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The recent developments in the various scientific fields have revealed the extraordinary effect of biotechnology on them, as these developments have presented new horizons in the field of biotechnology for everyday life. Biotechnology has a significant part to play in the various scientific fields which are discussed as below:
The fermentative or microbial production of amino acids is the primary topic of discussion as it is one of the primary objectives of biotechnology. Biotechnology has found its use in the industrial application such as the manufacturing of food, feed, pharmaceuticals, cosmetics and chemicals. It has established its share in the global market of 5-7% annually and is worth the amount of billion dollars. Scientists have banked on two main microbes Corynebacterium glutamicum and Escherichia coli for amino acid production, as their research criteria. The strains producing amino acids were developed through simple breeding methods such as mutation and selection. The amino acids development in terms of its production and diversification has led to the utilization of many fields for this purpose such as, physiology, biochemistry, molecular biology. The knowledge of these various field have helped in developing metabolic pathways, transport function and regulatory mechanisms. In addition the scientists have been able to develop means of identification of the target gene which is the control system of all the activities taking place in the organisms. This has been possible due to the usage of Genome sequencing, proteomics, genomics and metabolic flux analysis. This has proved to be a great success both qualitatively and quantitatively. But the intricate details of the metabolic pathways and networks are the primary part of the research (1) (2).
The production of Lysine through C. glutamicum and E. Coli, is one of the major breakthroughs. Previous attempts on C. Glutamicum with the help of classical genetics led to the less stable strains as compared to the wild type. But the recent researches have proved far more productive. The 3 gene targets mutant alleles for high lysine production such as lysC, hom and pyc for aspartokinase, homoserine dehydrogenase and pyruvate carboxylase respectively were made a part of the wild type. As a result the strains produced demonstrated a high lysine yield and was also able to sustain the robustness and significance of the wild type. Similar is the case of genome sequencing and annotations of the sequences which have led towards noteworthy indications of the missing metabolic steps in the lysine biosynthesis pathway. The mutation identifications have been possible owing to the comparative genomics that has benefitted the lysine synthesis. While point mutations identification have been possible owing to the wild type and the metabolically engineered comparative genome analysis which led to the construction of a superior strain for the production of lysine. Studies on the E.coli gene have made it possible for the mutant C. Glutamicum to use lactose, galactose or starch for the growth and development which was an additional point for it as the wild type was incapable of doing so. But the process of developing the production of amino acids by microbes and its diversification through modern genetics is still going on. C. Glutamicum is still the primary producer in case of lysine. . (3)(4)
Researchers have carried out novel studies on lysine in plants also. As was the case of corn, the primary food source for humans and animals alike. The corn was found to have low lysine content; hence a genetically engineered corn type was created such as opaque2 type corn. Besides lysine in corns, scientists are also working on increasing the amino acids content in plants also. (5)
Many needs and reasons have led to the biotechnological manufacturing of vitamins such as the increasing need of people taking vitamins in their daily routine. These vitamins are taken out from plants and animal sources but may not suffice the requirements of vitamins by the whole population. As vitamins deficiency in the diets of the people has increased pressure on the demand of vitamins. Another alternate of chemically manufacturing vitamins did not prove to be a great idea due to the health and environmental issues attached to this production.
Recently scientists have centralised their efforts towards the processes of fermentation and bioconversion as a biotechnological method for producing vitamins. But with these entire efforts one must admit the difficulty in the task of manufacturing microbial strains possessing the ability of hyper production of vitamins. The primary reasons are the requirement of further research in the comprehension of pathway elucidation and metabolic regulations in microbes. The biosynthesis of vitamins is a delicate and a complicated procedure owing to its limitation to the prokaryotic world. The presence of r-DNA in the commercial industry has improved the fermentation process and in addition has also immobilised bioconversions for the chiral vitamin compounds synthesis. The only issue yet left to be solved are the purification of vitamins from the fermentation mixtures (6) (7)
The unique capability of simultaneous production of antioxidant vitamins such as b-caretone, vitamin C and vitamin E is found present in a few prokaryotes such as flagellate alga Euglena gracilis Z. The vitamin content was found to increase in the cells when the species was developed in a 2 stages procedure of photoheterophical production and then photoautotrophic production. Therefore the yield of vitamins also increased all owing to the excess production of secondary metabolites in the various light conditions (8)
Bacillus, Methanobacterium, propionibacterium and Pseudomonas , the microbial genera for B12 have also been utilised in the industrial production of vitamins. The development on the vitamins production and its diversification was obtained by the usage of modern genetic engineering on Propionibacterium freudenreichii .this particular type contained 10 various genes associated with hem, cob and cbi genetic families by utilising different vectors to increase the production of vitamins B12. When the various gene types wee expressed, they produced different levels of vitamins such as cobA, cbiLF and cbiEGH, which showed an increase in the vitamin B12 by 1.5 fold, while the cobU and cobs indicated a slight increase. In case of a multigenetic combination of exogenous hemA gene and endogenous hemB and cobA, an extraordinary increase by 2.2 folds was observed as compared to the non recombinant type. This could be considered as a remarkable developed in the light of only 70 stages required in the chemical synthesis of vitamin B12 (9).
Plants have also been the focus of research to make the biosynthesis of vitamins possible in plants. The pathways of pro-vitaminA, C and E synthesis have been elaborated upon several times. Increased researches on the possibility of increased vitamins content in plants for increased health benefits and to have a source available for vitamins extraction are being carried out. The target gene identification is being aided by the facility of metabolic engineering (10)
The antibiotics are now been produced industrially which is in itself one of the outstanding examples of biotechnology. Therefore the antibiotics have been able to grasp a major part of the market both commercially and industrially. Antibiotics have a large role to play in therapeutics as compared to other drugs. Manufacturing and developing antibiotics has become a huge challenge in the light of new emerging diseases and the rapidly increasing occurrence of drug resistance in pathogens. The field of combinatorial chemistry, biochemistry, evolutionary developments of new genes and pathways are all contributing to a promising future. But the research will continue with the population boom and hence the rise in health problems (11).
The biotechnological researches were increased to identify the bacterial targets after the overall genome sequencing was done of the two pathogens Haemophilus influenza and Mycoplasma genitalium in 1995. The identification and precise characterisation of the bacteria was necessary to manufacture antibiotics targeting the diseases with accuracy. These efforts have led to the sequencing of 500 bacterial pathogens with their relevant drugs being produced (12).the antibiotics producing genes identification in various organisms has been made easy because of the genome analysis. The genome analysis done in the case of actinomycetes species has indicated numerous clusters encoding specific putative products. The identification of gene clusters associated with natural product biosynthesis was possible due to the high output genome scanning. This further aided the discovery of biosynthetic pathways encoding enediyne anti tumour antibiotics found in specific actinimycetes species. These certain methods and techniques will eventually help in locating the precise range of such products present thus increasing the possibilities of application of the product (13).
Penicillin, cephalosporin and cephamycins biosynthetic pathways have been genetically and biochemically characterised in detail. Ahead is an example involving Streptomyces clavuligens and Penicillium chrysogenu. The cloning process has been applied on genes producing b-lactam antibiotics except the isopenicillin N epimerase (cefD) gene. The increase in the amount of cephalosporin C production and the decrease in the amount of deacetoxycephalosporin production in fermentation broths was a result of amplification of specific genes. Transferring the expression of cefE gene from S. clavuligens to P. Chrysogenum, results in the formation of adipyl-7-aminodeacetoxycephalosporanic acid (adipyl 7-ADCA). Hence the exploitation of an inherent increased capacity of biosynthesis for P. Chrysogenum for the cheap manufacturing of these products and with the aim of fulfilling the demand. The antibacterial reaction of Adipyl 7-ADCA has an extremely strong effect on both the gram positive and the gram negative bacteria (14)
The separation of antibacterial content from the Myxobacteria, which has the potential of reacting against Tuberculosis producing pathogen, is another achievement. These are called as myxopronin, corallopronin and ripostatin. These have been characterised and separated to give results by restricting the bacterial RNA polymerase enzyme. They therefore attach to the location on the RNA polymerase not previously visible. Hence they have the potential to work against bacteria, against which other conventional antibiotics are useless. Although there is still time in the publishing of these researches and they will be recognised soon. The biotechnological achievements and efforts need to be recognised for developing the yield and diversity of the antibiotics (15)
The researches on bio fuels have been ongoing owing to the growing issues of increasing prices of crude oil, environmental issues, political turmoil, etc. Bio fuels have the potential of competing in the market with fossil fuels or scientists suggest they may even be able to replace them. This may only be possible if the bio fuel is able to provide a clean energy within the cost and may not result in the usage of natural source. Two main bio fuels have been produced so far which is ethanol from corn and biodiesel from soybeans. In future it is expected that the production of fuel from biomass will be done instead of food grains. Plant source can be converted into bio fuel by microbes, which can become the renewable energy source. Biomass conversion to valuable fuels and products is now been emphasized more with the advancements in genetics, process chemistry and bioengineering. Hence bio-refinery is the term coined for the whole process. Bio ethanol is the only bio fuel produced industrially today by microbial enzymes. But a further research is required in fuels such as bio ethanol, bio methane, biodiesel and biobutanol via microbes (16) (17) (18)
The production of biodiesel from crops, cooking oil or animal fat does not have commercial benefits although biodiesel is an efficient fuel option. Microalgae are one of the best sources of biodiesel according to research. Microalgae similar to plants use sunlight to produce oils but are found to be more efficient. The capacity of producing oil is more as compared to other oil producing plants. Other bio fuels it can produce are methane from anaerobic digestion of algal biomass, biodiesel production from microalgal oil and the production of bio hydrogen through the process of photobiology. Other than the above mentioned products, microalgae have the capacity to produce lipids, hydrocarbons and complex oils. Another source recently discovered is through oil producing heterotrophic microorganisms developed on the source of food such as the natural organic carbon present in sugar. The efficiency of this process is low due to the limited availability of organic carbon from the crop plants only. A large scale manufacturing of oil rich microalgal biomass would be needed for these processes. Hence the manufacturing of this biomass is not practical which negates its possibility to be adopted commercially (19).
Another important source for production of biodegradable compounds is the microbial fuel cells (MFCâ€™s). An anode is used as an electron acceptor the microbes generated an electrical output. The MFCâ€™s operate on various carbohydrates and other complex substrates present in the waste matters. The energy present in the bio convertible substrates is changed into electricity by them. When the microbes move from a natural electron acceptor such as the oxygen and the nitrate, to MFC anode which is the insoluble electron acceptor, the conversion into electricity is possible. This transfer is possible through membrane components and electron shuttles. The process of reduction takes place when the electrons flow to the cathode through a resistor. The MFC develops an electrical current and an off gas constituting of carbon dioxide, which is quite the opposite of the anaerobic digestion. The MFCâ€™s offer new opportunities for the fuels because of its benefits of large production of bio energy by the direct conversion of substrate energy into electrical energy. In addition it can operate at low temperatures and they do not need the treatment by gas or any other energy input. . (20)
The research work is certainly progressing at high speed although there is still time required in the commercial use of these processes. But up till now the scientific researchers are pointing out to promising prospects in future.