Modern Fermentation Technology and Metabolic Engineering
Chang Gung Biotechnology Corp. Ming-Chi Plant
Hirsutella sinensis Mycelia

Chang Gung Biotechnology Corp.
Modern Fermentation Technology and Metabolic Engineering

There is a broad range of applications for modern biotechnology, ranging from medicine to farming, forestry, fishing, animal husbandry, environmental protection, energy, and chemicals. However, biotechnology needs first to be industrialized in order for its benefits to become available to the general public.

Chang Gung Memorial Hospital and Chang Gung University have invested enormous manpower and resources over the years in clinical research, preclinical testing, drug delivery technology, new drug development, pharmacological research, tissue culture, biopharmaceutical and biomedicinal materials research, biological information applications, and biochip development. The Chang Gung-affiliated system of patient care and biomedical research has undoubtedly achieved much over the years, making it probably the best private bio-medical center of excellence in Asia. On the other hand, our Formosa Plastics Group, one of the largest industrial groups in the world, has long developed its own track record of industrial manufacturing excellence, dominating in all the product areas that it participates and attending to a single-minded focus on continued management improvements. It is based on this platform of excellence that we have sought to develop our own biotechnology efforts in formulating solutions to some of the more serious problems plaguing mankind today.

Let us begin with the serious issue of the soil and the poor state of affairs that all our natural foods find themselves today. Rampant overuse of chemical fertilizers, pesticides, and insecticides over the years has led to the rapid depletion of soil around the world. In addition, overly refined and processed food, polluted air and water, and excessive logging have all contributed to escalating ill effects on our health and the environment. The Chang Gung Biotechnology team has worked diligently over the years with various farming and food-harvesting methods in an effort to bring solutions to this food-related crisis. We have come to the conclusion that not only organic farming is required, but the general mineral health of the soil must be restored. Thus, in all our own food production, we emphasize strongly the inclusion of all the trace elements required by the body to restore health. This has required that we develop a comprehensive program of research on the effects of trace elements on body physiology as well as a proprietary technological platform enabling us to extract trace elements in a form that make them readily bio-available. In addition, we believe in the impact of positive intentionality in creating the harmonious coherence required to bring out the most beneficial foods. The effect of positive intentionality has been tested in a number of field experiments in Central America, all of which have confirmed this concept. The soil is truly a complex eco-system, and in order to capitulate further on this concept, we also work closely with a 15-acre agricultural and environmental biotechnology research center in La Ward, Texas, established by our Group to carry out field experiments on specialized farming methods and on the effects of various fungi and symbiotic microorganisms on plant health. In this respect, it can be said that our Agricultural and Environmental Research and Development Center is a world leader in the study and production of endomycorrhizal fungi, soil organisms with great importance to plant health.

The other crisis that can be mentioned is the modern concept of health. Most if not all the remaining diseases plaguing mankind are chronic in nature. Their very nature dictates a more pro-active, preventative approach on the part of the individual, as opposed to the more passive, hospital- and drug-based approach of the past. Certain foods have traditionally been known to help maintain health, given their enriched profile of metabolites and other active ingredients. These belong for example to the so-called ¡§superior¡¨ foods advocated by Traditional Chinese Medicine for several millennia that are known to have important medicinal properties. Many of these foods belong to fungi, bacteria, and herbal categories, but often their rarity has made their widespread use prohibitively expensive. It is our intention to mass-produce many such foods with medicinal properties in a safe and clean environment while attending to scientific details that allow us to enrich our foods for the same active ingredients that are only found normally in the wild form. In this way, we shall make such specialty foods affordable to the general public.

With a view towards developing metabolic engineering and related biotechnological downstream processes for bacteria, fungi, plant and animal cells, we have set up a modern fermentation facility in collaboration with the Ming Chi University of Technology. Our research focus is directed towards four main areas related to fermentation, sorted in this manner to accommodate for the resulting products:
(1) Biomass: propagation of bacteria, yeast, fungi, plant cells, insect cells, and animal cells.
(2) Metabolites: derivation of primary and secondary metabolites such as amino acids, nucleic acids, proteins, fats, vitamins, polysaccharides, and carbohydrates.
(3) Enzymes: production of purified industrial enzymes that include amylase, amylopectinase, peptidase, and various hormonally- or metabolically-relevant proteins and enzymes.
(4) Biotransformation: utilization of one or more types of enzymes in the above mentioned cells, with focus on one or multiple pathways, to obtain specific biochemical reactions that produce derivatives of economic value.

It can be seen that our biotechnology efforts are as comprehensive and integrated as are our multiple centers of excellence and interests. Thus, we have combined the industrial experience of our Formosa Group with the medical and basic science expertise of our Chang Gung Memorial Hospital-Medical Centers, Chang Gung University and Ming Chi University of Technology.

Based on the above considerations, we commenced full-scale planning and development of modern fermentation technologies in 1998, followed by building related research and commercial facilities. For purposes of continued research and development (R&D) in fermentation and metabolic engineering, we have set up several world-class laboratories and pilot-scale facilities.

Description of figures on the previous page

Figure 6. Animal Cell Culture Laboratory ¡V Protein purification equipment
Figure 7. Animal Cell Culture Laboratory
Figure 8. Animal Cell Culture Laboratory ¡V Protein electrophoresis equipment
Figure 9. Animal Cell Culture Laboratory ¡V Hollow fiber cell culture device
Figure 10. Animal Cell Culture Laboratory ¡V High Performance Liquid Chromatograph
Figure 11. Microbial Bioreactor Laboratory
Figure 12. Animal Cell Culture Laboratory ¡V Pilot-scale expanded bed
Figure 13. Animal Cell Culture Laboratory ¡V Cell culture reactor
Figure 14. Microbial Bioreactor Laboratory ¡V Online Instantaneous Magnetic Mass Spectrometer
Figure 15. Animal Cell Culture Laboratory ¡V Film imaging analyzer
Figure 16. Microbial Bioreactor Laboratory ¡V Elemental Analyzer (CNPS)
Background Microbial Bioreactor Laboratory ¡V Manufacturing Process Analysis Room

¡§Biotechnology Pilot Production Factory¡¨ and ¡§Purification and Separation Technology Laboratory¡¨, including Centrifuge and Filter Area, Homogenization Area, Condensing Area, Purification Area and Later-phase Processing Area. The total area exceeds 21,600 square feet.

Through years of diligent R&D, our company has become a worldwide leader in strain screening, strain improvement, genetic engineering of strains, molecular biological identification of strains, and strain manufacturing process development. We work with a wide spectrum of cells that include fungi, bacteria, plant and animal cells. We also lead in high cell density manufacturing technology, bioprocessing scale-up technology, process optimization, media design, bioprocess validation, process scale-down, simulation technology, and many other aspects of metabolic engineering.

Our company¡¦s technology can increase the concentration of wild-type Escherichia coli to 130 grams per liter or more (dry weight). For heterologous protein expressed in recombinant Escherichia coli, depending on different hosts and plasmid characteristics, the strain concentration can reach 70¡V100 grams per liter. Concentrations of Saccharomyces cerevisiae can reach 150 grams per liter or more and Pichia pastoris can reach 185 grams per liter or above.

In terms of strain screening and molecular identification of strains, our company has successfully screened and cultured a Hirsutella sinensis strain from natural Cordyceps sinensis specimens since 1999. In 2000, we utilized molecular identification technology to compare the 576 nucleotides found between the 18S and 25S genetic loci in rDNA samples derived from our strain and the wild form, through which efforts we were able to authenticate our Hirsutella sinensis to be identical to natural Cordyceps sinensis. This is a significant breakthrough since at that time all the strains sold on the market were of the wrong type! This effort also allows us to bring out this precious medicinal fungus¡Xonce reserved to emperors and the few members of the oligarchy who could afford it¡Xto the immediate reach of the general public.

Our company has sought further to simulate the natural habitat for Cordyceps sinensis, known to grow sparsely in only a few locations in the world at high altitude generally exceeding 4,000 meters. We have developed the proper culture media and growth conditions to optimize the product. We sought diligently to limit the accumulation of metabolic byproducts. More importantly, during culturing, specific nano-sized amorphous trace elements, extracted using our own proprietary materials technology, are carefully included. Most amazingly, as a result of these efforts and through high-performance liquid chromatography, we find that the Cordyceps sinensis product from our company has identical nutritional composition and metabolic products compared to natural Cordyceps sinensis!

Our efforts did not simply stop here. Our vision has always been to make all our products affordable to everyone. By utilizing numerous modern cell-growth optimization skills we have been able to optimize the cell yields and thereby reduce the production costs, savings which we intend to pass along to the end-user. In addition to making a precious food delicacy mass-affordable, we deem our offering to be superior to the precious wild form. That is, not only does our product have genetic fingerprint, nutritional composition and metabolite profile identical to natural Cordyceps sinensis, our product is in fact more stable and consistent (no bacterial pollutant or pesticide and no heavy metals).

The highest safety, hygiene, and quality benchmarks are three unyielding parameters that all Chang Gung Biotechnology products must surpass! Our company uses current Good Manufacturing Practice (cGMP) standards for all fermentation products. For example, for each lot of Cordyceps sinensis produced, five real-time dynamic parameters must be recorded in order for us to assure quality consistency on line.

Our fermentation research and production personnel are not only technologically proficient, but more importantly we are passionate about our company¡¦s vision and about our product offerings. Passion, positive intentionality, and professionalism go hand-in-hand in all aspects of our operation. There is not one product offered by us that we would not personally use or give to our own family members to use. We are certain that, with your sampling and getting to know us, you will also sense the level of our enthusiasm and goodwill as well as our product excellence.