The disclosure is in the technical field of synthetic biology and metabolic engineering. The disclosure provides engineered viable bacteria. In particular, the disclosure provides viable bacteria with mutated outer membrane biosynthetic pathway leading to disruption of the pathway, preferably substantially lacking lipopolysaccharide (LPS, endotoxin) within the outer membrane. The disclosure further provides methods of generating viable bacteria and uses thereof. The disclosure also provides compositions and methods for inducing immune responses and for researching and developing therapeutic agents. Furthermore, the disclosure is in the technical field of fermentation of metabolically engineered microorganisms producing bioproduct or metabolite.

This Invention discloses a method for production of N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt by microbial fermentation. The method is intended to manufacture N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt in higher efficiency and higher yield, by increasing the effects of N-Acetyl-D-Mannosamine Kinase.

A genetically engineered bacterial strain that produces N-acetylglucosamine, as well as a method of construction and use thereof. The genetically engineered bacterial strain can ferment N-acetylglucosamine under a condition of 40-50° C. Through knocking out genes for glucosamine 6-phosphate deaminase, N-acetylglucosamine-6-phosphate deacetylase and the N-acetylglucosamine transporter protein from a parental bacterium, an N-acetylglucosamine catabolism pathway is blocked. Moreover, overexpression genes for glucosamine 6-phosphate synthase and glucosamine 6-phosphate acetylase are introduced, enabling extra cellular accumulation of N-acetylglucosamine and high-temperature fermentation of N-acetylglucosamine at a temperature higher than 40° C.

Provided herein are perfusion fluids for enzymatically cleaving A-antigens from a donor organ, and methods, uses, associated therewith. In particular, the perfusion fluids comprise two enzymes, GalNAcDeacetylase and Galactosaminidase and the fluids may further comprise a buffered extracellular solution and/or a crowing agent. Furthermore, the compositions described herein were found to have activity at temperatures and pH levels suitable for cell viability.

Provided herein are enzymatic compositions for carbohydrate antigen cleavage, methods, uses, apparatuses and systems associated therewith. In particular, the composition comprises two enzymes, GalNAcDeacetylase and Galactosaminidase and the composition may further comprise a crowding agent. Furthermore, the compositions described herein were found to have activity a temperatures and pH levels suitable for cell viability.

This Invention discloses a method for production of N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt by microbial fermentation. The method is intended to manufacture N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt in higher efficiency and higher yield, by expression of vitreoscilla hemoglobin in microorganism.

The present invention discloses a process for producing N-acetyl-D-glucosamine and D-glucosamine salts by microbial fermentation. The invention includes a method to produce N-acetyl-D-glucosamine and/or D-glucosamine salts with higher efficiency and higher yield by increasing the effect of N-acetyl-D-aminomannose-6-phosphate epimerase in microorganisms.

This Invention discloses a method for production of N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt by microbial fermentation. The method is intended to manufacture N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt in higher efficiency and higher yield, by increasing the effects of N-Acetyl-D-Mannosamine Kinase.

The present invention discloses a process for producing N-acetyl-D-glucosamine and D-glucosamine salts by microbial fermentation. The invention includes a method to produce N-acetyl-D-glucosamine and/or D-glucosamine salts with higher efficiency and higher yield by increasing the effect of N-acetyl-D-aminomannose-6-phosphate epimerase in microorganisms.

This Invention discloses a method for production of N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt by microbial fermentation. The method is intended to manufacture N-Acetyl-D-Glucosamine and/or D-Glucosamine Salt in higher efficiency and higher yield, by expression of vitreoscilla hemoglobin in microorganism.