SINGLE-CELL PROTEIN PRODUCTION THROUGH Rhizo

pus oryzae CULTURE WITH THE MEDIUM OF TOFU W

ASTEWATER

Annisa Eka Nirwana

17734002

Journal by Leny Maryana1*, Syariful Anam1, Arsa Wahyu Nugrahani1 1Jurusan Farmasi, Fakultas MIPA, Universitas Tadulako,
Palu, Indonesia
Why I Choose this Topic?

The waste water of the food industry, especially from the tofu ind
ustry, contains high organic matter.

If it is disposed of into the environment without being processed

it will have a negative impact such as decrease in environmental
quality.

waste water of tofu also contain protein, fat, vitamins, and miner
als (Ca, Mg, Fe) so that it has the potential as a microorganism
growth medium
Single cell protein

Single cell protein is a high-protein biomass product derived from microbes.

Microbes producing single cell proteins generally grow in wastes that have elem
ents of carbon and nitrogen which are usually found in industrial waste.

The main components of single cell proteins are amino acids and mineral.

Single cell proteins can be used as a substitute for proteins from conventional so
urces such as agricultural, fishery and livestock products.

In addition, single cell proteins can produce highly nutritious foods called mycopr
oteins.
Materials

• Aquadest
• Sucrose
• Pure culture of Rhizopus oryzae
• 0.1% KH2PO4.3H2O
• Selenium
• concentrated sulfuric acid
• 5% NaOH
• indicator of methyl red
• hydrochloric acid 0.1001 N
• Potato Dextrose Agar medium (PDA) .
Methods

Making Medium From Tofu Waste water

– One liter of tofu waste water sample is filtered

– put into a beaker then pasteurized by means of waste
heated at 60 ° C for 30 minutes
– cooled in the refrigerator for 30 minutes
– left at room temperature 23 ° C for 24 hours.
– Do this process for three times
– Add KH2PO4.3H2O 0.1% and 2% granulated sugar to t
he pasteurized tofu waste water
– pH is adjusted by adding NaOH to reach pH 4, 5 and 6
and autoclaved at 121 ° C for 15 minutes
Methods

Medium Fermantation Process Tofu Waste Water

– In 150 ml sterile tofu waste water media inoculated wit

h 1 culture sample Rhizopus oryzae
– Incubated at room temperature on a 160 rpm speed sh
aker for 24; 48; and 72 hours.
– Every 24 hours analyzed the number of colonies growin
g using the surface plate method on Potato Dextrose A
gar (PDA) media calculated the cell dry weight and det
ermined the protein content.
Methods

Analysis of Cell Dry Weight

– The porcelain cup is cleaned and then put into the oven at 10
5 ° C for 1 hour
– The porcelain cup is cooled in a desiccator for 30 minutes the
n weigh to find out the weight.
– Weigh 10 grams of fermented waste medium in the porcelain
cup which is known to be of initial weight.
– Insert the waste medium that has been fermented into the ov
en at a temperature of 105 ° C until it is of constant weight.
– Cell dry weight is calculated from the weight difference in dry
cell mass (which has been oven) - the weight of the porcelain
cup.
Methods

Protein Analysis

– a fermented waste medium sample was weighed as much as 1 gram

and put in a 100 ml Kjeldahl flask
– Added 2 g selenium and 10 ml concentrated sulfuric acid.
– The mixture is put into turbo-digestion for 2 hours then left to cool f
or 30 minutes
– insert one by one into the vapodest Destilation to do the distillation
and titration process automatically.
– The results of this process will be known as the volume, and the blan
k volume.
– Protein levels will be calculated using the following equation:
% Protein =% N x conversion factor
Description:
%N : Percent Nitrogen
Conversion factor : For a mixture of protein compounds or for
which the composition of the constituents is not
known, then the multiplication factor of 6.25 is
used.
Results
Results
Conclusion

Total results showed the highest protein content obtained in the incubation period to 48 hour
s with an average percent about 0.47%, 0.47%, 0.46%

Mass of cell with an average value of 0.77%, 0.84%, and 0.91%.

The optimum pH of Rhizopus oryzae growth in the medium of tofu wastewater is about 5pH.

The wastewater of tofu potentially used as a growth substrate of Rhizopus oryzae.

By utilizing wastewater as a microbioal growth medium for single-cell protein can reduce resi
duals pollutant in the environment
Thank You