MIKROORGANISME:

Katalis Reaksi-Reaksi Kimia Air

Mikroorganisme: bakteri, jamur, dan ganggang

= katalis hidup yang menyebabkan sejumlah
proses kimia terjadi dlm air dan tanah.
Tetapi mikroba patogen dalam air yang diproses
untuk kubutuhan domestik harus dihilangkan.
Penelaahan umum tentang mikroba, tapi
perhatian khusus pd virus. Ukurannya kecil
(0,025 - 0,100 ), ketidakstabilan biologi, serta
sulit diisolasi dan dikulturkan. Maka virus
merupakan hal yang penting dalam pengolahan
dan penggunaan air

MIKROORGANISME:
Katalis Reaksi-Reaksi Kimia Air

Jamur dan bakteri diklasifikasikan sbg reduser .

Reduser memecahkan komponen kimia menjadi
spesi yg lebih sederhana, dan mendapatkan
energi yg dibutuhkan untuk pertumbuhan dan
metabolismenya.
Ganggang diklasifikasikan sbg produser, krn
memanfaatkan energi cahaya dan menyimpannya dlm energi kimia. Tetapi dlm ketiadaan sinar
matahari, ganggang menggunakan energi kimia
untuk kebutuhan metaboliknya.

MIKROORGANISME:
Katalis Reaksi-Reaksi Kimia Air

Lendir

Bakteri dapat berbentuk batang, bulat dan

spiral. Ukuranya antara 0,3 - 50 , sering
berkelompok 2 sampai jutaan sel.
Lendir untuk melindungi dari
serangan luar sel
Dinding sel
Diding sel menjadikan bentuk
Membran sitoplasma
sel dan kekakuan
Sitoplasma
membran sitoplasma untuk
kontrol pengangkutan bahan
Badan Inti
dari dan ke dalam sel
Sitoplasma sbg medium
Inklusi
proses metabolisme sel
Flagela
Badan Inti mengontrol proses
metabolisme dan reproduksi
Inklusi sbg cadangan makanan berisi karbohidrat, lemak
Flagela digunakan untuk bergerak

MIKROORGANISME:
Katalis Reaksi-Reaksi Kimia Air
Jamur/Fungi

(5 - 10 ) =
org. nonfotosintetik dan aerobik.
Strukturnya sangat beragam. Yg sederhana
(unisel) = ragi dan yg besar dan ruwet = jamur
payung.
Dapat hidup dlm media lebih asam drpd bakteri
Lebih toleran dlm konsentrasi logam berat yg
lebih tinggi drpd bakteri
Fungsi penting jamur = pemecah selulose dlm kayu
krn buangan sel jamur andalah enzim cellulase.
Walau jamur tidak tumbuh baik dlm air, tapi
sejumlah besar hasilannya punya peran penting
dlm air alami atau air limbah, misal asam humat

MIKROORGANISME:
Katalis Reaksi-Reaksi Kimia Air

Ganggang/algae
Organisme micro yg hidup dari makanan anorganik
menhasilkan bahan organik dari CO2 dng fotosintesa.
CO2 + H2O
{CH2O} + O2
Makanan umum yang dibutuhkan algae a.l. karbon
(CO2 dan HCO3-), nitrogen (NO3-), fosfor (ortofosfat) ,
belerang (SO42-) dan logam runutan termasuk Na, K,
Ca, Mg, Fe, Co, dan Mo.
Dalam ketiadaan sinar mekonsumsi bahan organik
dan O2..Apa akibatnya di perairan banyak algae pd
malam hari?

MIKROORGANISME:
Katalis Reaksi-Reaksi Kimia Air

Organisme Autotropik
tidak tergantung pd bahan organik untuk pertumbuhannya, dan tumbuh subur dlm medium anorganik
sepenuhnya.
Sebagai sumber karbon adalah CO2 atau CO3=. Sejumlah
sumber energi dapat dipakai, tgt pd spesies bakteri.
Contoh: Gallionella. Pd kondisi ada oksigen, bakteri ini
tumbuh dlm medium yg mengnadung NH4Cl, fosfat,
garam mineral, CO2 (sbg sumber C), dan FeS padat (sbg
sumber energi) dengan reaksi sbb:
4 FeS + 9 O2 + 10 H2O
4 Fe(OH)3 + 4 SO4= + 8 H+
Algae adalah autotropik yg memakai CO2 sbg sumber C
dan cahaya sbg sumber energi.

MIKROORGANISME:
Katalis Reaksi-Reaksi Kimia Air

Organisme Heterotropik

tergantung pd bahan organik baik sebagai sumber

karbon maupun sumber energi.
Jamur adalah heterotropik yg sumber C dan energinya
dari penguraian bahan organik

Bakteri aerobik

membutuhkan oksigen sbg penerima elektron:

O2 + 4 H + + 4 e 2 H2O

Bakteri anaerobik

dpt berfungsi hanya pada kondisi tanpa oksigen, krn

umumnya oksigen adalah racun bagi bakteri ini

Bakteri fakultatif

memanfaatkan oksigen bila ada, tapi bila tidak ada mk

menggunakan bahan lain sbg penerima elektron

MIKROORGANISME:
Katalis Reaksi-Reaksi Kimia Air

Pertumbuhan bakteri: dN/dt = kN atau N = Noekt

Fase Stasioner
Fase Kematian

Log Kons. Sel

bakteri hidup

Fase Log
(eksponensial
pertumbuhan)
waktu
Fase Lag

MIKROORGANISME:
Katalis Reaksi Redoks

Bakteri mendapatkan kebutuhan energinya untuk

metabolisme dan reproduksinya melalui reaksi redoks.
Reaksi redoks penting yg terjadi dlm air dan tanah a.l:
Respirasi aerobik
{CH2O} + H2O
CO2 + H+ + e (oksidasi) pE0=-8.20
O 2 + H+ + e
H2O (reduksi) pE0=-13.75
Oksidasi sulfida
O 2 + H+ + e
H2O (reduksi) pE0=-13.75
HS- + H2O
1/8 SO42- + 9/8 H+ + e (oksidasi) pE0=-3.75
Fermentasi
{CH2O} + H2O
HCOO- + 1 H+ + e ; pE0=-7.68
CH2O + H+ + e
CH3OH ; pE0=-3.01
Fiksasi N
{CH2O} + H2O
CO2 + H+ + e ; pE0=-8.20
1/6 N2 + 4/3 H+ + e
1/3 NH4+ ; pE0=-4.68

MIKROORGANISME:
Bakteri Metana
Bakteri

pembentuk metana mengkalisa reaksi:

1/8 CO2 + H+ + e
{CH2O} + H2O
{CH2O}

1/8 CH4 + H2O

CO2 + H+ + e

1/8 CO2 + 1/8 CH4

Bakteri

pembentuk metana adalah anaerobik a.l:

Methanobacterium
Methanobacillus
Methanococus
Methanosarcina
Pembentukan metana berguna untuk penguraian limbah
organik dalam jumlah besar baik di pengolahan limbah
ataupun di alam ( di perairan dalam atau di dalam tanah).
Bila metana memasuki daerah aerobik maka mudah bereaksi
dengan oksigen yang terlarut:
CH4 + 2 O2

CO2 + 2 H2O

MIKROORGANISME:
Bakteri Yang Memanfaatkan HC

Metana dioksidasi pada kondisi aerobik oleh banyak macam

bakteri a.l: Methanomonas. Hasil antara reaksi ini adalah
metanol, formaldehid, dan asam formiat.
Beberapa tipe bakteri dapat menguraikan HC yg lebih tinggi,
misalnya Mycrococcus, Pseudomonas, Mycobacterium, dan
Nocardia yang dapat mengurai alkana rantai panjang dengan
cara oksidasi. Pelajari reaksinya !!
Beberapa tipe mikroorganisma dapat memecahkan rantai
aromatik, misalnya jamur Cunninghamelia.Pelajari reaksinya !!

Persaingan Oksidasi Bahan Organik oleh

Ion Nitrat dan Agen Pengoksidasi lainnya

Suatu urutan yang menarik pada oksidasi bahan organik

oleh O2 terlarut, NO3- dan SO42- di sedimen atau perairan
hipolimnion. Reaksi yg mungkin terjadi:
{(CH2O)106(NH2)16P} + O2

CO2 + NO3- + H2PO4- + H2O + H+

{(CH2O)106(NH2)16P} + NO3- + H+
{(CH2O)106(NH2)16P} + SO42- + H+
Konsentrasi Oksidan

SO42-

O2

NO3-

Berat/volum Bahan Organik Terdegradasi

CO2 + N2 + H2PO4- + H2O

CO2 + NH4+ + H2PO4- + H2O + H2O

Nitrogen cycle

Nitrogen cycle

The Bio Redox Cycle for Nitrogen

Nitrification
NO2-

Nitrobacter

Protein Groups
Assimilation
NO3-

NO2-

Denitrification
Bacillus sp.
Pseudomonas sp.

NH2

Nitrosomonas

Am
mo
ni

f ic a

tion

on
i
t
ila
m
ssi
A
Protein Groups

NO
N2O

Nitrogen fixation
e.g. Azotobacter sp.
Rhizobium sp.
N2

Ass
imi
latio
n

NH2

o
m
Am

N2

NH3
n
o
i
t
ica
nif

Oxic
Anoxic
Nitrogen Fixation
e.g. Clostridium pasteurianum

Anammox
N2
Brocadia anammoxidans

Main N-cycle transformations

Assimilation

Assimilation

Mineralization

Denitrification

(algae + bacteria)

Org-N

NO2-

Assimilation

Nitrification 2

Nitrification 1

NH4+

(oxic bacteria)
Ammonification

Anammox

N2 - Fixation

NO3-

Denitrification

(anoxic bacteria)

- Soil bacteria
- Cyanobacteria
- Industrial activity
- Sulfur bacteria

(anoxic bacteria)
N2

N2O

NO2

+1

+2

gases

Oxidation state
-3

-2

-1

+3

+4

+5

Common species of nitrogen

Species
number

Formula

NitrateNO3-

+V

Nitrite NO2-

+III

Oxydation

Nitrous oxide gas

N2O

+I

Nitric oxide gas

Nitrogen gas N2

NO

+II

Ammonia (gas)

NH3

-III

Ammonium (ion)

NH4+

-III

Organic Amine

RNH2

-III

AMMONIFICATION

The formation of ammonia from dead organic

nitrogen containing compounds.

Rapidly recycled by microbes and plants!

AMMONIFICATION

Extracellular Enzymes in Ammonification

Substrates

Enzymes

Products

Proteins
Peptides

Proteinases, proteases Peptides, amino acids

Peptidases
Amino acids

Chitin
Chitobiose

Chitinase
Chitobiase

Chitobiose
N-acetylglucosamine

Peptidoglycan Lysozyme

N-acetylglucosamine &
N-acetylmuramic acid

DNA & RNA

Endonucleases &
Exonucleases

Nucleotides

Urea

Urease

NH3 & CO2

Fate of NH4+
NH4+ is a critical control point
Plant uptake
Microbial assimilation
Held on cation exchange sites
Fixed in interlayer of illite clays
Stabilized in soil organic matter
Volatilization of NH3
Nitrification

Assimilation

GDH
NAHPH
High NH4+

GS-GOGAT
ATP used
Low NH4+

Transaminases

Net NH4+ Production/Consumption

50 CO2-C
(Higher growth efficiency)
100 C

Fungi
(C:N ratio = 15)

N Requirement = 3.3
i.e., substrate C:N = 30

60 CO2-C
(Lower growth efficiency)
100 C

Bacteria
(C:N ratio = 5)

N Requirement = 8
i.e., substrate C:N = 12.5

Nitrification
1. Nitrification
Microbial oxidation of reduced nitrogen compounds (generally NH4+).
Catalysed

by 2 physiological groups of autotrophic (ie. use CO2 as carbon

source), aerobic bacteria called nitrifiers.

Autotrophic

NH4+ oxidisers (Nitrosomonas, Nitrosospira etc.):

NH4+ + 2 H2O
NO2- + 8 H+ + 6 e
1.5 O2 + 6 H+ + 6 e- 3 H2O
NH4+ + 1.5 O2
NO2- + H2O + 2 H+ + 275 kJ mol-1
Autotrophic

nitrite oxidisers (Nitrobacter, Nitrospira etc.)

NO2- + H2O NO3- + 2 H+ + 2 e0.5 O2 + 2 H+ + 2 e- H2O
NO2- + 0.5 O2 NO3- + 74 kJ mol-1
The

energy generated in the oxidation of the nitrogen compounds (Go) is

used by the bacteria for growth.

Chemolithotrophic Nitrifying Bacteria

Ammonia oxidizers
Nitrosomonas
Nitrosococcus
Nitrosospira
Nitrosolobus
Nitrosovibrio

Nitrite oxidizers
Nitrobacter
Nitrospina
Nitrococcus
Nitrospira

Nitrification
NO

NO
NO

NH4+

NH2OH

[HNO]

NO2NO2NHOH
N2O

NO3-

Ammonia Oxidation
Ammonia monooxygenase

Endergonic, cometabolic enzyme, inhibited by acetylene

NH3 + O2 + 2H+ 2e- NH2OH + H2O

Hydroxylamine oxidoreductase

Exergonic
NH2OH + H2O NO2- + 5H+ + 4eO2 + 2H+ + 2e- H2O

NO from intermediate oxidation of NH2OH

N2O produced by nitrite reductase
Net production of 2H+ per NH4+ oxidized

Nitrite Oxidation
Nitrite oxidoreductase (nitrite
dehydrogenase)

Exergonic, inhibited by chlorate

NO2- + H2O NO3- + 2H+ +2eO2 + 2H+ + 2e- H2O

About 1/3 the energy as ammonia oxidation

Heterotrophic Nitrification

Heterotrophic
bacteria and fungi
oxidize NH4+ or
organic N to NO2- or
NO3No energy produced
Important in some
habitats

Factors Affecting Nitrification

Nitrifiers present?

NO

NITRIFICATION
IMPROBABLE

YES
Aerobic conditions?

NO

YES
NH4+ availability

LOW

HIGH
Temperature, pH, nutrients, NO
inhibitors, etc.
YES

NITRIFICATION
PROBABLE

pH : 6.6 6.8
T : 30 -35 0C

Nitrification Inhibitors
N-serve (nitrapyrin)
DCD (dicyandiamide)
Dwell (etridiazole)
Wax-coated calcium carbide

Fate of NO3

NO3- is a critical control point

Plant uptake

Differ in preference for NH4+ and NO3-

Microbial assimilation

Regulated by NH4+ and organic N

Leaching

Anion, very mobile

Associated cations leach
10 mg NO3-N l-1 standard

Gaseous loss via denitrification

Dissimilatory reduction to NH4+

Nitrate Reduction
Process

Products

Energy
Conserved

Assimilatory
NO3- assimilation

NH4+

No

Dissimilatory
NO3- respiration

NO2-

Yes
Some
Yes

DNRA
Denitrification

NH4+>>N2O
N2>N2O>NO

Regulation

Soil
Condition

NH4+, organic N Low NH4+

O2
O2
O2

Anaerobic
Anaerobic
Anaerobic

Dissimilatory Reduction of NO3Process

Energy gained (G0, kcal mol-1)

e-

NO3-

e- accepted
per NO3-

DNRA
NO3- + 10H+ + 8e- NH4+ + 3H2O

-17.9

-143.3

Denitrification
2NO3- + 12H+ + 10e- N2 + 6H2O

-26.8

-133.9

DNRA

Wide range of bacteria

Aerobes: Bacillus, Pseudomonas

Microaerophiles: Campylobacter
Facultative anaerobes: Citrobacter, Klebsiella
Obligate anaerobes: Clostridium, Desulfovibrio

Little ATP generated; detoxify NO2- and/or

regenerate NAD
Associated with energy-rich (C-rich), low NO3environments

Denitrification

Anoxic process (ie. occurs in absence of oxygen)

Catalysed by facultative anaerobes called denitrifiers - a diverse group
of heterotrophic bacteria.
Electron acceptor: NO3-, NO2- and nitrogen oxides
Electron donor: Organic carbon source (e.g. acetate, glucose etc.) (i.e.
COD)
Biological reduction of NO3- and NO2- to gaseous N2 is a four-step
process:
NO3-
(+5)

NO2-
(+3)

NO (g)
(+2)

N2O (g)
(+1)

N2 (g)
(0)

Not all denitrifiers contain all the enzymes needed

entire reduction is carried out by different microorganisms.

Denitirification

Reduction of nitrate to nitrite

NO3- + 2 H+ + 2 e- NO2- + H2O (nitrate reductase)

Reduction of nitrite to nitric oxide

NO2- + 2 H+ + e- NO + H2O

(nitrite reductase)

Reduction of nitric oxide to nitrous oxide

2 NO + 2 H+ + 2 e- N2O + H2O
(nitric oxide reductase)

Reduction of nitrous oxide to dinitrogen gas

N2O + 2 H+ + 2 e- N2 + H2O (nitrous oxide reductase)

Most denitrifiers will use O2 (if available) to oxidise their carbon source,
in preference to NO3- as the electron acceptor. This is possiby because
slightly more energy is available from the reduction using oxygen as the
e- acceptor (Go = - 78.6 kJ mol-1) instead of nitrate (Go = - 72.1 kJ
mol-1).
Whether denitrification can occur in presence of O2 is unclear

Denitrification pathway

Denitrifying Enzymes
Dissimilatory NO3- reductase (Nar)
Mo & Fe; ATP production; regulated by O 2
Common to all that dissimilate NO3NO2- reductase (Nir)
Defining enzyme; regulated by O 2
Heme-Nir: 2/3 of denitrifiers (Pseudomonas, Alcaligenes,
Paracoccus, Thiobacillus)
Cu-Nir: 1/3 of denitrifiers (Bacillus, Rhizobium)

NO reductase (Nor)
Forms NN bond; regulated by O2; induced by N oxides
N2O reductase (Nos)
Cu; ATP production; regulated by O 2; induced by N oxides
Most sensitive to O2 and low pH

Denitrifier Genera
Genus
Organotrophs
Alcaligenes
Bacillus
Flavobacterium
Pseudomonas
Rhizobium

Characteristic
Common
Spore-former
Common
Common
N2-fixer

Phototrophs
Rhodopseudomonas

Anaerobe, reduce SO42-

Lithotrophs
Alcaligenes
Nitrosomonas
Paracoccus
Thiobacillus

H2 oxidizer, also organotroph

NH3 oxidizer
H2 oxidizer, also organotroph
S oxidizer

Factors Regulating Denitrification

Aeration (inhibited by O2 not ammonia)
NO3- availability

C availability
(4NO3 + C6H12O6 2N2 + 6 H2O)
pH
Temperature

Anammox
Anaerobic Ammonium Oxidation (Anammox)
The oxidation of ammonium to dinitrogen gas (N2) with nitrite as the electron
acceptor by autotrophic bacteria.
Discovered at the Kluyver Laboratory, Delft, The Netherlands in 1995.
For the first time, ammonium was discovered to be oxidised in the absence of
oxygen by a rare species of bacteria Planctomycetes, Candidatus Brocadia
anammoxidans.
NH4+ + NO2- N2 + 2 H2O (Go = -357 kJ mol-1)
Ammonium can be oxidised directly to dinitrogen gas, without the need for the
multi-step process of aerobic nitrification and heterotrophic denitrification.

Anammox
The electron donor is ammonium, the electron acceptor is nitrite.
Ammonium (ox. state -3) gets oxidised to N2 (0), and nitrite (+3) is
reduced to N2.
Autotrophic avoids the need for addition of a carbon source,
which is sometimes a cost in conventional systems.
All original attempts to isolate the responsible microorganism
failed; organism grows extremely slowly (max = 0.003 h-1),
probably lives in nature at the oxic/anoxic interface.
Advent of molecular microbiological techniques, eg. molecular
probing greater insight into natural habitats.

Anammox

4.3.1

Process only known from one species:

chemolitho-autotrophic planctomycete Brocadia anammoxidans
Energy generation:
NH4+ + NO2- N2 + 2 H2O
Carbon fixation:
CO2 + 2 NO2- + H2O CH2O + 2 NO3Measured overall growth at r = 0.0014 h-1 (Strous, 2000):
1 NH4+ + 1.32 NO2- + 0.066 HCO3- + 0.13 H+
1.02 N2 + 0.26 NO3- + 0.066 CH2O0.5N0.15 + 2.03 H2O
Known:
N2 comes from NH4+ and NO2N in biomass comes from NH4+

Cannon
CANON (Completely Autotrophic Nitrogen removal Over Nitrite)
Cooperation between aerobic and anaerobic ammonium oxidisers under
oxygen limitation.
Completely autotrophic promising opportunity for wastewaters with a
very low organic carbon content (eg. landfill leachates, aquaculture
waste).

Ammonium is oxidised to nitrite by aerobic ammonium oxidisers

(Nitrosomonas, Nitrosospira etc.);
NH4+ + 1.5 O2 NO2- + 2 H+ + H2O

The nitrite produced can be used by anammox;

NH4+ + 1.3 NO2- N2 + 2 H2O

Overall nitrogen removal by CANON:

1 NH4+ + 0.75 O2 0.5 N2 + 1.5 H2O + H+
Advantages of CANON system; low aeration costs (60% less than
traditional systems), requires no addition of a carbon source (process is
autotrophic) and the only end product is N2.