(This research work has been chosen as one of the promotion enterprise of Basic Research Activities for Innovative Biosciences by Bio-oriented Technology Research Advancement Institution (BRAIN) in 2001."Basic research on advanced use of filamentous fungi as protein factories")

Aspergillus oryzae is an industrially important filamentous fungi and considered as the "Japanese National Microorganism". It is being used for brewing of sake, bean paste, and soy sauce in Japan since several hundred years. During the brewing of sake, the starch of rice is efficiently converted into glucose by the amylases produced by Aspergillus oryzae, and in the later steps sake yeasts convert the glucose into alcohol. Amylase production capacity of Aspergillus oryzae is very high, and it is known that several grams of amylase is produced in 1 liter cultivation. The protein secretion capacity of the yeasts which have been researched well is only 1/100-1/1000 of Aspergillus oryzae. Among the eukaryotes, Aspergillus oryzae has the highest protein secretion capability. Furthermore, its safety is guaranteed by the long history of its safe usage for food manufacture. These aspects of Aspergillus oryzae globally attract much attention as a host for efficient protein production.

In this laboratory, we are breeding strains of Aspergillus oryzae with high protein production capabilities using advanced molecular biological techniques. In general, it is known that the protein production capability on solid-state cultivation is higher when compared with that of liquid cultivation. However, the mechanisms involved during such processes remain unknown.Therefore, using the information available on the Aspergillus oryzae EST and genome projects coupled with advanced molecular biology-techniques, we aim to construct a system to produce various, high purity, and large quantities of protein by liquid-cultivation. This makes the control of fermentation process easy. The advantage of this system would produce large quantities of proteins by using lower cultivation scale and saving energy. These objectives would improve faster productivity of various proteins for food, medical treatment, and environmental restoration.

We got the following high evaluationin the 3rd year .

" The filamentous fungal representative, Aspergillus oryzae, is used for the production of various enzymes and proteins by solid-state cultivation. This research analyzes an efficient enzyme production control mechanism and aims at the creation of the "Super Koji" which produces proteins efficiently in liquid culture medium. This research on the elucidation of the secretory apparatus which is important for the production of proteins efficiently in a culture medium is of international level and would lead to important clues in solving the protein secretion problems in fungi. Furthermore, the trials on comprehensive analysis of fungal expression mechanism in both solid and liquid-state cultivation is also highly esteemed. Further developments in these studies is expected to form the basis of research on eukaryotic microorganisms in Japan."

The following heterologous proteins are being expressed in Aspergillus oryzae

Animal

Human- Lysozyme,CLN2

Rubripes- Interleukin, Lectin

Jellyfish -Green Fluorescence Protein

Plant

Sweet Protein

Filamentous fungi

Hydrolase