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[edit] Introduction

[edit] Fungal Pharmaceuticals

The Kingdom Fungi is classified as eukaryotes. Fungi include a large group of organisms which are found in numerous forms, behaviours and life cycle patterns [1]. They are heterotrophic and spore-bearing, which make them unique organisms. Fungi also serve an important contribution towards the science and have several medical uses.

Over the past century fungi have been found to be extremely useful organisms in biotechnology. Fungi produce many unique complex molecules and several of its biologically active compounds have been discovered and used in the field of medicine. The role of fungi was established very early in history. During the beginning of civilization yeast has been used in the making of bread and alcohol. In more modern times the discovery of penicillin began the development of a new approach to treating microbial diseases in humans. Fungi and their metabolites have been immensely important in medicine and pharmaceutics. Since the discovery of penicillin by Alexander Fleming in 1928, there has been an increasing number of investigations seeking to exploit fungal metabolic pathways for potential novel drugs. In the following years, important antimicrobial pharmaceuticals such as Griseofulvin (derived from Penicillium griseofulvum) and Cephalosporin (Acremonium) were discovered [2]. The importance of fungi is being expanded way beyond its capacity to transform and protect.

A wide range of metabolites are discovered by examining fungi under a matrix of culture conditions. To date there are over 3000 different primary and secondary metabolites have been described from fungal sources [3]. Metabolites from chemical compounds are formed as part of the natural biochemical process of degrading. They serve an important ecological function such as antibiotics. Today the antibacterial market is dominated by three families of wide-spectrum drugs: the semi-synthetic β-lactams, largely in combination with the β-lactamase inhibitor clavulanic acid, e.g., Augmentin; the macrolides, such as clarithromycin; and the fluoroquinolones, e.g., ciprofloxacin. The beta-lactam antibiotics have been serving mankind for several years and still they continue to provide health to the world population by virtue of industrial production and discoveries of new secondary metabolite molecules with useful activities [4].

[edit] History

Ever since the 18th and 19th century fungi in pharmaceuticals have been vigorously studied. There are over 200 different fungi, however only 74% are used in antimicrobial activity. Basidiospores, have a long history of medical uses. The tendor polypore,Fomes Fomentarius, was used in hemostatic dressing and bandages. Dioscorisdes- a Greek physician discovered the use of Fomitopsis(Polyporus) officinalis that was used to treat tuberculosis. That very same polypore was found in the body of a 5300 year old Ice-Man. SInce bacteria were very successful organisms that are widespread and adaptable something needed to be done. Life for humans was not easy and the development of penicillium was a tremendous breakthrough. Penicillin production was triumph in both scientific and technological terms.[5]

It was in the year 1928, that Penicillium- most widely used fungi was discovered by accident at the St.Mary's Hospital in London by Alexander Fleming. Mr.Fleming left culture dishes of the pathogenic bacterium,Staphylococcus aureus on his work bench and upon his returned from a brief holiday, to his surprise a fungus colony had been found. Fleming identified the contaminating fungus-Penicillin. During the 1930's he continued to study the fungus but was unsuccessful at purifying and stabilizing it. However, he did not let them discourage him for he knew of the potential that penicillin could have both on the clinical and large scale.

Besides Alexander there were other "founding fathers" involved in the discovering and potential of penicillium. In 1940, Howard Flowery and Ernst Chain conducted a survey to show effective penicillin is in chemotherapeutic properties. In 1941, development of penicillin use in fermentation was discovered in the USA. Dorothy Hodgkin and Barbara Law established the β-lactam structure of penicillin by the use of X-ray crystallography. in 1956, John Sheehan used chemical synthesis to produced penicillin.

Some fungi have a history in Asian folk medicine. Ganoderma lucidum has been used in ancient times to treat hepatitis, hypertension, hypercholesterolemia, and gastric cancer, and found to possess antimicrobial and anti-HIV activities[6].

[edit] Fungal products found in pharmaceuticals

[edit] Taxol

Chemical structure of taxol.
Chemical structure of taxol.

Originally isolated in 1971, Taxol, or Paclitaxe as it is now referred to as, was found to have anti-tumour properties and is an important agent in chemotherapy [7]. It helps prevent the growth of tumours by disrupting microtubule formation, and preventing proper cell division from occurring[8]. Taxol was first harvested from yew trees, Taxus brevifolia, from the inner bark, however, the demand caused a marked rise in price. As a result, alternative means for producing taxol were explored; one being production of taxol from microbial cultures. The fungi, Pestalotiopsis microspora, collected from Taxus wallachiana, was found to produce taxol in culture [9]. P. microspora was able to produce 60-70 μg of taxol per litre of fungal culture. Its production peaked at roughly the 3 week mark and declined drastically at 5 weeks. Although the amount of taxol produced was one order of magnitude less than that harvested from T. brevifolia, the accessibly of growing a fungal culture in terms of space and time far exceeds that of harvesting from yew trees.

Biochemical pathway of cholesterol synthesis and effect of statins on pathway.
Biochemical pathway of cholesterol synthesis and effect of statins on pathway.

[edit] Statins

Statins are a group of secondary metabolites that are used to lower low-density lipoprotein (LDL) cholesterol. It is the most widely used pharmaceutical drug in the world. This ability is very important for combating coronary disease since two-thirds of the total cholesterol in an individual is synthesised in the body, with the other third from dietary sources[10]. Therefore, suppressing de novo synthesis of cholesterol is a significant method to lower cholesterol levels in an individual with hypercholesterolemia. Mevastatin was the first statin to be isolated in 1976 from Penicillium citrinum and P. brevicompactum. These compounds work by inhibiting 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, which is the rate-limiting enzyme for cholesterol synthesis[11]. Lovastatin was the first of these compounds to be approved for medical use in 1980. Since then, other statins that have been isolated or synthesized including fluvastatin, pravastatin and simvastatin, which all act in similar fashions to lower LDL cholesterol [12]. These pharmaceutical products have been extremely important to pharmaceutical companies, especially one statin derivative, atorvastatin, generating sales revenues of roughly 7 billion USD in 2005[13].

[edit] Cryptocandin

Some diseases that humans are susceptible to are those from fungi and can cause serious problems, especially since infections tend to target immunocompromised patients. To combat this, fungi themselves have been turned to as select strains produce antifungal agents that can be utilized against pathogenic fungi. One such agent is cryptocandin which was isolated from Cryptosporiopsis quercina[14]. This compound works most effectively against Trichophyton which a common fungi in skin infections such as athlete's foot and ringworm. [15]. Cryptocandin was also found to be as effective as amphotericin B, a common antifungal agent, at inhibiting Histoplasma capsulatum and Candida albicans, which can cause pulmonary and oral infection, respectively. [15]. Since it effects Trichophyton most heavily, cryptocandin is an effective antifungal agent for skin and nails diseases.

[edit] Cephalosporin

Chemical structure comparisons between Cephalosporin and Penicillin.
Chemical structure comparisons between Cephalosporin and Penicillin.

Cephalosporin was first discovered from Acremonium chrysogenum in 1953. Cephalosporin compounds were first isolated from cultures by an Italian scientist Giuseppe Brotzu in Sardinia in 1948. This fungal species served great purpose in the production of several antibiotics including Cephalosporin C, Cephalosporin P1-P5 and Penicillin N. This species of Cephalosporin is one of the most widely used and prescribed antibiotics which composes 29% of the antibiotic market [16]. The structure of Cephalosporin as well as its mode of action, is very similar to that of Penicillin such as their beta-lactam ring which contribute to their bactericidal action. Cephalosporin is very valuable due to their low toxicity and their broad spectrum of action against various diseases. This species will affect bacterial growth by inhibiting cell wall synthesis in gram-positive and negative bacteria [17]. The mode of action as stated previously of Cephalosporin are bactericidal agents. This species similar to Penicillin will disrupt the synthesis of the peptidoglycan layer of the bacterial cell walls, which in turn, causes the walls to break down and eventually the cell to die [18].

[edit] Mushrooms

Mushrooms have a high medical impact for pharmaceutical products[19]. Due to mushrooms having a high tolerance with the chemotherapy and radiotherapy the products obtained from it are used for cancer therapies. Mushroom fruiting bodies and their extracts are very effective and are an economically better option due to the faster growth of the fruiting body. There are few prominent mushrooms having pharmacological properties and they are: Agaricus brasiliensis, Ganoderma lucid,lentinula edodes'' and Grifola frondosa. Other biologically active substances found in mushroom includes immunosuppressants such as:nematicide,antimicrobial,antiviral, and hypocholesterolemic agents. Black tea fermented with dabaryomyces hansenii contains lots of major vitamins such as A,B1,B2,B12 and C. It also results in reduction of caffeine and tannins in significant amount.

This link provides a video from the National Geographic, involving magic mushrooms being used as a medical treatment to cluster headaches. (Warning, video contains disturbing scenes). Magical Mushroom

Reishi Mushroom Provides new fundamental medical treatments to various illnesses.

[edit] Laccases

Laccases belong to a large family of multicopper containing enzymes. They are mainly found in Fungi and some high order plants. First explained in 1883 makes them one of the oldest known enzymes. Their wide range of activities comes from their Electron Transfer abilities, they are best known for their importance in the delignification process of white rot fungi. Their diverse range of functions, especially in pathogenic fungi, make them an ideal enzyme to be studied in order to investigate the mechanism of how a pathogenic fungi behaves once inside its host environment. Another use for Laccases is in Drug Analysis. A new enzymatic method based on laccase has been developed to distinguish morphine from codeine simultaneously in drug samples, employing the enzyme's oxidizing abilities. Laccase is able to oxidize morphine but not codeine. This method is proven to be fast and efficient which makes it quite useful in the massive industry of pain killers. [20]

[edit] Cordyceps sinenis

Cordyceps sinenis is a Fungi popular among the old Chinese folk medicine, for hundreds of years people native to lands of China have used this fungus as a part of their herbal treatment processes to treat conditions like allergies and skin infections. It has been recently proven that an extract taken from mycelia of this fungus showed some very strong tumor suppressing characteristics. studies in Vitro and in VIVO conducted with mice proved the effectiveness of Ergosterols in its two forms 5α,8α-epidioxy-24(R)-methylcholesta-6,22-dien-3β-d-glucopyranoside and 5,6-epoxy-24(R)-methylcholesta-7,22-dien-3β-ol. [21]

[edit] Griseofulvin


Griseofulvin was first discovered in the early 1960's and was the first orally effective antibiotic that was used for dermatophytosis management and later broad-spectrum agents arrived. It is the only broadly useful antifungal agent and it's original source was Penicillium griseofulvin.This commercial production of Griseofulvin was derived from a much mutated strain of P. patulum. It is used for the treatment of dermatophytes, as it accumulates in the hair and skin following topical application. Furthermore, it is the only antibiotic that is effective against fungal infections of skin, athlete's foot, and ringworm. Griseofulvin is an antifungal antibiotic that will fight against infections caused by a fungus. This antifungal drug inhibits mitosis in fungal cells and weakly in mammalian cells by afecting mitotic spindle microtubule function [22]. Analysis of Griseofulvin has shown to have the ability to inhibit cell proliferation and mitosis as well as the inhibition of cell-cycle progression at prometaphase/anaphase of mitosis [23]. Griseofulvin only inhibits fungal growth but does not kill fungi, this antifungal agent affects a wide range of fungi but is limited to those with chitinous cell walls [24].

[edit] Cyclosporin

Tolypocladium inflatum.
Tolypocladium inflatum.
Cyclosporin's mode of action.
Cyclosporin's mode of action.

Cyclosporin is a drug obtained from a type of soil fungus found in Norway called Tolypocladium inflatum. Jean-Francois Borel, a microbiologist working for a labratory in Switzerland, discovered Cyclosporin in 1969 when he was vacationing in Norway [25]. This drug is fairly new and was brought to North America by the U.S. Food and Drug Administration in 1983 [26]. Cyclosporin is one of the most commonly used immunosuppressant drug in organ transplants. Its function is to suppress the action of certain cells in the body's immune system that can reject the transplanted organ, as well as, let the bulk of the body's immune system function normally and fight general infection [27]. Cyclosporin was found to inhibit the activity of white blood cells, the specific part of the immune system that starts the process of detecting and attacking foreign invaders. This prevents the attacks of cells called T-cells.

[edit] Cilofungin

Cilofungin is the first clinically applied member of antifungal drugs which was derived from a fungus in the genus Aspergillus [28]. This antibiotic is specifically produced to treat and effect Candida albicans. Cilofungin is a lipopeptide antifungal agent active against several species of Candida, and acts as a beta glucan synthase inhibitor [29]. This antibiotic utilizes phagocytosis and intracellular killing of Candida albicans blastospores by glass-adherent human neutrophils as well as interfering with an invading fungus' ability to synthesize the cell wall [30]. However, this drug was discontinued due to the incidence of metabolic acidosis.

[edit] Plectasin

Plectasin is an antimicrobial peptide with the potential for treating infections. It is a new fungal defensin, effective against drug-resistant bacteria isolated from a saprophytic ascomycete [31]. Plectasin has shown potent activity comparable to Vancomycin and Penicillin. An experiment performed reveals that plectasin tested against Streptococcus Pneumoniae proves to be highly effective against it. Protein therapeutics has the potential to generate immune response through the development of antidrug antibodies. Also, plectasin has potent "in vivo" activity and exhibits low systemic toxicity and a novel mode of action without cross-resistance to other antibiotics impediment. Therefore, its pharmacokinetic profile is favourable and it is within cost budget [32]. Hence, it can be used as antibacterial in future.

[edit] Processes Involved

[edit] Penicillium chrysogenum

Chemical structure of penicillin made from Penicillium chrysogenum.
Chemical structure of penicillin made from Penicillium chrysogenum.

Penicillium chrysogenum is a common species used in the production of penicillin because it has an improved product accumulation and improved cycles of mutations. The amino acids inolved in producing penicillin are cysteine, and valine and the third compound involved is aminoadipic acid. This makes penicillin a tripeptide. The primary enzyme involved in penicillin biosynthesis is ACV-synthetase, which helps to form the tripeptide ACV from the three fundamental amino acids. The second step in the biosynthesis process is the formation of β-lactam ring from ACV by the enzyme isopenicillin-N synthetase. It is important to note that oxygen is involved in the ring closure process but is not incorporated into the ring structure itself. The final step includes adding or exchanging side chains by acyteyl CoA. This is a two-step process which involves, firstly, the release of isopenicillin-N side chain to yield α-aminoadipate and 6-aminopenicillanic acid, and secondly, the addition of the new side chain and release of acetyl CoA. [33].

[edit] Industrial Processing

Antibiotics are secondary metabolites which are produced by fungi and many other organsisms. Secondary Metabolites are produced by many complex chemical pathways with many steps and reactions involving various enzymes, proteins, and other molecules. Different steps in the pathways can control how much of the product is produced, this is know as a rate-limiting step. Many factors go into the process of enhancing these rate-limiting steps in order to produce high yields of the secondary metabolite including pH, oxygen concentratoin, nutrients available, temperature, and the concentration of waste products.

penicillin production by fed batch submerged fermentation
penicillin production by fed batch submerged fermentation

The industrial production of secondary metabolites as antibiotics is known as industrial fermentation. An initial strain of an organism is grown in mass amounts. Fed-batch submerged fermentation is one of the most common forms of fermentation used for penicillin production.[34]. The specific strain of fungi to be used is first amplified through a number of intermediate seed stages prior to production. The production stage consists of large amounts of the fungi organism, which is called the inoculum, being placed along with a specific production medium, in large stirred rank reactors. Creating a high yield of the secondary metabolite is desired so penicillin precursors are added such as phenylacetic acid. However these precursors are toxic to the fungi thus they must be fed into the batch while maintaining a non-toxic concentration.[35]. Hence the name fed-batch fermentation. The batch is then filtered, getting rid of mould mycelium, cooled, and put in a holding tank where it will continue on to be purified.

[edit] Purification

There are many different forms of purification that are used even for just one secondary metabolite such as penicillin. Methods of purification for penicillin include solvent extraction [36], supported liquid membrane permeation [37], emulsification of liquid membrane[38], microfiltration [39], hollow-fibre membrane technique[40], and reactive liquid-liquid extraction[41].

[edit] Medicinal uses of Fungi in Society

Various Fungal Antibiotics
Various Fungal Antibiotics

[edit] Antibiotics

Fungal products are chemicals that are derived as part of the life process of one organism that can be used to kill or stop the growth of other microorganisms are referred to as antibiotics. An antibiotic is a substance produced by certain bacteria or fungi that kills other cells or interferes with their growth. In nature, these substances help some microbes survive by limiting the multiplication of other microbes that share the same environment [42]. Antibiotics that attack pathogenic (disease-causing) microbes without severely harming normal body cells are useful as drugs, the most common and most well known antibiotic is penicillin [43]. The discovery of penicillin marked the beginning of a new approach to treating human disease and established the importance of fungi in pharmaceuticals. Because of Fungi, the current top five best-selling antibiotics in the world are being produced as well as new treatments are being established to fight against diseases. Furthermore, new forms of medical interventions are being discovered all possible only because of the powerful fungi. Antibiotics are selectively toxic-that is, they damage some types of cells without harming others. Medically useful antibiotics attack infectious microbes or cancer cells without excessively hurting human cells. Antibiotics fight different types of illnesses in a variety of ways.

[edit] Fatty Acids

Mortierella alpina is a common species among the zygomycota used to produce arachidonic acid.
Mortierella alpina is a common species among the zygomycota used to produce arachidonic acid.

Arachidonic acid is one of the essential fatty acids that is required by most mammals. It is produced through a two-step fed-batch fermentation process by zygomycete fungi and is composed of a carboxilic acid with a 20-carbon chain. This essential fatty acid is found in the membranes of many animal cells which contribute to fluidity. Arachidonic acid is effective in lowering cholesterol and triacyglycerols in the blood, reducing the severity of arteriosclerosis and other cardiovascular diseases. [44]. It is also important in the development of the retina and the brain in newborn infants. This fatty acid is also a precursor to certain chemicals in the human body. Such precursors include leukotrienes, which mediate inflammation, prostaglandins (which help immune function), and Thromboxane, which is a strong vasoconstrictor. [45].

[edit] Statins

Statins have been recommended for wide spread use to control heart disease. Aspergillus terrreus and Aspergillus griseus produces these secondary metabolites statins that have been used to reduce or even remove low-density lipoproteins from blood vessels in humans. Statins acts on an enzyme in the liver that makes cholesterol. It works by blocking the enzyme, and the body removes cholesterol complexes from the inside of blood vessels. As a result, this reduces or removes blockages in the arteries and thereby, reducing the likelihood of heart attack, diabetes and strokes.[46]

[edit] Terms and Definitions

  • Bactericidal agents: Agents that will ultamitely kill bacteria and their cells, which can be done by heat of any chemical.
  • Crystallography: The branch of science concerned with the formation, properties, and structure of crystals.
  • Defensin: Protein molecules that are produced by animals to protect themselves against infection.
  • De novo synthesis: Refers to the synthesis of complex molecules from simple molecules such as sugars or amino acids, as opposed to their being recycled after partial degradation.
  • Dermatophytosis: An infection of the skin, hair, or nails caused by a dermatophyte and characterized by redness of the skin, small papular vesicles, fissures, and scaling.
  • Endophytic: Endophytic refers to fungi or bacteria that lives symbiotically with plants without causing any harmful effects to the plant.
  • Hypocholesterolemic agents: Hypocholesterolemia is a condition characterized by very low cholesterol levels without the assistance of medications used to lower cholesterol.
  • Metabolic acidosis: Acidosis and bicarbonate concentration in the body fluids resulting either from the accumulation of acids or the abnormal loss of bases from the body (as in diarrhea or renal disease
  • Peptidoglycan layer: The dense material consisting of cross-linked polysaccharide chains present in the cell wall of most bacteria.
  • Primary metabolism: The organism breaks down food in the environment to extract nutrients needed for the maintenance of cell structures
  • Secondary metabolism: It is driven by the competition for resources in a nutrient-poor environment.
  • T-cells are a type of lymphocyte that are produced or processed by the thymus gland and actively participating in the immune response.

[edit] Notes and References

  1. Sumbali, G. (2005). The Fungi. Retrieved from
  2. Amal, A., Debbab, A., Proksch, P. (2011) Fifty years of drug discovery from fungi. Fungal Diversity, 50: 3-19.
  3. Kettering M, Sterner O, et al. Antibiotics in the chemical communication of fungi. Z Natforsch C J Biosci 2004; 59(11–12):816–23
  4. Lubertozzi, D. Keasling, J. (2009). Developing Aspergillus as a host for heterologous expression. Elsevier, Biotechnology Advances, pg 53-57
  5. Joan,A., & King-Thom C. (2001) Alexander Fleming and the discovery of penicillin. Journal of Applied Microbiology. Volume 49:163-184.
  6. Aly, A. H., Debbab, A., & Proksch, P. (2011). Fifty years of drug discovery from fungi. Fungal Diversity, 50(1), 3-19.
  7. Wani, M., Taylor, H., Wall, M., Coggon, P., & McPhail, A. (1971). Plant antitumour agents. VI. The isolation and structure of taxol, a novel antileukemic and antitumor agent from Taxus brevifolia. Journal of the American Chemical Society, 93(9), 2325-2327.
  8. Horwitz, S. B. (1994). Taxol (paclitaxel): mechanisms of action. Annals of Oncology 5, S3-S6.
  9. Strobel, G., Yang, X., Sears, J., Kramer, R., Sidhu, R., & Hess, W. (1996). Taxol from Pestalotiopsis microspora, and enndophytic fungus of Taxis wallachiana. Microbiology, 142, 435-440.
  10. Manzoni, M. & Rollini, M. (2002). Biosynthesis and biotechnological production of statins by filamentous fungi and application of these cholesterol-lowering drugs. Journal of Applied Microbiology and Biotechnology. 58, 555-564.
  11. Manzoni, M., Bergomi, S., Rollini, M., & Cavazzoni, V. (1999). Production of statins by filamentous fungi. Biotechnology Letters, 21, 253-257.
  12. Endo, A. (2004). Statins is also used to treat diabetes, aging problems and heart disease. Just like many other medical drugs, too much of statin can be detrimental to ones health. Sideeffects of statins are headache,nausea,weakness, rash and even constipation.The origin of statins. International Congress Series, 1262, 3-8.
  13. Moorman, P. & Hamilton, R. (2007). Statins and cancer risk: What do we know and where do we go from here? Epidemiology, 18, 194-196
  14. Strobel, G., Daisy, B., Castillo, U. & Harper, J. (2004). Natural products from endophytic microorganisms. Journal of Natural Products. 67(2), 257-268.
  15. 15.0 15.1 Strobel, G., Miller, V., Martinez-Miller, C., Condron, M., Teplow, D. & Hess, W. (1999). Cryptocandin, a potent antimycotic from the endophytic fungus Cryptosporiopsis cf. quercina. Microbiology. 145, 1919-1936.
  16. Smith,A. Bailey, P. (1985). Production of Cephalosporin by fermentation. Glaxo Group Limited, 631-639
  17. Thompson, R L; Wright, A J. (February 1983). Cephalosporin antibiotics. National Library of Medicine;; 2 79-87
  18. O'Callaghan, C H; Sykes, R B; Staniforth, S E. (August 1976). National Library of Medicine. Antimicrobial agents and chemotherapy, 2: 245-248
  19. Wasser,S.P., & Weiss,A.L.(1999).Medicinal properties of substances occurring in higher Basidiomycetes mushroom:International Journal of Medicinal Mushrooms,1,31-62
  20. Mayera, Alfred M., and Richard C. Staples. "Laccase: new functions for an old enzyme." 60.6 (2002): 551-65.
  21. Zhen-Yuan, Zhua, Yaoa, Qiang, and lui yang. "Highly efficient synthesis and antitumor activity of monosaccharide saponins mimicking components of Chinese folk medicine Cordyceps sinensis." Journal of Asian Natural Products Research 14.5 (2012): 429-35.
  22. Panda, D. Rathinasamy, K. Santra, M. Wilson, L. (2005). Kinetic suppression of microtubule dynamic instability by griseofulvin. University of California, Santa Barbara, CA, 102, 9878-9883
  23. Panda, D. Rathinasamy, K. Santra, M. Wilson, L. (2005). Kinetic suppression of microtubule dynamic instability by griseofulvin. University of California, Santa Barbara, CA, 102, 9878-9883
  24. Haofan, J. Atsuya, Y. Shinya, M. Pinting, Y. Limin, H. (2008). Griseofulvin, an oral antifungal agent, suppresses hepatitis C virus replication in vitro. Hepatology Reseach, 38,9 909-918
  25. Henry,T. (1998). The discovery and development of Cyclosporin. Wolfson college, Cambridge, 12, 20-30.
  26. Henry,T. (1998). The discovery and development of Cyclosporin. Wolfson college, Cambridge, 12, 20-30.
  27. Kahan,B.(2003). Individuality: the barrier to optimal immunosuppression. Natures Review Immunology, 3, 831-838.
  28. Ko,Y. Ludescher,R. Frost,D. Wasserman,B. (1994). Use of cilofungin as direct fluorescent probe for monitoring antifungal drug-membrane interaction. Antimicrob Agents Chemother, 38, 1378-1385.
  29. Meshulam T, Levitz SM, Diamond RD, Sugar AM. (1989). Effect of cilofungin (LY121019), a fungal cell wall synthesis inhibitor, on interactions of 50 strains of Candida albicans with human neutrophils. US National Library of Medicine, National Institute of Health, 24, 741-745.
  30. Richardson,M. Scott,G. Shankland,G. (1992). Effect of cilofungin on phagocytosis and intracellular killing of Candida albicans by human neutrophils. European Journal of Clinical Microbiologyand Infectious Diseases, 11, 22-36.
  31. Yang et Al. (2011). Characterization of recombinant plectasin: Solubility, antimicrobial activity and factors that affect its activity. Process Biochemistry, pg 1050-1055
  32. Brinch et al. (2009). Influence of Antidrug Antibodies on Plectasin Efficacy and Pharmacokinetics. Antimicrobial Agents of Chemotherapy, 53(11), pg 4794-4800
  33. Moore,D., Robson,G.,& Trinci,T.(2013). 21st century guidebook to fungi. Cambridge Pr.
  34. Moore,D., Robson,G.,& Trinci,T.(2013). 21st century guidebook to fungi. Cambridge Pr.
  35. Moore,D., Robson,G.,& Trinci,T.(2013). 21st century guidebook to fungi. Cambridge Pr.
  36. Schugerl,K. (2005). Extraction of primary and secondary metabolites. Advances in Biochemical Engineering, 92, 1-48
  37. Kondo,K., Matsumoto,M., Ohtani,T.. (2007). Comparison of solvent extraction and supported liquid membrane permeation using an iconic liquid for concentrating penicillin G. Journal of Membrane Science, 289,92-96
  38. Lee,S.C.. (2000). Continuous Extraction of Penicillin G by emulsion liquid membranes with optimal surfactant compositions. The Chemical Engineering Journal",79,61-67.
  39. Adikane, H.V.,Nene,S.N., Singh, R.K.. (1999). Recovery of penicillin G from fermentation broth by microfiltration. Journal of Membrane Science, 162, 119-123
  40. Lazarova, Z., Schugerl,K., Syska,B.. Application of largescale hollow fiber membrane contractors for simultaneous extractive removal and stripping of penicillin G. Journal of Membrane Science,202,151-164
  41. Doherty, M.F., Malone, M.F., Pai, R.A.. Design of reactive extraction sysytems for biproduct recovery. AIChE Journa,48,3-12
  42. Noverr,M. Noggle,R. Toews,G. Huffnagle,G. (2004). Role of Antibiotics and Fungal Microbiota in Driving Pulmonary Allergic Responses. American Society for Microbiology, Infection and Immunity, 72, 4996-5003
  43. Noverr,M. Noggle,R. Toews,G. Huffnagle,G. (2004). Role of Antibiotics and Fungal Microbiota in Driving Pulmonary Allergic Responses. American Society for Microbiology, Infection and Immunity, 72, 4996-5003
  44. Moore,D., Robson,G.,& Trinci,T.(2013). 21st century guidebook to fungi. Cambridge Pr.
  45. Moore,D., Robson,G.,& Trinci,T.(2013). 21st century guidebook to fungi. Cambridge Pr.
  46. Betty A.F., Daniel F., & Alice S.W.(2002). Bailey & Scott's Diagnostic Microbiology,Eleventh Editon:Mosby,Inc,St.Louis,Missouri.

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