Recent Patents on Biotechnology - Volume 4, Issue 2, 2010

Plant transformation technology plays an ever-increasing role in improving crop plant traits and addressing research questions. In recent years, a number of important patents on plant transgenic technologies have been published. These patents represent significant new progresses, many representing novel approaches in the field of plant transgenic technology. This review highlights selected as well as the most significant advancements made mostly within the past 2 years in plant transgenic research. These advancements include the improvements of plant transformation systems, development of new plant selectable markers, engineering for plant stress tolerance, production of vaccines in transgenic plants, etc. Of these, the engineering for plant stress tolerance accounts for a high percentage of the transgenic field of inventions. Some results are drastic and unexpected, having a great potential in their applications towards a wider range of plant species. This review also tries to avoid terms and languages pertaining to the patent and legal documents to ease readings.

Vaccinium is a genus of shrubs several of which, including cranberry, lingonberry, and blueberry, produce edible fruit. Consumption of the fruit of these plants and juices pressed from it has long been known to have healthful effects. A number of functional compounds have been extracted from the fruits and seeds of these plants. Anthocyanidins and related compounds have been reported to possess antimicrobial, antioxidant, and anti-inflammatory properties. Extracts of the fruits have been applied to the inhibition of non-enzymatic glycosylation in anti-aging preparations. The oil of the cranberry seed is high in antioxidants and is a source of omega-3, -6, and -9 fatty acids making it an attractive cosmetics component. The inhibition of tumor growth, angiogenesis, and metastasis by extracts of these fruits has been described. Extracts of the plants have also been applied to the development of antimicrobial paints and coatings. Here, we provide an overview of the wide range of applications described for various compounds from and preparations of these plants and survey the recent patents related to those applications.

As single cell systems, protoplasts have been used in physiological, biochemical and molecular studies aiming towards the investigation, improvement or modification of plants. In grapevine, protoplasts have been isolated from several source tissues but not from grape berry, a major challenge given the uniqueness of grape fruit for human diet and wine production. Also, as the ripe grape berry has long been considered a ‘small bag of sugary water’ without cell compartmentation and/or membrane integrity, the isolation of intact cells from the mesocarp is of special scientific significance. Protoplasting from grape berry mesocarp cells was achieved with cellulase and pectolyase digestion, followed by differential and gradient centrifugations; however, given the special characteristics of berry tissue, cell wall digestion and protoplast purification were performed in a special environment to maintain their integrity and viability. Light and epifluorescence microscopy revealed the spatial organization of the cytoplasm, where an intricate acidic vacuolar apparatus predominates supporting the idea that berry softening during ripening is not strictly associated with loss in compartmentation and/or membrane integrity. Following the worldwide economical and social importance of wine in modern days, grape berry protoplasts are a major advance for both basic research of fruit ripening and biotechnological applications.

Multifunctional nanoprobes, such as nanocrystals, nanoshells, and luminescent nanomaterials, have been developed for imaging biological processes, such as cell signaling, neuroimaging, protein conformation probing, DNA conformation probing, gene transcription, virus infection and replication in cells, protein dynamics, tumor diagnosis, and therapy evaluation. With the application of nanotechnology for CNS-active agents' delivery, nanostructured materials are emerging as a powerful means for diagnosis of CNS disorders, including brain tumors, because of their unique optical size, and surface properties. This review summarizes the recent patents on imaging nanoprobes for brain tumor diagnosis and therapy. The future development in this active cross-disciplinary field will be discussed as well.

Recent advances in agricultural biotechnology have enabled the field of plant biology to move forward in great leaps and bounds. In particular, recent breakthroughs in molecular biology, plant genomics and crop science have brought about a paradigm shift of thought regarding the manner by which plants can be utilized both in agriculture and in medicine. Besides the more well known improvements in agronomic traits of crops such as disease resistance and drought tolerance, plants can now be associated with topics as diverse as biofuel production, phytoremediation, the improvement of nutritional qualities in edible plants, the identification of compounds for medicinal purposes in plants and the use of plants as therapeutic protein production platforms. This diversification of plant science has been accompanied by the great abundance of new patents issued in these fields and, as many of these inventions approach commercial realization, the subsequent increase in agriculturally-based industries. While this review chapter is written primarily for plant scientists who have great interest in the new directions being taken with respect to applications in agricultural biotechnology, those in other disciplines, such as medical researchers, environmental scientists and engineers, may find significant value in reading this article as well. The review attempts to provide an overview of the most recent patents issued for plant biotechnology with respect to both agriculture and medicine. The chapter concludes with the proposal that the combined driving forces of climate change, as well as the ever increasing needs for clean energy and food security will play a pivotal role in leading the direction for applied plant biotechnology research in the future.

The heterologous protein expression system of Pichia pastoris is now widely used for expression of many human proteins, because the efficiently expressed proteins will be correctly folded in Pichia pastoris cells and also efficiently secreted from the cells. Recombinant human serum albumin (rHSA) is efficiently secreted from Pichia pastoris. Nowadays, the expression of rHSA exceeds 10g in 1 L fermentor culture broth, and the protein is completely purified. Recombinant HSA expressed in Pichia pastoris was approved as a medicine by the authorities in 2007, and launched in 2008 in Japan. One of the insulin precursors (IP) was also successfully expressed in Pichia pastoris, and secreted up to 3.6g in 1 L medium using a multi-copy transformant. The insulin precursor could be efficiently converted to insulin, the final product, in vitro. Human growth hormone was also expressed in Pichia pastoirs, and secreted up to 49 mg in 1 L medium. These proteins are also important for clinical applications. Midkine and pleiotrophin may be two of the candidates for clinical applications. Secretion signals, the copy number of an expression cassette in transformants, and culture conditions for fermentation were examined for efficient expression of these proteins in Pichia pastoris. The best signal was selected, and other factors were optimized. The amounts of native midkine and native pleiotrophin expressed were approximately 0.36g and 0.26g in 1 L medium, respectively. Expression of bile-salt stimulated lipase (BSSL) had been extremely low in the beginning of a fermentor culture experiment. However, approximately 1 g rBSSL in 1 L medium was finally expressed in a fermentor by unlimited feeding of glycerol for cell growth and optimization of other factors. BSSL from human milk and rBSSL from Pichia cells are glycosylated. The structure differences between these glycans are obvious. When humanization of Pichia glycans is established by genetic engineering, the Pichia pastoris expression system will become indispensable for the production of therapeutic proteins.

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