Biomedical subjects

Tadahisa Iwata

Publications and source records attributed to Tadahisa Iwata.

At least 19 recordsLinked to original sources

PubMed2006

Adsorption of biopolyester depolymerase on silicon wafer and poly[(R)-3-hydroxybutyric acid] single crystal revealed by real-time AFM.

The adsorption behavior of PHB depolymerase from R. pickettii T1 on a silicon wafer and on P(3HB) single crystals has been studied by real-time and AFM in air and a buffer solution. First, the morphology of PHB depolymerase adsorbed on a silicon wafer was characterized to show that one molecule of PHB depolymerase has dimensions of 2.2 +/- 0.7 nm height and 16 +/- 5 nm width. The observation of PHB depolymerase adsorbed on a P(3HB) single crystal indicated that the dimensions of enzyme on the crystalline surface in air were 1.2 +/- 0.5 nm high and 28 +/- 7 nm wide, while enzyme molecules with dimensions of 2.1 +/- 0.6 nm height and 16 +/- 7 nm width were detected in a buffer solution. Comparison of the dimensions of PHB depolymerase in air with those in a buffer solution showed that the enzyme was squashed in air, but not in a buffer solution. In addition, the influence of enzymatic adsorption on the molecular state of the P(3HB) crystalline surface was investigated. The AFM images of P(3HB) single crystals after enzymatic adsorption and washing with ethanol indicated that the adhesion of PHB depolymerase changed the molecular state and generated holes on the crystalline surface.

Adsorption↗

PubMed2005

The crystal structure of polyhydroxybutyrate depolymerase from Penicillium funiculosum provides insights into the recognition and degradation of biopolyesters.

Polyhydroxybutyrate is a microbial polyester that can be produced from renewable resources, and is degraded by the enzyme polyhydroxybutyrate depolymerase. The crystal structures of polyhydroxybutyrate depolymerase from Penicillium funiculosum and its S39 A mutant complexed with the methyl ester of a trimer substrate of (R)-3-hydroxybutyrate have been determined at resolutions of 1.71 A and 1.66 A, respectively. The enzyme is comprised of a single domain, which represents a circularly permuted variant of the alpha/beta hydrolase fold. The catalytic residues Ser39, Asp121, and His155 are located at topologically conserved positions. The main chain amide groups of Ser40 and Cys250 form an oxyanion hole. A crevice is formed on the surface of the enzyme, to which a single polymer chain can be bound by predominantly hydrophobic interactions with several hydrophobic residues. The structure of the S39A mutant-trimeric substrate complex reveals that Trp307 is responsible for the recognition of the ester group adjacent to the scissile group. It is also revealed that the substrate-binding site includes at least three, and possibly four, subsites for binding monomer units of polyester substrates. Thirteen hydrophobic residues, which are exposed to solvent, are aligned around the mouth of the crevice, forming a putative adsorption site for the polymer surface. These residues may contribute to the sufficient binding affinity of the enzyme for PHB granules without a distinct substrate-binding domain.

Amino Acid Sequence↗

PubMed2005

Hydrolytic behavior of enantiomeric poly(lactide) mixed monolayer films at the air/water interface: stereocomplexation effects.

The Langmuir film balance technique was used to determine the hydrolytic kinetics of monolayers of the stereocomplex formed from mixtures of enantiomeric polylactides, poly(L-lactide) (L-PLA) and poly(D-lactide) (D-PLA), spread at the air-water interface. The present study investigated parameters such as degradation medium, mixture composition, and time on the relative degradation rate. The pi-A isotherms of monolayers of the mixtures provide clear evidence for the presence of a stereocomplex; the isotherms of monolayers of individual polyenantiomer show a transition at about 8.5 mN/m, whereas the transition of monolayers containing a stereocomplex formed from the equimolar mixture shifted to higher surface pressure, about 11 mN/ m. The rate of hydrolysis was recorded by a change in occupied area when the monolayer is maintained at a constant surface pressure. The hydrolysis of the mixture monolayers under basic conditions was slower than that of individual polyenantiomer monolayers, depending on the composition or the degree of complexation. In the presence of proteinase K, the enzymatic hydrolysis rate of mixture monolayers with >50 mol % l-PLA was much slower than that of the single-component L-PLA monolayer. The monolayers formed from mixtures with < or =50 mol % L-PLA did not show any change of occupied areas. This result is explained by the inactivity of D-PLA and stereocomplexed chains to the enzyme. From both results, it can be concluded that the retardation of the hydrolysis of mixture monolayers is mainly due to a strong interaction between D- and L-lactide unit sequences, which prevents the penetration of water or enzyme into the bulk.

Journal Article↗

PubMed2005

Uniaxial drawing of poly[(R)-3-hydroxybutyrate]/cellulose acetate butyrate blends and their orientation behavior.

Miscible blends of PHB and CAB were prepared by the solvent-casting method with various blend compositions, and their orientation behavior was investigated during uniaxial drawing. X-ray analysis revealed that the orientation of the crystallizable PHB component in the drawn PHB/CAB blends was changed from c-axis-orientation to a-axis-orientation with increasing CAB content. The a-axis-orientation was a result from the a-axis-oriented crystal growth caused by the intramolecular nucleation and the confined crystal growth. For quantitative assessment of the chain orientation, the Hermans orientation functions of the two respective components were obtained from the polarized FT-IR measurements. The orientation function of pure PHB stretched to 5 times of its initial length was approximately 0.8. However the value decreased rapidly with increasing CAB content, and it turned to a negative value from 30 wt.-% CAB content. This indicates that the PHB chains were aligned perpendicular to the drawing direction. On the contrary, the value of the CAB component remained almost unchanged at about 0.1 regardless of the blend composition and the annealing time, indicating that the CAB chains were constantly oriented parallel to the drawing direction without any chain relaxation. In addition, SAXS analysis suggested that the lamellar stacking direction also changed from parallel to perpendicular in the stretching direction with increasing CAB content.

Algorithms↗

PubMed2005

Crystal structure and morphology of poly(16-hexadecalactone) chain-folded lamellar crystals.

Solution-grown, chain-folded lamellar crystals of poly(16-hexadecalactone) (PHDL) were crystallized isothermally from 1-hexanol at 70 degrees C. The morphology of lozenge-shaped crystals was studied by TEM and AFM. The lamellae are ca. 10 nm thick and the chains run orthogonal to the lamellar surface with folding along (110) and (110) planes. The crystal structure of PHDL was determined by XRD and election diffraction of single crystals. The chains are in the 2(1) helix conformation close to all-trans and the structure consists of an orthorhombic unit cell with a P2(1)2(1)2(1) space group with the lattice constants a = 0.746 +/- 0.001 nm, b = 0.504 +/- 0.001 nm, and c (chain axis) = 4.116 +/- 0.003 nm. There are two chains per unit cell, which exist in an antiparallel arrangement. Molecular packing structure has been studied in detail, taking into account both diffraction data and energy calculations. The setting angles, with respect to a axis, were +/-40 degrees for the corner and center chains, respectively. By using the electron and XRD data, the best molecular packing model was refined to R-factors of 0.168 and 0.196, respectively. A brief comparison of chain-packing structure is also made with related polymer structures.

Crystallization↗

PubMed2005

Strong fibers and films of microbial polyesters.

Poly[(R)-3-hydroxybutyrate] (P(3HB)) and its copolymers are accumulated by a wide variety of microorganisms as intracellular carbon and energy material, and are extensively studied as biodegradable and biocompatible thermoplastics. However, these microbial polyesters have not been recognized as practical because of their stiffness and brittleness. Recently, by new drawing techniques, we succeeded in obtaining strong fibers and films from microbial polyesters, produced by both wild-type and recombinant bacteria. The improvement in mechanical properties of the fibers and films is due not only to the orientation of molecular chains, but also to the generation of a zigzag conformation and network structure, formed by fibrillar and lamellar crystals. The structure of strong fibers with a tensile strength over 1.0 GPa was analyzed by micro-beam X-ray diffraction with synchrotron radiation. The strong fibers and films were completely degraded in natural, river freshwater or by extracellular polyhydroxybutyrate depolymerases. In this feature article, the processing, mechanical properties, highly ordered structure and biodegradability of strong fibers and films produced from microbial polyesters are presented.

3-Hydroxybutyric Acid↗

PubMed2004

In-situ atomic force microscopy observation of enzymatic degradation in poly(hydroxyalkanoic acid) thin films: normal and constrained conditions.

The enzymatic degradation of lamellar crystals in poly(hydroxyalkanoic acid) thin films has been visualized by using in-situ dynamic force mode (tapping mode) atomic force microscopy (AFM) in buffer solution. It was found that poly(hydroxybutyric acid) (PHB) depolymerase from Ralstonia pickettii T1 degraded the thin surface layers formed at room temperature first, and that lamellar crystals formed at the crystallization temperature (110 degrees C) were eroded from the crystallographic a-axis to show splintered morphologies at the tips of the crystals. In some cases, lamellar crystals were hydrolyzed from the crystallographic b-axis, resulting in the formation of small crevices. These results suggest that disordered molecular chain-packing regions exist in the crystal along the crystallographic a- and b-axes, and that enzymatic degradation predominantly occurs from these defective regions. In addition, cantilever-tip-induced enzymatic degradation was carried out in the presence of PHB depolymerase. A concave area was artificially formed on the stacked lamellar crystals by the AFM tip. In-situ AFM observation has revealed that enzymatic degradation proceeds along both the longitudinal and lateral directions of the lamellae. At the same time, the PHB depolymerase preferentially eroded the concave area along the crystallographic c-axis. These results demonstrated that the PHB depolymerase predominantly degrades the less-ordered molecular chain-packing regions in the crystals.

3-Hydroxybutyric Acid↗

PubMed2004

Crystal structure, thermal behavior and enzymatic degradation of poly(tetramethylene adipate) solution-grown chain-folded lamellar crystals.

Solution-grown chain-folded lamellar single crystals of poly(tetramethylene adipate) (PTMA) were prepared from a dilute solution of 2-methyl-1-propanol by isothermal crystallization. PTMA crystals were hexagonal-shaped and polyethylene decoration of the crystals resulted in a "six cross-sector" surface morphology and showed that the average direction of chain folding is parallel to the crystal growth planes of [110] and [010]. Chain-folded lamellar crystals gave well-resolved electron diffraction diagrams corresponding to all the equatorial reflections of the X-ray fiber diagram obtained from stretched PTMA melt-quenched film (beta structure). The unit cell parameters of the beta structure of PTMA were determined as a = 0.503 nm, b = 0.732 nm and c (fiber axis) = 1.442 nm with an orthorhombic crystal system. The fiber repeat distance is appropriate for an all-trans backbone conformation for the straight stems. The setting angle, with respect to the a axis, is +/-46 degrees for the corner and center chains. Thermal behavior of lamellar crystals has been investigated by means of transmission electron microscopy (TEM) and atomic force microscopy (AFM). The lamellar thickness at the edges of the crystal increased after thermal treatment with taking the molecular chains into recrystallization parts; the holes then opened up at the thickening front of the crystal. The morphological changes of lamellar crystals after enzymatic degradation by Lipase type XIII from Pseudomonas sp. and water-soluble products were characterized by TEM, AFM, gel permeation chromatography, high performance liquid chromatography and fast atom bombardment mass spectrometry. The degradation progressed mainly from the edges of the lamellar crystals without decreasing the molecular weights and the lamellar thicknesses. The central portion of single crystals was often degraded by enzymatic attacks. This result combined with thermal behavior indicates that the loosely chain-packing region exists inside the single crystal, and that molecular chains in this region have higher mobility against thermal and enzymatic treatments.

Adipates↗

PubMed2002

Crystal structure of the (R)-specific enoyl-CoA hydratase from Aeromonas caviae involved in polyhydroxyalkanoate biosynthesis.

The (R)-specific enoyl coenzyme A hydratase ((R)-hydratase) from Aeromonas caviae catalyzes the addition of a water molecule to trans-2-enoyl coenzyme A (CoA), with a chain-length of 4-6 carbons, to produce the corresponding (R)-3-hydroxyacyl-CoA. It forms a dimer of identical subunits with a molecular weight of about 14,000 and is involved in polyhydroxyalkanoate (PHA) biosynthesis. The crystal structure of the enzyme has been determined at 1.5-A resolution. The structure of the monomer consists of a five-stranded antiparallel beta-sheet and a central alpha-helix, folded into a so-called "hot dog" fold, with an overhanging segment. This overhang contains the conserved residues including the hydratase 2 motif residues. In dimeric form, two beta-sheets are associated to form an extended 10-stranded beta-sheet, and the overhangs obscure the putative active sites at the subunit interface. The active site is located deep within the substrate-binding tunnel, where Asp(31) and His(36) form a catalytic dyad. These residues are catalytically important as confirmed by site-directed mutagenesis and are possibly responsible for the activation of a water molecule and the protonation of a substrate molecule, respectively. Residues such as Leu(65) and Val(130) are situated at the bottom of the substrate-binding tunnel, defining the preference of the enzyme for the chain length of the substrate. These results provide target residues for protein engineering, which will enhance the significance of this enzyme in the production of novel PHA polymers. In addition, this study provides the first structural information of the (R)-hydratase family and may facilitate further functional studies for members of the family.

Aeromonas↗

PubMed2002

Nanoparticles of amorphous ruthenium sulfide easily obtainable from a TiO2-supported hexanuclear cluster complex [Ru6C(CO)16]2-: a highly active catalyst for the reduction of SO2 with H2.

TiO(2)-supported ruthenium-metal particles were derived from an anionic hexanuclear carbido carbonyl cluster [Ru(6)C(CO)(16)](2-) and compared with those prepared conventionally by impregnation of TiO(2) with a solution of RuCl(3) followed by reduction with H(2). The average sizes of the metal particles in both systems are similar, that is, 12 A for molecular cluster-derived particles and 15 A for those derived from the RuCl(3) precursor, although the size distribution is sharper in the former case. These supported particles efficiently promote the reduction of SO(2) with H(2) to give elemental sulfur. Their active form is ruthenium sulfide as confirmed by EXAFS and X-ray diffraction measurements. The nanoscale ruthenium sulfide particles, which originated from the cluster complex, have an amorphous character and show activity even at low temperature (463 K), whereas ruthenium sulfide formed from RuCl(3)-derived metal dispersion is a pyrite-type RuS(2) crystallite and needs a temperature above 513 K to effect the same catalysis. Amorphous ruthenium sulfide maintains its nano-sized scale (approximately 14 A) regardless of the reaction temperature, while RuS(2) crystallite aggregates to form larger nonuniform particles.

Catalysis↗

PubMedDate not supplied

Nonhydrolytic fragmentation of a poly[(R)-3-hydroxybutyrate] single crystal revealed by use of a mutant of polyhydroxybutyrate depolymerase.

This paper reports the initial process of the enzymatic degradation of solution-grown lamellar single crystals of bacterial poly[(R)-3-hydroxybutyrate] (P(3HB)) with an extracellular polyhydroxybutyrate (PHB) depolymerase purified from Alcaligenes faecalis T1. We used a hydrolytic-activity-disrupted mutant of the PHB depolymerase in order to avoid the influence of hydrolytic reaction in the system. The effect of addition of the mutant enzyme upon the P(3HB) single crystals was investigated by turbidimetric assay, high-performance liquid chromatography (HPLC), and atomic force microscopy (AFM). Suspension turbidity of the P(3HB) single crystals increased after addition of the mutant enzyme having no hydrolytic activity. No soluble product from the P(3HB) single crystals with the mutant enzyme was detected by HPLC. AFM observation of the P(3HB) single crystals adsorbed on highly ordered pyrolytic graphite revealed that the mutant enzyme yielded a lot of lengthwise crystal fragments from the P(3HB) single crystals. On the basis of these results, we concluded that the mutant enzyme disturbs the molecular packing of the P(3HB) polymer chain around the loose chain packing region in the single crystal, resulting in the fragmentation. Therefore, it is suggested that the enzymatic degradation of P(3HB) single crystals with a wild-type PHB depolymerase progresses via three steps: (1) adsorption of the enzyme onto the surface of the single crystal; (2) disturbance of the molecular packing of P(3HB) polymer chain in the single crystal by the adsorbed enzyme; and (3) hydrolysis of the disturbed polymer chain by the adsorbed enzyme.

Alcaligenes↗

PubMedDate not supplied

Crystallization, stability, and enzymatic degradation of poly(L-lactide) thin film.

Poly(L-lactide) (PLLA) thin film with 100 nm thickness was crystallized at 160 degreesC for 20 min from the melt obtained at 220 degreesC. Hexagonal crystals with three types of growth (derivative growth lamellae, overgrowth multistacked lamellae, and undergrowth multistacked lamellae) were simultaneously observed by atomic force microscopy (AFM). These phenomena are due to the differences of the formative points of secondary crystal nuclei against the basal lamella. Enzymatic degradation of PLLA thin film revealed two types of amorphous regions. These regions were identified as the free amorphous region around the crystals and the restricted amorphous region between the crystal and glass substrate. In situ observation of thermal behavior of lamellar crystals was performed to understand the correlation between the chain folding and stability of the crystal by using temperature-controlled AFM. The morphology of the sectors with [100] growth plane had changed to a comblike morphology despite the fact that the [110] growth plane remained unchanged, suggesting that the stability of the chain folding and the chain-packing state affected the thermal behavior.

Crystallization↗

PubMedDate not supplied

Oxidative degradation of cis- and trans-1,4-polyisoprenes and vulcanized natural rubber with enzyme-mediator systems.

Oxidative degradation of cis- and trans-1,4-polyisoprenes by two types of enzyme-mediator systems, lipoxygenase/linoleic acid and horseradish peroxidase/1-hydroxybenzotriazole, was investigated at 37 degrees C in aqueous media and analyzed by gel permeation chromatography. Lipoxygenase and horseradish peroxidase activate their substrates, linoleic acid and 1-hydroxybenzotriazole, respectively, for scission of main chains of both 1,4-polyisoprenes. Molecular weights of 1,4-polyisoprenes decreased during the treatment under both enzyme-mediator systems, and the depolymerization was completely inhibited by the addition of butylated hydroxytoluene. When the enzyme or the mediator from a reaction system was omitted, depolymerization did not progress, indicating that the scission of polymer chain is induced by the radicals generated only in the presence of both enzyme and mediator. Fenton reagent with linoleic acid was also effective against the degradation of both 1,4-polyisoprenes. Vulcanized natural rubber latex gloves were treated under these three methods, and surface degradation with hole formation was observed with a scanning electron micrograph.

Chromatography, Gel↗

PubMedDate not supplied

Synthesis, solid-state structure, and surface properties of end-capped poly(L-lactide).

End-capped poly(L-lactide) (PLLA) samples with dodecyl or 2-(2-(2-methoxyethoxy)ethoxy)ethyl (MEEE) ester were synthesized by ring-opening polymerization of L-lactide in the presence of zinc dodecanoxide or zinc 2-(2-(2-methoxyethoxy)ethoxy)ethoxide as a catalyst, respectively. On the basis of NMR analysis, it was confirmed that the carboxylic acid chain ends of PLLA molecules were selectively substituted by dodecyl or MEEE ester groups. To evaluate the wettability on the surface of end-capped PLLA films, the advancing contact angle (thetaa) with water was measured. The amorphous PLLA films showed relatively similar thetaa values regardless of the chemical structure of the polymer chain end. In contrast, the thetaa values of semicrystalline films were varied over a wide range, dependent on the chemical structure of the chain end. In addition, the thetaa values of dodecyl ester end-capped PLLA film with low molecular weight increased with an increase in the crystallization temperature. Both the crystallinity and lamellar thickness of dodecyl ester end-capped PLLA films increased with the crystallization temperature. These results suggest that the segregation of the chain ends on the PLLA film surface was strongly affected by the crystallization conditions.

Absorption↗

PubMedDate not supplied

Metastability and transformation of polymorphic crystals in biodegradable poly(butylene adipate).

Polymorphism phenomenon of melt-crystallized poly(butylene adipate) (PBA) has been studied by wide-angle X-ray diffraction (WAXD), small-angle X-ray scattering (SAXS), and differential scanning calorimetry (DSC). It has been found that the isothermal crystallization leads to the formation of PBA polymorphic crystals, simply by changing the crystallization temperature. The PBA alpha crystal, beta crystal, and the mixture of two crystal forms grow at the crystallization temperatures above 32 degrees C, below 27 degrees C, and between these two temperatures, respectively. The relationship between PBA polymorphism and melting behaviors has been analyzed by the assignments of multiple melting peaks. Accordingly, the equilibrium melting temperatures Tm degrees of both alpha and beta crystals were determined by Hoffman-Weeks and Gibbs-Thomson equations for the purpose of understanding the structural metastability. The Tm degrees of the PBA alpha crystal was found to be higher than that of the beta crystal, indicating that the PBA alpha crystal form is a structurally stable phase and that the beta crystal form is a metastable phase. The analysis of growth kinetics of PBA polymorphic crystals indicates that the metastable PBA beta crystal is indeed the kinetically preferential result. Based on the thermal and kinetic results, the phenomenon of stability inversion with crystal size in melt-crystallized PBA was recognized, in terms of the growth mechanisms of PBA alpha and beta crystals and the transformation of beta to alpha crystals. The PBA beta --> alpha crystal transformation takes place at a sufficiently high annealing temperature, and the transformation has been evident to be a solid-solid-phase transition process accompanied by the thickening of lamellar crystals. The molecular motion of polymer chains in both crystalline and amorphous phases has been discussed to understand the thickening and phase transformation behaviors.

Biocompatible Materials↗

PubMedDate not supplied

Crystal structure and morphology of poly(12-dodecalactone).

Poly(12-dodecalactone) (PDDL) crystals in the form of chain-folded lamellae were prepared by isothermal crystallization from a 1-hexanol solution. The lozenge-shaped crystals with and without spiral growth have been studied by transmission electron microscopy and atomic force microscopy. Wide-angle X-ray diffraction data, obtained from PDDL lamellae sedimented to form oriented mats and annealed solvent-cast film, were supplemented with morphological and structural data from electron microscopy. PDDL crystallizes as an orthorhombic form with a P2(1)2(1)2(1) space group and lattice constants of a = 0.746 +/- 0.001 nm, b = 0.500 +/- 0.001 nm, and c (chain axis) = 3.281 +/- 0.003 nm. There are two chains per unit cell, which existed in an antiparallel arrangement. The fiber repeat distance is appropriate for an all-trans backbone conformation for the straight stems. Molecular packing of this structure has been studied in detail, taking into account both diffraction data and energy calculations. The setting angles, with respect to the a axis, were +/-43 degrees for the corner and center chains according to intensity measurements and structure factor calculations. The optimized shift along the crystallographic c axis is 0.1c (0.328 nm). A final model was obtained to yield R = 0.180 with X-ray diffraction data and R = 0.162 with electron diffraction data. A brief comparison is also made with related polymer structures.

Biocompatible Materials↗

PubMedDate not supplied

Enzymatic degradation processes of lamellar crystals in thin films for poly[(R)-3-hydroxybutyric acid] and its copolymers revealed by real-time atomic force microscopy.

Enzymatic degradation processes of flat-on lamellar crystals in melt-crystallized thin films of poly[(R)-3-hydroxybutyric acid] (P(3HB)) and its copolymers were characterized by real-time atomic force microscopy (AFM) in a phosphate buffer solution containing PHB depolymerase from Ralstonia pickettii T1. Fiberlike crystals with regular intervals were generated along the crystallographic a axis at the end of lamellar crystals during the enzymatic degradation. The morphologies and sizes of the fiberlike crystals were markedly dependent on the compositions of comonomer units in the polyesters. Length, width, interval, and thickness of the fiberlike crystals after the enzymatic degradation for 2 h were measured by AFM, and the dimensions were related to the solid-state structures of P(3HB) and its copolymers. The width and thickness decreased at the tip of fiberlike crystals, indicating that the enzymatic degradation of crystals takes place not only along the a axis but also along the b and c axes. These results from AFM measurement were compared with the data on crystal size by wide-angle X-ray diffraction, and on lamellar thickness and long period by small-angle X-ray scattering. In addition, the enzymatic erosion rate of flat-on lamellar crystals along the a axis was measured from real-time AFM height images. A schematic glacier model for the enzymatic degradation of flat-on lamellar crystals of P(3HB) by PHB depolymerase has been proposed on the basis of the AFM observations.

Biophysical Phenomena↗

PubMedDate not supplied

Uniaxial drawing and mechanical properties of poly[(R)-3-hydroxybutyrate]/poly(L-lactic acid) blends.

Blends of poly(L-lactic acid) (PLLA) with two kinds of poly[(R)-3-hydroxybutyrate] (PHB) having different molecular weights, commercial-grade bacterial PHB (bacterial-PHB) and ultrahigh molecular weight PHB (UHMW-PHB), were prepared by the solvent-casting method and uniaxially drawn at two drawing temperatures, around PHB's T(g) (2 degrees C) for PHB-rich blends and around PLLA's T(g) (60 degrees C) for PLLA-rich blends. Differential scanning calorimetry analysis showed that this system was immiscible over the entire composition range. Mechanical properties of all of the samples were improved in proportion to the draw ratio. Although PLLA domains in bacterial-PHB-rich blends remained almost unstretched during cold drawing, a good interfacial adhesion between two polymers and the reinforcing role of PLLA components led to enhanced mechanical properties proportionally to the PLLA content at the same draw ratio. On the contrary, in the case of UHMW-PHB-rich blends, the minor component PLLA was found to be also oriented by cold drawing in ice water due to an increase in the interfacial entanglements caused by the very long chain length of the matrix polymer. As a result, their mechanical properties were considerably improved with increasing PLLA content compared with the bacterial-PHB system. Scanning electron microscopy observations on the surface and cross-section revealed that a layered structure with uniformly oriented microporous in the interior was obtained by selectively removal of PLLA component after simple alkaline treatment.

Calorimetry, Differential Scanning↗