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We report matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and off-line coupling of size-exclusion chromatography with MALDI-TOFMS analysis (SEC/MALDI-TOFMS) methods for the detailed characterization of poly[(R,S)-3-hydroxybutyrate-co-L-lactic acid], P[(R,S)-3HB-co-LA], and poly[(R,S)-3-hydroxybutyrate-co-epsilon-caprolactone], P[(R,S)-3HB-co-CL], copolymer samples which are expected to be used in special medical application as scaffolds for cartilage and soft tissue engineering. The novel copolyesters contained randomly distributed (R,S)-3-hydroxybutyrate structural units, were synthesized by transesterification of the corresponding homopolymers, i.e. atactic poly[(R,S)-3-hydroxybutyrate], a-PHB, and poly(L-Lactide) (PLLA) or poly(epsilon-caprolactone) (PCL), respectively. The MS methods used for the characterization of the resulting polydisperse copolyester samples were supported by classical methods (NMR, SEC). The structures of individual copolyester macromolecules, including end-group chemical structures, were established using initially MALDI-TOFMS and then SEC/MALDI-TOFMS. The compositions of the copolyesters were determined by two methods, namely based on 1H NMR and MALDI-TOF spectra. The two sets of values showed good agreement. The sequence distribution was determined using the signal intensities of individual copolyester macromolecules, which appeared in MALDI-TOF mass spectra. Furthermore, sequence analysis gave information about the degree of transesterification. The copolyesters synthesized, with only one exception, were demonstrated to be almost random, which implies that the ester-ester exchange was close to completion.
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PMID:Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with size-exclusion chromatographic fractionation for structural characterization of synthetic aliphatic copolyesters. 1647 Jul 27

The biodegradability of poly(tetramethylene succinate) (PTMS), a synthetic aliphatic polyester with a high melting point, was evaluated. The ecological study showed that the distribution of PTMS-degrading microorganisms in soil environments was quite restricted compared with the distribution of microorganisms that degrade poly((epsilon)-caprolactone) (PCL), a polyester with a low melting point. However, in soil samples in which the formation of a clear zone was observed, PTMS-degrading microorganisms constituted 0.2 to 6.0% of the total number of microorganisms, which is very close to the percentage (0.8 to 8.0%) observed for PCL-degrading microorganisms. Five strains were isolated from colonies which formed distinct clear zones on agar plates with emulsified PTMS. In liquid cultures of the isolates with ground PTMS powder, strain HT-6, an actinomycete, showed the highest PTMS degrading activity. It assimilated about 60% of the ground PTMS powder after 8 days of cultivation. When a PTMS emulsion was used, a higher degradation rate was observed and more than 90% of the PTMS was assimilated in 6 days. PTMS degradation products were analyzed by gas chromatography, and it was found that 1,4-butanediol, 4-hydroxy n-butyrate, and succinic acid accumulated during cultivation. Degradation of PTMS film by the strain occurred in two steps: fragmentation and then the formation of hemispherical holes on the surface of the film. Strain HT-6 was also able to assimilate PCL and poly((beta)-hydroxybutyrate) (PHB). The crude enzyme showed a wide range of substrate specificity, being able to degrade low-molecular-weight PTMS, PCL, PHB, and even high-molecular-weight PTMS.
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PMID:Microbial degradation of an aliphatic polyester with a high melting point, poly(tetramethylene succinate). 1653 23

Thermophilic actinomycetes were isolated from sediment of the Chingshuei hot spring in north Taiwan, and the strain HS 45-1 was selected from colonies which formed distinct clear zones on agar plate with emulsified polyethylene succinate (PES). The film of PES disappeared within 6 days in liquid cultures at 50 degrees C. The strain HS 45-1 was also able to degrade poly (epsilon-carpolactone) (PCL) and poly (3-hydroxybutyrate) (PHB) films completely within 6 days in liquid cultures. Basing on the results of phynotypic characteristics, phylogenetic studies and DNA-DNA hybridization, strain HS 45-1 should be assigned to Micorbispora rosea subsp. taiwanensis.
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PMID:Degradation of polyethylene succinate (PES) by a new thermophilic Microbispora strain. 1710 89

Thermophilic actinomycetes strains were isolated from various environment in Taiwan and screened for degradation of poly(ethylene succinate) (PES), poly(epsilon-caprolactone) (PCL) and/or poly(beta-hydroxybutyrate) (PHB) by the clear-zone method. Out of 341 strains of thermophilic actinomycetes, 105 isolates were PHB-degraders (30.8%), 198 isolates were PCL-decomposers (58.1%), and 99 isolates could degrade PES (29.0%). Furthermore, 77 isolates could degrade both PHB and PCL (22.6%), 35 isolates could degrade both PHB and PES (10.3%), 81 isolates could degrade both PES and PCL (23.8%) and 31 isolates could degrade the three polyesters used in this study (9.1%). Base on the morphological and chemical characteristics, these 31 isolates belonging to Actinomadura (12.9%), Microbispora (25.8%), Streptomyces (48.4%), Thermoactinomyces (9.7%) and Saccharomonospora genus (3.22%).
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PMID:Polyester-degrading thermophilic actinomycetes isolated from different environment in Taiwan. 1765 12

The detection of phase separation and identification of miscibility in biopolymer blends is an important aspect for the improvement of their physical properties. In this article, the phase separation in blends of poly(3-hydroxybutyrate) (PHB) with poly(L-lactic acid) (PLA) and poly(epsilon-caprolactone) (PCL), respectively, has been studied as a function of the blend composition by FT-IR imaging spectroscopy. For both polymer blend systems, a miscibility gap has been found around the 50:50% (w/w) composition of the two components. Furthermore, the separating phases have been identified as blends of the two polymer components and their compositions could be determined from calibrations based on the spectra of the blends in the compositional range of miscibility. The data derived from FT-IR spectroscopic imaging were corroborated by additional DSC analyses and mechanical stress-strain measurements of polymer blend films, which exhibited a characteristic fracture behavior as a function of PHB composition.
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PMID:FT-IR imaging spectroscopy of phase separation in blends of poly(3-hydroxybutyrate) with poly(L-lactic acid) and poly(epsilon-caprolactone). 1816 80

Two component biodegradable carriers for biofilm airlift suspension (BAS) reactors were investigated with respect to development of biofilm structure and oxygen transport inside the biofilm. The carriers were composed of PHB (polyhydroxybutyrate), which is easily degradable and PCL (caprolactone), which is less easily degradable by heterotrophic microorganisms. Cryosectioning combined with classical light microscopy and CLSM was used to identify the surface structure of the carrier material over a period of 250 days of biofilm cultivation in an airlift reactor. Pores of 50 to several hundred micrometers depth are formed due to the preferred degradation of PHB. Furthermore, microelectrode studies show the transport mechanism for different types of biofilm structures, which were generated under different substrate conditions. At high loading rates, the growth of a rather loosely structured biofilm with high penetration depths of oxygen was found. Strong changes of substrate concentration during fed-batch mode operation of the reactor enhance the growth of filamentous biofilms on the carriers. Mass transport in the outer regions of such biofilms was mainly driven by advection.
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PMID:Application of two component biodegradable carriers in a particle-fixed biofilm airlift suspension reactor: development and structure of biofilms. 1837 27

Enzymatic modification of a microbial polyester was achieved by the ring-opening polymerization of epsilon-caprolactone (CL) with low-molecular weight telechelic hydroxylated poly[( R)-3-hydroxybutyrate] (PHB-diol) as initiator and Novozym 435 (immobilized Candida antarctica Lipase B) as catalyst in anhydrous 1,4-dioxane or toluene. The ring-opening polymerization was investigated at different conditions with two different types of PHB-diols: PHB-diol(P), containing a primary OH and a secondary OH end groups, and PHB-diol(M), consisting of 91% PHB-diol(P) and 9% PHB-diol containing two secondary OH end groups. The reactions were followed by GPC analyses of the resulting polymers at different time points, and the optimal conditions were established to be 70 degrees C at a weight ratio of CL/enzyme/solvent of 8:1:24. The ring-opening polymerization of CL with PHB-diol(M) (Mn of 2380, NMR) at the molar ratio of 50:1 under the optimal conditions in 1,4-dioxane gave the corresponding poly[HB(56 wt %)-co-CL(44 wt %)] with Mn (NMR) of 3900 in 66% yield. Polymerization of CL and PHB-diol(P) ( Mn of 2010, NMR) at the same condition in toluene gave the corresponding poly[HB(28 wt %)-co-CL(72 wt %)] with Mn (NMR) of 7100 in 86% yield. Both polymers were characterized by (1)H and (13)C NMR and IR analyses as di-block copolyesters containing a PHB block with a secondary OH end group and a poly(epsilon-caprolactone) (PCL) block with a primary OH end group. NMR analyses and control experiments suggested no formation of random copolymers and no change of the PHB block during the reaction. The enzymatic ring-opening polymerization was selectively initiated by the primary OH group of PHB-diol, whereas the secondary OH group remained as an end group in the final polymers. The thermal properties of the di-block poly(HB-co-CL)s were analyzed by DSC, with excellent T g values for the elastomer domain: poly[HB(56 wt %)- co-CL(44 wt %)] with M n (NMR) of 3900 demonstrated a T g of -57 degrees C, Tm of 145, 123, and 53 degrees C; and poly[HB(28wt%)-co-CL(72wt%)] with Mn (NMR) of 7100 gave a Tg of -60 degrees C, Tm of 147 and 50 degrees C. Thus, the selective enzymatic ring-opening polymerization with PHB-diol as macro-initiator provides a new method for the preparation of PHB-based block copolymers as biomaterials with good thermoplastic properties and novel structures containing functional end groups.
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PMID:Enzymatic preparation of novel thermoplastic di-block copolyesters containing poly[(R)-3-hydroxybutyrate] and poly(epsilon-caprolactone) blocks via ring-opening polymerization. 1854 Jun 75

Poly(ester-urethane)s (PUs) were synthesized using hexamethylene diisocyanate (HDI) or toluene diisocyanate (TDI) to join short chains (M(n) = 2000) of poly(R-3-hydroxybutyrate) (PHB) diols and poly(epsilon-caprolactone) (PCL) diols with different feed ratios under different reaction conditions. The multiblock copolymers were characterized by nuclear magnetic resonance spectrometer (NMR), gel permeation chromatography (GPC), differential scanning calorimetry (DSC), thermogravimetric analyses (TGA), X-ray diffraction (XRD), and scanning electron microscope (SEM). XRD spectra and second DSC heat thermograms of the multiblock copolymers revealed that the crystallization of both PHB and PCL segments was mutually restricted, and, especially, the PCL segment limited the cold crystallization of the PHB segment. The SEM of platelet adhesion experiments showed that the hemocompatibility was affected to some extent by the chain flexibility of the polymers. Hydrolysis studies demonstrated that the hydrolytic degradation of PUs was generated from the scission of their ester bonds or/and urethane bonds. Simultaneously, the rate of ester bond scission was determined to some extent by the crystallization degree, which was further affected by the configuration of polymer chains. These highly elastic multiblock copolymers combining hemocompatibility and biodegradability may be developed into blood contact implant materials for biomedical applications.
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PMID:Characterization, biodegradability and blood compatibility of poly[(R)-3-hydroxybutyrate] based poly(ester-urethane)s. 1867 Dec 59

Star block copolymers based on poly[(R)-3-hydroxybutyrate] (PHB) and poly(epsilon-caprolactone) (PCL), termed SPHBCL, were successfully synthesized with structural variation on arm numbers and lengths via coupling reactions and ring opening polymerizations. Arm numbers 3, 4 and 6 of SPHBCL were synthesized by using different multifunctional cores, such as trimethyol propane, pentaerythiritol and dipentaerthritol, respectively. Gel permeation chromatography (GPC) and (1)H and (13)C nuclear magnetic resonance were used to characterize the structure of SPHBCL. GPC failed to produce accurate molecular weights of the SPHBCL due to the discrepancy of star copolymer structures. The melting temperature of SPHBCL decreased with increasing degree of branching. Thermal decomposition temperature was revealed to be lower than that of linear block copolymer LPHBCL counterparts based on PHB and PCL. Films made from various SPHBCL copolymers had different porous or networking surface morphology, and all possessed improved biocompatibility in terms of less blood clotting and more osteoblast cell growth compared with their corresponding homopolymers PHB and PCL. Among them, it was found, however, that the 4-arm star block copolymer 4SPHBCL-25 showed unique surface properties, i.e. a regular nanoravine structure was observed by scanning electron microscopy and atomic force microscopy. This 4-arm star copolymer also showed the best biocompatibility.
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PMID:Synthesis, characterizations and biocompatibility of novel biodegradable star block copolymers based on poly[(R)-3-hydroxybutyrate] and poly(epsilon-caprolactone). 1967 52

In the present study, variable-temperature Fourier transform infrared (FT-IR) spectra of a poly(epsilon-caprolactone) (PCL) homopolymer and a poly(3-hydroxybutyrate) (PHB)-poly(epsilon-caprolactone) (PCL) blend were analyzed by generalized two-dimensional correlation spectroscopy (2DCOS) and perturbation-correlation moving-window two-dimensional (PCMW2D) correlation spectroscopy. The C=O stretching vibration bands of PCL and PHB were employed to explore the structural changes in the PCL homopolymer and the PHB-PCL blend during the heating process. For the melting of PCL homopolymer in the temperature range of 50 to 70 degrees C, we observed in the synchronous and asynchronous 2D correlation spectra one crystalline (1724 cm(-1)) and one amorphous (1737 cm(-1)) C=O stretching vibration band, which are also detectable in the one-dimensional FT-IR spectra. This result was also confirmed by PCMW2D correlation spectroscopy. During the heating process of the PHB-PCL blend in the temperature range 30-200 degrees C, the PCMW2D correlation analysis provided detailed information. Thus, in the synchronous PCMW2D correlation spectrum the melting of PCL was observed in the temperature region between 30 and 70 degrees C. The recrystallization of PHB in the blend in the temperature range 70-120 degrees C was accompanied by a shift of the C=O stretching band from higher wavenumber (1724 cm(-1)) corresponding to an imperfect crystalline state to the lower wavenumber (1721 cm(-1)) characteristic of a well-ordered crystalline state. In the temperature range 120-200 degrees C the melting process of PHB in the blend is captured by a sharp transition from the crystalline (1722 cm(-1)) to the amorphous (1747 cm(-1)) C=O stretching band.
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PMID:Variable-temperature Fourier transform infrared spectroscopic investigations of poly(3-hydroxyalkanoates) and perturbation-correlation moving-window two-dimensional correlation analysis. Part II: Study of poly(epsilon-caprolactone) homopolymer and a poly(3-hydroxybutyrate)-poly(epsilon-caprolactone) blend. 1979 86


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