In an effort to contribute to human health and a
sustainable society, the Biomolecular Engineering
Laboratory is devoted to research and education in
fermentation and novel bioprocesses with eukaryotic
microbes including Saccharomyces cerevisiae and
Aspergillus oryzae, as well as developing
high-throughput methods for functional analyses of
genes. See the list below to know who we are,
what we have done and what we are trying.
BEL Research objectives
Engineering and production of proteins for
industrial / diagnostic applications
Directed evolution
Bacterial expression
Yeast extracellular production
"difficult-to-express" proteins of which
industrial applications are promising
Receptor proteins
Fungal laccases
Peptides mimicking bioactive chemical compounds
Gene expression regulation and function
Molecules involved in chromosomal segregation
A quantitative reporter for yeast assays
(fungal secretory beta-galactosidase)
Peptide transporters
Protein-protein interaction analysis
Identifications of interaction domains
domain-based antagonist
BEL Publications
see below for detail
Classes (Educational
duties in charge)
For the 1st year undergrads
Guides to Food Sciences (in partial)
For the 2nd year undergrads
Basic Molecular Biology
For the 3rd year undergrads
Genetic Engineering (Techniques in DNA
recombination)
Exercise : Genetic Engineering
Subcloning of a recombinant gene (engineered
jellyfish GFP)
Current topics in Biotechnology
For the graduates' course
study, we are responsible for
Advanced course for Molecular Biology (which
covers advanced technologies used in molecular
biology)
Technical consultation for gene cloning, plasmid
construction and recombinant protein production, etc
BEL/LGE
History in brief
LGE (Lab. Genetic Engineering) Established in 1987
by emeritus professor Keiichi Takeishi
Dr. Horie joined (1988).
The first seniors graduated (1991).
Dr. Takeishi, elected to Dean (1999-2003).
Dr. Takeishi retired (2006).
Dr. Kawarasaki joined (2006).
LGE 20th anniversary (2007).
LGE,
reorganized to BEL (2008).
Dr.
Horie left (2009).
Dr. Ito
K joined (2010).
Dr. Ito
K promoted (2016).
Dr.
Tanaka joined (2017).
Dr. Tanaka promoted, moved to TUAT (2021).
BEL People (at
present)
Yasuaki Kawarasaki Ph.D.
Undergrads
Graduate student
Dr. M. Tanaka, Visiting researcher.
Visiting researchers from corporations
BEL In-house facilities
Thermal cyclers for PCR x4
Takara qPCR x1
Shimazu HPLC x1
Bioshakers for cultivation x3
Cultivation chambers x4
Microplate shakers X3
Auto-biophotorecorders x4
UV/Vis spectrophotometer x1
VersaFluor (Fluorometer) x1
Microspectrophotometer x1
Cell counter x1
Microscopes x3
UV crosslinker x1
Sonicator for cell lysis x1
Safety cabinets x2
Gel/Image analyzer w/ UV x1
Centrifuges (Swing-type, for 96well plates,
Angled-type for microtubes x3)
Luminometer x1
Fridges/Freezers x3
Electroporation x2
Milli Q (ultrapure-water dispenser)
Marubishi Jar-fermentor (2L) w/ cooling unit
x1
Luminescent-image capture x1
Shared facilities
(available nearby)
DNA sequencers
Mass spectrometer
LC-TOF-MS
Conforcal microscope
Ultracentrifuge (Hitachi)
Benchtop U-centrifuge (Beckman)
Cell sorter
Lumino/Fluoro imagers
Colony picker
NMR
96-well plate reader (UV/Vis)
Genechip reader
Peptide synthesizer
BIAcore, etc
Weekly Events
Reading: textbooks in English (Essential cell
biology)
Brief reports on research progresses (every Monday)
Journal club (every Friday)
Clean up (every Friday)
Guest talk (Tuesday, bi-week)
TGIF (occasionally, on Friday)
Annual Events and
Meetings
Jan., ACB annual symposium
Feb., UG 3rds recruiting as cadets
Feb., Thesis submission & qualification
Mar., Shizuoka Young Scientists' Conference
(regional conference)
Mar., Graduation/Farewell party
Mar., JSBBA annual meeting
Apr., Enrollment: "welcome to squadron" party
July, short talks about the researches (end of
semester)
Sept., Entrance exam. for the graduate course
Sept., SBJ annual meeting
Oct., JSBBA regional meeting
(Research excursion, somewhere in autumn)
Dec., MBSJ annual meeting
Dec., Talks about the researches, including summary,
introduction, and achievements
The research and educational activities in BEL constitute the
JABEE program for the School of Food and Nutritional Sciences
Papers published in international
journals (reviewed publications)
Hideo Nakano,
Tadaaki Tanaka, Yasuaki Kawarasaki and Tsuneo
Yamane*: An Increased Rate of Cell-free Protein
Synthesis by Condensing Wheat-germ Extract with
Ultrafiltration Membranes. Bioscience,
Biotechnology and Biochemistry, 58, 631-634 (1994)
Yasuaki Kawarasaki, Hideo Nakano and Tsuneo
Yamane*: Prolonged Cell-free Protein Synthesis in a
Batch System Using Wheat Germ Extract. Bioscience,
Biotechnology and Biochemistry, 58, 1911-1913
(1994)
Tsuneo Yamane*, Yasuaki Kawarasaki and
Hideo Nakano: In Vitro Protein Biosynthesis Using
Ribosome and Foreign mRNA--An Approach to Construct a
Protein Biosynthesizer. Annals of the New York
Academy of Sciences, vol. 750, Enzyme Engineering
XII, pp146-157 (1995)
Yasuaki Kawarasaki, Takahiro Kawai, Hideo Nakano
and Tsuneo Yamane*: A Long-lived Batch Reaction System
of Cell-free Protein Synthesis. Analytical
Biochemistry, 226, 320-324 (1995)
Hideo Nakano, Tadaaki Tanaka, Yasuaki Kawarasaki
and Tsuneo Yamane*: Highly Productive Cell-free Protein
Synthesis System Using Condensed Wheat-germ Extract, Journal
of Biotechnology, 46, 275-282 (1996)
Yasuaki Kawarasaki, Hideo Nakano and Tsuneo
Yamane*: Purification and Some Properties of Wheat Germ
Acid Phosphatases, Plant Science, 119, 67-77
(1996).
Hideo Nakano, Yasuaki Kawarasaki and Tsuneo
Yamane *: Recent Advances in Cell-free Protein Synthesis
towards a Protein Biosynthesizer Annals of
the New York Academy of Sciences, vol. 799 Enzyme
Engineering XIII, pp 406-412 (1996)
Yasuaki Kawarasaki, Hideo Nakano, and Tsuneo
Yamane*: Phosphatase-immunodepleted Cell-free Protein
Synthesis System, Journal of Biotechnology, 61,
199-208 (1998)
Yugo Iwasaki, Tatsuya Nishiyama, Yasuaki Kawarasaki,
Hideo Nakano, and Tsuneo Yamane*: Importance of
Disulfide Bridge Formation on Folding of Phospholipase D
from Streptomyces antibioticus, Journal of
Bioscience and Bioengineering, 89, 506-508 (2000)
Yasuaki Kawarasaki*, Satoru Kasahara, Natsuko
Kodera, Tomoya Shimbata, Satoshi Sekiguchi, Hideo
Nakano, and Tsuneo Yamane: A trimmed viral
cap-independent translation enhancing sequence for a
rapid in vitro gene expression, Biotechnology
Progress., 16, 517-521 (2000)
Yasuaki Kawarasaki*, Yoko Sasaki, Akinori
Ikeuchi, Shoji Yamamoto and Tsuneo Yamane: A method for
functional mapping of protein-protein binding domain by
preferential amplification of the shortest amplicon
using PCR, Analytical Biochemistry, 303, 34-41
(2002)
Suang Rungpragayphan, Yasuaki Kawarasaki, Takao
Imaeda, Katsunori Kohda, Hideo Nakano and Tsuneo
Yamane*: High-throughput, Cloning-independent Protein
Library Construction by Combining Single-molecule DNA
Amplification with in Vitro Expression, J. Mol.
Biol., 318, 395-405 (2002)
Yasuaki Kawarasaki*, Yasuhiro Yamada, Maki
Ichimori, Tomoya Shinbata, Katsunori Kohda, Hideo
Nakano, Tsuneo Yamane: Stabilization of affinity tagged
recombinant protein during/after its expression in
cell-free protein synthesis system using wheat-germ
extract., J. Biosci. Bioeng., 95, 209-214 (2003)
Akinori Ikeuchi, Yasuaki Kawarasaki*, Tomoya
Shinbata, and Tsuneo Yamane: Chimeric Gene Library
Construction by a Simple and Highly Versatile Method
Using Recombination-Dependent Exponential Amplification.
Biotechnol. Progr., 19, 1460-1467 (2003)
Yasuaki Kawarasaki, Karl E. Griswold, James D.
Stevenson, Tzvia Selzer, Stephen J. Benkovic, Brent
Iverson, and George Georgiou*: Enhanced Crossover
SCRATCHY: Construction and High-throughput Screening of
a Combinatorial Library Containing Multiple
Non-Homologous Crossovers. Nucl. Acid. Res., 31,
e126, (2003)
Akinori Ikeuchi, Yoko Sasaki, Yasuaki Kawarasaki*,
and Tsuneo Yamane: Exhaustive identification of
interaction domains using a high-throughput method based
on two-hybrid screening and PCR-convergence: Molecular
dissection of a kinetochore subunit Spc34p. Nucl.
Acid. Res., 31, 6953-6962, (2003)
Denfu Chen, Yasuaki Kawarasaki, Hideo Nakano,
and Tsuneo Yamane*: Cloning and in vitro and in vivo
expressions of plant glutathione S-transferase Zeta
class genes. J. Biosci. Bioeng., 95, 594-600
(2003)
Jeong K.J, Kawarasaki Y, Gam J, Harvey BR,
Iverson BL, Georgiou G.*: Periplasmic fluorescent
reporter protein and its application in high-throughput
membrane protein topology analysis. J. Mol. Biol.,
341, 901-9 (2004)
Griswold K., Kawarasaki Y., Ghoneim N., Benkovic
S., Iverson B.L., and Georgiou G.*: Evolution of Highly
Active Enzymes by Homology-Independent Recombination, Proc.
Natl. Acad. Sci. USA, 102, 10082-10087 (2005)
Kojima T., Takei Y., Ohtsuka M., Kawarasaki Y.,
Yamane T., Nakano T.*: PCR amplification from single DNA
molecules on magnetic beads in emulsion: application for
high-throughput screening of transcription factor
targets., Nucl. Acid. Res. 33, e150 (2005)
Zhao M., Zhou L., Kawarasaki Y., and Georgiou
G.*: Regulation of RraA, a protein of RNase E-mediated
RNA decay., J. Bacteriol., 188, 3257-3263,
(2006)
Tullman-Ercek D., DeLisa M.P., Kawarasaki Y.,
Iranpour P., Ribnicky B., Palmer T., and Georgiou G*.,
J. Biol. Chem. 282, 8309-16 (2007)
Kamiya T, Ojima T, Sugimoto K, Nakano H,
*Kawarasaki Y. Quantitative Y2H screening:
Cloning and signal peptide engineering of a fungal
secretory LacA gene and its application to yeast
two-hybrid system as a quantitative reporter. J.
Biotechnol. 146，151-159, (2010)
Ikeuchi A., Kamiya T., Sugimoto K.,
Yamane T., Nakano H., and *Kawarasaki Y., A
method for reverse interactome analysis: High-resolution
mapping of Interdomain interaction network in Dam1
complex and its specific disorganization based on
interaction domain expression. Biotech. Progr.
26, 945-953 (2010)
Ito K., *Ito S., Shimamura T., Kawarasaki Y.,
Abe K., Misaka T., Kobayashi T., and Iwata S.,
Crystallization and preliminary X-ray analysis of a
glucansucrase from the dental caries pathogen
Streptococcus mutans. Acta Cryst. F66, 1086-1088 (2010)
Ito K., *Ito S., Shimamura T., Weyand S., Kawarasaki
Y., Misaka T., Abe K., Kobayashi T., Cameron A.,
and Iwata S. Crystal structure of glucansucrase from
dental caries pathogen Streptococcus mutans. J. Mol.
Biol., 408, 177-186 (2011)
Kojima T, Nagao N, Ando D, Ojima T, Kawarasaki Y,
Kobayashi I, Nakajima M, *Nakano H. Emulsion culture: A
miniaturized library screening system based on
micro-droplets in an emulsified medium. J. Biosci.
Bioeng.112, 299-303 (2011)
Kimata K., Yamaguchi M., Saito Y.,
Hata H., Miyake K., Yamane T., Nakagawa
Y., Yano A., Ito K., and *Kawarasaki Y.,
High cell-density expression system: A novel method for
extracellular production of "difficult-to-express"
proteins. J. Biosci. Bioeng. 113, 154-159 (2012)
*Ito K, Hikida A, Kitagawa S, Misaka T,
Abe K, Kawarasaki Y., Soy Peptides Enhance
Heterologous Membrane Protein Productivity during the
Exponential Growth Phase of Saccharomyces cerevisiae.,
Biosci Biotechnol Biochem 76, 628-31 (2012).
Hikida A., *Ito K., Motoyama T., Kato R.,
Kawarasaki Y.: Systematic analysis of a dipeptide
library for inhibitor development using human dipeptidyl
peptidase IV produced by a Saccharomyces cerevisiae
expression system. Biochem. Biophys. Res. Commun., 430,
1217-22 (2013).
*Ito K,Hikida A, Kawai S, Lan
VTT, Motoyama T, Kitagawa S, Yoshikawa Y, Kato R,
Kawarasaki Y.: Analysing the substrate
multispecificity of a proton-coupled oligopeptide
transporter using a dipeptide library. Nat. Commun.,
4:2502 doi: 10.1038/ncomms3502, (2013)
Kurose T., Saito Y., Kimata K.,
Nakagawa Y., Yano A., Ito K., and *Kawarasaki
Y.: Secretory expression of Lentinula edodes
intracellular laccase by yeast high-cell-density system:
a sub-milligram production of “difficult-to-express”
secretory protein. J. Biosci. Bioeng. 117, 659-663
(2014) 10.1016/j.jbiosc.2013.11.014).
Lan V.T.T.,Ito K., Ohno M.,
Motoyama T., Ito S., Kawarasaki Y.: Analyzing a
dipeptide library to identify human dipeptidyl peptidase
IV inhibitor. Food Chem. 175, 66-73 (2015)
Mori A., Hara S., Sugahara T., Kojima T., Iwasaki Y., Kawarasaki
Y., Sahara T., Ohgiya S., Nakano H.: Signal
peptide optimization tool for the secretion of
recombinant protein from Saccharomyces cerevisiae. J
Biosci Bioeng. 120, 518-525 (2015)
Lan V.T.T., Ito K., Ohno M., Motoyama T., Ito S., Kawarasaki Y.:
Analyzing a dipeptide library to identify human dipeptidyl peptidase IV
inhibitor. Food Chem. 175, 66-73 (2015)
Mori A., Hara S., Sugahara T., Kojima T., Iwasaki Y., Kawarasaki Y.,
Sahara T., Ohgiya S., Nakano H.: Signal peptide optimization tool for
the secretion of recombinant protein from Saccharomyces cerevisiae. J
Biosci Bioeng. 120, 518-525 (2015)
Ohno M., Ito K., Lan V.T.T., Kusubata M., Tometsuka C., Koyama Y.I.,
Motoyama T., Ito S., Kawarasaki Y.: Synergistic inhibition of human
dipeptidyl peptidase IV by combinations of peptides. Peptides 69,
115-117, (2015)
Hashidume T, Sakano T, Mochizuki A, Ito K, Ito S, Kawarasaki Y,
*Miyoshi N.: Identification of soybean peptide leginsulin variants in
different cultivars and their insulin-like activities. Sci Rep. 2018 Nov
15;8(1):16847. doi: 10.1038/s41598-018-35331-5.
Su H, Ito K, Kawarasaki Y, Morita H, Nose H, Ikeda K,
Nakadouzono F, Gokuden M, Kamiyama S, Tokaji A, Rikitake Y, Kawaguchi T,
Umekita K, Oishi S, Abe F, Kanda T, Kawabata H, Ando S, Ohashi N.
Insight of diagnostic performance using B-cell epitope antigens derived
from triple P44-related proteins of Anaplasma phagocytophilum. Diagn.
Microbiol. Infect. Dis. 2019 Oct;95(2):125-130 (2019). doi:
10.1016/j.diagmicrobio.2019.05.008. Epub 2019
Ito K. Koike M., Kuroda Y., Yamazaki-Ito T., Terada Y., Ishii T., Nakamura Y., Watanabe T., Kawarasaki Y.:
Bitterness-masking peptides for epigallocatechin gallate identified
through peptide array analysis. Food Science and Technology Research 27
(2), 221-228 (2021)
Ito K., Hosoya T., Yamazaki-Ito T., Terada Y., Kawarasaki Y.
Dipeptidyl peptidase IV inhibitory dipeptides contained in hydrolysates
of green tea grounds. Food Science and Technology Research 27 (2),
329-334 (2021)
Tanaka M., Ito K., Matsuura T., Kawarasaki Y., Gomi K.
Identification and distinct regulation of three di/tripeptide
transporters in Aspergillus oryzae. Biosci. Biochem. Biotechnol., 85
(2), 452-463 (2021)
*Kawarasaki Y., Ohashi S., Watabe R., Tanaka M., Kurose T., Ito K.:
High Cell-Density Expression System: Yeast Cells in a Phalanx
Efficiently Produce a Certain Range of “Difficult-to-Express” Secretory
Recombinant Proteins. In: Garcia Fruitós E., Arís Giralt A. (eds)
Insoluble Proteins. Methods in Molecular Biology, vol 2406. Humana, New
York, NY. (2022) doi: 10.1007/978-1-0716-1859-2_16.
Underlined; the BEL lab members
*asterisk; corresponding author
