Laboratory of Enzyme Chemistry
(Prof. Asano's Laboratory)

Biotechnology Research Center, Toyama Prefectural University, Japan

Sep 1, 1998 OPENED
Nov 26, 2014 UPDATED!
Member Name E-mail
Professor Yasuhisa ASANO asano@
Associate Professor Hidenobu KOMEDA hkomeda@
     
Assistant Kayo MURATA z02034@st.
Assistant Kumiko SHIMA z0201e@st.
     
ERATO Sub-Leader Kimiyasu ISOBE isobe@
ERATO Senior fellow Yasumasa KUWAHARA kuwahara@
     
ERATO Post Doctoral Fellow Yuko ISHIDA y-ishida@
ERATO Post Doctoral Fellow Daisuke MATSUI d-matsui@
ERATO Post Doctoral Fellow Mohammad DADASHIPOUR  
ERATO Post Doctoral Fellow Atsutoshi INA ina@
ERATO Post Doctoral Fellow Seiji OKAZAKI okazaki@
ERATO Post Doctoral Fellow Masafumi KAMEYA mkameya@
ERATO Post Doctoral Fellow Kazunori YAMAMOTO k-yamamoto@
ERATO Post Doctoral Fellow Takuya YAMAGUCHI yamaguchi@
ERATO Post Doctoral Fellow Shogo NAKANO snakano@
ERATO Post Doctoral Fellow Ken-ichi FUHSHUKU fuhshuku@
ERATO Post Doctoral Fellow Richard METZNER metzner@
ERATO Post Doctoral Fellow Atsushi IZUMI izumi@
ERATO Master's Fellow Yufeng MIAO miao@
     
ERATO Researcher by Collaboration Yasuaki TAKAKURA yasuaki_takakura@st.
     
ERATO Technical Staff Ryoko UEDA ueda@
ERATO Technical Staff Yuko OKU y-oku@
ERATO Technical Staff Sayaka SHICHIDA shichida@
     
ERATO Assistant Sayuri OHNO sayuri@
ERATO Assistant Yoko FUKAGAWA fukagawa@
ERATO Assistant Kaori KISHIHARA kishihara@
ERATO Assistant Yukiko SHINNAKA shinnaka@
ERATO Assistant Etsuko AKAE akae@
     
ERATO Headquarter Motoki MATSUDA m-matsuda@
ERATO Headquarter Yasuko ITAMOTO itamoto@
ERATO Headquarter Hiroko TAUCH tauchi@
ERATO Headquarter Tomoyo YOSHIKAWA yoshikawa@
ERATO Office Megumi HASHI hashi@
     
Doctor Course 2nd year Student Nobuhiro KAWAHARA t376001@st.
Doctor Course 1st year Student
Graduate Course Student by Academic Exchange Agreement with Prince of Songkla University, Thailand
Aem NUYLERT  
     
Master Course 2nd year Anna TERAI t356008@st.
Master Course 2nd year Shingo NAGAMORI t356011@st.
Master Course 2nd year Kana HONOKI t356014@st.
Master Course 2nd year Miho MURAKOSHI t356015@st.
     
Master Course 1st year Keiko OIKE t016007@st.
Master Course 1st year Atsushi OHNO t016010@st.
Master Course 1st year Toshiaki MORI t016037@st.
     
4th year student Yusuke OKAYAMA t116010@st.
4th year student Nobuyoshi KUMADA t116014@st.
4th year student Kanae SAWAMORI t116017@st.
4th year student Koutarou SIBATA t116018@st.
4th year student Natsuki NISHINO t116028@st.
     
3rd year student Takahiro OISHI t216008@st.
3rd year student Ryota OHASHI t216009@st.
3rd year student Kazuma KANAMORI t216015@st.
3rd year student Yuzo FUKUDA t216037@st.
3rd year student Ryota MISAWA t216039@st.
3rd year student Yukio WATANABE t216045@st.

E-mail address: place "pu-toyama.ac.jp" after @, or @st.


He has been Editors of:

Journal of Molecular Catalysis B: Enzymatic (Elsevier),
Bioscience, Biotechnology and Biochemistry (Tokyo),
Journal of Bioscience and Bioengineering (Osaka),
Frontiers of Chemical Engineering in China (Springer),
ChemCatChem (Wiley), and
Industrial Biotechnology (Mary Ann Liebert, Inc.).

He has been awarded:

The Progress Award in Synthetic Organic Chemistry (1990)
The Japan Bioscience, Biotechnology and Agrochemistry Society Award for the Encouragement of Young Scientists (1991)
Toyama Award (1993)
The Chemical Society of Japan Award for Technical Development for 2004 (2005)
The Japan Bioscience, Biotechnology and Agrochemistry Society Award (The Best Award of Japan Society of Bioscience, Biotechnology, and Agrochemistry) (2008)
The Japan Bioindustry Association Award (2008)
A Medal with Purple Ribbon (2011)
ERATO Asano Active Enzyme Molecule Project (2012)
The Toyama Newspaper "Culture Award" (2013)
Enzyme Engineering Award (2013)
Biocat Award – Science 2014(2014)

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 in the enantioselective synthesis of natural and unnatural L amino acids; microdetermination of L-Phe in blood samples of neonates utilizing PheDH (1/3 of Japanese babies are tested with the enzyme); characterization, gene cloning, structure elucidation, and X-ray structure determination 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 and X-ray structure determination; discovery, characterization, structure determination of D-stereospecific peptidases and D-amidases and uses in the syntheses of D-amino acids and D-amino acid containing peptides; discovery and characterization of transphosphorylase producing inosine 5'-phosphate, evolutionary mutagenesis of the enzyme for industrial application (3000 or more tons/year by Ajinomoto Co. from 2003) and X-ray structure determination, and occurrence of aldoxime dehydratase in nitrile degraders, etc.

An amino acid amide racemase activity was discovered in a-amino-e-caprolactam racemase and the dynamic kinetic resolution of amino acid amide with an amino acid amidase was realized. >From 2003, a new project on protein chips using PheDH was started. Since 2003, A “multi enzyme chip” is being developed for enzyme assay system for diagnosis of phenylketonuria (PKU), galactosemia (GAL), maple syrup urine disease (MSUD) and homocystinuria (HCU), in the Toyama Medical Bio-Cluster Project.

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. On Aug 1, 2006, a scientific exchange agreement was made in between The Faculty of Agro-Industry, Prince of Songkla University, Thailand, and the Graduate School of Engineering, Toyama Prefectural University. They and Drs. H-Kittikun and Hongpattarakere’s groups in Thai side are promoting activities in collaborative research on the development of resources of useful microorganisms and plants, and exchange of scientists and students.

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 15 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 Professor, 1 Lecturer, 1 Assistant Professor, 1 postdoctoral fellow, 1 postmaster fellow, 1 special researcher from a company, 4 technicians, 4 postgraduate research students, and 1 undergraduate student. 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.


RESEARCH PROJECTS

1. RECENT 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 and other amino acids

1.3 Screening for new hydroxynitrile lyases of plant origin and thier application to stereoselective cyanohydrin synthesis, and thier gene expression in heterologous hosts

1.4 Discovery of amino acid amide racemase and its use in the dynamic kinetic resolution of amino acid amides

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)
Amidases

6. 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 and Department of Biotechnology; JST, ERATO
Toyama Prefectural University
5180 Kurokawa, Imizu,
Toyama 939-0398 JAPAN
Tel: +81-766-56-7500 ext 530; Fax: +81-766-56-2498
Mail to: asano@

E-mail address: place "pu-toyama.ac.jp" after @