Abstract: This disclosure is in the technical field of synthetic biology and metabolic engineering. The disclosure provides engineered viable bacteria having a reduced or abolished synthesis of poly-N-acetyl-glucosamine (PNAG), Enterobacterial Common Antigen (ECA), cellulose, colanic acid, core oligosaccharides, Osmoregulated Periplasmic Glucans and Glucosylglycerol (O), glycan, and trebalose. The disclosure further provides methods for the production of bioproduct by the viable bacteria and uses thereof. Furthermore, the disclosure is in the technical field of fermentation of metabolically engineered microorganisms producing bioproduct.

Abstract: 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.

Abstract: The present invention is in the technical field of synthetic biology and metabolic engineering. More particularly, the present invention relates to a method to determine the expression stability of a heterologous gene at a chromosomal location in a cell undergoing burden and to produce mutated cells or organisms transformed with a heterologous gene at a chromosomal location, wherein the expression of said heterologous gene is not influenced by a burden or wherein the expression of said heterologous gene is reduced by a burden. The present invention describes methods to locate interesting chromosomal knock-in locations in a cell. Such engineered cells and organisms are applied for the production of bioproducts, such as but not limited to carbohydrates, lipids, proteins, organic acids, amino acids, alcohols, antibiotics and peptides. Preferably, the invention is applied in the technical field of fermentation of metabolically engineered microorganisms.