Preparation of Esterified Starches with Different Amylose Content and Their Blending with Polybutylene Succinate.

Three types of starch with different amylose content were esterified and blended with polybutylene succinate (PBS) to obtain esterified manioc starch/PBS (EMS/PBS), esterified corn starch/PBS (ECS/PBS), and esterified waxy corn starch/PBS (EWS/PBS) composites. The EMS/PBS and ECS/PBS composites with high amylose content displayed typical V-type crystal structures. The original crystals of EWS, which had low amylose content, were disrupted during the esterification process. EWS exhibited the strongest interaction with PBS and the most favorable interface compatibility. The pyrolysis temperature was in order of EMS/PBS < ECS/PBS < EWS/PBS. The elongation at break of the three blends was higher than that of pure PBS. The esterification and plasticization of the EWS/PBS composite were the most comprehensive. The EWS/PBS composite showed the lowest storage modulus (G') and complex viscosity (η*). The interfacial bonding force of the composite materials increased with more amylopectin, decreasing intermolecular forces and destroying crystal structures, which decreased G' and η* and increased toughness. The EWS/PBS composite, with the least amylose content, had the best hydrophobicity and degradation performance.

Optimization of medium composition for 3-hydroxycarboxylic acid production by Pseudomonas mendocina-biodegraded polyhydroxybutyrate.

We optimized the culture medium for 3-hydroxycarboxylic acid production by Pseudomonas mendocina DS-04-T-biodegraded polyhydroxybutyrate (PHB) using the Plackett-Burman design, steepest ascent method, and Box-Behnken design. The optimized concentrations of the constituents of the culture medium were as follows: PHB (7.57 g/L), NH4 Cl (5.0 g/L), KH2 PO4 (2.64 g/L), Na2 HPO4 ·12H2 O (12 g/L), MgSO4 ·7H2 O (0.5 g/L), and CaCl2 ·2H2 O (5 mg/L). The yield of 3-hydroxycarboxylic acid obtained using the optimized culture medium was 56.8 ± 1.64%, which was 2.5-fold higher than that obtained when the unoptimized culture medium was used.

A polylactic acid degrading lipase from Bacillus safensis: Characterization and structural analysis.

A polylactic acid degrading triacylglycerol lipase (TGL) was identified from Bacillus safensis based on genome annotation and validated by real-time quantitative PCR. TGL displayed optimal activity at pH 9.0 and 55 °C. It maintained stability at pH 9.0 and temperatures 45 °C. The activity of TGL was found to benefit from the presence of potassium sodium ions, and low concentrations of Triton X-100. The TGL could erode the surface of polylactic acid films and increase its hydrophilicity. The hydrolysis products of polylactic acid by TGL were lactate monomer and dimer. TGL contains a classical catalytic triad structure of lipase (Ser77, Asp133, and His156) and an Ala-X-Ser-X-Gly sequence. Compared with some lipases produced by the same genus Bacillus, TGL is highly conserved in its amino acid sequence, mainly reflected in the amino acid residues that exercise the enzyme activity, including the catalytic activity center and the substrate binding sites.

Biodegradation of polylactic acid by a mesophilic bacteria Bacillus safensis.

A mesophilic bacterial strain, Bacillus safensis PLA1006, was isolated from landfill soil and was tested for growth on polylactic acid (PLA) emulsion medium. The strain formed clear zones on the medium and produced protease and lipase. The macroscopic morphology of the PLA films was not changed significantly after treatment with Bacillus safensis PLA1006 but the films were whitened. Weight loss of PLA films was about 8% after 30 days of incubation with Bacillus safensis PLA1006 in mineral salt medium. Scanning electron microscopy revealed etching on the surface of PLA film treated by Bacillus safensis PLA1006. This also caused an increase in hydrophilicity of the PLA films surface. Attenuated total reflectance - Fourier transform infrared spectroscopy analysis of PLA films after treated by Bacillus safensis PLA1006 showed no new absorption peaks but a decrease in the intensity of all absorption peaks. The hydrolysis products of PLA by the strain contained monomers and oligomers of lactic acid. Zymogram detection showed that proteases may play a role in the degradation of PLA.

Purification and characterization of an extracellular poly(3-hydroxybutyrate-co-3-hydroxyvalerate) depolymerase from Acidovorax sp. HB01.

The poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV)-degrading strain Acidovorax sp. HB01 was isolated from an activated sludge sample. A novel PHBV depolymerase with a molecular weight of 43.4 kDa was purified to homogeneity from the culture supernatant of the HB01 strain. The optimum pH and temperature of the PHBV depolymerase were 7.0 and 50 °C, respectively. The PHBV depolymerase can also degrade polyhydroxybutyrate, poly (3-hydroxybutyrate-co-4-hydroxybutyrate), and poly(caprolactone); however, the PHBV degradation activity of the depolymerase is higher than its activity against the other polymers. Effect of metal ions and various inhibitors on the PHBV depolymerase activity was examined. The addition of Na(+), K(+), and Ca(2+) markedly increased the hydrolysis rate, whereas the enzyme activity was inhibited by Zn(2+), Mg(2+), Mn(2+), and particularly by Cu(2+) and Fe(2+). Ethylenediaminetetraacetic acid was found to have a significant inhibitory effect. The main degradation product of depolymerase was identified as the 3-hydroxybutyric acid monomer and 3-hydroxyvaleric acid monomers via mass spectrometry.

Biodegradation of polybutylene adipate-co-terephthalate by Priestia megaterium, Pseudomonas mendocina, and Pseudomonas pseudoalcaligenes following incubation in the soil.

Soil that contained polybutylene adipate-co-terephthalate (PBAT) was incubated with Priestia megaterium, Pseudomonas mendocina, and Pseudomonas pseudoalcaligenes to improve the biodegradative process of this polymer. The mixture of Pr. megaterium and Ps. mendocina was highly effective at biodegrading the PBAT, and after eight weeks of soil incubation, approximately 84% of the PBAT film weight was lost. Mixtures of the other two species also positively affected the synergistic degradation of PBAT film in the soil, but the mixture of three species had a negative effect. The residual PBAT film microstructure clearly demonstrated the degradation of PBAT, and the degree of degradation was related to the different species. Cleavage of the PBAT film ester bond after soil microbial action affected its properties. The incubation of PBAT in soil that contained these species affected soil dehydrogenase and soil lipase in particular. The secretion of lipase by these species could play an important role in the degradation of PBAT in the soil.

Preparation of porous materials by selective enzymatic degradation: effect of in vitro degradation and in vivo compatibility.

Poly(butylene succinate) (PBS) and poly(lactic acid) (PLA) were melt-blended and formed into a film by hot press forming. The film was selectively degraded by cutinase and proteinase K to form a porous material. The porous materials were characterized with respect to their pore morphology, pore size, porosity and hydrophilicity. The porous materials were investigated in vitro degradation and in vivo compatibility. The results show that the pore size of the prepared porous materials could be controlled by the proportion of PBS and the degradation time. When the PBS composition of PBS/PLA blends was changed from 40 wt% to 50 wt%, the mean pore diameter of the porous materials significantly increased from 6.91 µm to 120 µm, the porosity improved from 81.52% to 96.90%, and the contact angle decreased from 81.08° to 46.56°. In vitro degradation suggests that the PBS-based porous materials have a good corrosion resistance but the PLA-based porous materials have degradability in simulated body fluid. Subcutaneous implantation of the porous materials did not cause intense inflammatory response, which revealed good compatibility. The results of hematoxylin and eosin and Masson's trichrome staining assays demonstrated that the porous materials promote chondrocyte production. Porous materials have great potential in preparing implants for tissue engineering applications.

Enzymatic hydrolysis of polyester: Degradation of poly(ε-caprolactone) by Candida antarctica lipase and Fusarium solani cutinase.

Poly(ε-caprolactone) (PCL) particles were melt-pressed into films using a hot press and then subjected to degradation by lipase from Candida antarctica and cutinase from Fusarium solani, respectively. The differences in weight loss, degradation modes, thermal stability, and crystallization were investigated after degradation by two kinds of enzymes. The result showed that mass loss of PCL films degraded by lipase was higher than that degraded by cutinase at the same enzyme concentrations. The degradation mode of PCL films is layered for cutinase degradation and penetrated for lipase degradation. Crystallinity of PCL had no obvious decrease after degradation by cutinase, but it markedly decreased after lipase-degradation. PCL films occurred one-step decomposition during heating and the cutinase-degraded products had similar thermal stability. Whereas the thermal stability of lipase-degraded PCL decreased significantly and the weight loss of the PCL occurred in several steps with increasing lipase hydrolysis time.

Selective enzymatic degradation and porous morphology of poly(butylene succinate)/poly(lactic acid) blends.

Poly(butylene succinate) (PBS) and poly(lactic acid) (PLA) were melt-blended in different proportions and selectively degraded by cutinase and proteinase K, respectively. The selective enzymatic degradation process was systematically investigated. The degraded PBS/PLA blends were analyzed via scanning electron microscopy, Fourier-transform infrared spectroscopy, powder X-ray diffraction, and differential scanning calorimetry. The results of the weight loss of PBS/PLA blends degraded by cutinase and proteinase K suggested that PLA hindered the cutinase-catalyzed degradation of PBS, whereas the addition of PBS in the blends accelerated the degradation of PLA within a specific PBS/PLA ratio. The change in crystallinity after degradation was closely related to the different way of degradation. The characterization of PBS/PLA blends after degradation showed that selective enzymatic degradation could not completely degrade PBS or PLA component. After degradation, the pores formed by proteinase K were more uniform and larger than those formed by cutinase. This work provides a new insight into the selective enzymatic degradation processes of the porous materials, which will be used in tissue engineering or oil-water separation in the future.

Enzymatic degradation of poly(butylene succinate) with different molecular weights by cutinase.

Poly(butylene succinate) (PBS) films with different molecular weights were enzymatically degraded by cutinase. Changes in the properties of the films before and after enzymatic degradation were studied through scanning electron microscopy, differential scanning calorimetry, thermogravimetry, X-ray powder diffraction, proton nuclear magnetic resonance, and gel-permeation chromatography analysis. The weight loss of the films initially decreased and then increased with increasing molecular weight. Crystallinity was inversely proportional to weight loss and tended to decrease with prolonged degradation time. Crystalline and amorphous regions were simultaneously degraded. The thermal stability of PBS films decreased after enzymatic degradation. PBS was the main component of the enzymatically degraded polymers. The molecular weights of the films did not considerably change before and after degradation by cutinase.

Preparation, characterization, and biodegradation of poly(butylene succinate)/cellulose triacetate blends.

Poly(butylene succinate) (PBS) and cellulose triacetate (CT) were blended using chloroform as solvent. The solid-state properties of PBS/CT blends were confirmed by Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), X-ray diffraction (XRD), thermogravimetric analysis, scanning electron microscopy (SEM), and water contact angle measurements. FTIR results show that PBS and CT were physically blended. Tensile strength was not distinguished when the weight percent of CT was <15%, and Young's modulus increased gradually with increasing CT. DSC and XRD results show that the crystals were homogeneous, and crystallinity had no apparent decrease when <10% CT was added to the PBS matrix. However, the addition of more CT components could destroy the crystal behavior of PBS. SEM showed that no phase separation occurred between the two materials. The addition of CT increased the hydrophilicity of PBS/CT1-15 blends. The weight loss was nearly 90% after 16h of degradation for PBS/CT10. The appropriate proportion of PBS to CT was 90:10.

Quantitative detection of Streptococcus mutans and bacteria of dental caries and no caries groups in permanent teeth from a north China population.

BACKGROUND: Streptococcus mutans (S. mutans) is the prime pathogen of dental caries. There are few reports that studied the relationship between S. mutans, bacteria and dental caries in permanent teeth when compared to those in primary teeth. This study aimed to detect S. mutans and bacteria of dental caries and non-caries groups in permanent teeth from a north China population by real-time polymerase chain reaction (PCR) and compare the relationship between the number of these bacteria and the prevalence of dental caries in permanent teeth. METHODS: Human saliva samples were collected from 142 subjects with permanent teeth. According to their dental tooth (DT), 142 subjects were divided into a dental caries group (DT ≥ 1) and a non-caries group (DT = 0). With specific primers for S. mutans and 16S rRNA, the total number of S. mutans and total bacteria of 142 saliva samples were detected by real-time PCR and statistically analyzed. RESULTS: There was no significant difference between the detection rates of S. mutans (P = 0.118) and medians of S. mutans (P = 0.115). The ratio of S. mutans to total bacteria in people with dental caries was significantly higher than in those without caries (P < 0.001), but the total number of bacteria in people with dental caries was significantly lower than in those without caries (P < 0.001). CONCLUSIONS: S. mutans had different effects on caries in the permanent teeth of several individuals from a north China population. The ratios of S. mutans to total bacteria in saliva detected by real-time PCR with Sm479F/R and 16S RNA primers were closely associated with the prevalence of dental caries in the same population. These assays may be useful for the assessment of an individual's risk of dental caries.

[Quantitative detection of Streptococcus mutans in different people with dental caries by real-time polymerase chain reaction].

OBJECTIVE: To establish a quantity detection method of Streptococcus mutans (Sm) and bacteria and compare the relationship between the number of these bacteria and the prevalence of dental caries in different people. METHODS: With specific primers for a unique sequence in a 14 kb HaeIII restriction fragment consistently presenting during detecting Sm by chromosomal DNA fingerprints, the total number of Sm and bacteria of 99 saliva samples were detected by real-time polymerase chain reaction (PCR) and statistically analyzed. RESULTS: The primers were specific for Sm and the minimum detectable level by real-time PCR was 0.1 microg/L. The total number of bacteria in the dental caries and people without caries was 51.4 x 10(8) cell copies/L and 221.6 x 10(8) cell copies/L respectively, in which the ratio of Sm to bacteria was 0.0193 and 0.0059 respectively. The differences were significantly different between the people with dental caries and those without caries in the total number of bacteria and the ratio of Sm to bacteria. CONCLUSIONS: The primers can be used to detect the Sm by real-time PCR. The ratio of Sm to bacteria was closely associated with the prevalence of dental caries.