Supporting Information for "Comprehensive biodegradation analysis of chemically modified poly(3-hydroxybutyrate) materials with different crystal structure"

This work presents a comprehensive analysis of the biodegradation of polyhydroxybutyrate (PHB) and chemically modified PHB with different chemical and crystal structures in a soil environment. A polymer modification reaction was performed during preparation of the chemically modified PHB films, utilizing 2,5-dimethyl-2,5-di(tert-butylperoxy)-hexane as a free-radical initiator and maleic anhydride. Films of neat PHB and chemically modified PHB were prepared by extrusion and thermocompression. The biological agent employed was natural mixed microflora in the form of garden soil. The course and extent of biodegradation of the films was investigated by applying various techniques, as follows: a respirometry test to determine the production of carbon dioxide through microbial degradation; scanning electron microscopy (SEM); optical microscopy; fluorescence microscopy; differential scanning calorimetry (DSC); and X-ray diffraction (XRD). Next-generation sequencing was carried out to study the microbial community involved in biodegradation of the films. Findings from the respirometry test indicated that biodegradation of the extruded and chemically modified PHB followed a multistage (2–3) course, which varied according to the spatial distribution of amorphous and crystalline regions and their spherulitic morphology. SEM and polarized optical microscopy (POM) confirmed that the rate of biodegradation depended on the availability of the amorphous phase in the interspherulitic region and the width of the interlamellar region in the first stage, while dependence on the size of spherulites and thickness of spherulitic lamellae was evident in the second stage. X-ray diffraction revealed that orthorhombic α-form crystals with helical chain conformation degraded concurrently with β-form crystals with planar zigzag conformation. The nucleation of PHB crystals after 90 days of biodegradation was identified by DSC and POM, a phenomenon which impeded biodegradation. Fluorescence microscopy evidenced that the crystal structure of PHB affected the physiological behavior of soil microorganisms in contact with the surfaces of the films.

Supporting Information contains: FTIR spectra for the studied films; Macroscopic photographs of the extruded PHB and chemically modified PHB films before and after specific biodegradation periods in the soil environment; Biodegradation curves (soil burial tests at 55% soil humidity and 25 °C) for PHB in powder, extruded and chemically modified forms; Fluorescence micrographs of PHB-degrading microbial consortia in the process of degrading PHB; Diversity of the bacterial community on the surfaces of the materials at 3 intervals, described by Shannon and Simpson indexes; Scatter plot of principal component analysis, applying Nonmetric Multidimensional Scaling (NMDS) and Bray–Curtis dissimilarity to highlight resemblances between the bacterial communities on the investigated samples

Full publication can be found here: https://zenodo.org/records/10201374