Bioprocessing in Industrial Production A Summary of the Current Research and Trends in Japan
Yaeko Mitsumori Senior Science & Technology Officer British Embassy January 2004
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Executive summary
Worldwide, in an effort to “green” chemical production, companies and governments are exploring the substitution of chemical processes with bioprocesses. Some U.S. companies such as DuPont and Cargill have been aggressively implementing bioprocessing in their manufacturing. However in Japan the commercial use of bioprocessing has so far been curtailed. Nevertheless some Japanese companies are actively exploring the implementation of bioprocess in their commercial scale production, particularly in the fine chemicals area. To increase the use of bioprocessing in industrial production in Japan, there are several challenges to be met. In particular, the time needed to implement bioprocessing production alternatives, the fixed costs and the prices of glucose sources all need to be reduced. In order to promote further uptake of bioprocessing in Japan, the New Energy and Industrial Technology Development Organisation (NEDO) an independent administrative institution under the Ministry of Economy, Trade and Industry (METI), has been promoting several governmentally assisted projects and in FY2003 NEDO appropriated a total of 3.87 billion yen (approximately £22million) to five leading national projects on bioprocessing. Outside these NEDO projects, other universities and research laboratories have been promoting their own bioprocess studies, including the Japan Marine Science & Technology Centre (JAMSTEC) project on new deep sea micro-organisms, Keio University’s E-cell project, and Prof. Imanaka of Kyoto University hyperthermophilie’s project. It is hoped that recent breakthroughs will encourage Japanese companies to increase the use of bioprocessing in their manufacturing projects in the future.
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Bioprocessing in Industrial Production A Summary of the Current Research and Trends in Japan
1. INTRODUCTION
Clean or green chemical synthesis is a major priority for many countries around the world as a potential contributor to improving the global environment. The use of bioprocessing, a manufacturing system fully utilising the metabolism of cells and enzymes, is one method being explored to achieve “green” chemical synthesis. Global environmental issues have been winning more attention among citizens. This, coupled with recent advances in molecular biology and a decrease in the price for glucose is driving the increased use of bioprocesses in industrial production. In the US, several companies, for example DuPont and Cargill, are implementing “bioprocesses” into their manufacturing processes. However in Japan the commercial implementation of “bioprocessing” has so far been comparatively less. This report summarises the current status of industrial efforts to implement bioprocessing techniques in Japan, and highlights some of the current research efforts being promoted by the government and others to enhance the use of bioprocessing.
2. CURRENT STATUS OF BIOPROCESS MARKETS IN JAPAN
There is no well-established market research on the use of “bioprocessing” in Japan. But, in 2002, the Dia Research Martech Inc, a leading Japanese market research firm, undertook a survey of all the bio related business markets in Japan. This survey concluded that the total value of the bio related business market in Japan was 1.43 trillion yen, broken down by product as follows: Table 1. Major bioprocess products in Japan Major bio process products Medicine Washing agents Acrylamide Isomerized sugar Trehalose Bio remediation Total Market scale (unit: bil yen) 391 140 5 73 8 2 1,433
The study also looked at the extent to which bioprocesses were used in various chemical ________________ Page 3
manufacturing processes, to assess the economic impact of bioprocessing in Japan. The study concluded that in Japan, as in the rest of the world, the use of bioprocessing techniques is growing, but the economic impact in Japan is still fairly modest. The results are summarised in the table below: Industries Petrochemical industries Products Production Bio process (bil yen) ratio (%) petrochemical 3.8 8.3 0.2 0.32 5.2 1.3 2.3 0.2 3.9 7.8 0.47 1.1 0.68 1.7 1.8 1.0 28.0 3.0 80 16 15 6.4 20 30 12 40 30 10 26 15 5 30 1.7 16
Basic chemicals Basic products Ethanol Methanol Other petrochemical products Synthetic resins for fibre products Fine chemicals Surface active agent/cosmetics Dyes Other final chemical products Medical, agricultural industries Medicines Agricultural chemical Foods industry Sugar Starch/saccharide Seasoning Pulp industry Pulp Electronic & machines industries Medical devices & machines, sensor Energy industry Petroleum products Environment industry Disposal of wastes
According to NEDO, the major reasons the industrial uptake of bioprocessing in Japan has been less compared to other countries is: · · · The length of time it takes to implement bio processes, for example screening micro-organisms, improving the function of the screened micro-organisms, optimising culture conditions. Higher costs for facilities The high price of glucose sources in Japan.
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3. INDIVIDUAL COMPANY ACTIVITIES
Despite comparatively low take up of bioprocessing in Japan, a number of Japanese companies are now actively investigating the conversion of their chemical process into bioprocesses, to reduce energy consumption, the amount of harmful industrial wastes produced and production costs. Examples of Japanese companies using bioprocessing in industrial production processes are shown in the table below:
Category Widely use products Pharmaceutical intermediates
Products chemical Acrylamide
Manufacturers Mitsubishi Rayon Sankyo Tanabe Mitsubishi Rayon Ajinomoto Kyowa Hakko Fuji Pharmaceutical Mitsubishi Chemical Tosoh Kaneka Corp Mitsubishi Chemical Ajinomoto Kaneka Corp Mitsubishi Chemical Mitsui Chemical Toyota
Pravastatin Diltiazem Pyridine-3-carboxamide L-Dopa Trans-4-Hydroxyproline D-Pantothenic acid Foods products, food L-Asparatate intermediates Aspartame Acethlthio methyl propionate Erythritol Fine chemicals D-Phenylalanine Acetoxy Azetidine Aminomethyl Chloropyridine Bio degradable plastic Poly Lactic Acid Lactic Acid Source: NEDO
Companies that are particularly active in bioprocessing include: Daicel Daicel Chemical Industries, a leading chemical company, has been producing a variety of chiral products using bioprocessing techniques. Three major chiral products on the market are: (R)-1,3-Butanediol (BDO) (an intermediate for Penem antibiotics); Ethyl (S)-4-Chloro-3-Hydroxybutanoate (SECHB) (an intermediate for HMG-CoA reductase inhibitors); and L-Homophenylalanine (HPHE) (an intermediate for ACE inhibitors). In addition, Daicel is producing two chiral products at pilot plants and five in laboratories. Daicel is aiming to increase sales revenues from its chiral business division from current 2.5 billion yen to 20 billion yen in FY2010. Mitsubishi Mitsubishi Chemical, a leading petrochemical company, has been producing L-Robose, an ________________ Page 5
intermediate for anti-virus medicine using bioprocesses. The production scale is about several tons per year and the price several tens of thousands of yen per kilogram. The firm is targeting to sell the product to world mega pharmaceutical companies such as GSK, Merck, Pfizer and Roche. Mitsubishi Chemical also has active bioprocessing projects in the areas of food & food intermediates such as L-Aspararate and some fine chemicals such as Ainomethyl Chloropyridine.
4. NATIONAL BIOPROCESS PROJECTS
Despite the low uptake by industry to date, there is a considerable stream of public funds flowing into research on developing and applying bioprocessing technology to clean chemical synthesis processes. NEW ENERGY AND INDUSTRIAL TECHNOLOGY DEVELOPMENT ORGANISATION (NEDO) Over the last few years, the New Energy and Industrial Technology Development Organisation (NEDO) has launched several national projects to develop technologies to reduce the environmental impact of chemical processes using environmentally friendly “bioprocesses.” These include the following: · The Bioprocess development project ( 2000- 2005) · The Plant production process controlling technologies development project (2002 – 2010); · Analysis on bio decomposition & processing mechanism, & controlling technologies project (2002 – 2006) · Construction of unknown micro-organisms libraries (2002 – 2007). · Plant function improving technologies development project (2003 – 2005) In FY2003, NEDO appropriated a total of 3.87 billion-yen (approximately £22million) to theses five leading national projects. The annual budget for each of these projects since their launch is shown below: Project (Unit: million yen) FY99 Bio process development Plant production process control Bio decomposition & processing mechanism New micro-organisms library Plant function improving 500 technologies FY00 300 FY01 2,000 FY02 1,890 970 770 500 500 550 485 FY03 1,513 882 659 430 391 FY04 1,250 860 640 420 246
Further details on each of these projects is as follows: I. Bioprocess development project
Leader: Prof. Sakayu Shimizu, Kyoto University Period: 2000-2005 ________________ Page 6
Aims and Objectives: This project is aimed at identifying and developing microorganism strains useful for industrial production. Along with development of real host cells, the project is promoting modelling of cells in sillico. In addition, the group is establishing libraries of micro-organisms and genes in order to aid industries in implementing bioprocesses. Scope: The project is being conducted in two-phases. Phase I is focused on the development of host cells for wide use in a variety of bioprocesses, targeting E. coli, Bacillus subtilis and yeast. Under the leadership of Professor Shimizu, three sub groups are working on: the production of new host cells in a wet lab, gene analysis, and the development of highly productive host cells by deleting unnecessary genes. (the Minimum Genome Factory Project); the study of host cells in sillico to produce an ideal micro-organism for industrial purposes; and the construction of libraries for useful micro-organisms, including useful micro-organisms from Japan as well as overseas, cloned genes involved in transformation and organic solvency, and analysis of the functions of genetic products. During the second phase, the project will focus on basic technologies for developing host cells ability to survive under harsh environments. II. Plant production process controlling technologies development project
Leader: Prof. Atsuhiko Shinmyo, Nara Institute of Science & Technology Period: 2002-2010 Aims and Objectives: The project is aimed at developing technologies that control material production in plants. Scope: The project is targeted at identifying the enzymes and genes of model plants involved in plant metabolism, analysis of their production controlling mechanisms and the construction of a database based on the collected data. By fully utilising the database, the project will analyse the production system for a targeted product, and establish technologies that let plants produce the targeting industrial materials efficiently. The project will deal with both model plants and selected plants useful for industrial production. III. Bio decomposition & processing mechanism
Leader: Prof. Yasuo Igarashi, Tokyo University Period: 2002-2006 Aims and Objectives: Focusing on anaerobic bacteria, the project will analyse the structure and function of bacteria involved in bio-decomposition, monitor key bacteria, and develop technologies to control these bacteria. Scope: The project has two key targets a. the production of methane b. the decomposition of materials in soil which are normally difficult to decompose. The project will focus on identifying factors that accelerate the decomposing process. IV. Construction of micro-organisms libraries
Leader: Shigeaki Harayama, Director of Biotechnology Development Centre (NBDC), National Institute of Technology and Evaluation ________________ Page 7
Period: 2002-2007 Aims and Objectives: To identify new micro-organisms, analyse them and construct micro-organisms libraries. Scope: The project is aiming to collect, culture and preserve 10,000 micro organisms which are easily cultured in normal environmental conditions, and 28,000 micro-organisms which are difficult to culture. The project is aiming to identify 3,530 genes from these micro-organisms and construct libraries. V. Plant function improving technologies development project
Leader: Prof. Atsuhiko Shinmyo, Nara Institute of Science & Technology Period: 2003-2005 Aims and Objectives: To identify the genes involved in production, stress control and expression of function in plants, animals and micro-organisms. It also aims to develop technologies for introducing these genes into plants to improve a variety of plant functions. Scope: The project will develop technologies (1) to use a plant’s metabolism for producing industrial materials (2) to increase a plant’s tolerance to stress (3) to introduce multiple genes into plants. The project is aiming to identify 30 genes involved in production and to create recombinant plants through introducing these genes. A full list of the participants in each of these programmes is at Annex 1.
5. OTHER GOVERNMENT PROJECTS
In addition to NEDO funded research, there are several other projects both government and independent to promote bioprocessing in Japan. I. “Bioventure Centre for Extremophiles” The “Bioventure Centre for Extremophiles” project is a government-industry collaboration effort to utilise deep-sea bio resources. The Frontier Research System for Extremophiles at the Japan Marine Science & Technology Centre (JAMSTEC), an independent administrative institution under the Ministry of Education, Culture, Sports, Science and Technology (MEXT) launched the project three years ago. Currently a dozen Japanese companies including foods processing companies, IT companies, pharmaceutical companies, paper companies, and toiletry companies are participating in the joint programme. Utilising the “Shinkai 6500” (a manned research submersible which reaches depths of up to 6,500 meters) and “Kaiko” (a remote operated vehicle which reaches depths of up to 11,000), the participants of the project have been studying extremophiles collected by these two submarines. The project has already identified twelve new, potentially useful micro-organisms and has obtained 10 patents based on the project findings. In addition, another 18 patents have been applied for. ________________ Page 8
II.
“E-cell”
The “E-cell” project, an in-sillico cell modelling project, was launched in 1995 and is headed by Masaru Tomita, Professor at the Institute for Advanced Bio-sciences at Keio University. Currently a group of about 70 scientists are working on the development of E-cell Version 3. It is hoped Version 3 will serve as a universal simulation platform that can integrate any sets of different simulation algorithms. It is designed to carry out simulation efficiently with different time scales and spacial resolutions. The group has also developed Systems Biology Markup Language (SBML). The project involves several Japanese companies including Mitsui Knowledge Industry Co, Intec Web, Genome Informatics Corp., and Mitsubishi Space Software. III. “Thermococcus kodakaraensis KOD1”
Dr. Tadayuki Imanaka, Professor of Synthetic Chemistry and Biological Chemistry at Kyoto University is studying hyperthermophilies from Kodakara Island, South of Kagoshima Prefecture, and so far has identified one potentially very useful thermophile, Pyrococcus kodakaraensis KOD1. The archaeal protein is extremely thermostable with an optimal temperature of 95 C degree. Professor Imanaka has identified the entire genome sequence for the KOD1 and analysed the functions of the thermoresistance protein. Prof. Imanaka has also found that enzymes produced from KOD1 are extremely stable, and maintain their function for a long time even under extreme circumstances. These newly discovered hyperthermophilies are expected to be used for a variety of industry processes. It is hoped that the breakthroughs arising from these programmes will enhance the uptake of bioprocesssing techniques across Japanese industry.
6. REFERENCES
“21st Bio Technology Symposium Proceeding” by Bio Technology Development Research Cooperative (11 Nov 2003) “New Energy and Industrial Technology Development Organization (NEDO) Annual report 2003 – 2004” by NEDO (2003) “Green Biotechnology” edited by Hajime Unno, Yoshio Okahata published by Kodansha (20 May 2003) “BioScience & Industry” by Japan Bioindustry Association (Vol 61 Dec 2003) “BioScience & Industry” by Japan Bioindustry Association (Vol 61 Nov 2003) “New Bioresources Modification Study Group Symposium Proceeding” by Japan Bioindustry Association (5 Dec 2003) ________________ Page 9
“New Bioprocess Technology Research” by Dia Research Martech (March 2003) “System Biology and Manufacturing using Bio Genetic Information – New Development of Fermentation Industry Targeting Further Development of Green Chemistry Proceeding” compiled by the Japan Bioindustry Association (17 Dec 2003) “E-Cell Project” web site: http://www.e-cell.org “Frontier Research System for Extremophiles” brochure compiled by Japan Marine Science & Technology Center (2003)
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Annex I
Participants and research topics of the NEDO Projects
(1) Bio process development project Project leader: Prof. Sakayu Shimizu, Kyoto University Participants of the project: (Host cells development group) Kyowa Hakko Kogyo Kao Corp Asahi Glass Co Mitsubishi Chemical Suntry Ltd Research Institute of Innovative Technology for the Earth (RITE) (cell modelling group) Keio University Osaka University Kyushu Institute of Technology (Micro-organism genetic resource library construction group) Mercian Corp Daicel Chemical Industries Marine Biotechnology Institute Co Japan Energy (Nippon Mining Holdings Group) Kaneka Corp (2) Plant production process controlling technologies development project Project leader: Prof. Atsuhiko Shinmyo, Nara Institute of Science & Technology Participants of the project: Ajinomoto Co Takara Bio Inc Nippon Paper Industries Co (Nippon Unipac) Oji Paper Hitachi Zosen Corp Bridgestone Corp Tokiwa Phytochemical Co Planted Research Institute Marine Biotechnlogy Institute Co ToyoboCo Kazusa DNA Research Institute National Institute of Advanced Industrial Science and Technology (AIST) Research Institute of Innovative Technology for the Earth (RITE) (3) Analysis on bio decomposition & processing mechanism, & controlling technologies Project leader: Prof. Yasuo Igarashi, Tokyo University Participants of the project: Ebara Corp Fujikasui Engineering Co ________________ Page 11
Marine Biotechnology Institute Co Kurita Water Industries Mitsui Engineering & Shipping Co Kubota Corp Central Research Institute of Electric Power Industry Research Institute of Innovative Technology for the Earth (RITE) (4) Construction of unknown micro-organisms library Project leader: Shigeaki Harayama, Director of Biotechnology Development Centre (NBDC), National Institute of Technology and Evaluation Participants of the project: National Institute of Technology and Evaluation Marine Biotechnology Institute Co Xanagen Japan Bioindustry Association (JBA) (5) Plant function improving technologies development project Project leader: Prof. Atsuhiko Shinmyo, Nara Institute of Science & Technology Participants of the project Oji Paper Suntry Ltd Taisei Corp Hitachi Zosen Corp Mitsui Chemicals Inc Toyota Motor Corp Toyobo Co Ltd Research Institute of Innovative Technology for the Earth (RITE)
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