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1.
Methods Enzymol ; 648: 391-421, 2021.
Article in English | MEDLINE | ID: mdl-33579413

ABSTRACT

The enzymatic degradation of polyethylene terephthalate (PET) results in a hydrolysate consisting almost exclusively of its two monomers, ethylene glycol and terephthalate. To biologically valorize the PET hydrolysate, microbial upcycling into high-value products is proposed. Fatty acid derivatives hydroxyalkanoyloxy alkanoates (HAAs) represent such valuable target molecules. HAAs exhibit surface-active properties and can be exploited in the catalytical conversion to drop-in biofuels as well as in the polymerization to bio-based poly(amide urethane). This chapter presents the genetic engineering methods of pseudomonads for the metabolization of PET monomers and the biosynthesis of HAAs with detailed protocols concerning product purification.


Subject(s)
Fatty Acids , Polyethylene Terephthalates
2.
Soft Matter ; 17(11): 3191-3206, 2021 Mar 21.
Article in English | MEDLINE | ID: mdl-33621291

ABSTRACT

Various studies have described remarkable biological activities and surface-active properties of rhamnolipids, leading to their proposed use in a wide range of industrial applications. Here, we report on a study of the effects of monorhamnolipid RhaC10C10 and dirhamnolipid RhaRhaC10C10 incorporation into model membranes of varying complexity, including bacterial and heterogeneous model biomembranes. For comparison, we studied the effect of HAA (C10C10, lacking a sugar headgroup) partitioning into these membrane systems. AFM, confocal fluorescence microscopy, DSC, and Laurdan fluorescence spectroscopy were employed to yield insights into the rhamnolipid-induced morphological changes of lipid vesicles as well as modifications of the lipid order and lateral membrane organization of the model biomembranes upon partitioning of the different rhamnolipids. The partitioning of the three rhamnolipids into phospholipid bilayers changes the phase behavior, fluidity, lateral lipid organization and morphology of the phospholipid membranes dramatically, to what extent, depends on the headgroup structure of the rhamnolipid, which affects its packing and hydrogen bonding capacity. The incorporation into giant unilamellar vesicles (GUVs) of a heterogeneous anionic raft membrane system revealed budding of domains and fission of daughter vesicles and small aggregates for all three rhamnolipids, with major destabilization of the lipid vesicles upon insertion of RhaC10C10, and also formation of huge GUVs upon the incorporation of RhaRhaC10C10. Finally, we discuss the results with regard to the role these biosurfactants play in biology and their possible impact on applications, ranging from agricultural to pharmaceutical industries.


Subject(s)
Phospholipids , Rhamnose , Glycolipids , Lipid Bilayers , Spectrometry, Fluorescence , Unilamellar Liposomes
3.
Genome Announc ; 5(20)2017 May 18.
Article in English | MEDLINE | ID: mdl-28522721

ABSTRACT

Clostridium thermoalcaliphilum is an obligate anaerobic and rod-shaped bacterium isolated from sewage sludge. It is an alkaliphilic thermotolerant organism and utilizes sucrose, glucose, fructose, maltose, cellobiose, amino acids, and Casamino Acids as substrates. The draft genome comprises 2.031 Mbp and 2,027 predicted protein-coding genes.

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