Bio-Process Engineering Group

PI: Koreyoshi IMAMURA, Professor(kore@cc.okayama-u.ac.jp)
Graduate School of Natural Science and Technology
Faculty of Engineering
Original website (Japanese)

Main Research focus:
A – Physicochemical characteristics of proteins stabilized by an amorphous sugar matrix (ASM)
B – Proteins adsorption onto metal surfaces:
Controlled by application of external electric field
Applications to surface cleaning
C - Atomic force microscopy investigations of protein conformation changes:
Hydrophobic interplays between bio-molecules and surfaces
D – Genetic engineering of proteins: immobilization techniques on functionalised surfaces
Applications to tomorrow’s biosensing systems
E – Specific designs of low-cost, versatile and eco-friendly drug delivery systems
Biomaterials fibers as innovative drug delivery vectors

A few examples …… (confidential topics have been omitted)
Flavours
Amorphoussugar matrix

Drugs

Cells

Proteins

Emulsion

Advanced stabilization ofproteins and enzymes.

A - Research topic:**Physicochemical characteristics ofproteins stabilized by an amorphoussugar matrix (ASM)
Key feature of ASM to stabilize
proteins:
èNo hydration of the protein
thanks to sugar (hydroxylgroup)/protein interactionèKeep protein conformation andselectivity.
èAddition of sugar surfactant may
help to maximize the function ofthe sugar amorphous matrix andachieve superior stabilization

D - Research topic:
Coll. Pr. H. Imanaka** Monitoring protein immobilization
to design advanced biosensing devices
Peptide - protein
Cushion -> control
immobilization andorientation
Affinity tag -> keep
activity / selectivity(4)

Denaturation
Wrong orientation

Advanced biomolecule engineering

Functional biomolecule
immobilization method:èEngineered “tag” peptides withsolid surface-affinity to control
the orientation of biomolecules
è“Tag” helps to maintain theirfunction and attach them to thesurface of solid materialsèElucidation of“tag”/biomolecules/surface
interactions

matrix

SolutionDrugreleasehydrophobicdrugdrug
solubilizer

E – Research topic:
Coll. Dr. O. Tramis

Solvent

Cell
scaffold

Body
fluid

stirring

Biomaterial
fiber

Seasoning
Aroma
etc….

Outer shell

Drug-loadedcore

Drug-loaded fibre

Advanced low-cost production of DDS**Design and production of innovative drugdelivery systems (DDS)Fiberization of drug-loaded
biomaterials:
èLow-cost device using ecofriendly production principleèHydrophobic drug is loaded
inside a “core-shell” structureèHydrophobic drug is sustainably
released when fibre is dissolvedin waterMajor Equipments
A – Microscopy

Inverted optical microscope Olympus CK2 (Sato Shouji Inc., Japan)
Atomic force microscope (Nanoscope IIIa; Veeco Instruments, USA)*B – Spectroscopy

UV-Vis V-730 spectrophotometer (Jasco Inc., Japan)
Fourier transform IR spectrometer (Nicolet 4700; Thermo Scientific Inc., Madison, WI)*FTIR spectrometer (FT/IR 4600, JASCO Co.) with ATR (ATRPRO400-S, JASCO Co., Tokyo, Japan)

C – Protein synthesis

PCR GoTaqGreen Master Mix (Promega Co., Madison, WI)
Purifier Quantum Prep PlasmidMiniprep Kit (Bio-Rad Laboratories, Inc. (Hercules, CA)
Purifier High Pure Plasmid Isolation Kit (Roche)
DNA sequencer ABI PRISM 310 Genetic Analyzer (Milpitas, CA).

**D – Physical chemistry

Modulated DSC Q2000 (TA Instruments Co., New Castle, DE)
Contact angle measurement Kruss EasyDrop (Kruss, Germany)
Zeta-potential analyser (ELSZ; Otsuka Electronics, Japan)

E – Other specific equipment**Underwater ellipsometer (Mizojiri Optical Co. Ltd., Tokyo, Japan)
Ellipsometer (LTE; Gaertner Scientific, Germany)Freeze-dryer (UT-80, EYELA TOKYO RIKAKIKAI Co., Tokyo, Japan)
Vacuum dryer (GLD-100, ULVAC Japan, Ltd., Tokyo, Japan)
Spray dryer (ADL-311-A, Yamato Scientific Co., Ltd., Tokyo, Japan)***Advanced drug delivery production (homemade)***International

Collaborations

Group in France (Tarbes, INP-ENIT, Toulouse “III” university)è Protein Immobilisation on surfaces / solid dispersion of hydrophobic drugs- Tramis, O., Iizuka, R., Nakao, H., Imanaka, H., Ishida, N., & Imamura, K. (2020). Immobilization of surface non-affinitive protein onto a metal surface by an external
electric field. Journal of bioscience and bioengineering, 129(3), 348-353.
- Sekitoh, T., Okamoto, T., Fujioka, A., Tramis, O., Takeda, K., Matsuura, T., ... & Imamura, K. (2020). Sole-amorphous-sugar-based solid dispersion of curcumin and theinfluence of formulation composition and heat treatment on the dissolution of curcumin. Drying Technology, 1-10.

Group in Australia (RSPE, Australian National University) è AFM -- interaction forces between surfaces in air and liquid- Ishida, N., & Craig, V. S. (2019). Direct measurement of interaction forces between surfaces in liquids using atomic force microscopy. KONA Powder and Particle
Journal, 36, 187-200.
- Ishida, N., Matsuo, K., Imamura, K., & Craig, V. S. (2018). Hydrophobic attraction measured between asymmetric hydrophobic surfaces. Langmuir, 34(12), 3588-3596.

Young researchers supervision
Ms. Ei Ei Htwe, Myanmar, Ph.D. student 2013-2016 (3 years)
Dr. Tramis Olivier, France (Toulouse), Research Fellow, 2017-2018 (5 months) and 2019-2020 (6 months)
Dr. Daniel Jonathan, France (Bordeaux), Visiting researcher, 2018 (2 months)
Thomas Ringler, Sorbonne University, Master Intern. from February to August 2023

Large Scale International Projects
MSCA – I-F ------ European-funded “Global” Fellowship –Application submitted in September 2021 ---- collaborations with:** Strasbourg University (France, Dr. Tramis), collaboration with IMS (Institut du Médicament de Strasbourg)** Paris “Sud” University and** Singapore “SUTD” University

Selected publications
Yokota, H., Kadowaki, M., Matsuura, T., Imanaka, H., Ishida, N., & Imamura, K. (2020). The Use of a Combination of a Sugar andSurfactant to Stabilize an Au Nanoparticle Dispersion against Aggregation during Freeze-Drying. Langmuir, in press
Sekitoh, T., Okamoto, T., Fujioka, A., Tramis, O., Takeda, K., Matsuura, T., ... & Imamura, K. (2020). Sole-amorphous-sugar-basedsolid dispersion of curcumin and the influence of formulation composition and heat treatment on the dissolution of curcumin.
Drying Technology, in press
Takeda, K., Sekitoh, T., Fujioka, A., Yamamoto, K., Okamoto, T., Matsuura, T., ... & Imamura, K. (2019). Physical Stability of an
Amorphous Sugar Matrix Dried From Methanol as an Amorphous Solid Dispersion Carrier and the Influence of Heat Treatment.
Journal of pharmaceutical sciences, 108(6), 2056-2062.
Htwe, E. E., Nakama, Y., Yamamoto, Y., Tanaka, H., Imanaka, H., Ishida, N., & Imamura, K. (2018). Adsorption characteristics ofvarious proteins on a metal surface in the presence of an external electric potential. Colloids and Surfaces B: Biointerfaces, 166,262-268.
Shimizu, T., Korehisa, T., Imanaka, H., Ishida, N., & Imamura, K. (2017). Characteristics of proteinaceous additives in stabilizing
enzymes during freeze-thawing and-drying. Bioscience, biotechnology, and biochemistry, 81(4), 687-697.
Takeda, K., Gotoda, Y., Hirota, D., Hidaka, F., Sato, & Imamura, K. et al. (2017). Surfactant-free solid dispersions of hydrophobic
drugs in an amorphous sugar matrix dried from an organic solvent. Molecular Pharmaceutics, 14(3), 791–798.
Imanaka, H., Yamadzumi, D., Yanagita, K., Ishida, N., Nakanishi, K., & Imamura, K. (2016). The use of a proteinaceous “cushion”
with a polystyrene-binding peptide tag to control the orientation and function of a target peptide adsorbed to a hydrophilicpolystyrene surface. Biotechnology progress, 32(2), 527-534.

BPE lab members FY 2019