheric

nitrogen is in the form of N≡N.

Some procaryotescan reduce this to a bi l i ll il blbiologically available form.

Ammonium nitrite Ammonium, nitrite, nitrate have single bonds and are more easily available.

OOxygen

Readily fixed from O and water Readily fixed from O2 and water. Anaerobic procaryotes use oxygen from

d ti id f t ti d tsugar reduction, acid fermentation and water. Photosynthetic organisms produce oxygen

f i f b di idfrom conversion of carbon dioxide.

Ph hPhosphorous

Major component in cell Major component in cell wall, nucleic acid and cell energy transferscell energy transfers.

Main constituent in ATP and ADPand ADP.

Primary natural source is ferric and calciumis ferric and calcium phosphate from rock.

S lfSulfur Sulfur can be used Sulfur can be used

as oxidant or reducing agent.g g

Procaryotes can readily oxidize

l t l lf telemental sulfur to sulfate.

S l iSelenium

Found in sulfur bearing formations Found in sulfur bearing formations. Toxic in large doses (puts the loco in

l d)locoweed). Procaryotes use selenium compounds to

d id t d i ll breduce oxidants and repair cell membranes damaged by peroxides.S l i i t f d t b ild Selenium is part of an enzyme used to build DNA from RNA. Regulates the rate of DNA formation and repairformation and repair.

M j C tiMajor Cations

K+ potassium regulates pH and K+, potassium, regulates pH and electrochemical gradients.N + di d i l t t t d Na+, sodium, used in solute transport and flagellar motors.M 2+ t bili ib d ll Mg2+ stabilizes ribosomes and cell membranes.C 2+ d i ti ti d Ca2+ used in enzyme activation and membrane stabilization. Current research into use in signalinginto use in signaling.

M j A iMajor Anion

Chlorine occurs in procaryotes as chloride Chlorine occurs in procaryotes as chloride, Cl-.It b t t t d t d Its use by procaryotes not yet understood.

The quantity of natural emissions of hl i t d i i h t th thchlorinated organics is much greater than the

emissions from industrial production.

A i A idAmino Acids

Combination of Combination of carboxylic acid and an amine groupan amine group.

Basic building block of proteinsof proteins.

Twenty amino acids are encoded withare encoded with genetic code.

NAD+/NADHNAD+/NADH

Coenzyme found in all Coenzyme found in all living cells.B ilt f i id Built from amino acids.

NAD+ is an oxidizer, tiaccepting

electrons/forms NADH.NADH i d ti b NADH is reductive by donating electrons.

ATPATP

Transports energy in cells for metabolism Transports energy in cells for metabolism. Split of one or two phosphates provides the

t t t ll di i ienergy to start cell division. Cells can regenerate ATP by replacing

h h tphosphates.

RNARNA

Constructed from a Constructed from a nucleobase, a ribose sugar and phosphatesugar and phosphate.

Directs synthesis of proteins catalyzesproteins, catalyzes biological reactions and delivers aminoand delivers amino acids to the ribosome.

DNADNA

Constructed from Constructed from long polymers of nucleotides withnucleotides with backbones of alternating sugar andalternating sugar and phosphate joined by ester bonds.

Forms into a circular double helix indouble helix in procaryotes.

B O id tiBenzene Oxidation

Procaryotes use NADH to Procaryotes use NADH to oxidize benzene, breaking one double-breaking one double-bond and adding two hydroxideshydroxides.

Iron at a +4 oxidation state is used to pass thestate is used to pass the extra electrons.

BBenzene

T t hl th lTetrachloroethylene

• Reduction of PCE to TCE• Reduction of PCE to TCE.• Can only be accomplished under anaerobic

conditionsconditions.

T i hl th lTrichloroethylene• Five aerobicFive aerobic

pathways for TCE.• Three paths using a p g

toluene-based enzyme.y

• Reductive de-chlorination in anaerobic conditions leads to i l hl idvinyl chloride.

T i hl th lTrichloroethylene

• Process uses iron and sulfur catalysts• Process uses iron and sulfur catalysts.• The epoxide can pass through cell walls

to complete the breakdownto complete the breakdown.

Ch Fi tiChrome Fixation• 88 species have been isolated that reduce CrVI88 species have been isolated that reduce CrVI.• Anaerobic conditions required and elevated

pH(10) for some species.p ( ) p

SSummary

Identified elements required for a healthy bio Identified elements required for a healthy bio-community.R i d th t i t t b ildi bl k Reviewed the most important building blocks of life.L k d f d d ti th Looked a few degradation pathways.

Source: http://umbbd.msi.umn.edu/Univ. of Minnesota Biocatalysts/Biodegradation Database.

Q ti ?Questions?