Biotechnology Research Center, Toyama Prefectural University, Japan
RECENT PUBLICATIONS
Enzyme Engineering XIX
JAPANESE
Where is Toyama located? and A view of Biotechnol Res Ctr Toyama Prefectural University Welcome to Toyama Prefecture, Japan List of Staff, members and students
Member Name Professor Yasuhisa ASANO asano@ Lecturer Hidenobu KOMEDA hkomeda@ Post Doctoral Fellow Shinjiro TACHIBANA z02015@st. Assistant Sanae IGARASHI Assistant Kazue AOTA Assistant Kaori NAKATA z02013@st. Assistant Asako TAKAGOSHI z0201j@st. Assistant Maiko TSUCHIDA z0201k@st. Doctor Course Student from Thailand Techawaree UEATRONGCHIT z0201k@st. Doctor Course Student Yasuhisa FUKUDA s67002@st. Master Course Student Kazuyuki YASUKAWA s556006@st. Junior College 3rd year Student Yukiko SHIBATA s0667005@st. Junior College Student Manami NISHINO s527026@st. Junior College Student Kazuyo NITTA s527027@st.
Research interests of Prof. Asano Prof. Asano's group in the Biotechnology Research Center of Toyama Prefectural University carries out an extensive screening for novel enzymes catalyzing new reactions, with their deep knowledge in microbiology, biochemistry, enzymology, synthetic organic chemistry, and molecular genetics, etc. They have been pioneers in the exploitation of novel enzymes, as can be seen in Prof. Asano's discovery of nitrile hydratase, which is now utilized in the industrial production of acrylamide, nicotinamide and 5-cyanovaleramide in multi-tons scale, and in the first crystallization and characterization of phenylalanine dehydrogenase followed by its successful use in the mass-screening of phenylketonuria in Japan. Not only the exploring studies, but also intensive basic studies are carried out to understand the enzymological characteristics by the use of synthetic chemicals. The structure of the enzymes are analyzed by the techniques in molecular cloning, nucleotide and protein sequencing. Site-directed mutagenesis of the genes are routinely carried out by random and designed manners.
Highlights of current work include; discovery of novel phenylalanine dehydrogenases (PheDH) from nature, use of PheDHs and opine dehydrogenase in the enantioselective synthesis of natural and unnatural L-amino acids; practical microdetermination of L-Phe in blood samples of neonates in Japan utilizing PheDH; characterization, gene cloning, structure elucidation, and expression of a new enzyme opine dehydrogenase as one of the first examples of D-stereospecific amino acid dehydrogenase; discovery of 3-methylaspartate ammonia-lyase from facultative anaerobes for chiral synthesis; discovery, characterization, structure determination of, alkaline D-peptidase and D-amino acid amidase and uses in the syntheses of D-amino acids and D-amino acid containing peptides; discovery of phenylacetaldoxime dehydratase in nitrile degraders; discovery of amino acid amide racemase; discovery of new hydroxynitrile lyases of plant origin and thier application to stereoselective cyanohidrin synthesis, etc. They are also successful in developing a new enzymatic method of selective phosphorylation of nucleosides by screening, optimization of the novel acid phosphatase by a molecular evolutionary approach, and its industical use.
They have been collaborating with Prof. Cooper's group of Biochemistry Department at the Weill Medical College of Cornell University, USA. on the use of PheDH. They are also cooperating with Profs. Engel's and Mayhew's groups of Univ. College Dublin, Ireland, to explore roles of some amino acid residues in PheDH and opine dehydrogenase by the site-directed mutagenesis, and the structural analyses by X-ray crystallography with Prof. Rice's group of Univ. of Sheffield, U.K. Their laboratory is the first one opened in the Center in 1992. Three laboratories were opened later in 1995, and the expansion of the Center was completed in 1996, when they started enrolling graduate students. Although they have only 10 years of history in the Center, some of the projects were started in the Sagami Chemical Research Center, Kanagawa, Japan in 1984. Their laboratory currently consists of 1 Lecturer, 3 postdoctoral fellows, 5 technicians, 6 postgraduate research students, and 2 junior college students. Financial supports have been provided by the Government of Toyama Prefecture, the Ministry of Education of Japan, Japan Society for the Promotion of Sciences, and various private scientific foundations in Japan.
He has been Editors of Journal of Molecular Catalysis B: Enzymatic, Bioscience, Biotechnology and Biochemistry (Tokyo), and Journal of Bioscience and Bioengineering (Osaka).
RESEARCH PROJECTS
1. NEW TOPICS
1.1 A new enzymatic method of selective phosphorylation of nucleosides, optimization of the enzymatic reaction by directed evolution, and its industrial use
1.2 Development of chip technology to determine blood L-phenylalanine
1.3 Screening for new hydroxynitrile lyases of plant origin and thier application to stereoselective cyanohydrin synthesis
1.4 Discovery of amino acid amide racemase
2. AMINO ACID DEHYDROGENASES
2.1 Phenylalanine dehydrogenase (EC 1.4.1.20) from Sporosarcina ureae, Bacillus sphaericus, Bacillus badius, Micrococcus sp.
L-Phenylalanine determination in the blood of neonates
Synthesis L-amino acids from their keto analogs.
2.2 Opine dehydrogenase from Arthrobacter sp.
Synthesis of secondary amine dicaroboxylic acids
X-ray crystallography and reaction mechanism (with Prof. D. Rice)
3. LYASES
3.1 D-Malate hydro-lyase
Synthesis of D-malate from maleate
Synthesis of citramalate from itaconate
3.2 3-Methylaspartate ammonia-lyase
Discovery in Enterobacteria
Synthesis of aspartate analogs from fumarate
X-ray crystallography (with Prof. D. Rice)4. D-AMINO ACID CONTAINING PEPTIDE HYDROLASES
4.1 D-Aminopeptidase (EC 3.4.11.19) from Ochrobactrum anthropi
Synthesis of D-alanine N-Alkylamides and D-Ala oligomers
Similarity of the primary structure with b-lactamases and PBP
4.2 Alkaline D-peptidase (a new enzyme)
Synthesis of D-Phe oligomers
Similarity of the primary structure with b-lactamases and PBP
4.3 D-Amino acid amidase
4.4 Microbial degradation of (D-Glu)8 and (D-Asp)8
5. ALDOXIME AND NITRILE METABOLISM IN MICROORGANISMS
Nitrile hydratase (a lyase)
Aldoxime dehydratase (a new lyase)
Amidases6. STEREOSPECIFIC ACYLATION OF GLYCEROL BY LIPASES
7. OTHERS
Stereospecific cis-1,2-indandiol oxidation to yield (R)-2-hydroxy-1-indanone
New epoxide hydrolases
JAPANESE
Professor Yasuhisa Asano
Biotechnology Research Center
Toyama Prefectural University
5180 Kurokawa, Kosugi,
Toyama 939-0398 JAPAN
Tel: +81-766-56-7500 ext 530; Fax: +81-766-56-2498
Mail to: asano@pu-toyama.ac.jp