Sustainable production of microbial polyhydroxyalkanoate (PHA) biopolyesters on a larger scale has to consider the "four magic e": economic, ethical, environmental, and engineering aspects. Moreover, sustainability of PHA production can be quantified by modern tools of Life Cycle Assessment. Economic issues are to a large extent affected by the applied production mode, downstream processing, and, most of all, by the selection of carbon-rich raw materials as feedstocks for PHA production by safe and naturally occurring wild type microorganisms. In order to comply with ethics, such raw materials should be used which do not interfere with human nutrition and animal feed supply chains, and shall be convertible towards accessible carbon feedstocks by simple methods of upstream processing. Examples were identified in carbon-rich waste materials from various industrial braches closely connected to food production. Therefore, the article shines a light on hetero-, mixo-, and autotrophic PHA production based on various industrial residues from different branches. Emphasis is devoted to the integration of PHA-production based on selected raw materials into the holistic patterns of sustainability; this encompasses the choice of new, powerful microbial production strains, non-hazardous, environmentally benign methods for PHA recovery, and reutilization of waste streams from the PHA production process itself.

Polyhydroxyalkanoates (PHAs) are very promising materials that might serve as an environmentally friendly alternative to petrochemical plastics. The main obstacle preventing PHAs from entering the market massively is the final cost of the polymer material, a significant portion of which is attributed to carbon substrate. Hence, the researchers have been intensively seeking cheap substrates for sustainable production of PHAs. Lignocellulose represents a very promising substrate for PHAs production -it is abundant, cheap, and it does not compete with human food chain. On the other hand, utilization of lignocellulose materials as substrates for biotechnological processes represents a challenge due to many factors, such as necessary hydrolysis of the biomass to yield fermentable sugars and presence of numerous antimicrobial agents. Therefore, this work summarizes recent advances in biotechnological conversion of lignocellulose materials into PHAs. The review not only deals with the process of fermentation, but it also considers different approaches of lignocellulose hydrolysis and detoxification.

The methylotrophic yeast Pichia pastoris (Komagataella spp.) is a popular microbial host for the production of recombinant proteins. Previous studies have shown that mis-sorting to the vacuole can be a bottleneck during production of recombinant secretory proteins in yeast, however, no information was available for P. pastoris. In this work the authors have therefore generated vps (vacuolar protein sorting) mutant strains disrupted in genes involved in the CORVET (class C core vacuole/endosome tethering) complex at the early stages of endosomal sorting. Both Δvps8 and Δvps21 strains contained lower extracellular amounts of heterologous carboxylesterase (CES) compared to the control strain, which could be attributed to a high proteolytic activity present in the supernatants of CORVET engineered strains due to rerouting of vacuolar proteases. Serine proteases were identified to be responsible for this proteolytic degradation by liquid chromatography-mass spectrometry and protease inhibitor assays. Deletion of the major cellular serine protease Prb1 in Δvps8 and Δvps21 strains did not only rescue the extracellular CES levels, but even outperformed the parental CES strain (56 and 80% higher yields, respectively). Further deletion of Ybr139W, another serine protease, did not show a further increase in secretion levels. Higher extracellular CES activity and low proteolytic activity were detected also in fed batch cultivation of Δvps21Δprb1 strains, thus confirming that modifying early steps in the vacuolar pathway has a positive impact on heterologous protein secretion.

Background The yeast Pichia pastoris is a widely used host for the secretion of heterologous proteins. Despite being an efficient producer, we observed previously that certain recombinant proteins were mistargeted to the vacuole on their route to secretion. Simultaneous disruption of one vacuolar sorting pathway together with vacuolar proteases prevented this mis-sorting and resulted in higher levels of secreted heterologous protein. Inspired by the positive results, we now set out to investigate the influence of further parts of the vacuolar pathway, namely the Cvt-pathway and the homotypic fusion and protein sorting (HOPS) complex. Results Strains impaired in the Cvt pathway (∆ atg11 , ∆ atg8 ) had no effect on secretion of the model protein carboxylesterase (CES), but resulted in lower secretion levels of the antibody fragment HyHEL-Fab. Disruption of genes involved in the HOPS complex led to vacuole-like compartments of the B category of vps mutants, which are characteristic for the deleted genes YPT7, VPS41 and VAM6 . In particular ∆ ypt7 and ∆ vam6 strains showed an improvement in secreting the model proteins HyHEL-Fab and CES. Additional disruption of the vacuolar protease Pep4 and the potential protease Vps70 led to even further enhanced secretion in ∆ ypt7 and ∆ vam6 strains. Nevertheless, intracellular product accumulation was still observed. Therefore, the secretory route was strengthened by overexpression of early or late secretory genes in the vacuolar sorting mutants. Thereby, overexpression of Sbh1, a subunit of the ER translocation pore, significantly increased HyHEL-Fab secretion, leading up to fourfold higher extracellular Fab levels in the ∆ ypt7 strain. The beneficial impact on protein secretion and the suitability of these strains for industrial applicability was confirmed in fed-batch cultivations. Conclusions Disruption of genes involved in the HOPS complex, especially YPT7 , has a great influence on the secretion of the two different model proteins HyHEL-Fab and CES. Therefore, disruption of HOPS genes shows a high potential to increase secretion of other recombinant proteins as well. Secretion of HyHEL-Fab was further enhanced when overexpressing secretion enhancing factors. As the positive effect was also present in fed-batch cultivations, these modifications likely have promising industrial relevance. Electronic supplementary material The online version of this article (10.1186/s12934-019-1155-4) contains supplementary material, which is available to authorized users.