BY: SAI MANOGNA (MSIWM014)

Introduction:

An essential compound for general cellular metabolism is vitamin B6 (vitB6), or pyridoxine. Vit B6 has been involved in over 140 biochemical reactions in the cell as a cofactor. This vitamin was first discovered in 1934 by György and colleagues. While most of the co-catalyzed vitB6 reactions are linked to amino acid biosynthesis and catabolism, vitB6 also contributes to the biosynthesis of fatty acids and animals’ breakdown and plants of certain storage compounds, as well as in animals and plants.

Plant hormones, neurotransmitters, and organelle-specific compounds such as chlorophyll are biosynthesized. Moreover, reactive oxygen species ( ROS) can be quenched by vitB6. VitB6 is advantageous for photosynthesis because of its function in ROS scavenging and chlorophyll synthesis and is discussed as a potential factor to relieve biotic and abiotic stress.

Components of Vit B6:

A group of six chemically related compounds, all of which contain a pyridine ring as their center, constitute the vitamin. They differ from each other in the 4 ‘location of the pyridine variable group, which can either be an aminomethyl group (pyridoxamine (PM)), a pyridoxine (PN) hydroxyl methyl group, or an aldehyde group (pyridoxal (PL). Once the various derivatives are phosphorylated, they can serve as cofactors, with vitB6 being the biologically active form of pyridoxal 5’-phosphate (PLP).

Fig: a. Pyridoxal, b. Pyridoxal 5’-phosphate

       c. Pyridoxamine, d. Pyridoxamine 5’-phosphate

       e. Pyridoxine, f. Pyridoxine 5’-phosphate

Production of Vit B6:

Several studies on the production of vitamin B6 and vitamin B6 are known to be provided by various microorganisms belonging to the Saccharomyces, Pichia, Klebsiella, Achromobacter, Bacillus, and Flavobacterium genera. It is possible to manufacture high-yielding vitamin B6. Microorganisms belonging to the genus Rhizobium will accumulate in the culture broth a significant amount of vitamin B6 that can be recovered from it in the desired purity or under aerobic conditions by extracting the resulting vitamin B6 from the fermentation broth in an aqueous culture medium.

1. Saccharomyces carlsbergensis will assay the quality of vitamin B6 in a fermentation broth.

2. High-performance liquid chromatography may also separately calculate the quality of vitamin B6 components such as pyridoxine, pyridoxal, and pyridoxamine in a fermentation broth.

3. The aqueous culture medium, which contains assimilable sources of carbon, digestible sources of nitrogen, inorganic salts, and other nutrients necessary for microorganism development, microorganisms belonging to the genus Rhizobium are incubated.

4. Glucose, fructose, sucrose lactose, galactose, maltose,  dextrin, starch, or glycerol may be used as carbon sources.

5. Peptone, soybean powder, steep corn liquor, meat extract, ammonium sulfate, ammonium nitrate, urea, or mixtures can be used as a source of nitrogen.

6. Also, calcium, magnesium, zinc, manganese, cobalt, and iron may be used in inorganic salts, sulfates, hydrochlorides, or phosphates. Moreover, it is also possible to incorporate conventional nutrient factors or an anti-foaming agent such as vegetable oil, animal oil, or mineral oil.

7. Appropriately, the pH of the culture medium is about 5.0-9.0, ideally about 6.5-7.5.

8. The temperature of cultivation is appropriate from about 10 °- 40 ° C., ideally from about 26 ° to about 30 ° C.

9. The cultivation time is appropriate for 1 to approximately 14 days, ideally for 2 to approximately seven days.

10. Aeration and agitation usually give favorable results in cultivation.

11. The presence in the medium of a compound chosen from pyruvate, D-glyceraldehyde, glycolaldehyde, glycine, 1-deoxy-D-threopentulose, 4-hydroxy-L-threonine, and a suitable combination thereof gives the vitamin B6 titer with more desirable results and is therefore favored.

12. As a supplement for the development of vitamin B6, the combination of 1-deoxy-D-threopentulose and 4-hydroxy-L-threonine is particularly significant.

13. The synthesis of vitamin B6 can also be accomplished by incubating pyruvate, D-glyceraldehyde, glycolaldehyde, glycine, 1-deoxy-D-threopentulose and 4-hydroxy-L-threonine cells of microorganisms belonging to the genus Rhizobium, separated from the culture broth in a buffer of the correct pH value.

14. Vitamin B6 produced may be separated from the culture broth after cultivation and purified.

15. For this reason, by using different vitamin B6 properties, a method used to separate the product from the culture broth can be applied. Thus, for instance, the filtrate’s desired material is purified using an ion exchange resin or similar means after the cells have been separated from the culture broth.

16. The ideal substance is recrystallized from alcohol upon elution.

The microorganism is used in conjunction with all strains belonging to the genus Rhizobium that are capable of producing vitamin B6 and are deposited for availability in a public depository, i.e., culture collection.

First Oxygen producing Species:

According to researchers, vitamin B6 may have given birth to the Earth’s first oxygen-producing species. The earth witnessed a significant increase in atmospheric oxygen levels around 2.4 billion years ago. Scientists have long held that this surge in oxygen, called the Great Event of Oxygenation, was related to the first photosynthetic organisms. Oxygen is a by-product of photosynthesis that converts carbon dioxide into sugar foods using sunlight. Nevertheless, no one understood why these species producing oxygen appeared in the first place.

Researchers found that about 2.9 billion years ago, at the same time that the enzyme manganese catalase appeared, the oldest oxygen-based method involved the manufacture of pyridoxal, a type of vitamin B6. Manganese catalase splits hydrogen peroxide into water and oxygen. When attempting to cope with environmental hydrogen peroxide, early species may have come across this enzyme, which some geochemists claim was plentiful at the time in Earth’s glaciers and was released by the bombardment of solar radiation. By breaking down the glacial hydrogen peroxide with manganese catalase, the species ultimately obtained the oxygen they required to create pyridoxal.

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