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AN<br />overview project<br />on<br />MAGENTA PIGMENT ISOLATION FROM FUNGUS<br />                                                <br />FROM<br />               SHRM BIOTECH PVT. LIMITED KOLKATA<br />(JULY-AUGUST 2010)<br />                                submitted by<br />ARKABRATA Bandyapadhyay<br />                                  3rd YEAR<br />B.Sc (H) IN BIO-TECHNOLOGY<br />Under the supervision of<br />Mr. Ashish Roy<br />(Project Coordinator)<br />REGISTRATION NUMBER: 1586 of 2008-2009<br />                                  ROLL NUMBER: BUR D BT 2009/O5<br />                                                 <br /> CONTENTS <br />                                                                                                                                                                                                                     <br />                                                                                                                                                                                             Pageno.<br />ABSTRACT………………………………………………………………………… <br />INTRODUCTION………………………………………………………………….. <br />        WHY MAGENTA PIGMENT? ...................................................................................................<br />        PHISICS OF COLUR………………………………………………………………………………….. <br />        PIGMENT………………………………………………………………………………………………<br />        MAGENTA……………………………………………………………………………………………..<br />        HUE……………………………………………………………………………………………………..<br />        CMYK…………………………………………………………………………………………………...<br />VARIATION OF MAGENTA……………………………………………………....<br />        MAGENTA DYE………………………………………………………………………………………..<br />        PROCESS MAGENTA………………………………………………………………………………….<br />        ELECTRIC MAGENTA ………………………………………………………………………………..<br />REVIEW OF LITERATURE……………………………………………………….<br />        HISTORY OF MAGENTA….………………………………………………………<br />        MAGENTA IN POPULAR CULTURE……………………………………………<br />        COMICS BOOK………………………………………………………………………………………..<br />        BUSINESS……………………………………………………………………………………………...<br />       ASTRONOMY………………………………………………………………………………………….<br />       FILM ……………………………………………………………………………………………………<br />       MUSIC…………………………………………………………………………………………………..<br />       PARAPSYCHOLOGY………………………………………………………………………………….<br />       POLITICS……………………………………………………………………………………………….<br />MATERIALS AND METHOD……………………………………………………...<br />MATERIALS………………………………………………………………………….<br />          INSRUMENT…………………………………………………………………………………………….<br />         GLASS WARES………………………………………………………………………………………….<br />         FIBERS……………………………………………………………………………………………………<br />         CULTURE ………………………………………………………………………………………………..<br />         CHEMICALS……………………………………………………………………………………………...<br />METHODS……………………………………………………………………………..<br />         PREPARATION (100ml.) OF POTATO DEXTROSE AGAR (PDA)…………………………………...<br />         PREPARATION(100ml.) OF CZAPEK’S BROTH MEDIA……………………………………………..<br />         COLLECTING OF FUNGUS……………………………………………………………………………..<br />         MAGENTA PIGMENT PRODUCTION PROCESS……………………………………………………..<br />        QUANTITATIVE ANALYSIS OF    PIGMENT………………………………………………………...<br />PURIFICATION OF PIGMENT……………………………………………………..<br />         PREPARATION OF SILICA GEL FOR TLC…………………………………………………………….<br />         PREPARATION OF MOBILE PHASE……………………………………………………………………<br />         PROCEDURE………………………………………………………………………………………………<br />         RETARDATION FACTOR……………………………………………………………………………….<br />RESULT AND DISCUSSION…………………………………………………………<br />         OBSERVATION OF MANGO TREE’S FUNGUS……………………………………………………….<br />         OBSERVATION OF NEEM TREE’S FUNGUS………………………………………………………….<br />         OBSERVATION OF JACK FRUIT TREE’S FUNGUS………………………………………………….<br />CONCLUTION…………………………………………………………………………<br />ADVANTAGES(Health and safety aspects of natural dyes)………………………...<br />LIMITATION…………………………………………………………………………...<br />REFERENCES…………………………………………………………………………..<br />EXTARNAL LINKS…………………………………………………………………….<br />ACKNOWLEDGEMENT<br />I owe a great many thanks to a great many people who helped and supported me during my project paper.<br />My deepest thanks to the project coordinator Mr.ASHISH ROY and also my collage H.O.D Dr. Bidyut Bandhopadhaya the Guide of the project for guiding and correcting various documents of mine with attention and care. He has taken pain to go through the project and make necessary correction as and when needed.<br />I express my thanks to the Principal of University of  Burdwan, for extending his support.<br />My deep sense of gratitude to Mr. Ashish Roy my project coordinator and collage H.O.D. Mr. Bidyut Bndhopadhaya,  SHRM BIO- TECH pvt. ltd. Kolkata support and guidance. Thanks and appreciation to the helpful people at SHRM BIO- TECH pvt. ltd. Kolkata, for their support.<br />I am greatly obliged to the director of SHRM who accommodated me for the training of the organization.<br />I would also thank my Institution and my faculty members without whom this project would have been a distant reality. I also extend my heartfelt thanks to my family, my parents(Goutam Bandyapadhayay and Anulekha Bandyapadhyay),my sister(Angana Bandyapadhyay) my friends and well wishers.<br />DECLARATION<br />I DO HERE BY DECLARE THAT THIS DISSERTATION REPORT SUBMITTED BY ME ABOUT MAGENTA PIGMENT ISOLATION FROM FUNGUS, IS OF MY OWN AND NOT SUBMITTED TO ANY OTHER INSTITUTION OR PUBLISHED ANYWHERE ELSE.<br />ARKABRATA BANERJEE<br />REG NO.<br />ROLL NO.<br />DATE:<br />ABSTRACT<br />         The fungus producing magenta was collected from different tree like mango,neem,jack fruit etc. in nature and culture these fungus from different trees on different PDA(potato dextrose agar) media plate. The fungal strain that produced magenta pigment was closely related to Phoma herbarum. The type of fibers added to Czapek’s medium influenced which pigments were produced. The pigment was purified through TLC chromatography. The pigment structure was partially determined. This is the first report of the production of magenta pigment by a microorganism specifically in the presence of nylon-6 fibers, via an unknown mechanism. This phenomenon raises the question of why and how the fungus disperses the pigment inside the fiber and suggests that fabrics can be dyed using microorganisms.<br /> <br />INTRODUCTION<br />WHY MAGENTA PIGMENT?<br />           <br />         Demand for natural instead of synthetic pigments for coloring fabrics, foods and cosmetics is increasing. Unlike pigments that are synthetic, those from natural sources allow subtle differences in tone because such pigments generally comprise various color components. Microbes have recently received focus as sources of natural pigments . Many fungal species produce pigments. Monascus fungi produce red or yellow pigments that are used as food colorants. Hahella produces red, prodigiosin, Ashbya produces yellow riboflavin , Phoma produces orange aza-anthraquinone and Chromobacterium produces blue violacein. We discovered a fungus that produces magenta pigment. We found that the produced colors were magenta, pink or green depending on the type of fiber added to the culture medium. These findings indicated that various fibers influence the metabolic pathways involved in the production of secondary pigment metabolites.<br />PHYSICS OF COLOUR :<br />             Electromagnetic radiation is characterized by its wavelength and its intensity. When wavelength is with in visible spectrum the range of wavelength human can perceive, approximately from 380 nm to 740 nm. It is known visible light. The science of colour is called as chromatics.<br />PIGMENT :<br />            A pigment is the material that changes the colour of light .it reflects as the result of selective colour absorption. This physical process differs from Fluorescence, phosphorescence and other forms of Luminescence in which the material itself emits light. Many materials absorb certain wavelength of light that humans have chosen and develop for use as pigments usually have special properties that make them ideal for coloring other materials. A pigment has a high tinting strength relative to the materials it colors. It must be stable form at ambient temperature.<br />         Archaeologists have uncovered evidence that early humans used paint for aesthetic purpose such as body decoration. Pigments and paint grinding equipment believed to be between 350,000 and 400,000 years old have been reported in a cave at twin Rivers, near Lusaka, Zambia. The earliest known pigments were natural minerals.<br />         Microbes have recently received focus as sources of natural pigments, many fungal species produce pigments. Monascus fungi produces Red or yellow pigments that are used as food colorants. Hahella produces red, Prodigiosin, Ashbya produces Yellow Riboflavin, and Phoma produces Pink or Orange pigment.<br />MAGENTA :<br />          Magenta is a colour made up of equal part of red and blue light. This would be the precise definition of the colour as defind for computer display. It is pure chroma on the colour wheel mid way between violet and rose. In HSV colour space, Magenta has a Hue of 300˚C. <br />         Magenta is not a spectral but an extra spectral colour. It can’t be generated by light of single wavelength. Human being can only see as far as 380 nm in to the spectrum i.e. as far as violet. The Hue Magenta is the complement of green. Magenta pigment absorb green light, thus Magenta and green are opposite colour.<br />         Magenta actually contains more purple pigment than Fuchsia that is actually used to create Magenta. Magenta was invented in 1980. The colour originally called as Fushin made from coal tar dyes in 1859. Besides being called original magenta, it is also called Rich magenta. It is mixture of red and blue wavelength.<br />HUE :<br />       Hue is one of the main properties of a colour defined technically as the degree to which a stimulus can be described as red, green, blue, and yellow.<br />CMYK : <br />        The best places to start when trying to understand the colour spectrum is with the CMYK colour model which is composed of cyan that is a bluish green colour, magenta is a purplish red colour, yellow that is medium yellow and key which is black. The CMYK colour model is newer to the industry compared to the more traditional RGB or red, green, blue model.<br />VARIATION OF MAGENTA<br />    Magenta is a purplish pink colour evoked by light with less power in yellowish green wavelength then in blue and red wavelength. The hue magenta is the complement of green. Magenta pigment absorb green light, thus magenta and green are opposite colour.<br />          <br /> The three variation of magenta pigment are;<br />MAGENTA DYE (ORIGINALVARIATION)<br />PROCESS MAGENTA (PIGMENTMAGENTA) (PRINTER MAGENTA)<br />ELECTRIC MAGENTA (ADDTIVE SECONDARY MAGENTA)<br />    1. MAGENTA DYE :<br />              Before printer’s magenta was invented in the 1890 for CMYK printing and electric magenta was invented in 1980 for computer display. Magenta dye colour is also called Rich Magenta. Magenta was one of the first aniline dyes discovered shortly after the Battle of Magenta (1859). The colour was originally called Fuchine or Roseine, but for marketing purpose in 1860 the colour name was changed to magenta.<br />    2. PROCESS MAGENTA :<br />               In colour printing, the colour called Process Magenta or Pigment Magenta. It is one of the primary pigment colours which along with yellow and cyan constitute the 3 subtractive primary colours of pigment. The secondary colours of pigment are blue, green, and red. As such the CMYK printing process was invented in 1890. Process Magenta is not an RGB colour, and there is no fixed conversion from CMYK to RGB.<br /> 3. ELECTRIC MAGENTA :<br />                     For computer colour radiation, that specific Hue of magenta composed of equal part of red and blue light was termed the WEB colour fuchsia and was assigned as an alias for the RGB code of magenta on the list of standardized web colours “ELECTRIC” Magenta and fuchsia are exactly same colour. Some time Electric Magenta is called “ELECTONIC” Magenta.<br />Review of literature<br />        Dyeing has almost ceased to exist as a traditional art. In this 20th century the importance of colour is our lives seems to be realizing less and less. It has been forgotten that strong and beautiful colour, such as used to abound in all everyday things is an essential to the full joy of life.<br />               A DYE can generally describe as a colored substance that has an affinity to the substrate which it is being applied. Both dye and pigment appear to be colored because; they absorb some wavelength of light. The first human made (synthetic) organic dye, Mauveine, was discovered by William Henry Perkin in 1856. Many thousand of synthetic dyes have since been prepare. Synthetic dyes quickly replaced the natural traditional dyes.<br />One aspects of the invention relates to a process for preparing a magenta or Azo pigment involving the steps of coupling at least one diazonium component of one or more aromatic amine where R is a lower alkyl group or a Hydro carboxyl group.<br />                The present invention relates to metabolized Azo blue shade red to magenta pigment suitable for use as a coloring agent. Methods of making and using the metalized azo magenta pigment and plastic coating and inks. According to the present invention magenta exhibit high tinctorial strength while simultaneously exhibiting good heat stability, light fastness.<br />                In the middle of nineteenth century all dyes available to man came from natural sources. Most of these were vegetable extract and few were animal product. Today however practically all dyes are synthetic. They are prepared from aromatic compound. If a natural dye did not have the chemical grouping necessary to react with the chemical grouping of a particular fabric, which the fabric couldn’t dyed with that material. According to present invention contain few (no more than about one) Halogen atom or pigment molecule. So the metalized azo magenta pigment is environmentally friendly. The azo pigment of present invention are prepared by initially diazotizing one or more aromatic amines suitable for use in this invention and there after coupling diazonium component with a coupling component suitable for use in this invention to form described dye.<br />HISTORY OF MAGENTA<br />Magenta is the first aniline dye discovered shortly after the battle of Magenta (1859).This occurred near the town of magenta in Northern Italy. The colour was originally called Fuchsine or rosine but for marketing purpose in 1860 the colour name was change to Magenta after the battle.<br />The history of Magenta dye is very intresting. Egyptian mummies have been found wrapped in cloth dyed from the madder plant.<br />Alexander the great is supposed to have deceived the Persians into thinking that his armies were wounded by sprinkling his solder with a red dye, probably madder juice, which contain the dye alizarian.<br /> The dark blue indigo dye has been known for over 4000 years, when Romans invaded England, they found that the country was inhabited by the ancient people called picts. They tattooed and painted themselves with indigo. Latin meaning of Briton is painted man. <br />Mark Antony’s flight from the crucial naval battle of Actium was especially conspicuous because he fled in Cleopatra’s barge, which was carrying snails, known for their colour Royal purple. <br />Magenta in Human culture<br />   <br />COMICS BOOKS :   <br />Magenta is a fictional character in the DC Comics' series Teen Titans.<br />BUSINESS :<br />               <br />Magenta is a color trademark of Deutsche Telekom AG, parent company of T-Mobile<br /> ASTRONOMY :<br />Fig:Artist's vision of a spectral class T brown dwarf<br />  Astronomers have reported that spectral class T brown dwarves (the ones with the coolest temperatures) are colored magenta because of absorption by sodium and potassium atoms of light in the green portion of the spectrum.<br />FILM :<br /> Magenta is a fictional character (quot;
a domesticquot;
) in the Rocky Horror Picture Show.<br />MUSIC :<br />There is a song called Magenta on the 1998 album Blue Wonder Power Milk by Hoover phonic.<br />Magenta is the title of a spoken word song by Ken Nordine on his 1967 Colors album.<br />PARAPSYCHOLOGY :<br />To psychics who claim to be able to observe the aura with their third eye, someone who has a magenta aura is usually described as being artistic and creative. It is reported that typical occupations for someone with a magenta aura would be such professions as artist, art dealer, actor, author, costume designer, or set designer.<br />POLITICS :<br />The color magenta is used to symbolize anti-racism by the Amsterdam-based anti-racism Magenta Foundation.<br />In Spain, magenta is used as the official color of the UPyD quot;
Union, Progress and Democracyquot;
 political party.<br />MATERIALS AND METHOD<br />                                            : MATERIALS :<br />1.     INSTRUMENTS:<br />      Chemical balance <br />      pH meter<br />     Autoclave<br />     Laminar airflow for aseptic condition<br />     Incubator<br />     Refrigerator for keeping stock solution and chemicals<br />     Hot air oven<br />     Shaker<br />     Microscope<br />    <br />     2.   GLASS WARES:<br />    Test Tubes<br />    Volumetric flacks and beaker for media preparation<br />    Conical flasks and Petri dishes (culture vessels)and Forceps<br />3.    FIBERS:<br />    Cotton<br />    Silk and polyester (Polyethyleneterepthalate)<br />    Filter paper<br />  <br />   4.    CULTURE:<br />    For production for pigment fungus was cultivated in the following media.<br />    Potato dextrose agar (PDA)<br />    Czapek’s agar<br />   5.    CHEMICALS:<br />   Detergent<br />  Ethanol, Methanol,Isopropanol and Ethyl acetate and n-Hexane.<br />                                               : METHODS :<br />,[object Object], <br />Potato -- 20g.<br />Dextrose -- 2g.<br />Distilled water -- as required as for 100ml. media<br />Agar -- 2g.<br />Now peel off the potato and cut into short pieces then boil it with 50ml. of distilled Water and after boiled filter it with cheese cloth to separate the water and potato.Then measured the boiled water in measuring cylinder and then add the extra distilled water In the measuring cylinder upto 100ml.Then add Dextrose and Agar.<br />Media are commonly sterilized by autoclaving culture media in glass container sealed    with cotton plug or aluminium foil. Then autoclaved at 15 lbs and 121ºC for 15 to 40 mins.   The pressure shouldn’t exceed 20 psi as higher pressure may lead to the decomposition of  carbohydrate and the component of the medium.<br />2.   PREPARATION(100ml.) OF CZAPEK’S BROTH MEDIA:<br />,[object Object]
Dipotassium hydrogen phosphate -- 0.1g.
Magnesium sulphate -- 0.05g.
Potassium chloride -- 0.05g.
Ferrous sulphate -- 0.001g.
Sucrose -- 3g.Distilled water -- 100 ml.<br />At first added sodium nitrate, dipotasium hydrogen phosphate,magnesium  chloride,ferrous sulphate,sucrose into 40ml. of distilled water and potassium chloride into 10ml.of distilled water and Then both of these solution mixed in measuring cylinder and then added distilled water upto 100ml.<br />Media are commonly sterilized by autoclaving culture media in glass container sealed with cotton plug or aluminium foil. Then autoclaved at 15 lbs and 121ºC for 15 to 40 mins.The pressure shouldn’t exceed 20 psi as higher pressure may lead to the decomposition of  carbohydrate and the component of the medium.<br />Fig: Czapek’s Broth.<br />,[object Object]
Fungus was collected from plant like Mango tree,Neem tree,Jack fruit tree .
MAGENTA PIGMENT PRODUCTION PROCESS:
The collected fungus from 3 different trees were grow on three different Petri plates with         PDA(potato dextrose agar) media for screening of fungus having the capability to produce Magenta Pigment.Then the three plates were store in the dark room at 25ºC for 7 days.The spreading of fungus on media plate have to perform in laminar air flow for aseptic condition.<br />,[object Object],After 7 days fungus that produce Magenta Pigment on agar plate were isolated     and Cultured in different Czapek’s broth(100ml.) media having different fibers like cotton,silk and polyester(all this fibers were added in same amount),and incubated for 15 days.<br />,[object Object]
Fig: Magenta Pigment produced in Czapek’s broth using cotton and silk.
The fibers were cut into small pieces and sterilized with the help of Autoclave.
After 15 days the pigment was collected differently in glass container from three   fibers (cotton,silk,polyester) by pressing the fibers. <br />  5. CHEAKING OF PRODUCTIVITY or QUANTITATIVE ANALYSIS OF    PIGMENT:<br />-220980611505 Collected three different pigment from three different fibers were treated with 200ml. of  Methanol and then dried out in air to obtained the powder dye(Magenta Pigment).<br />        Fig: Extraction of magenta pigment by using methanol<br /> Point Noted: all the petri plates were used after store these plates in hot air oven for minimum 10 mins. And when the pigment was added in the Czapek’s broth media,this task  have to perform in Laminar air flow for aseptic condition.<br />Sterilizing the culture vessel and instruments(Glass culture vials, metal instruments can be sterilized by properly wrapped with aluminium foil before sterilization and autoclaving under steam at a pressure of 15 lbs and temperatures of 121ºc for 15 mins. The instrument used for aseptic manipulation is usually made of metal. Normally instruments such as forceps, scalpels, needles and spatula are sterilized by dipping in 95% ethanol followed by flaming and cooling.)<br />6.PURIFICATION OF PIGMENT:<br /> TLC (thin layer chromatography) is used for the purification of magenta pigment.<br /> Thin layer chromatography (TLC) is a simple, quick and inexpensive procedure that gives the chemist a quick answer as to have many component are in a mixture. <br />Thin layer chromatography is also used to support to identity of a compound in a mixture when the Rf of a compound is compared with the Rf of a known compound (run on the same TLC plate).<br />A TLC plate is a thin of glass, metal or plastic which is coated with a Thin Layer of a solid adsorbent (usually silica, alumina).  A small amount of the mixture to be analyzed is spotted near the bottom of this plate.<br />The TLC plate is then placed in a shallow pool of a solvent in a developing chamber so that only the very bottom of plate is in the liquid. This liquid, or the eluent is the mobile phase and it slowly rises up to the TLC plate by capillary action.<br />As the solvent moves past the spot that was applied. Equilibrium is established for each component of mixture between the molecule of that component which is adsorbed on solid and molecule which are in solution. When the solvent has reached the top of the plate, the plate is removed from developing chamber, dried and the separated component of the mixture. If the compounds are colour visualization is straight forward.<br />Usually the compounds are not coloured so, UV lamp is used to visualize the plate.<br />The retardation factor or Rf is defined as the distance travelled by the compound divided by the distance travelled by the solvent.<br />  PREPARATION OF SILICA GEL FOR TLC PLATE:<br />Take one spoon of silica gel in a beaker and add 5o ml of water in to it. Then  add a                              <br />   small pinch of calcium carbonate (CaCO3). Mix the solution thoroughly and then spread it       <br />   on to the glass plate by a glass rod.<br />      (b)   PREPARATION OF MOBILE PHASE:<br />,[object Object],   Butanol,water and glacial acetic acid. At first take 12 ml of Butanol then 1 ml of water and <br />   one drop of Glacial acetic acid in the chromatographic chamber and cover it.<br />     (c) PROCEDURE:           <br />                 For purification of pigment TLC (Thin Layer Chromatography) is used. The crude   <br />   pigment isolated was then loaded on the TLC plate and the pure pigment was observed. For    <br />  his procedure at first TLC plate and Mobile phase (Butanol, water and Glacial acetic acid) <br />  was prepared. Then the pigment which was washed with Methanol was loaded on the TLC <br />  plate with the capillary tube and TLC plate was put in the mobile phase and kept it for some <br />  time until the mobile phase run up to one third of the TLC plate and then removed he plate  <br />  from developing<br />              chamber (MOBILE PHASE) and dried. The main magenta band on the TLC plate was analyzed H-NMR or by spraying Ninhydrin solution over the TLC plate and observed the magenta band by analyzing the Rf values.<br />                                                               <br />                                                                                 Fig: Pure Magenta<br />                                                                                                   <br />      (d) RETARDATION FACTOR (Rf):<br />,[object Object],        The Retardation factor or Rf is defined as the distance traveled by the compound(solute) divided by the distance traveled by solvent.                                                                                                   <br />           Rf = Distance traveled by solute / distance traveled by Mobile phase.<br />RESULT AND DISCUSSION<br />1. OBSERVATION OF MANGO TREE’S FUNGUS:<br />,[object Object]
This time the cotton give more magenta pigment than other.
Silk gives magenta pigment less than cotton.
Polyester gives magenta pigment more than silk but less than cotton. 2. OBSERVATION OF NEEM TREE’S FUNGUS:<br />,[object Object],3.  OBSERVATION OF JACK FRUIT TREE’S FUNGUS:<br />,[object Object]
This time the cotton gives more pigment than other.
Silk gives magenta pigment less than cotton.
Polyester gives magenta pigment more than silk but less than cotton.CONCLUSION             <br /> Thus, this study concludes that pigment production depends on the nature of medium, the culture condition along with its temperature and the source of fungus also.<br /> Physiological test also suggests some of the property of the isolated pigment. It was identified that the fungus produced magenta and other pigments on Czapek’s agar in the presence of various fibers<br />ADVANTAGES(Health and safety aspects of natural dyes)<br />Though all Natural dyes are not 100% safe, they are less toxic than their synthetic counter parts. Many of the Natural dyes like turmeric, annatto and saffron are permitted as food additives. Many natural dyes have pharmacological effects and possible health benefit.<br />They are obtained from renewable sources. Natural dyes cause no disposal problems, as they are biodegradable.<br />Practically no or mild reactions are involved in their preparation.<br />They are unsophisticated and harmonized with nature.<br />Many natural dyes have the advantage that even though they have poor wash fastness ratings, they don’t stained the adjacent fabrics in the washing process because of the non subtractive nature of the dye towards the fabric.<br />An exception to this is turmeric, which shows substantvity for cotton.<br />Natural dyes are cost effective.<br />It is possible to obtain a full range of colours using various mordents.<br />LIMITATION:<br />,[object Object],Availability<br />Colour yield<br />Complexity of textile dyeingprocess<br />Reproducibility of colour shade.<br />Beside these there are other technical drawbacks of natural dyes,<br />Limited number of suitable dye<br />Great difficulty in blending.<br />Non standardized<br />In adequate degree of fixation.<br />Water pollution by heavy metals and large amount of organic substances.<br />REFERENCES<br />Birch, A.J., Butler, D.N., and Rickards, R.W. (1964) the structure of the aza-anthraquinone phomazarin. Tetrahedron Lett., 28, 1853-1858.<br />Felsenstein, J. (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution, 39, 783-791.<br />Holger, B.D.S, Stefan, W., and Marcus, W. (2000) the role of fungal appressoria in plant infection. Microbe. Infect., to, 1631-1641.<br />Luft, J.H. (1961) improvement in epoxy resin embedding methods. J.Biophys. Biochem.Cytol., 9, 409-414.<br />Perera, T.H.S., Gregory, D.W., Marshal, D. and Gow, N.A.R. (1997) contact-sensing by hyphae of dermatophytic and saprophytic fungi. J.Med.Vet.Mycol. 35,289-293.<br />Nakashima, T., Kurachi, M., Kato,Y.,Yamaguchi, k.,and Oda, T.(2005) Characterization of bacterium isolated from the sediment at costal area of Omura Bay of in Japan and several biological activity of pigment produced by this isolates. Microbiol. Immunol., 49, 407-415.<br />Mapari, S.A.S., Nielsen, K.F., Larsen, T.O., Frisvad, J.C., Meyer, A.S., and Thrane, U. (205) exploring fungal biodiversity for the production of water soluble pigment as potential natural food colourants. Curr.Opin.Biotechnol. 16,231-238.<br />Thompson, J.D., Higgins, D.G., and Gibson, T.J. (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res., 22, 4673-4680.<br />Tseng, Y.Y., Chen, M.T. and Lin, C.F.(2000) growth, pigment production and protease activity of Monascus purpureus as affected by salt, Sodium Nitrate, Poly Phosphate and various Sugars. J.Appl. Microbiol., 88, 31-37.<br />White, T.J., Bruns, T.Lee, S., and Taylor, J. (1990) Amplification and direct sequence of fungal ribosomal RNA genes for phylogenetics. In PCR protocols, a guide to methods and application, Ed. By Innis, M. A., Gelfand, D. H., Sninsky, J. J., and White, T.J. , Academic press, Inc., San Diego, pp. 315-322.<br />Rttori, D. and Duran, N. (1998) Production, extraction and purification of violacein: An antibiotic pigment produced by Chromobacterium violaceum. World J. Microbiol.Biotechnol. , 14, 685-688.<br />Shin, C.S., Kim, H.J., Kim, M.J., and Ju, J.Y.(1998) Morphological change and enhanced pigment production of Monascus when cocultured with Saccharomyces cerevisiae or Aspergillus oryzae. Biotechnol. Bioeng., 59, 576-581.<br />Stahmenn, K.P., Arst, H.N., Jr., Althofer, H., Revuelta, J.L., Monschau, N., Schlupen, C., Gatgens, C., Wiesenburg, A., and Schlosser, T. (2001) Riboflavin, overproduced during sporulation of Ashbya gossypii, protects its hyaline spores against ultraviolet light. Environ. Microbiol., 3,545-550.<br />EXTERNAL LINKS<br />How the color magenta was discovered in 1859 through research into coal tar (aniline) dyes.<br />Pictures of actual aniline dye samples in various shades of magenta.<br />Color Tribe List of colors<br />Magenta is a product of the brain rather than a spectral frequency<br />Magenta Ain't A Colour<br />
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus
Magenta Pigment Isolation From Fungus

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