The Role Played by Acetyl Group Distribution Patterns in Substituted Starch Toward Enzymatic De-acetylation and Chain Fragmentation

The nature and distribution of the acetylated groups were evaluated by ¹³C-NMR and ¹H-NMR. The starch substrate with a DS of 1.5 comprises only two patterns: -(14)-d-glucopyranose and 2,3,6-tri-O-acetyl--(14)-d-glucopyranose. The starch with a DS of 3.0 also comprises two patterns: 2,3,4,6-tetra-O-acetyl--(14)-d-glucopyranose and 2,3,6-tri-O-acetyl--(14)-d-glucopyranose; whereas starch (DS = 1.9) contains 4 patterns: 2,3,6-tri-O-acetyl--(14)-d-glucopyranose, 2,3,4,6-tetra-O-acetyl--(14)-d-glucopyranose terminal, 2,6-di-O-acetyl--(14)-d-glucopyranose, and 3,6-di-O-acetyl--(14)-d-glucopyranose. Using esterase from Viscozyme, it has been possible to hydrolyze up to 7% of the DS 3.0 starch. An -amylase (Fungamyl 800) was then added to these acetylesterases. With a 2.4 FAU/mL fraction of -amylase and 2.4 U/mL from the Viscozyme's acetylesterase, 28% of the acetylated end groups were hydrolyzed for the starch substrates with DS 3.0. Moreover, a synergic action between -amylase and acetylesterase was noticed, allowing fragmentation of 32% for DS 1.5, 30% for DS 1.9, and 11% for DS 3.0.     

Physical properties and biodegradability of blends containing poly(ε-caprolactone) and tropical starches

In order to assess feasibility of tropical starches (sago and cassava starches) as biodegradable plastic materials, blending with poly(-caprolactone) (PCL), a biodegradable polymer, was carried out. It was confirmed that the physical properties (tensile strength and elongation) of PCL/sago and PCL/cassava blends were similar to those of PCL/corn blend, suggesting that sago and cassava starches can also be blended with PCL for production of biodegradable plastic. However, the properties of all PCL/starch blends were still low compared with those of polyethylene. Enzymatic degradability evaluation showed that lipase degradation of PCL and-amylase degradation of starch increased as the starch content in the blend increased. Burial test of the blends for 1, 3, and 5 months was carried out and the rate of degradation of the PCL/sago blend was confirmed to be slower than those of PCL/corn and PCL/cassava blends. Observation of the film blends structure by scanning electron microscope revealed that the starch was dispersed in a PCL continuous phase. Furthermore, changes in the film surface before and after enyzme treatments were observed.     

Solid State Reactions of Potato Starch with Urea and Biuret

Reaction of granular potato starch with urea and biuret resulted in the formation of products, which were soluble neither in cold nor boiling water. The net reaction was a monosubstitution of the hydrogen atom in one hydroxyl group in each D-glucose unit of starch with the either CO–NH₂ or CO–NH–CO–NH₂ moiety, respectively. Properties of the products, particularly these with urea, depended on the mode of reaction. Reactions were carried out in the microwave oven as well as with convection heating. The products retained the granular form of starch but a vast majority of granules were damaged. -Amylolysis of those materials revealed that their susceptibility to the enzyme increasing in the order: starch-amylolysis with simultaneous insolubility in water make these products suitable as ruminant fodder and, eventually, biodegradable material.     

Effect of Starch Granule Size on Viscosity of Starch-Filled Poly(hydroxy ester ether) Composites

The effect of starch granule size on the viscosity of starch-filled poly(hydroxy ester ether) (PHEE) composites was characterized using size-fractionated potato starch, as well as unfractionated starches (rice, corn, wheat, and potato). Potato starch was separated using an air classifier into four particle size fractions: <18 m, 18-24 m, 24-30 m, and >30 m. The starch was dried to a moisture content of 0.5% to minimize moisture effects on composite rheology. PHEE and potato starch were extruded with starch volume fractions of 0.46 and 0.66. Stress relaxation, frequency and strain sweep, and temperature-dependence measurements were carried out. Although small variations in viscosity were seen with the different potato starch fractions, differences were not significant at a volume fraction of 0.46. Viscosity differences between the different particle size fractions were more pronounced at a volume fraction of 0.66. The temperature dependence could be described by an Arrhenius relation, with an apparent activation energy of 84 kJ/mole. At a volume fraction of 0.46, the starch/PHEE viscosities increased in the order potato starch < wheat starch corn starch < rice starch.     

Preparation and Biodegradation of Starch/Polycaprolactone Films

Starch granules were modified with trisodium trimetaphosphate (TSTP) and characterized by P³¹-NMR, FTIR and DSC. Seventy-micron films were prepared from modified starch and polycaprolactone blends by solvent casting technique. Three different types of films—PCL (100% polycaprolactone), MOD-ST/PCL (50% modified starch and 50% polycaprolactone blend) and NONMOD-ST/PCL (50% nonmodified starch and 50% polycaprolactone blends)—were prepared, and their thermal, mechanical, and morphologic properties were investigated to show the increased performance of PCL with the addition of starch and also the effect of modification. It was observed that with the addition of starch the Young's modulus of polycaprolactone was increased and became less ductile, whereas tensile strength and elongation at break values decreased. Biodegradation of these films was inspected under different aerobic environments with the presence of Pseudomonas putida, activated sludge, and compost. It was observed that whereas P. putida had almost no effect on degradation during 90 days, with the presence of activated sludge, considerable deformation of films was observed even in the first 7 days of degradation. In a compost environment, degradation was even faster, and all polymer films were broken into pieces within first 7 days of degradation and no film remained after 15 days.     

Microwave-Assisted Solvent-Free or Aqueous-Based Synthesis of Biodegradable Polymers

Microwave radiation was used as the energy source for various types of chemical derivatizations of polysaccharides and for the synthesis of biodegradable polyesters in solvent-free or aqueous-based reaction systems. A medium to high degree of substitution was obtained for starch acetates, starch succinates, carboxymethyl konjac, aminated starch, and aminated chitosan. Ring-opening polymerization of lactide and -caprolactone proceeded rapidly even at low power output in the presence of tin octanoate catalyst. Complete monomer conversion and high molecular weight were achieved in less than 6 minutes under nonisothermal conditions. The yield rapidly increased with increasing power output and showed no significant change in a wide range of batch sizes. Polycaprolactone was successfully grafted from starch and konjac acetate in 3 minutes, yielding as high as 24% grafting efficiency and 25% grafting degree.     

Picocyanobacteria – Sensitive Bioindicators of Contaminant Stress in an Alpine Lake (Traunsee Austria)

The abundance and photosynthetic activity ofpicocyanobacteria in the oligotrophic alpine lake Traunseewere measured at a station located close to the outlet ofindustrial soda waste and at a mid-lake reference stationduring spring, 1999 through to autumn, 2000.Picocyanobacterial numbers measured by flow cytometry inTraunsee (0.7–13.2 × 10⁴ ml^-1) were comparable tothose of other oligotrophic lakes, and there was nosignificant difference between the contaminated and thereference sampling location. Picoplankton (<2 m)photosynthetic rates measured in vitro by the ¹⁴C-technique were significantly reduced at the contaminated siterelative to the reference station at low photosyntheticallyavailable radiation (10 E m^-2 s^-1), while nodifference between these two stations was found at moderatelyhigh light intensity (100 E m^-2 s^-1). Theinvestigation was complemented by laboratory experiments withcultured picocyanobacteria. Three Synechococcus spp.strains were exposed to water taken from either of the twoTraunsee stations and from a control station located inneighbouring Attersee. Cell-specific photosynthetic activitymeasured by 4-h in vitro incubations revealed no significantdifference among the three stations investigated. Growthrates of the same three Synechococcus spp. strains weremeasured by flow cytometry over several days in thelaboratory. One strain, in particular, was sensitive to watertaken from the contaminated site; growth rate of this strainwas significantly reduced, relative to when exposed to watertaken from the reference station. Taken together, our resultsdemonstrate that picocyanobacteria are highly sensitivebioindicators of contaminant stress. The overall impact ofthe emissions from the industrial outlet on thepicocyanobacteria was, however, relatively minor.     

Measurement of the biodegradation of starch-based materials by enzymatic methods and composting

The aim of this study was to evaluate the suitability of in vitro enzymatic methods for assaying the biodegradability of new starch-based biopolymers. The materials studied included commercial starch-based materials and thermoplastic starch films prepared by extrusion from glycerol and native potato starch, native barley starch, or crosslinked amylomaize starch. Enzymatic hydrolysis was performed using excessBacillus licheniformis -amylase andAspergillus niger glucoamylase at 37°C and 80°C. The degree of degradation was determined by measuring the dissolved carbohydrates and the weight loss of the samples. Biodegradation was also determined by incubating the samples in a compost environment and measuring the weight loss after composting. The results indicated that the enzymatic method is a rapid means of obtaining preliminary information about the biodegradability of starch-based materials. Other methods are needed to investigate more accurately the extent of biodegradability, especially in the case of complex materials in which starch is blended with other polymers.     

Electrical resistance characteristics of starch foams

The insulative character of expanded polystyrene loose-fill packing material supports the immobile triboelectric charge on its surface, causing static cling. One beneficial property of starch-based loose-fill is its antistatic behavior, which prevents the buildup of electrostatic charges on the foam surface, resulting in no static cling. This investigation explores the electrical resistance characteristics of plasticized starch materials such as commercial loose-fill. Electrical resistance standards used in this study to measure surface resistance and static decay properties are ASTM D 257-78, EOS/ESD S-11, and EIA 541. Following these established testing protocols, the electrical resistance of starch-based and expanded polystyrene loose-fill is quantified. Surface resistivity, measured at 12% RH, of starch-based loose-fill products is less than 1.0×10¹² per square characteristic of inherently static dissipative materials.Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by the USDA implies no approval of the product to the exclusion of others that may be suitable.Presented at the 1995 International Chemical Congress of Pacific Basin Societies, Symposium onEnvironmental Polymer Biodegradation, December 17–22, 1995, Honolulu, Hawaii, USA.     

Characterization and enzymatic degradation of a cross-linked bacterial polyester

The bacterial polyester, poly(-hydroxybutyrate-co--hydroxyvalerate) (PHB/V), was cross-linked with 1, 5, 7, 10, 20, and 30 wt% benzoyl peroxide by thermal decomposition reactions. Solvent extractions were carried out to determine the cross-linked fractions of the films. The sol/gel data were used to estimate cross-link densities. Films of PHB/V cross-linked with 10% benzoyl peroxide were placed in contact with purified depolymerase A secreted byP. lemoignei. These samples exhibited weight loss rates which were half that of un-cross-linked PHB/V, but the network was degraded completely by the enzyme. The results of this study suggest that anendo-type enzymatic degradation may occur, in addition to theexo-type activity, which is normally presumed to occur with theP. lemoignei depolymerase system.     

Enzymatic degradation of poly([R,S] β-hydroxybutyrate)

The synthetic analogue of a bacterially produced polyester, poly(-hydroxybutyrate) (PHB) was synthesized from racemic -butyrolactone using anin situ trimethyl aluminum-water catalyst. The polymer was fractionated into samples differing in molecular weight and isotactic diad content. The latter was closely related to degree of crystallinity. The biodegradation of these fractions were examined by monitoring mass loss over time in the presence of anAlcaligenes faecalis T1 extracellular bacterial poly(-hydroxybutyrate) depolymerase. The fraction with high isotactic diad tacticity content showed little or no degradation over a 50 hour incubation period, whereas the fraction of intermediate isotactic diad content degraded in a continuous steady fashion at a rate that was less than that for bacterial PHB. The low isotactic diad fraction underwent a rapid initial degradation, followed by no further mass loss. The presence of stereoblocks in the polymer structure of the various fractions was an influence on the degree of susceptibility towards degradation and is related to sample crystallinity.     

Air Pollution Effects on Forest Trees in Balikesir,Turkey

Air pollution has become an actual problem in Balikesir because of rapid urbanisation and increasing of the polluting sources. Air pollutionproblems occur under the unfavorable meteorological conditions which increase in frequency of occurrence in winter due to use of low quality lignite coals for heating. The `Protection of Air Quality' in Turkey dated 1986 is based on the human health criteria. But the critical limit values are lethal for the forest trees as far as SO₂ is concerned. The field observations showed that leaves of the trees have yellow spots indicating acid burns in Balikesir. This study aims at determining the relationships between the SO₂ concentrationsin the air under selected climatic conditions, and the effects of air pollution on forest trees in Balikesir. Samples of leaves were collected from the City Parks in Balikesir, Deirmenboaz forested area located 10 km far fromBalikesiron the Balikesir-Bursa highway, and from the forestation areas near the Balikesir-Edremit highway. Sulfur contents of the leaves were very high especially in the city (2650–5300 g m^-3).Samples from other areas had values lower than this range but above the usual values (850–3612 g m^-3). The diameter increments of the trees were found to be lower and these results clearly showed the adverse impacts. Negative correlations were found between these sulfur concentrations and the daily mean temperatures and wind speeds. Positive correlation was seen between these concentrations and pressures, humidities. Emission sources must be controlled in order to safeguard the forests which protects the soil, facilitate water production, assimilate carbon dioxide in air and produce oxygen. It must be considered that the problems are not only regional but have a global identity.     

Synthesis and biodegradation of poly(γ-butyrolactone-co-l-lactide)

Biodegradable polyesters were synthesized by ring-opening copolymerization of -butyrolactone (BL) and its derivatives withl-lactide (LLA). Although tetraphenyl tin was the main catalyst used, other organometallic catalysts were used as well.¹H and¹³C NMR spectra showed that poly(BL-co-LLA)s were statistical and that their number-average molecular weights were as high as 7×10⁴. The maximum BL content obtained from copolymerization BL/LLA was around 17%. TheT _m andT _g values of the copolymers showed a gradual depression with an increase in BL content. NoT _m was obtained for the copolymers containing more than 13 mol% BL. The biodegradability of the copolyesters was evaluated by enzymatic hydrolysis and nonenzymatic hydrolysis tests. The enzymatic hydrolysis was carried out at 37°C for 24 h using lipases fromRhizopus arrhizus andR. delemar. Hydrolyses by both lipases showed that an increase in BL content of the copolymer resulted in enhanced biodegradability. Nonenzymatic accelerated hydrolysis of copolymers at 70°C was found to increase proportionally to their exposure time. The hydrolysis rate of these copolymers was considerably faster than that of PLLA. The higher hydrolyzability was recorded for the BL-rich copolymers. The copolymerization of -methyl--butyrolactone (MBL) or -ethyl--butyrolactone (EBL) with LLA resulted in relatively LA-rich copolymers.     

Liquid crystallinity of levan/water/starch solutions

Transmitted light measurements performed with a UV-visible spectrophotometer were used to characterize how starch affects the position of boundaries on the phase diagram for dilute aqueous solutions of levan (a branched polymer of fructose). Data were collected in the range 15 to 70°C; the minimum concentrations required for separation of a nematic phase and the minimum concentration required for a fully nematic solution were identified within this range. While hard interactions (repulsion between rod-like molecular segments) dictate the formation of a liquid crystalline phase at and above ambient temperature in the absence of starch, soft interactions become more significant as solutions are cooled toward ambient when starch is present. Small amounts of starch might be used as a filler to modify the mechanical properties (while retaining the process-related benefits) of levan films cast from liquid crystalline solution.     

Degradation of poly(β-hydroxyalkanoates) and polyolefin blends in a municipal wastewater treatment facility

Six types of plastics and plastic blends, the latter composed at least partially of biodegradable material, were exposed to aerobically treated wastewater (activated sludge) to ascertain their biodegradability. In one study, duplicate samples of 6% starch in polypropylene, 12% starch in linear low-density polyethylene, 30% polycaprolactone in linear low-density polyethylene, and poly(-hydroxybutyrate-co-hydroxyvalerate) (PHB/V), a microbially produced polyester, were exposed to activated sludge for 5 months, and changes in mass, molecular weight average, and tensile properties were measured. None of the blended material showed any sign of degradation. PHB/V, however, showed a considerable loss of mass and a significant loss of tensile strength. In a second study, PHB/V degraded rapidly, but another type of microbial polymer which forms a thermoplastic elastomer, poly(-hydroxyoctanoate), did not degrade. These results illustrate the potential for disposal and degradation of PHB/V in municipal wastewater.     

Humidity- and Age-Tolerant Starch-Based Sponge for Loose-Fill Packaging

Loose-fill packaging sponges were extruded from mixtures of 54–62% hydroxypropylated (HP 5%) amylomaize V (50% amylose) and wheat starches, 17–24% synthetic polymer, 13% water, 7% blowing agent(s), and 0.5% nucleating agent. One product made from 28% HP wheat starch, 28% HP amylomaize V starch, 12% ethylene vinyl alcohol (EVOH) copolymer, 8% polystyrene (PS), and 3% polystyrene maleic anhydride (PSMA) copolymer, plus the other raw materials, had a compressibility and resilience that matched those of expanded polystyrene (EPS), although its bulk density was four times higher. The starchy sponge showed 16% shrinkage in volume at 90% relative humidity and was 2% soluble in excess water, both at 25°C. After aging for 18 months near 25°C, the HP starchy sponge gave only a trace of fines in a simulated shipping test, compared to 20% fines from a biodegradable, starch-based, loose-fill sponge of commerce.     

Synthesis of copolyesters containing poly(ethylene terephthalate) and poly(ε-caprolactone) units and their susceptibility toPseudomonas sp. lipase

Copolyesters containing poly(ethylene terephthalate) (PET) and poly(-caprolactone) (PCL) were synthesized from PET and PCL homopolymers by transesterification reaction at 270°C in the presence of catalyst. The copolyesters were characterized by¹³C-NMR and differential scanning calorimetry (DSC). The degradation behavior of PCL byPseudomonas sp. lipase in buffer solution (pH 7) and tetrahydrofuran (THF) was investigated by gel permeation chromatography (GPC) and¹H-NMR. From these experiments, it was found thatPseudomonas sp. lipase acted endoenzymatically on PCL. Using this lipase, degradation tests for PET/PCL copolyesters whose PCL content was below 50% by weight were also performed in buffer solution (pH 7). However, evenPseudomonas sp. lipase with high degradation activity on PCL did not easily degrade the PCL unit in PET/PCL copolyesters.     

Purification of Aspergillus fumigatus (Pdf1) Poly(β-hydroxybutyrate) (PHB) Depolymerase Using a New, Single-Step Substrate Affinity Chromatography Method: Characterization of the PHB Depolymerase Exhibiting Novel Self-Aggregation Behavior

The extracellular poly(-hydroxybutyrate) (PHB) depolymerase of Aspergillus fumigatus Pdf1 was purified by a new, simple, one-step affinity chromatography method using the substrate PHB. The purified enzyme was glycosylated, with the molecular mass of 40 KD, and exhibited a novel self-aggregation behavior by means of hydrophobic interaction that was resolved by Triton X-100 (TX-100) pretreatment of enzyme and also TX-100 incorporation in the native gel. The apparent K _m value of purified enzyme for PHB was 119 g/mL and 3-hydroxybutyrate was detected as the main endproduct of PHB hydrolysis. The depolymerase was insensitive to phenylmethyl sulfonyl fluoride (PMSF), sodium azide, ethylenediaminetetraacetic acid (EDTA), and para-chloromercuric benzoic acid (PCMB), but was inactivated by dithioerythritol (DTT) and showed specificity for short chain-length poly(-hydroxyalkanoates) (PHAs) such as PHB, poly(hydroxyvalerate) (PHV), and copolymers of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). Medium-chain-length PHA failed to get hydrolyzed. The enzyme, however, exhibited strong cross reactivity with the Comamonas sp. PHB depolymerase antibodies, but not with PHV depolymerase antibodies of Pseudomonas lemoignei. Southern hybridization and dot blot analysis of A. fumigatus Pdf1 genomic DNA with alkaline phosphatase labeled probes of P. lemoignei PHB and PHV depolymerase genes revealed no homology, although the enzyme hydrolyzed both PHB and PHV.     

Molecular Weight Changes and Polymeric Matrix Changes Correlated with the Formation of Degradation Products in Biodegraded Polyethylene

The molecular weight changes in abiotically and biotically degraded LDPE and LDPE modified with starch and/or prooxidant were compared with the formation of degradation products. The samples were thermooxidized for 6 days at 100°C to initiate degradation and then either inoculated with Arthobacter paraffineus or kept sterile. After 3.5 years homologous series of mono- and dicarboxylic acids and ketoacids were identified by GC-MS in abiotic samples, while complete disappearance of these acids was observed in biotic environments. The molecular weights of the biotically aged samples were slightly higher than the molecular weights of the corresponding abiotically aged samples, which is exemplified by the increase in from 5200 g/mol for a sterile sample with the highest amount of prooxidant to 6000 g/mol for the corresponding biodegraded sample. The higher molecular weight in the biotic environment is explained by the assimilation of carboxylic acids and low molecular weight polyethylene chains by microorganisms. Assimilation of the low molecular weight products is further confirmed by the absence of carboxylic acids in the biotic samples. Fewer carbonyls and more double bonds were seen by FTIR in the biodegraded samples, which is in agreement with the biodegradation mechanism of polyethylene.     

Chromatographic fingerprinting as a means to predict degradation mechanisms

The degradation products of polymers are identifiable by chromatography. The degradation product patterns (or fingerprints) formed depend on the type of polymer, the degradation mechanism(s), and also the type of additive present in the material. The chromatographic fingerprint of biotically aged degradable low-density polyethylene (i.e., LDPE+starch+prooxidant) shows, in particular, the absence of low molecular weight carboxylic acids, which suggests an assimilation of these carboxylic acids by the microorganisms. The degradation products of natural polymers are usually intermediates that are used again in the anabolic cycles. It is possible to transfer the terminology from the natural polymers, where the catabolism of natural polymers consists of three stages, and apply this also to the degradable synthetic polymers. During stage I the natural polymers degrade to their major building blocks (e.g., amino acids, glycerol, hexoses, pentoses, etc.), during stage II these products are collected and converted to a smaller number of even simpler molecules [e.g., acetyl-coenzyme A (CoA)]. In stage III, finally, the acetyl-CoA enters the citric acid cycle, where energy is gained in parallel with the release of CO₂ and H₂O.Presented at the international workshop,Polymers from Renewable Resources and their Degradation, Stockholm, Sweden, November 10–11, 1994.