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The First Transgenic Crop

An Application of Biotechnology
The transfer of foreign genes onto plants was first worked on by four different research groups on 1983, as summarized in the table below.
Group of Researchers headed by Mary-Dell Chilton Jeff Schell and Marc van Montagu Robert Fraley, Stephen Rogers, and Robert Hosch headed by John Kemp and Timothy Hall Location Washington University, St. Louis, Missouri Belgium Monsanto Wisconsin Plant modified cells of Nicotiana plumbaginifolia, a close relative of ordinary tobacco tobacco plants petunia plants sunflower plant Effect resistant to kanamycin

resistant to kanamycin and to methotrexate resistant to kanamycin inserted a bean gene

Most of the research groups modified their plants to be resistant to kanamycin, which is an antibiotic. This resistance is useful in the experiments, for we can easily determine if the transfer of genes is successful. We just have to grow the test plants and see if they can survive in an environment containing kanamycin. The transfer of foreign genes is made possible by Agrobacterium tumefaciens, which causes crown gall disease in plants. This bacterium causes disease by inserting one of its genes into the host plant, which when following the genetic instruction produces tumors and prolific masses. The bacterium then eats the tumors. To make this bacterium useful, it must first be disarmed (i.e., the disease-causing gene must be removed). This was done by traditional DNA recombination.
References: Department of Soil and Crop Sciences. (2004, January 29). History of Plant Breeding. (Colorado State University) Retrieved July 7, 2013, from http://cls.casa.colostate.edu/transgeniccrops/history.html Gasser, C. S., & Fraley, R. T. (1992). Transgenic Crops. Scientific American, 62-69.

-Cyclodextrin
An Application of Enzymes
Cyclodextrins (CDs) are cyclic oligosaccharides, or a ring of connected glucose units. The rings may contain 6, 7, or 8 glucose units, and they are called -, -, and -cyclodextrin, respectively. The figure below shows the structure of the cyclodextrins.

from http://www.chemiedidaktik.uni-wuppertal.de/disido_cy/cyen/info/pic_info/abc_cyclodextrine.gif

They have a hydrophobic cavity and hydrophilic rims, as shown in the 3D image below. This highlyorganized structure makes them a candidate for delivery of nonpolar compounds through polar environments, such as drugs inside the human body. Most important of all, CDs are nontoxic and thus can be ingested or injected into humans without adverse effects. The structure of cyclodextrin was discovered back in 1935, but misunderstandings in the toxicity of these compounds impeded the studies for its application on drug delivery for decades.

CDs are produced from starch using a family of enzymes known as cyclodextrin glycosyl transferases (CGTases), which are produced by bacterial strains such as Bacillus macerans, Klebsiella oxytoca and Bacillus circulans. CGTases break the helical polysaccharide chains from starch in locations as shown below. The most abundant cyclodextrin has seven glucose units, since it is the most stable.

The enzymes are first obtained from cultured bacteria, isolated, and reacted with a 30% starch suspension. However, since the reaction mixture is viscous, -amylase first hydrolyses the starch into simpler polysaccharides. Once the starch is broken down to desired chain lengths, the mixture is heated to degrade the amylase. CGTases are then added after cooling the mixture, to produce a mixture of the three cyclodextrins. The three CDs are separated by the changes in the size of their cavities: different solvents, shown in the figure below, are used to form precipitates with each.

The CGTase responsible for each type of CD have been isolated, thus enabling manufacturers to produce pure -, -, or -cyclodextrin as the need arises. References:
Wuppertal University. (2007, July 4). From Corn to Cyclodextrin. (Wacker Chemie AG) Retrieved August 9, 2013 from http://www.chemiedidaktik.uni-wuppertal.de/disido_cy/cyen/info/01_manufacture_cy.htm Kurkov, S. V, & Loftsson, T. (2013). Cyclodextrins. International journal of pharmaceutics, 453(1), 167 80. doi:10.1016/j.ijpharm.2012.06.055. Srinivasan, K., Stalin, T., & Sivakumar, K. (2012). Spectral and electrochemical study of host-guest inclusion complex between 2,4-dinitrophenol and -cyclodextrin. Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy, 94, 89100. doi:10.1016/j.saa.2012.03.066

Poly--hydroxybutyrate
An Industrial Application of Microorganisms
Poly--hydroxybutyrate (PHB) is an aliphatic polyester found inside many prokaryotes. These microorganisms use PHB to store carbon and energy during unbalanced growth conditions. Therefore, to manufacture PHB from these microbioreactors, the bacteria are grown in a carbon-rich environment which is depleted of other nutrients (such as nitrogen). PHB is an attractive replacement for petrochemical plastics, since it is a biodegradable thermoplastic. The Imperical Chemical Industries began working on the biopolymer in the 1980s, genetically engineering bacteria to produce PHB as 92% of their dry weight, and adding copolymers to improve its flexibility and toughness. ICI then commercialized PHB as Biopol, to be used in textile spinning. PHBs have found use in medical applications, such as the filaments for surgical sutures and controlled drug release, due to its biodegradability. However, the manufacture of PHB is still more costly to replace the petroleum-derived plastics. References Hamieh, A., Olama, Z., & Holail, H. (2013). Microbial production of polyhydroxybutyrate , a biodegradable plastic using agro-industrial waste. Global Advanced Research Journal of Microbiology, 2(3), 54 64. Retrieved from http://garj.org/garjm/index.htm Griffin, M., & Magor, A. (1986). Possible uses of micro-organisms in the manufacture of plastics and synthetic fibres. Biotechnology and Genetic Engineering , 4. Retrieved from http://www.tandfonline.com/doi/pdf/ 10.1080/02648725.1986.10647829 Dawes, E. A. (1988). Polyhydroxybutyrate: an intriguing biopolymer. Bioscience reports, 8(6), 537 47. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/3242641