GROUP 2 WINDY LOU BAGAY PRECIOUS DELA CRUZ CYRENE D. ESTRELLANA REYJONDEL MARASIGAN CARL DAVE S. MENDOZA MCB 150 C-3L
BIOGEOCHEMICAL CYCLE
Oxidation and reduction of substances that results in the cycling of elements within and between different environments Carried out by microorgsnisms cycles and their many feedback loops are essential for plant agriculture and the overall health of sustainable planet life
BIOGEOCHEMICAL CYCLE
Carbon cycle Nitrogen Cycle Sulfur Cycle Iron Cycle Phosphorus, Calcium, and Silica Cycles
NITROGEN CYCLE
involves the reduction and oxidation reactions, change in form and location of nitrogen on earth Highy dependent on the action of microorganisms
NITROGEN CYCLE
OBJECTIVES
To demonstrate steps in nitrogen cycle using control organisms and soil sample; To describe the enzymatic activities of organisms involved in the nitrogen cycle; To be able to define the importance of microorganisms in biogeochemical cycles.
AMMONIFICATION
Decomposition of organic nitrogen such as amino acids and nucleotides to ammonia Ammonia (NH3 )exist as ammonium ion (NH4+ ) at neutral pH In alkaline soils, ammonia is lost by vaporization because it is volatile
AMMONIFICATION
Ammonifying bacteria:
Bacillus
Clostridium Proteus
Pseudomonas
Streptomycin
AMMONIFICATION (EXERCISE)
Peptone
broth
substrates for ammonification
Provides
Two
test:
Qualitative
Quantitative
AMMONIFICATION (EXERCISE)
Qualitative test
Nesslers
AMMONIFICATION (EXERCISE)
Qualitative test
0.1N
sulfuric acid (titrant) and Methyl red (indicator) Methyl red: red=pH below 4.4 yellow=pH above 6.0 high volume of acid used= high amount of NH3 (basic)
AMMONIFICATION (EXERCISE)
mg of NH3 in 100 ml
Samples/Cultures
mg of NH3/100 ml
112 22.4
B. cereus P. fluorescens
P. vulgaris
Non-acid, waterlogged soil Water logged soil non-
+
+
11.2
700
Acid soil
Control 0
NITRIFICATION
Process by which ammonium or ammonia is oxidized into nitrite (NO2-), which is further oxidized to nitrate (NO3-) Occurs only under aerobic condition Carried out by nitrifiers (strict aerobe)
NITRIFICATION
action
Oxygen pH
concentration
condition only
fastest when pH is from 7.5 to 8.5
Aerobic
Reactions
Inhibiting
Metals
substances
(inhibits the first step in nitrificatio)
Nitrapyrin
By nitrosofying bacteria
3 sub-processes
NH3 is oxidized into hydorxyl amine (NH2OH) by the enzyme mono-oxygenase NH3 + O2 + 2H + + 2e- NH2OH + H2O NH2OH is oxidized into NO2NH2OH + H2O NO2- + 5H+ + 4e Electrons and oxygen and free hydrogen ions are converted to water O2 + 2H + + 2e- H2O
By nitrifying bacteria
Soil sample
Nitrite Formation
N/A
Waterlogged soil
Acid soil Control
incubation period is needed for full conversion of NO2- to NO3 Reduced growth rate of nitrifying bacteria due to acid produced during nitrification (nitrification stops at a pH below 6.0)
DENITRIFICATION
Biological reduction of nitrate to gaseous nitrogen compounds by facultative heterotrophic bacteria or denitrifiers
Bacillus,
occurs when oxygen levels are depleted (under strict anaerobic conditions or reduced oxygen tension)
DENITRIFICATION
nitrate respiration When NO3- ions act as terminal electron acceptors on the absence of oxygen NO3- is converted into variety of reduced products Enzyme: dissimilatory nitrate reductase
Inhibited
by oxygen
DENITRIFICATION (EXERCISE)
Nitrate broth
Test for pH, gas production and presence of NH3, NO2 and NO3
Soil Sample
pH
9
8.5
+
+
DENITRIFICATION (EXERCISE)
(-) for NH3 = may have been vaporized due to alkaline condition (-) for nitrate = may have been reduced to other N gas compounds (+) for nitrite = nitrate was utilized as electron acceptor and converted to nitrite (+) for gas = nitrite is converted to N2 or N2O
NITROGEN FIXATION
Process where N2 gas is converted into NH3 by diazotrophic bacteria Involves a sequence of reduction steps that require major energy expenditures due to stability of the dinitrogen triple bond Enzyme: nitrogenase
Oxygen
sensitive
NITROGEN FIXATION
removal of oxygen by respiration Biofilm formation Compartmentalization (heterocyst formation) Conformational protection (complexing with specific protein
NITROGEN FIXATION
Occurs in environments with abundant carbon source and low nitrogen Two types:
Symbiotic
Microscopic observation
directly
for the cultivation of Rhizobium sp. and for studying root nodulation
- fermentable sugar alcohol source Magnesium - provides cations essential for the growth of Rhizobia Yeast extract - amino acids, vitamin B complex and accessory growth factors for Rhizobia; provides oxidation-reduction potential of medium in the range favorable for Rhizobia; serves as hydrogen donor in respiratory process
In the exercise:
N-free
glucose medium - Observed for biofilm formation N-free Iron-free Glucose agar -presence of pigment
Biofilm formation: to retard oxygen diffusion that can inactivate the nitrogenase enzyme
Thank you!!!
END