Media balancing experiments carbon and nitrogen as variables (in alcohol fermentation)

Aim: Media balancing experiments carbon and nitrogen as variables (in alcohol fermentation)

BACKGROUND

Various economically significant substances with uses in the production of energy, pharmaceuticals, chemicals, and food are produced using fermentation technology on a large scale. Although fermentation technologies have been utilized for decades, the quest for sustainable product production that meets market demands while being cost-effective has been hard.

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The most crucial factor for any fermentation-based product is the availability of fermented product that meets market demand. There have been reports of a variety of microorganisms producing a small amount of primary and secondary metabolites.

Several high yielding approaches have been developed in the past to suit market demand, and they have been effectively applied in a variety of processes, includes oil extraction, biotransformation, the creation of new primary or secondary metabolites, etc.

Media balancing experiments

Prior to any large-scale metabolite production, medium optimization is still one of the most critically explored phenomena. It also poses several problems.

Prior to the 1970s, media optimization was done using traditional approaches, which were costly, time-consuming, included numerous experiments, and had subpar accuracy.

However, with the development of contemporary mathematical and statistical tools, media optimization has improved in terms of its vibrancy, effectiveness, efficiency, economy, and robustness in producing results.

The best fermentation conditions (such as pH, temperature, agitation speed, etc.) and the optimal medium components (such as carbon, nitrogen, etc.) must be determined in order to build a production medium.

Furthermore, maximal product concentration might be attained by optimizing the aforementioned medium properties.

REQUIREMENTS

Sterile forceps

Sterile spatula

Petri dish

Potato Dextrose Agar (PDA)

Erlenmeyer flask

Inoculation loop

Sterile distilled water

Fermentation medium (yeast extract 0.5 g/L, Peptone 3.0 g/L, (NH4)2SO4 1.5 g/l, K2HPO4 4.0 g/L, MgSO4.7H2O 0.3 g/L, CaCl2.7H2O 0.3 g/L)

PROCEDURE

1. Collect a soil from the garden, weigh 1 gm of soil and added to 9 ml sterile distilled water.
2. Add 1 ml to petri dish, pour molten PDA medium and incubate at 30c for 3 to 5 days.
3. Select A. niger spp. by staining method, take loopful A. niger and spread it on PDA and incubate at 30˚C for 3-5 days.
4. Prepare fermentation medium, distribute in different flask for variable study of carbon and nitrogen sources, add glucose and nitrogen in various concentration to the fermentation medium like 0.2, 0.4, 0.6, 0.8 and 1.0 g/L, and autoclave at 121˚C for 30 min.^[2]
5. Add loopful A. niger culture to each flask, incubate it at 30˚C and calculate ethanol concentration of each sample by using following equation:^[1]

Ethanol concentration (g/L) = 10 × alcohol degree (◦) × ρ

where is the ethanol density at 20°C, which is 0.789 g/L.

The following equation was used to compute the ethanol productivity.

Ethanol productivity Q (g/L/h) = (Ef – Ei)/ (tf − ti)

            where Ef and tf are the same parameters at the end of fermentation, and Ei and ti are the initial ethanol concentration and time, respectively.

6. Purify the ethanol by using distillation method. ^[2]

CONCLUSION

The variable of carbon and nitrogen study was successfully done, by calculation ethanol concentration.

REFERENCES

1. Fahad M. Alminderej, Zeineb Hamden, Yassine El-Ghoul, Bechir Hammami, Sayed M. Saleh and Hatem Majdoub, “Impact of Calcium and Nitrogen Addition on Bioethanol Production by S. cerevisiae Fermentation from Date By-Products: Physicochemical Characterization and Technical Design”, Fermentation, MDPI journal, 28 october 2022.
2. Vijay Jha, Nirupa Kumari, Birendra Prasad and Tushar Ranjan, “Ethanol production by Aspeergillus niger US4MTCC9931 and Saccharomyces cerevisiae MTCC174 using different Lignocellulosic biomass feed stocks”, Bioresources, 14 (4), 2019.
3. K Manikandan and T Viruthagiri,” Optimization of C/N ratio of the medium and Fermentation conditions of Ethanol Production from Tapioca Starch using Co – Culture of Aspergillus niger and Sachormyces cerevisiae“, International Journal of ChemTech Research, Vol. 2, No.2, April-June 2010.
4. Vineeta Singh, Shafiul Haque, Ram Niwas, Akansha Srivastava, Mukesh Pasupuleti and C.K.M. Tripathi, “Strategies for fermentation medium optimization: An in-depth review”, Frontiers in microbiology, Vol.7, January 2017.
5. Bernardo Onagar Yépez Silva-Santisteban, Attilio Converti and Francisco Maugeri Filho, “Intrinsic Activity of Inulinase from Kluyveromyces marxianus ATCC 16045 and Carbon and Nitrogen Balances”, Food Technol. Biotechnol., 44 (4), 2006.
6. Nathalia Dias Silva, Mariana Resende Alvim, Carlos Gonzalo Alvarez Rosario, Denise Crocce Romano Espinosa and Marcela dos Passos Galluzzi Baltazar, “Nutrients’ Supplementation Impacts on Alcoholic Fermentation of Mixed Wort Containing Corn and Sugarcane”, Research Square, January 2022.

Also read:

• Strain improvement of the industrially important isolate A. niger using EtBr for higher yield of the product
• Fermentation process of alcohol production
• Preparation of Baker’s Yeast from molasses
• Determination of Growth curve of yeast and compute growth rate & growth yield
• Determination of TDP & TDT of E.coli for designing of a sterilizer
• Screening for citric acid producing organisms
• Effect of inhibitors on enzyme activity

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