Valorizing a hydrothermal liquefaction aqueous phase through co-production of chemicals and lipids using the oleaginous yeast Yarrowia lipolytica.

Hydrothermal liquefaction is a promising technology to upgrade wet organic waste into a biocrude oil for diesel or jet fuel; however, this process generates an acid-rich aqueous phase which poses disposal issues. This hydrothermal liquefaction aqueous phase (HTL-AP) contains organic acids, phenol, and other toxins. This work demonstrates that Y. lipolytica as a unique host to valorize HTL-AP into a variety of co-products. Specifically, strains of Y. lipolytica can tolerate HTL-AP at 10% in defined media and 25% in rich media. The addition of HTL-AP enhances production of the polymer precursor itaconic acid by 3-fold and the polyketide triacetic acid lactone at least 2-fold. Additional co-products (lipids and citric acid) were produced in these fermentations. Finally, bioreactor cultivation enabled 21.6 g/L triacetic acid lactone from 20% HTL-AP in mixed sugar hydrolysate. These results demonstrate the first use of Y. lipolytica in HTL-AP valorization toward production of a portfolio of value-added compounds.

Anaerobic digestion of organic fraction from hydrothermal liquefied algae wastewater byproduct.

The wastewater stream from hydrothermal liquefaction (HTL) process used in biofuel production, contains a large amounts of organic compounds where several can be regarded as environmentally hazardous and requires significant treatment before disposal. In this study, semi-continuous anaerobic digestion is used to degrade the organic fraction of wastewater streams from HTL of the algae Tetraselmis (AgTet) and Chlorella (AgChlr). Results indicated high methane yields at 20-30% (v/v) HTL wastewater together with clarified manure, producing 327.2mL/gVSin (or volatile solids in feed) for AgTet and 263.4mL/gVSin for AgChlr. There was a significant reduction in methane production at concentrations higher than 40% (v/v) HTL wastewater in the feed, possibly due to the accumulation of chloride salts or inhibitory compounds such as pyridines, piperidines and pyrrolidines. This was further confirmed by comparing COD, salt and the ammonia concentrations of the effluents after anaerobic digestion at different concentrations of wastewater in manure.

Biological conversion of the aqueous wastes from hydrothermal liquefaction of algae and pine wood by Rhodococci.

In this study, R. opacus PD630, R. jostii RHA1, R. jostii RHA1 VanA-, and their co-culture were employed to convert hydrothermal liquefaction aqueous waste (HTLAW) into lipids. After 11days, the COD reduction of algal-HTLAW reached 93.4% and 92.7% by R. jostii RHA1 and its mutant VanA-, respectively. Woody-HTLAW promoted lipid accumulation of 0.43glipid/gcell dry weight in R. opacus PD630 cells. Additionally, the total number of chemicals in HTLAW decreased by over 1/3 after 7days of coculture, and 0.10g/L and 0.46g/L lipids were incrementally accumulated in the cellular mass during the fermentation of wood- and algal-HTLAW, respectively. The GC-MS data supported that different metabolism pathways were followed when these Rhodococci strains degraded algae- and woody-HTLAW. These results indicated promising potential of bioconversion of under-utilized carbon and toxic compounds in HTLAW into useful products by selected Rhodococci.

Hydrothermal liquefaction of biomass: developments from batch to continuous process.

This review describes the recent results in hydrothermal liquefaction (HTL) of biomass in continuous-flow processing systems. Although much has been published about batch reactor tests of biomass HTL, there is only limited information yet available on continuous-flow tests, which can provide a more reasonable basis for process design and scale-up for commercialization. High-moisture biomass feedstocks are the most likely to be used in HTL. These materials are described and results of their processing are discussed. Engineered systems for HTL are described; however, they are of limited size and do not yet approach a demonstration scale of operation. With the results available, process models have been developed, and mass and energy balances determined. From these models, process costs have been calculated and provide some optimism as to the commercial likelihood of the technology.

The ubiquitous protein domain EAL is a cyclic diguanylate-specific phosphodiesterase: enzymatically active and inactive EAL domains.

The EAL domain (also known as domain of unknown function 2 or DUF2) is a ubiquitous signal transduction protein domain in the Bacteria. Its involvement in hydrolysis of the novel second messenger cyclic dimeric GMP (c-di-GMP) was demonstrated in vivo but not in vitro. The EAL domain-containing protein Dos from Escherichia coli was reported to hydrolyze cyclic AMP (cAMP), implying that EAL domains have different substrate specificities. To investigate the biochemical activity of EAL, the E. coli EAL domain-containing protein YahA and its individual EAL domain were overexpressed, purified, and characterized in vitro. Both full-length YahA and the EAL domain hydrolyzed c-di-GMP into linear dimeric GMP, providing the first biochemical evidence that the EAL domain is sufficient for phosphodiesterase activity. This activity was c-di-GMP specific, optimal at alkaline pH, dependent on Mg(2+) or Mn(2+), strongly inhibited by Ca(2+), and independent of protein oligomerization. Linear dimeric GMP was shown to be 5'pGpG. The EAL domain from Dos was overexpressed, purified, and found to function as a c-di-GMP-specific phosphodiesterase, not as a cAMP-specific phosphodiesterase, in contrast to previous reports. The EAL domains can hydrolyze 5'pGpG into GMP, however, very slowly, thus implying that this activity is irrelevant in vivo. Therefore, c-di-GMP is the exclusive substrate of EAL. Multiple-sequence alignment revealed two groups of EAL domains hypothesized to correspond to enzymatically active and inactive domains. The domains in the latter group have mutations in residues conserved in the active domains. The enzymatic inactivity of EAL domains may explain their coexistence with GGDEF domains in proteins possessing c-di-GMP synthase (diguanulate cyclase) activity.