propionic acid productiiiiiiiiiiion.pptx

propionic acid fermentation

1. PROPIONIC ACID FERMENT A TION Monday, January 28, 2019 Sir. Stymass Kasty 1

2. INTRODUCTION Fermentation is the chemical transformation of organic substances into simpler compounds by the action of enzymes, complex organic catalysts, which are produced by microorganisms such as molds, yeasts, or bacteria. Fermentation in Food Processing is the process of converting carbohydrates to alcohol or Organic acids using Microorganisms such as Yeast or Bacteria under Anaerobic Conditions. Necessary Because allows cells to generate ATP without Oxygen 2

3. INTRODUCTION 3 Propionic acid (CH3-CH3-COOH) is produced by several anaerobic bacteria among which are the Coryneform Propionibacterium and Veillonella , Clostridium and Selenomonas . The main Propionic acid fermenters are Propionibacterium acidipropionici and Propionibacterium freudenreichii . Propionibacteria possess cytochromes and catalase and can tolerate some amount of oxygen.

4. USES OF PROPIONIC ACID 4 IN FOOD INDUSTRY. Propionic acid and its sodium, calcium and potassium salts are incorporated to suppress the growth of mould and rope in breads and cakes, on the surface of cheese, malt, fruits, vegetables and tobacco, grain and silage preservation and to prevent the blowing of canned frankfurters without affecting their flavour. Dipping containers, caps and wrappers in solutions of these salts is also effective.

5. USES OF PROPIONIC ACID 5 IN FOOD INDUSTRY. The association of propionic acid with lactic and acetic acids has been recommended for the preservative of food. This recommendation was reinforced by the works which demonstrated the synergistic effect of these acids on the inhibition of Listeria monocytogenes growth in foods.

6. USES OF PROPIONIC ACID 6 IN FOOD INDUSTRY. The Food and Drug Administration (FDA) lists the acid, the Na+, Ca++ and K+ salts as preservatives in their summary of Generally Recognized As Safe (GRAS) additives and no upper limits are imposed, except for bread, rolls and cheeses (0.30 to 0.38%). The strain Propionibacterium subsp. Chermanii is used in Cheese making to create CO2 bubbles that become “ Eyes ” (Round holes in the cheese) Propionate is metabolized like other fatty acids in the mammalian body

7. USES OF PROPIONIC ACID 7 IN ANIMAL THERAPY Sodium propionate has been used in: Wound infections Dermatoses (Antifungal agents) Anti-arthritic drugs and Conjunctivitis

8. USES OF PROPIONIC ACID 8 OTHER USES Production of Cellulose plastics Used in textile, filters, reverse osmosis membranes, lacquer formulations and molding plastics. Used as Herbicides Used in the manufacture of Ester solvents and fruit flavors. Example, Citronellyl propionate and Geranyl propionate. Perfume bases and Butyl rubber to improve processability and scorching resistance.

9. MIXED-ACID FERMENTATION 9 The Biological process by which a six-carbon sugar like Glucose is converted in to a complex and variable mixture of acids These fermentations generally generate Acids such as Acetic acid, lactic acid and succinic acid. Sometimes also generate neutral products such as Butanediol These are the characteristics of most members of Enterobacteriaceae

10. PROPIONIC ACID FERMENTATION 10 The first works on propionic acid fermentation resulted in the formulation of the Fitz equation : 3 lactic acid ------> 2 propionic acid + 1 acetic acid + 1 CO2 + 1 H20 OR 1.5 glucose ------> 2 propionic acid + 1 acetic acid + 1 CO2 + 1 H20

11. PROPIONIC ACID FERMENTATION 11 Theoretical maximum yields are 54.8% (w/w) as propionic acid and 77% as total acids. Formation of propionic acid is accompanied by the formation of acetate for stoichiometric reasons and to maintain hydrogen and redox balances. Propionibacteria have been the primary candidates for the development of a biological process due to their unique metabolism. Dairy species such as P. acidipropionici strains, have been explored as potential propionic acid producers due to their tendency to produce propionate at a higher titre and yield. Propionibacterium uses Wood-Werkman cycle to produce propionate. Wood-Werkman cycle is energetically the most efficient propionate fermentation route currently known.

12. PROPIONIBACTERIUM Gram- positive Facultative Anaerobic Non-sporulating Rod shaped Facultative parasites and Commensals of human and other animals Living in and around the Sweat glands, Sebaceous glands and other areas of the skin. They may cause Acne (p. acnes) Selective media for their Growth is Sodium Lactate Members of the Genus Propionibacerium are widely used in: Production of Vitamin B12 Tetrapyrrole compounds Probiotic and Cheese industries PROPIONIC ACID PRODUCTION 12

13. P R OPIONIC ACID FERM E NTA T I O N PATHWAY Pyruvate reacts with methyl malonyl coA to form Propionyl coA and Oxaloacetate. Oxaloacetate give rise to malate, fumarate and succinate by reverse TCA cycle. Propionyl coA transfer its coA to succinate to form succinylcoA and propionate. 13

14. THE PROPIONIC ACID FERMENTATION OF PROPIONIBACTERIUM 14

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16. PROPIONIC ACID FERMENTATION ROUTES 16 B. DEGRADATION OF AMINO ACIDS TO PRODUCE PROPIONATE. This involves the degradation of Valine, Threonine , Isoleucine and Methionine . These can lead to the production of Propionate and ATP via Propionyl-CoA. Escherichia coli has been engineered to produce high yield of Propionyl-CoA. The combination of the Valine or Isoleucine and Threonine Pathways are synergistic given the redox and energy balances and can, together improve the propionate yield

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19. SCHEMATIC FLOW SHEET DIAGRAM OF THE PROCESS cel l s a r e from the and recy c l e d The s e par a ted medium back to the bioreactor using an ultrafiltration unit ( UF Unit ) It relies on the Continuous production of Propionic acid by increasing the Viable Cells to the bioreactor rather than Batch Fermentation 24

20. INDUSTRIAL PROPIONIC ACID PRODUCTION 20

21. CHALLENGES FOR INDUSTRIAL PROPIONIC A C ID PRODU C TION 21 Culturing Propionibacterium is limited to: Low growth rates Low productivities Acid stress and The co-generation of by-products which increase downstream processing costs and limit the economic feasibility of the fermentation process.

22. CHALLENGES FOR INDUSTRIAL PROPIONIC A C ID PRODU C TION 22 Targeted genetic engineering of propionibacteria to increase the production of propionate remain challenging because of different reasons. Propionibacteria have high GC content, which complicates genetic manipulation and contributes to poor gene annotations . There are few closed genomes and a small number of cloning vectors are available.

23. CHALLENGES FOR INDUSTRIAL PROPIONIC A C ID PRODU C TION 23 The ability of strains to readily develop spontaneous antibiotic resistance. Presence of strong restriction modification systems which contribute to the low transformation efficiency of propionibacteria But, A number of recent studies have reported the modification of P. freudenreichii subsp. shermanii (P. shermanii) and P. jensenii while only a couple of contentious studies have reported modification of the high-producing, genetic modification resistant strain, P. acidipropionici

24. CHALLENGES FOR INDUSTRIAL PROPIONIC A C ID PRODU C TION According to R.A.G.Garcia et al (2017), “ The expression of methylation components of restriction modification systems in host organisms has resulted in large improvements in the transformation efficiencies of non-model organisms and may be a critical step to improving the transformation efficiency in propionibacteria , particularly P. acidipropionici . Despite promising progress in the rational design of P. shermanii and P. jensenii , these modified strains still fall short of the natively high-producing P. acidipropionici. Therefore, a second line of research has focussed on random-mutagenesis strategies to enhance propionic acid production in P. acidipropionici .” Monday, January 28, 2019 Sir. Stymass Kasty 29

25. CON C LUSION 25 Propionic acids and their salts have many uses in different fields of Biotechnology apart from Food Industries, therefore there is a need of increasing the production of Propionate by taking the advantage of all non-pathogenic Bacteria which are capable of ferment different substances such as Sugar, Amino acid and others to produce large amount of Propionate.