The present invention relates to genetically modified bacteria and methods of optimizing genetically modified bacteria for the production of a metabolite.

The present invention relates to genetically modified bacteria and methods of optimizing genetically modified bacteria for the production of a metabolite.

The invention provides non-natural microbial organisms containing enzymatic pathways and/or metabolic modifications for enhancing carbon flux through acetyl-CoA, or oxaloacetate and acetyl-CoA. Embodiments of the invention include microbial organisms having a pathway to acetyl-CoA and oxaloacetate that includes phosphoketolase (a PK pathway). The organisms also have either (i) a genetic modification that enhances the activity of the non-phosphotransferase system (non-PTS) for sugar uptake, and/or (ii) a genetic modification(s) to the organism's electron transport chain (ETC) that enhances efficiency of ATP production, that enhances availability of reducing equivalents or both. The microbial organisms can optionally include (iii) a genetic modification that maintains, attenuates, or eliminates the activity of a phosphotransferase system (PTS) for sugar uptake. The enhanced carbon flux through acetyl-CoA and oxaloacetate can be used for production of a bioderived compound, and the microbial organisms can further include a pathway capable of producing the bioderived compound.

Residual lignin recovered from biological conversion processes and pulp and paper industry wastes can be converted to water-soluble products by alkaline oxidation under oxygen pressure at high solids loading within the temperature range 130-180° C. No problems associated with repolymerization reactions are encountered at solids loading between 10-30% where the initial molar ratio of hydroxide base to lignin residual is at last 0.4, possibly because of enhanced reactivity in the oxidation reaction of aromatic groups in self-associate structures. The water-soluble oxidation products can be fractionated to recover a low molecular weight fraction in which pH can be adjusted using C02 without forming precipitates. Sodium carbonate byproduct can be recovered from the pH adjusted reaction mixture using acetone precipitation. The low molecular weight fraction of the pH adjusted oxidation products can be used as feed for anaerobic digestion to biomethane.

A method for improved anaerobic digestion is presented. The method includes mixing a volume of waste material with water to form a feedstock mixture. The volume of waste material includes an initial amount of biomass and the feedstock mixture includes methanogenic bacteria either naturally present in the waste material or introduced artificially. The method also includes introducing one or more promoter substances to the feedstock mixture. The one or more promoter substances are capable of modifying the methanogenic bacteria. Modifying includes stimulating novel enzyme production in the methanogenic bacteria.

Provided are an anaerobic immobilized bacterial agent, a preparation method for same, and applications thereof. The preparation method for the bacterial agent is: selecting four different anaerobic functional bacterial strains, utilizing a pure bacteria culturing technique to produce corresponding culture broths, then mixing the four culture broths according to a certain volume ratio to acquire a compound functional broth, subsequently concentrating into a functional flora precipitation, then dissolving the functional flora precipitation into a polyvinyl alcohol aqueous solution, dripping the solution into a first buffer solution to produce polyvinyl alcohol gel beads, and placing the gel beads produced into a second sulfate-containing buffer solution to produce sulfate-modified polyvinyl alcohol gel beads, that is, the anaerobic immobilized bacterial agent.

It has been found that wild-type ferulic acid decarboxylase derived from Saccharomyces has high catalytic activity for the production of unsaturated hydrocarbon compounds. Further, it has been found that in the ferulic acid decarboxylase, substitute of the amino acid at position 398 with glutamine, methionine, asparagine, phenylalanine, histidine, or threonine more improves the catalytic activity, making it possible to provide a method capable of producing an unsaturated hydrocarbon compound such as butadiene with high productivity, and an enzyme used in the method.

It has been found that wild-type ferulic acid decarboxylase derived from Saccharomyces has high catalytic activity for the production of unsaturated hydrocarbon compounds. Further, it has been found that in the ferulic acid decarboxylase, substitute of the amino acid at position 398 with glutamine, methionine, asparagine, phenylalanine, histidine, or threonine more improves the catalytic activity, making it possible to provide a method capable of producing an unsaturated hydrocarbon compound such as butadiene with high productivity, and an enzyme used in the method.

The disclosure relates to bioreactors, for example for biological treatment and, more specifically to bioreactor insert apparatus including biofilms and related methods. The bioreactor insert apparatus provides a means for circulation of reaction medium within the bioreactor, a biofilm support, and biological treatment of an inlet feed to die reactor/insert apparatus. The bioreactor insert apparatus has a high relative surface area for biofilm attachment and is capable of generating complex flow patterns and increasing treatment efficiency/biological conversion activity in a biologically-active reactor. The high surface area structure incorporates multiple biofilm support structures such as meshes at inlet and outlet portions of the structure. The biofilm support structures and biofilms thereon can increase overall reaction rate of the bioreactor and/or perform some solid/liquid separation in the treatment of the wastewater or other influent.

A system of controllable separation between recyclable organic waste and graywater sewage is described; a respective method for controllable separating recyclable organic waste from graywater sewage is further described; the system includes: a garbage disposal unit, and a separation module; the method includes: draining both the graywater sewage and the recyclable organic waste; processing the organic waste into a semiliquid mixture or slurry of round organic matter and fluid; discharging the semiliquid mixture or slurry of ground organic matter and fluid and the graywater sewage; and releasing the semiliquid mixture or slurry of ground organic matter and fluid; releasing the graywater sewage; and separating the semiliquid mixture or slurry of ground organic matter and fluid from the graywater sewage.