Poly(3-hydroxybutyrate) (PHB)—a renewable and biodegradable polymer—is a promising alternative to nonbiodegradable synthetic plastics that are derived from petrochemicals. The methods currently employed for PHB production are costly, in part, due to the expensive cultivation feedstocks and the need to sterilize the culture medium, which is energy-intensive. This study investigates the feasibility of nonsterile PHB production from several saline organic wastes using a salt-tolerant strain, Zobellella denitrificans ZD1 (referred to as strain ZD1). Factors such as the pH, salinity, carbon/nitrogen (C/N) ratio, nitrogen source, and electron acceptor that might affect the growth of strain ZD1 and its PHB production were determined. Our results showed successful nonsterile PHB production by growing the strain ZD1 on nonsterile synthetic crude glycerol, high-strength saline wastewater, and real municipal wastewater-activated sludge under saline conditions. The PHB production was significantly enhanced when the levels of salts and nitrate-nitrogen in the culture medium were increased. This study suggested a promising low-cost nonsterile PHB production strategy from organic wastes using strain ZD1.

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Polyhydroxybutyrate (PHB) is biodegradable and renewable and thus considered as a promising alternative to petroleum-based plastics. However, PHB production is costly due to expensive carbon sources for culturing PHB-accumulating microorganisms under sterile conditions. We discovered a hyper PHB-accumulating denitrifying bacterium, Zobellella denitrificans ZD1 (referred as strain ZD1 hereafter) capable of using non-sterile crude glycerol (a waste from biodiesel production) and nitrate to produce high PHB yield under saline conditions. Nevertheless, the underlying genetic mechanisms of PHB production in strain ZD1 have not been elucidated. In this study, we discovered a complete pathway of glycerol conversion to PHB, a novel PHB synthesis gene cluster, a salt-tolerant gene cluster, denitrifying genes, and an assimilatory nitrate reduction gene cluster in the ZD1 genome. Interestingly, the novel PHB synthesis gene cluster was found to be conserved among marine Gammaproteobacteria. Higher levels of PHB accumulation were linked to higher expression levels of the PHB synthesis gene cluster in ZD1 grown with glycerol and nitrate under saline conditions. Additionally, a clustered regularly interspaced short palindromic repeat (CRISPR)-Cas type-I-E antiviral system was found in the ZD1 genome along with a long spacer list, in which most of the spacers belong to either double-stranded DNA viruses or unknown phages. The results of the genome analysis revealed strain ZD1 used the novel PHB gene cluster to produce PHB from non-sterile crude glycerol under saline conditions.

Lignocellulosic biomass, packed with sugars, is one of the most available renewable resources for biofuels and bioproducts production. To release the sugars for the production, enzymatic hydrolysis (saccharification) of pretreated lignocellulosic biomass are required. However, the saccharification process is costly, inefficient, and requires multi-step operations. This is in part due to the high cost and the limited selection of commercial enzymes which commonly have different optimal pH and temperatures. Here we reported a one-step saccharification of pretreated lignocellulosic biomass using immobilized biocatalysts containing five different saccharifying enzymes (SEs) with a similar optimum pH and temperature. The five SEs - endo-1,4-β-d-glucanase (an endoglucanase, eglS), cellobiohydrolase (an exoglucanase, cbhA), and β-glucosidase (bglH), endo-1,4-β-xylanase (an endoxylanase, xynC) and β-xylosidase (bxlB) - were successfully expressed and produced by E. coli BL21. Better saccharification of pretreated corn husks was observed when using the five crude SE enzymes than those using two commonly used SEs, endo-1,4-β-d-glucanase and β-glucosidase. The five SEs were cross-linked in the absence or the presence of magnetic nanoparticles (hereafter referred as SE-CLEAs and M-SE-CLEAs, respectively). By using SE-CLEAs, the highest amount of reduced sugar (250 mg/g biomass) was measured. The activity of immobilized SEs is better than free crude SEs. The M-SE-CLEAs can be reused at least 3 times for effective saccharification of pretreated lignocellulosic biomass.

Microbial and emerging chemical contaminants are unwanted constituents in reclaimed wastewater, due to the health concerns of using the water for agricultural irrigation, aquifer recharges, and potable water. Removal of these contaminants is required but it is currently challenging, given that there is no simple treatment technology to effectively remove the mixture of these contaminants. This study examined the effectiveness of ZnO-assisted photocatalytic degradation of several constituents, including 1,4-dioxane, trihalomethanes (THMs), triclosan (TCS), triclocarban (TCC), antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARGs), under low intensity of UV exposure. E. coli with an ARGs-carrying circular plasmid (pUC19) was used as a model antibiotic resistant bacterium. Our results show that commercial zinc oxide (C-ZnO) assisted photodegradation of 1,4-dioxane, and dehalogenation of THMs, TCS, and TCC, while tetrapodal zinc oxide (T-ZnO) enhanced the dehalogenation of TCS and TCC. Additionally, T-ZnO assisted the photocatalytic inactivation of the E. coli within 6 h and caused structural changes in the plasmid DNA (pUC19) with additional UV exposure, resulting in nonfunctional AGR-containing plasmids. These results also suggest that higher UV dose is required not only to inactivate ARB but also to damage ARGs in the ARB in order to decrease risks in promoting ARB population in the environment. Overall, our results implicated that, under