Lignocellulose is a major biopolymer in plant biomass with a complex structure and composition. It consists of a significant amount of high molecular aromatic compounds, particularly vanillin, syringeal, ferulic acid, and muconic acid, that could be converted into intracellular metabolites such as polyhydroxyalkanoates (PHA) and hydroxybutyrate (PHB), a key component of bioplastic production. Several pre-treatment methods were utilized to release monosaccharides, which are the precursors of the relevant pathway. The consolidated bioprocessing of lignocellulose-capable microbes for biomass depolymerization was discussed in this study. Carbon can be stored in a variety of forms, including PHAs, PHBs, wax esters, and triacylglycerides. From a biotechnology standpoint, these compounds are quite adaptable due to their precursors' utilization of hydrogen energy. This study lays the groundwork for the idea of lignocellulose valorization into value-added products through several significant dominant pathways.

• Klíčová slova
• bioplastics production, consolidated bioprocessing, kitchen waste, lignocellulose depolymerization, polyhydroxyalkanoates (PHA), renewable aromatics,
• MeSH
• biomasa MeSH
• biopolymery chemie   metabolismus   MeSH
• lignin * chemie   metabolismus   MeSH
• odpadní produkty MeSH
• polyhydroxyalkanoáty chemie   biosyntéza   metabolismus   MeSH
• potraviny MeSH
• ztráty potravin a plýtvání s nimi MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• biopolymery MeSH
• lignin * MeSH
• lignocellulose MeSH Prohlížeč
• odpadní produkty MeSH
• polyhydroxyalkanoáty MeSH
• ztráty potravin a plýtvání s nimi MeSH

Polyhydroxyalkanoates (PHAs) are intracellular biopolymers that microorganisms use for energy and carbon storage. They are mechanically similar to petrochemical plastics when chemically extracted, but are completely biodegradable. While they have potential as a replacement for petrochemical plastics, their high production cost using traditional carbon sources remains a significant challenge. One potential solution is to modify heterotrophic PHA-producing strains to utilize alternative carbon sources. An alternative approach is to utilize methylotrophic or autotrophic strains. This article provides an overview of bacterial strains employed for PHA production, with a particular focus on those exhibiting the highest PHA content in dry cell mass. The strains are organized according to their carbon source utilization, encompassing autotrophy (utilizing CO2, CO) and methylotrophy (utilizing reduced single-carbon substrates) to heterotrophy (utilizing more traditional and alternative substrates).

• Klíčová slova
• PHA, PHB, autotrophy, heterotrophy, methylotrophy,
• MeSH
• Bacteria * metabolismus   MeSH
• polyhydroxyalkanoáty * biosyntéza   metabolismus   MeSH
• uhlík metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• polyhydroxyalkanoáty * MeSH
• uhlík MeSH

Aneurinibacillus thermoaerophilus CCM 8960 is a thermophilic bacterium isolated from compost in Brno. The bacterium accumulates polyhydroxyalkanoates (PHAs), a biodegradable and renewable alternative to petrochemical polymers. The bacterium reveals several features that make it a very interesting candidate for the industrial production of PHA. At first, due to its thermophilic character, the bacterium can be utilized in agreement with the concept of next-generation industrial biotechnology (NGIB), which relies on extremophiles. Second, the bacterium is capable of producing PHA copolymers containing a very high portion of 4-hydroxybutyrate (4HB). Such materials possess unique properties and can be advantageously used in multiple applications, including but not limited to medicine and healthcare. Therefore, this work focuses on the in-depth characterization of A. thermoaerophilus CCM 8960. In particular, we sequenced and assembled the genome of the bacterium and identified its most important genetic features, such as the presence of plasmids, prophages, CRISPR arrays, antibiotic-resistant genes, and restriction-modification (R-M) systems, which might be crucial for the development of genome editing tools. Furthermore, we focused on genes directly involved in PHA metabolism. We also experimentally studied the kinetics of glycerol and 1,4-butanediol (1,4BD) utilization as well as biomass growth and PHA production during cultivation. Based on these data, we constructed a metabolic model to reveal metabolic fluxes and nodes of glycerol and 1,4BD concerning their incorporation into the poly(3-hydroxybutyrate-co-4-hydroxybutyrate (P(3HB-co-4HB)) structure. KEY POINTS: • Aneurinibacillus sp. H1 was identified as Aneurinibacillus thermoaerophilus. • PHA metabolism pathway with associated genes was presented. • Unique monomer composition of produced PHAs was reported.

• Klíčová slova
• 4-hydroxybutyrate, Aneurinibacillus species H1, De novo assembly, Next-generation industrial biotechnology, PHA, Plasmid pAT1,
• MeSH
• Bacillales MeSH
• butylenglykoly MeSH
• glycerol MeSH
• kyselina 3-hydroxymáselná MeSH
• polyestery metabolismus   MeSH
• polyhydroxyalkanoáty * MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• butylenglykoly MeSH
• glycerol MeSH
• kyselina 3-hydroxymáselná MeSH
• polyestery MeSH
• polyhydroxyalkanoáty * MeSH

A novel model of biodegradable PHA copolymer films preparation was applied to evaluate the biodegradability of various PHA copolymers and to discuss its biomedical applicability. In this study, we illustrate the potential biomaterial degradation rate affectability by manipulation of monomer composition via controlling the biosynthetic strategies. Within the experimental investigation, we have prepared two different copolymers of 3-hydroxybutyrate and 4-hydroxybutyrate-P(3HB-co-36 mol.% 4HB) and P(3HB-co-66 mol.% 4HB), by cultivating the thermophilic bacterial strain Aneurinibacillus sp. H1 and further investigated its degradability in simulated body fluids (SBFs). Both copolymers revealed faster weight reduction in synthetic gastric juice (SGJ) and artificial colonic fluid (ACF) than simple homopolymer P3HB. In addition, degradation mechanisms differed across tested polymers, according to SEM micrographs. While incubated in SGJ, samples were fragmented due to fast hydrolysis sourcing from substantially low pH, which suggest abiotic degradation as the major degradation mechanism. On the contrary, ACF incubation indicated obvious enzymatic hydrolysis. Further, no cytotoxicity of the waste fluids was observed on CaCO-2 cell line. Based on these results in combination with high production flexibility, we suggest P(3HB-co-4HB) copolymers produced by Aneurinibacillus sp. H1 as being very auspicious polymers for intestinal in vivo treatments.

• Klíčová slova
• Aneurinibacillus sp. H1, PHA copolymers, biodegradation, polyhydroxyalkanoates (PHA), simulated body fluids,
• Publikační typ
• časopisecké články MeSH

Production of polyhydroxyalkanoates (PHA), microbial biopolyesters, employing extremophilic microorganisms is a very promising concept relying on robustness of such organisms against microbial contamination, which provides numerous economic and technological benefits. In this work, we took advantage of the natural susceptibility of halophilic and thermophilic PHA producers to hypotonic lysis and we developed a simple and robust approach enabling effective isolation of PHA materials from microbial cells. The method is based on the exposition of microbial cells to hypotonic conditions induced by the diluted solution of sodium dodecyl sulfate (SDS) at elevated temperatures. Such conditions lead to disruption of the cells and release of PHA granules. Moreover, SDS, apart from its cell-disruptive function, also solubilizes hydrophobic components, which would otherwise contaminate PHA materials. The purity of obtained materials, as well as the yields of recovery, reach high values (values of purity higher than 99 wt.%, yields close to 1). Furthermore, we also focused on the removal of SDS from wastewater. The simple, inexpensive, and safe technique is based on the precipitation of SDS in the presence of KCl. The precipitate can be simply removed by decantation or centrifugation. Moreover, there is also the possibility to regenerate the SDS, which would substantially improve the economic feasibility of the process.

• Klíčová slova
• Halomonas halophila, PHA isolation, Schlegelella thermodepolymerans, extremophiles, halophiles, polyhydroxyalkanoate (PHA), sodium dodecyl sulfate (SDS), thermophiles,
• Publikační typ
• časopisecké články MeSH

Polyhydroxyalkanoates (PHA) are microbial polyesters that have recently come to the forefront of interest due to their biodegradability and production from renewable sources. A potential increase in competitiveness of PHA production process comes with a combination of the use of thermophilic bacteria with the mutual use of waste substrates. In this work, the thermophilic bacterium Tepidimonas taiwanensis LMG 22826 was identified as a promising PHA producer. The ability to produce PHA in T. taiwanensis was studied both on genotype and phenotype levels. The gene encoding the Class I PHA synthase, a crucial enzyme in PHA synthesis, was detected both by genome database search and by PCR. The microbial culture of T. taiwanensis was capable of efficient utilization of glucose and fructose. When cultivated on glucose as the only carbon source at 50 °C, the PHA titers reached up to 3.55 g/L, and PHA content in cell dry mass was 65%. The preference of fructose and glucose opens the possibility to employ T. taiwanensis for PHA production on various food wastes rich in these abundant sugars. In this work, PHA production on grape pomace extracts was successfully tested.

• Klíčová slova
• Tepidimonas taiwanensis, grape pomace, polyhydroxyalkanoates, thermophiles,
• Publikační typ
• časopisecké články MeSH

Actinobacteria belonging to the genus Rubrobacter are known for their multi-extremophilic growth conditions-they are highly radiation-resistant, halotolerant, thermotolerant or even thermophilic. This work demonstrates that the members of the genus are capable of accumulating polyhydroxyalkanoates (PHA) since PHA-related genes are widely distributed among Rubrobacter spp. whose complete genome sequences are available in public databases. Interestingly, all Rubrobacter strains possess both class I and class III synthases (PhaC). We have experimentally investigated the PHA accumulation in two thermophilic species, R. xylanophilus and R. spartanus. The PHA content in both strains reached up to 50% of the cell dry mass, both bacteria were able to accumulate PHA consisting of 3-hydroxybutyrate and 3-hydroxyvalerate monomeric units, none other monomers were incorporated into the polymer chain. The capability of PHA accumulation likely contributes to the multi-extremophilic characteristics since it is known that PHA substantially enhances the stress robustness of bacteria. Hence, PHA can be considered as extremolytes enabling adaptation to extreme conditions. Furthermore, due to the high PHA content in biomass, a wide range of utilizable substrates, Gram-stain positivity, and thermophilic features, the Rubrobacter species, in particular Rubrobacter xylanophilus, could be also interesting candidates for industrial production of PHA within the concept of Next-Generation Industrial Biotechnology.

• Klíčová slova
• Rubrobacter spartanus, Rubrobacter xylanophilus, extremophiles, polyhydroxyalkanoates, stress conditions,
• Publikační typ
• časopisecké články MeSH

Extremophilic microorganisms are considered being very promising candidates for biotechnological production of various products including polyhydroxyalkanoates (PHA). The aim of this work was to evaluate the PHA production potential of a novel PHA-producing thermophilic Gram-positive isolate Aneurinibacillus sp. H1. This organism was capable of efficient conversion of glycerol into poly(3-hydroxybutyrate) (P3HB), the homopolyester of 3-hydroxybutyrate (3HB). In flasks experiment, under optimal cultivation temperature of 45 °C, the P3HB content in biomass and P3HB titers reached 55.31% of cell dry mass and 2.03 g/L, respectively. Further, the isolate was capable of biosynthesis of PHA copolymers and terpolymers containing high molar fractions of 3-hydroxyvalerate (3HV) and 4-hydroxybutyrate (4HB). Especially 4HB contents in PHA were very high (up to 91 mol %) when 1,4-butanediol was used as a substrate. Based on these results, it can be stated that Aneurinibacillus sp. H1 is a very promising candidate for production of PHA with tailored material properties.

• Klíčová slova
• Aneurinibacillus sp., P(3HB-co-3HV-co-4HB), P(3HB-co-4HB), polyhydroxyalkanoates, thermophiles,
• Publikační typ
• časopisecké články MeSH