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THE PRODUCED FERTILIZER EFFECT ON THE GROWTH OF PS.NR.

22

As materials for the composting process, food waste such as potatoes, fruit peels, coffee
ground, egg shells and tea leaves are used. In addition to dry leaves, to reduce the excess
moisture, dust, shredded paper and newspaper are added to the compost. They are the
materials that can absorb water due to their ability of absorption. Therefore, cooked food,
meat or fish should not be included in the compost because they contain pathogen, which
may contaminate the compost. Hard items like bones or sticky and greasy items like cheese
should not be included in the composting process as well.

Greens and browns are required as the basic source as composting materials in the
composting process. They are used to being referred to as sources of carbon and nitrogen. We
mean the cooking waste by greens, while browns say dry leaves, sawdust, shredded paper and
dirt.

Table 1: List of Carbon and Nitrogen source

Greens Browns
(High in Nitrogen- N) (High in Carbon- C)

Discarded vegetables or vegetable peels Dry leaves


Fruit peels Soil
Coffee and tea grounds Shredded paper and newspaper

Table 2: List of Carbon to Nitrogen ratio of each material

Materials C/N ratio Reference


Discarded vegetables or 24:1 Adhikari, Barrington,
vegetable peels Martinez, & King, (2009)
Fruit peels 35:1 Rim Toumi, (2017)
Coffee and tea grounds 20:1 Angima, (2005)
Dry leaves 60:1 Rim Toumi, (2017)
Soil 10:1 Batjes, (2014)
Shredded paper 170:1 Rim Toumi, (2017)
Shredded newspaper 170:1 Batjes, (2014)
The mature phase for organic composting is that the organic materials continue to decompose
and convert to humic substance that is biologically stable. The increase in the nitrogen value
at the end of the composting period may occur due to the microorganism's use of nitrogen to
create cells, thus decreasing nitrogen, and some of the organisms eventually die, which is
reused as nitrogen and thus contributes to the increase. The nitrogen content in food waste
fertilizer is approximately 0.9% which is quite high for an organic fertilizer [1].

Many species will use nitrate, ammonia, and ~-amino acids as alternative sources of nitrogen
within the group of bacteria called the "pseudomonads." Before assimilation to amino
nitrogen, nitrate is reduced to ammonia by nitrite (Painter, 1970). Two routes can assimilate
ammonia. The first includes glutamate dehydrogenase (GDH) and is prevalent in these
bacteria although it possibly only functions in ammonia assimilation when the substrate
concentration is high (Brown, Macdonald-Brown and Stanley, 1972). The second route
involves the two enzymes glutamine synthetase (GS) and L-glutamine:2-oxoglutarate
aminotransferase (GOGAT) and is of particular importance in low environmental ammonia
(Tempest, Meers and Brown, 1970; Meers and Pedersen, 1972; Brown et al., 1972). When
amino acids are used as a source of both nitrogen and carbon, some type of deamination may
occur and this would seem to require a catabolically operating GDH. [2]

Different research has shown that compost fertilizer can be used to isolate the resistant
bacteria such as Ps.NR.22 with a high ability to decompose organic compounds. [3] Ps.
NR.22 requires additional nitrogen sources for essential biochemical processes and associated
enzymes, including lipase. Therefore, with the high amount of nitrogen content is food waste
fertilizer produced which is approximately 0.9% will support the growth of Ps.NR.22 and
increase the lipase production [4]. From the research paper of Novel Treatment of Heavily
Oiled Wastewater Using Ps.NR.22 Producing Usable Free Fatty Acid (FFA) done by Nik
Him NIK RAIKHAN and Ahmad Robert KHAIRUL IZWAN, with the addition of sterile
nitrogen compound with oil removal rate of 82.1±0.01% with lipase activity estimated at
18.8±0.01U / mL, the best oil removal was achieved compared to lower sources of nitrogen
compounds. Figure 1 shows the oil removal (%) values and the lipase activity correlated with
each of the values recorded from the wastewater containing the different nitrogen sources.
Figure 1: Percentage of oil removal by 9.1g/L of various nitrogen sources in oily wastewater
using direct catalysis by Ps.NR.22 species after 36 hours of fermentation (35°C, 150rpm);

According to Figure 1, the best oil removal values included urea, ammonium nitrate, yeast
extract and peptone, along with the three (3) groups of nitrogen sources; nitrogen compound,
nitrogen soil, and nitrogen fertilizer (78.7±0.03%-82.1±0.01%). [4]

The nitrogen content in produced fertilizer will increase the seepage of enzyme lipase by
Ps.NR.22. Enzyme lipase is essential in hydrolysis process to degrade lipids from oil-waste in
waste water and producing several types of useful free fatty acid (FFA) and glyceride.

Generally speaking, fat lipase hydrolysis results in free LCFAs and water-soluble glycerol.
From the Hydrolysis process of enzyme lipase degrades waste-oil in waste water, it has
developed six forms of unsaturated FFA: lauric acid (C12:0), myristic acid (C14:0), palmitic
acid (C16:0), linoleic acid (C18:2), oleic acid (C18:1) and caprylic acid (C8:0). GC in
conjunction with a mass spectrometer (MS) or a flame ionization detector (FID) is the most
commonly used FA and TFA determination technique (Figure 2). [4]
Figure 2: FTIR spectra of oily wastewater sample: A. Before treatment; B. After treatment

Several hydrolysis experiments (both for FL and IL) were performed under optimum
conditions to quantify the amount of free output of LCFA and the results are shown in Figure
3. [5]

Figure 3: Free LCFA formation during hydrolysis.

Therefore, the more the nitrogen content inside produced fertilizer, the more lipase enzyme
produced and the more free fatty acid (FFA) will be produced from hydrolysis of lipid in
waste water.

The kinetic mechanism of lipases is believed not to depend on the form of catalyzed reaction
(hydrolysis, acidolysis, transesterification, etc.).

The reaction starts with a nuclear attack on the carbon in the serine residue of the active site
from the ester bond of the susceptible substratum by hydroxyl group, forming an acyl-
enzyme complex and releasing alcohol from the lipid. Subsequently, the complex of acyl-
enzymes is hydrolyzed, releasing regenerated lipase. Figure 2 shows the stages of the reaction
catalyzed by the lipase and its intermediates.
Figure 2: Mechanism of the hydrolysis reaction of ester bonds catalyzed by esterases and
lipases. The catalytic triad and water are shown in black; the oxyanion hole residues are in
blue; the substrate is in red. (a) Nucleophilic attack of the serine hydroxyl on the carbonyl
carbon of the susceptible ester bond; (b) tetrahedral intermediate; (c) acyl-enzyme
intermediate and nucleophilic attack by water; (d) tetrahedral intermediate; (e) free enzyme.
REFERENCE

1) Hazren A. Hamid*, Lim Pei Qi, Hasnida Harun, Norshuhaila Mohamed Sunar,
Faridah Hanim Ahmad, Mimi Suliza Muhamad, Nuramidah Hamidon. Development
of Organic Fertilizer from Food Waste by Composting in UTHM Campus Pagoh.

2) C. M. BROWN a, DEBORAH S. MACDONALD=BROWN AND S. O. STANLEY


~ The mechanisms of nitrogen assimilation in Pseudomonads.

3) Abooalfazl Azhdarpoor1, Bagher Mortazavi2* and Gholamreza Moussavi2 Oily


wastewaters treatment using Pseudomonas sp. isolated from the compost fertilizer

4) Nik Him NIK RAIKHAN*, Ahmad Robert KHAIRUL IZWAN NOVEL


TREATMENT OF HEAVILY OILED WASTEWATER USING PSEUDOMONAS
AERUGINOSA NR.22 PRODUCING USABLE FREE FATTY ACIDS (FFA).

5) Jeganaesan Jeganathan, George Nakhla ∗, Amarjeet Bassi Hydrolytic pretreatment of


oily wastewater by immobilized lipase.

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