A hot topic: thermophilic plastic biodegradation.

Biological degradation of plastic waste is an environmentally and economically friendlier alternative to current recycling practices and enables the cycling of plastic monomers back into virgin-quality plastics. However, due to slow reaction rates, there is a lack of an industrially viable biodegradation strategy for most plastics. Here, we highlight the applicability of a thermophilic biodegradation strategy over a mesophilic approach, to enhance enzyme accessibility and catalyze plastic biodegradation. Thus, at reactions closer to the melting temperature or glass transition temperature of plastics, thermophilic reactions can offer an alternative direction to conventional plastic biodegradation strategies.

Mild fractionation of sugarcane bagasse into fermentable sugars and ß-O-4 linkage-rich lignin based on acid-catalysed crude glycerol pretreatment.

Acid-catalysed crude glycerol (ACG) pretreatment was carried out at 110 °C and 130 °C for mild fractionation of sugarcane bagasse into fermentable sugars and high-quality lignin. ACG pretreatment at 110 °C led to sugar yields of 71%-74%, comparable to those with acid-catalysed reagent-grade glycerol (AG). ACG pretreatment removed more lignin (53%-75%) than AG pretreatment (38%-49%), likely due to the presence of organic impurities in ACG. Hence, 28% more lignin was recovered from ACG pretreatment hydrolysate than with the AG pretreatment. NMR analysis revealed that recovered lignin was modified by glycerol through etherification of ß-aryl ethers and esterification of hydroxycinnamic acids, which prevented lignin condensation and led to the generation of ß-O-4 linkage-rich lignin at mild conditions (110 °C for 3 h and 5 h). This study suggests that crude glycerol is a suitable low-cost solvent for mild fractionation of lignocellulosic biomass into fermentable sugars and high-quality lignin for value-adding applications.

Scale-up of two-step acid-catalysed glycerol pretreatment for production of oleaginous yeast biomass from sugarcane bagasse by Rhodosporidium toruloides.

Two-step dilute acid and acid-catalysed glycerol pretreatment was developed to maximise sugar yield from sugarcane bagasse. At the laboratory scale, dilute acid pretreatment at 130 °C followed by acid-catalysed glycerol pretreatment at 170 °C led to a total sugar (C5 + C6) yield of 82%, 31% higher than that from one-step acid-catalysed glycerol pretreatment. At the pilot scale, the two-step dilute acid and acid-catalysed glycerol pretreatment led to a maximum sugar yield of 74%, 13% higher than that from one-step pretreatment with 52% reduction in glycerol usage. The enzymatic hydrolysate containing glucose and residual glycerol were used to produce microbial oils by a Rhodosporidium toruloides strain. A fed-batch cultivation strategy led to the production of 44.8 g/L cell mass, including 26.6 g/L oil, 8.6 g/L protein and 12.7 mg/L carotenoid. The cell mass and oil yields were 19% higher than those from batch cultivation as feedstock inhibition and catabolite repression were alleviated.

Making a Bacterial Thermophilic Enzyme in a Fungal Expression System.

This unit describes production of a bacterial thermophilic xylanase enzyme in an industrially exploited filamentous fungus, Trichoderma reesei. Successful expression of a gene of interest in a heterologous host involves front-end design of the expression constructs using bioinformatics tools, making the constructs in the laboratory, and introducing them into the expression host. This is followed by synthesis and characterization of the gene product on a laboratory scale and optimization of the cultivation parameters in a controlled, scaled-up fermentation. The thermophilic xylanase B (XynB) enzyme from the bacterium Dictyoglomus thermophilum discussed here can be easily purified by heat-precipitation from the culture supernatant of the mesophilic host. A functional XynB can also be produced in Escherichia coli, but at a lower yield compared to that obtained in T. reesei. The protocol provided here can be adapted to various other proteins and filamentous fungal hosts. © 2018 by John Wiley & Sons, Inc.

Improved fermentation efficiency of S. cerevisiae by changing glycolytic metabolic pathways with plasma agitation.

Production of ethanol by the yeast Saccharomyces cerevisiae is a process of global importance. In these processes, productivities and yields are pushed to their maximum possible values leading to cellular stress. Transient and lasting enhancements in tolerance and performance have been obtained by genetic engineering, forced evolution, and exposure to moderate levels of chemical and/or physical stimuli, yet the drawbacks of these methods include cost, and multi-step, complex and lengthy treatment protocols. Here, plasma agitation is shown to rapidly induce desirable phenotypic changes in S. cerevisiae after a single treatment, resulting in improved conversion of glucose to ethanol. With a complex environment rich in energetic electrons, highly-reactive chemical species, photons, and gas flow effects, plasma treatment simultaneously mimics exposure to multiple environmental stressors. A single treatment of up to 10 minutes performed using an atmospheric pressure plasma jet was sufficient to induce changes in cell membrane structure, and increased hexokinase 2 activity and secondary metabolite production. These results suggest that plasma treatment is a promising strategy that can contribute to improving metabolic activity in industrial microbial strains, and thus the practicality and economics of industrial fermentations.

Draft Genome Sequence of Caloramator mitchellensis, a Thermoanaerobe Isolated from the Waters of the Great Artesian Basin.

The genome sequence of Caloramator mitchellensis strain VF08, a rod-shaped, heterotrophic, strictly anaerobic bacterium isolated from the free-flowing waters of a Great Artesian Basin (GAB) bore well located in Mitchell, an outback Queensland town in Australia, is reported here. The analysis of the 2.42-Mb genome sequence indicates that the attributes of the genome are consistent with its physiological and phenotypic traits.