Vitamins Production Methods PDF

**Production of vitamins (10 marks)** Vitamins are organic compounds that perform specific biological functions for normal maintenance and optimal growth of an organism. These vitamins cannot be synthesized by the higher organisms, including man, and therefore they have to be supplied in small amou...

**Production of vitamins (10 marks)** Vitamins are organic compounds that perform specific biological functions for normal maintenance and optimal growth of an organism. These vitamins cannot be synthesized by the higher organisms, including man, and therefore they have to be supplied in small amounts in the diet. Microorganisms can be successfully used for the commercial production of many of the vitamins e.g. thiamine, riboflavin, pyridoxine, folic acid, pantothenic acid, biotin, vitamin B~12~, ascorbic acid, β-carotene (pro-vitamin A), ergosterol (pro-vitamin D). However, from economic point of view, it is feasible to produce vitamin B~12~, riboflavin, ascorbic acid and β-carotene by microorganisms. Several microorganisms can be employed for the production of vitamin B~12~, with varying yields. Glucose is the most commonly used carbon source. The most commonly used microorganisms are --- *Propionibacterium freudenreichii, Pseudomonas denitrificans, Bacillus megaterium and Streptomyces olivaceus.* Glucose is the most commonly used carbon source for large scale manufacture of vitamin B~12~. Other carbon sources like alcohols (methanol, ethanol, isopropanol) and hydrocarbons (alkanes, decane, hexadecane) with varying yields can also be used. Fermentation temperature is 27^0^ C and incubation time is 4-5 days. **Production of Vitamin B~12~ Using Propionibacterium sp:** *Propionibacterium freudenreichii* and *P. shermanii,* and their mutant strains are commonly used for vitamin B~12~ production. The process is carried out by adding cobalt in two phases. **Anaerobic phase:** This is a preliminary phase that may take 2-4 days. In the anaerobic phase 5′-deoxyadenosylcobinamide is predominantly produced. **Aerobic phase:** In this phase, 5, 6-dimethyl- Benzimidazole is produced which finally form coenzyme of vitamin B-~12~ namely 5′-deoxyadenosylcobalamin. The bulk production of vitamin B~12~ is mostly done by submerged bacterial fermentation with beet molasses medium supplemented with cobalt chloride. **Recovery of vitamin B~12~:** The cobalamins produced by fermentation are mostly bound to the cells. They can be solubilized by heat treatment at 80-120°C for about 30 minutes at pH 6.5-8.5. The solids and mycelium are filtered or centrifuged and the fermentation broth collected. The cobalamins can be converted to more stable cyanocobalamins. This vitamin B~12~ is around 80% purity and can be directly used as a feed additive. However, for medical use (particularly for treatment of pernicious anemia), vitamin B~12~ should be further purified (95-98% purity). **Production of Vitamin B~12~ using Pseudomonas sp:** *Pseudomonas denitrificans* is also used for large scale production of vitamin B~12~ in a cost-effective manner. Addition of cobalt and 5, 6-dimethyl Benzimidazole to the medium is essential. The yield of vitamin B~12~ increases when the medium is supplemented with betaine (usual source being sugar beet molasses). #### Commercial Production of Riboflavin: There are three processes employed for the large scale production of riboflavin. Several microorganisms (bacteria, yeasts and fungi) can be employed for the production of riboflavin. In the acetone-butanol fermentation, employing the organisms *Clostridium aceto­butylicum* and *Clostridium butylicum*, riboflavin is formed as a byproduct. Commercial production of riboflavin is predominantly carried out by direct fermentation using the ascomycetes. The two plant pathogens namely *Ashbya gossypii* and *Eremothecium ashbyii* are most commonly employed due to high yield. Among these two organisms, *A. gossypii* is preferred as it is more stable with a high producing capacity of riboflavin. Industrial production of riboflavin is mostly carried out with the organism, Ashbya gossypii by using simple sugars such as glucose and corn steep liquor. Glucose can be replaced by sucrose or maltose for the supply of carbon source. It is essential to carefully sterilize the medium for good yield of riboflavin. The initial pH of the culture medium is adjusted to around 6-7.5. The fermentation is conducted at temperature 26-28°C with an aeration. The process is carried out for about 5-7 days by submerged aerated fermentation. Candida sp can also produce riboflavin. Besides sugars, other carbon sources have also been used for riboflavin production. A pure grade of riboflavin can be prepared by using *Saccharomyces sp*, utilizing acetate as sole carbon source. Methanol-utilizing organism *Hansenula polymorpha* was found to produce riboflavin. The other carbon sources used used with limited success for riboflavin production are aliphatic hydrocarbons (organism *Pichia guilliermoudii*) and n-hexadecane (organisms --- *Pichia miso*). Riboflavin is found in fermentation broth and in a bound form to the cells. The latter can be released by heat treatment i.e. 120°C for about 1 hour. The cells can be discarded after filtration or centrifugation. The filtrate can be further purified and dried, as per the requirements **Single cell protein (5marks)** Single cell protein refers to edible protein extracted from pure microbial cultures or dried cell biomass. Microorganisms like algae, fungi, yeast, and bacteria have very high protein content in their biomass. These microorganisms can be grown using inexpensive substrates like agricultural waste viz. wood shavings, sawdust, corn cobs etc. and even human and animal waste. The microorganisms utilize the carbon and nitrogen present in these materials and convert them into high-quality protein which can be used a supplement in both human and animal feed. The single cell proteins can be readily used as fodder for achieving fattening of calves, pigs, and poultry and even in fish breeding. The important microorganism used as single cell proteins are Chlorella, Scenedensmus, Chlamydomonas, Spirulina (algae),Saccharomyces, Candida, Torulopsis (fungi) Methylomonas , Alcaligenes etc. Like any other microbial culture, production of pure microbial cultures for desired protein products requires a carbon source, nitrogen source, and a source of other nutrients like phosphorus to support optimal growth of the culture. Contamination is prevented by maintaining strict sterile conditions throughout the process. The components of the culture media are either heat sterilized or filtered through micro-porous membranes. The selected microorganism is then inoculated in pure conditions. Most of the processes are highly aerobic, hence a good supply of oxygen is an indispensable requirement. After multiplication of the biomass, it is recovered from the medium and purified further for enhanced usefulness and storability. **Advantages- Refer text.** **Probiotics** Probiotics are defined as live microorganisms, which when administered in adequate amounts, confer a health benefit on the host. Health benefits have mainly been demonstrated for specific probiotic strains of the following genera: *Lactobacillus*, *Bifidobacterium*, *Saccharomyces*,*Enterococcus*, *Streptococcus*, *Pediococcus, Leuconostoc*, *Bacillus*, *Escherichia coli*. **Probiotics** are live [microorganisms](https://en.wikipedia.org/wiki/Microorganism)  provide health benefits when consumed, generally by improving or restoring the [gut flora](https://en.wikipedia.org/wiki/Gut_flora). Probiotics are considered [generally safe to consume](https://en.wikipedia.org/wiki/GRAS), but may cause [bacteria](https://en.wikipedia.org/wiki/Bacteria)-[host](https://en.wikipedia.org/wiki/Host_(biology)) interactions and unwanted [side effects](https://en.wikipedia.org/wiki/Side_effect) in rare cases. Some *Lactobacilli* are used for the production of yogurt, cheese, sauerkraut, pickles, sourdough, wine and other fermented products. In all cases, sugars are metabolised into lactic acid; thus creating a hostile environment for spoilage microorganisms and enabling food preservation. **Types of Probiotics** ----------------------- Many types of bacteria are classified as probiotics. They all have different benefits, but most come from two groups. **Lactobacillus**. This may be the most common probiotic. It\'s the one found in yogurt and other fermented foods. Different strains can help with [diarrhea](https://www.webmd.com/digestive-disorders/digestive-diseases-diarrhea) and may help people who can\'t digest lactose, the sugar in milk. **Bifidobacterium**. Found in some dairy products. It may help reduce the symptoms of [irritable bowel syndrome](https://www.webmd.com/ibs/default.htm) ([IBS](https://www.webmd.com/ibs/video/drossman-what-is-ibs)) and some other conditions. ***Saccharomyces boulardii* **is a yeast found in probiotics. It appears to help fight diarrhea and other digestive problems. Live probiotic [cultures](https://en.wikipedia.org/wiki/Microbiological_culture) are part of [fermented dairy products](https://en.wikipedia.org/wiki/Fermented_dairy_product), other [fermented](https://en.wikipedia.org/wiki/Fermented) foods, and probiotic-fortified foods. Some [fermented products](https://en.wikipedia.org/wiki/List_of_fermented_foods) that contain [lactic acid bacteria](https://en.wikipedia.org/wiki/Lactic_acid_bacteria) (LAB) include: vegetables such as [pickled vegetables](https://en.wikipedia.org/wiki/Pickling), [sauerkraut](https://en.wikipedia.org/wiki/Sauerkraut), soy products such as [tempeh](https://en.wikipedia.org/wiki/Tempeh), [miso](https://en.wikipedia.org/wiki/Miso), and [soy sauce](https://en.wikipedia.org/wiki/Soy_sauce); and dairy products such as [yogurt](https://en.wikipedia.org/wiki/Yogurt), [kefir](https://en.wikipedia.org/wiki/Kefir), and [buttermilk](https://en.wikipedia.org/wiki/Buttermilk). More precisely, sauerkraut contains the bacteria *Leuconostoc mesenteroides, Lactobacillus plantarum, Pediococcus pentosaceus, Lactobacillus brevis, Leuconostoc citreum, Leuconostoc argentinum, Lactobacillus paraplantarum, Lactobacillus coryniformis*, and* Weissella* spp.    Kefir contains *Lactobacillus acidophilus, Bifidobacterium bifidum, Streptococcus thermophilus, Lactobacillus delbrueckii* subsp.*bulgaricus, Lactobacillus helveticus, Lactobacillus kefiranofaciens, Lactococcus lactis*, and *Leuconostoc *species.Buttermilk contains either *Lactococcus lactis* or *L. bulgaricus*. **Benefits of probiotics** - probiotics help balance the friendly bacteria in our digestive system. - probiotics can help prevent and treat diarrhea. - probiotic supplements improve some mental health conditions. - certain probiotic strains can help keep our heart healthy. - probiotics may reduce the severity of certain allergies and eczema. **Probiotic foods** Yogurt. Yogurt is one of the best sources of probiotics, which are friendly bacteria that can improve your health. **Yogurt**  is a food produced by [bacterial](https://en.wikipedia.org/wiki/Bacteria) [fermentation](https://en.wikipedia.org/wiki/Fermentation_(food)) of milk.The bacteria used to make yogurt are known as *yogurt cultures*. The fermentation of [lactose](https://en.wikipedia.org/wiki/Lactose) by these bacteria produces [lactic acid](https://en.wikipedia.org/wiki/Lactic_acid), which acts on [milk protein](https://en.wikipedia.org/wiki/Milk_protein) to give yogurt its [texture](https://en.wikipedia.org/wiki/Texture_(food)) and characteristic tart flavor. [Cow](https://en.wikipedia.org/wiki/Cow)\'s milk is most commonly used to make yogurt. Milk from [water buffalo](https://en.wikipedia.org/wiki/Water_buffalo), [goats](https://en.wikipedia.org/wiki/Goat), [ewes](https://en.wikipedia.org/wiki/Sheep), [mares](https://en.wikipedia.org/wiki/Mare), [camels](https://en.wikipedia.org/wiki/Camel), and [yaks](https://en.wikipedia.org/wiki/Yak) is also used to produce yogurt where available locally. The milk used may be [homogenized](https://en.wikipedia.org/wiki/Milk#Creaming_and_homogenization) or not, even [pasteurized](https://en.wikipedia.org/wiki/Pasteurized) or [raw](https://en.wikipedia.org/wiki/Raw_milk). Each type of milk produces substantially different results. Yogurt is produced using a culture of [*Lactobacillus delbrueckii* subsp.*bulgaricus*](https://en.wikipedia.org/wiki/Lactobacillus_delbrueckii_subsp._bulgaricus) and [*Streptococcus thermophilus*](https://en.wikipedia.org/wiki/Streptococcus_thermophilus) bacteria. In addition, other [lactobacilli](https://en.wikipedia.org/wiki/Lactobacillus) and bifidobacteria are sometimes added during or after culturing yogurt To produce yogurt, milk is first heated, usually to about 85 °C (185 °F), to[denature](https://en.wikipedia.org/wiki/Denaturation_(biochemistry)) the milk proteins so that they do not form [curds](https://en.wikipedia.org/wiki/Curd). After heating, the milk is allowed to cool to about 45 °C (113 °F). The bacterial culture is mixed in, and that temperature of 45 °C is maintained for 4 to 12 hours to allow fermentation to occur. Kefir- Kefir is a fermented probiotic milk drink.It is a [fermented milk drink](https://en.wikipedia.org/wiki/Fermented_milk_products) similar to a thin [yogurt](https://en.wikipedia.org/wiki/Yogurt) that is made from **kefir grains**, a specific type of mesophilic [symbiotic culture](https://en.wikipedia.org/wiki/SCOBY).  The kefir grains initiating the fermentation consist of a symbiotic culture of [lactic acid bacteria](https://en.wikipedia.org/wiki/Lactic_acid_bacteria) and [yeasts](https://en.wikipedia.org/wiki/Yeast) embedded in a matrix of [proteins](https://en.wikipedia.org/wiki/Protein), [lipids](https://en.wikipedia.org/wiki/Lipid), and[polysaccharides](https://en.wikipedia.org/wiki/Polysaccharide). The matrix is formed by microbial activity and resemble small [cauliflower](https://en.wikipedia.org/wiki/Cauliflower) grains, with color ranging from white to creamy yellow. A complex and highly variable community can be found in these grains, which can include lactic acid bacteria, [acetic acid bacteria](https://en.wikipedia.org/wiki/Acetic_acid_bacteria), and yeasts. While some microbes predominate, [*Lactobacillus*](https://en.wikipedia.org/wiki/Lactobacillus) species are always present. The microbe flora can vary between batches of kefir due to factors such as the kefir grains rising out of the milk while fermenting or curds forming around the grains, as well as temperature. During fermentation, changes in the composition of ingredients occur. [Lactose](https://en.wikipedia.org/wiki/Lactose), the sugar present in milk, is broken down mostly to [lactic acid](https://en.wikipedia.org/wiki/Lactic_acid) (25%) by the lactic acid bacteria, which results in acidification of the product. Propionibacteria further break down some of the lactic acid into [propionic acid](https://en.wikipedia.org/wiki/Propionic_acid) . Other substances that contribute to the flavor of kefir are [pyruvic acid](https://en.wikipedia.org/wiki/Pyruvic_acid), [acetic acid](https://en.wikipedia.org/wiki/Acetic_acid), diacetyl and acetoin, [citric acid](https://en.wikipedia.org/wiki/Citric_acid), [acetaldehyde](https://en.wikipedia.org/wiki/Acetaldehyde), and [amino acids](https://en.wikipedia.org/wiki/Amino_acids) resulting from protein breakdown. Other probiotic foods include Sauerkraut,Tempeh,Kimchi. ,Miso and Pickles.