Riboflavin (Vitamin B2) Production: Step-by-Step Process
Introduction:
Riboflavin (Vitamin B2) is an essential vitamin required for energy production, cellular function, and growth. It is widely used in food supplements, pharmaceuticals, and animal feed additives. Commercial production of riboflavin primarily involves microbial fermentation.
Microorganisms Used:
- Bacteria: Bacillus subtilis
- Fungi: Ashbya gossypii, Eremothecium ashbyii, Candida famata
Raw Materials:
- Carbon sources: Glucose, molasses
- Nitrogen sources: Ammonium salts, peptone
- Other nutrients: Salts (MgSO₄, KH₂PO₄), vitamins
Steps in Riboflavin Production:
1. Inoculum Preparation:
- A pure culture of the selected microorganism is grown in a nutrient-rich medium to prepare a dense inoculum.
2. Media Formulation:
- The production medium contains:
- Carbon source: Glucose or molasses
- Nitrogen source: Ammonium sulfate or yeast extract
- Trace elements: Magnesium, iron, and phosphate
3. Sterilization:
- The culture media and fermenter are sterilized to prevent contamination.
4. Fermentation Process:
- Inoculation: The prepared inoculum is transferred to the bioreactor.
- Fermentation Conditions:
- Temperature: 28-32°C
- pH: 5.5-6.5
- Aeration: High oxygen supply is maintained.
- The microorganism produces riboflavin as it metabolizes the nutrients in the medium.
5. Monitoring and Control:
- Parameters: pH, temperature, aeration, and substrate concentration are carefully monitored to optimize riboflavin yield.
6. Harvesting:
- After fermentation (typically 3-5 days), the broth contains riboflavin.
7. Recovery and Purification:
- Cell Removal:
- Microbial cells are separated using centrifugation or filtration.
- Extraction:
- Riboflavin is extracted from the broth using acid precipitation or solvent extraction.
- Crystallization:
- The extracted riboflavin is purified and crystallized.
- Drying:
- The purified riboflavin crystals are dried to obtain the final product.
8. Quality Control:
- The final product is tested for purity, potency, and compliance with regulatory standards.
Summary of Steps:
- Inoculum preparation
- Media formulation
- Sterilization
- Fermentation (under controlled conditions)
- Monitoring and control
- Harvesting
- Recovery and purification (extraction, crystallization, drying)
- Quality control
Applications of Riboflavin:
- Food Industry: Fortification of cereals, dairy products, and beverages.
- Pharmaceuticals: Vitamin supplements and multivitamin tablets.
- Animal Feed: Added to improve animal health and growth.
- Cosmetics: Used in skincare products.
Conclusion:
Riboflavin production through microbial fermentation is an efficient, sustainable method for producing this essential vitamin. Advances in biotechnology continue to enhance production yields, ensuring a reliable supply for various industries.