Development of a plasmid addicted system that is independent of co-inducers, antibiotics and specific carbon source additions for bioproduct (1-butanol) synthesis in .

Synthetic biology approaches for the synthesis of value-based products provide interesting and potentially fruitful possibilities for generating a wide variety of useful compounds and biofuels. However, industrial production is hampered by the costs associated with the need to supplement large microbial cultures with expensive but necessary co-inducer compounds and antibiotics that are required for up-regulating synthetic gene expression and maintaining plasmid-borne synthetic genes, respectively. To address these issues, a metabolism-based plasmid addiction system, which relies on lipopolysaccharide biosynthesis and maintenance of cellular redox balance for 1-butanol production; and utilizes an active constitutive promoter, was developed in .

Altered residues in key proteins influence the expression and activity of the nitrogenase complex in an adaptive CO2 fixation-deficient mutant strain of Rhodobacter sphaeroides.

Previously, the RubisCO-compromised spontaneous adaptive Rhodobacter sphaeroides mutant, strain 16PHC, was shown to derepress the expression of genes that encode the nitrogenase complex under normal repressive conditions. As a result of this adaptation, the active nitrogenase complex restored redox balance, thus allowing strain 16PHC to grow under photoheterotrophic conditions in the absence of an exogenous electron acceptor. A combination of whole genome pyrosequencing and whole genome microarray analyses was employed to identify possible loci responsible for the observed phenotype.

Acetate-dependent photoheterotrophic growth and the differential requirement for the Calvin-Benson-Bassham reductive pentose phosphate cycle in Rhodobacter sphaeroides and Rhodopseudomonas palustris.

Rhodobacter sphaeroides ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO)-deletion strain 16 was capable of photoheterotrophic growth with acetate, while Rhodopseudomonas palustris RubisCO-deletion strain 2040 could not grow under these conditions. The reason for this difference lies in the fact that Rba. sphaeroides and Rps.

Redox homeostasis phenotypes in RubisCO-deficient Rhodobacter sphaeroides via ensemble modeling.

Photosynthetic bacteria are capable of carrying out the fundamental biological processes of carbon dioxide assimilation and photosynthesis. In this work, ensemble modeling (EM) was used to examine the behavior of mutant strains of the nonsulfur purple photosynthetic bacterium Rhodobacter sphaeroides containing a blockage in the primary CO(2) assimilatory pathway, which is responsible for cellular redox balance. When the Calvin-Benson-Bassham (CBB) pathway is nonfunctional, spontaneous adaptive mutations have evolved allowing for the use of at least two separate alternative redox balancing routes enabling photoheterotrophic growth to occur.

Integrative control of carbon, nitrogen, hydrogen, and sulfur metabolism: the central role of the Calvin-Benson-Bassham cycle.

Nonsulfur purple (NSP) photosynthetic bacteria use the Calvin-Benson-Bassham (CBB) reductive pentose phosphate pathway for the reduction of CO(2) via ribulose 1,5-bisphosphate (RuBP) carboxylase-oxygenase (RubisCO), as a means to build cell mass during chemoautotrophic or photoautotrophic conditions. In addition, the CBB pathway plays an important role in maintaining redox balance during photoheterotrophic growth conditions. In this communication we describe protein-protein interactions between two transcriptional regulators CbbR and RegA and the possible role of the CbbX protein in regulating the CBB pathway in Rhodobacter sphaeroides.

Identification of the DNA bases of a DNase I footprint by the use of dye primer sequencing on an automated capillary DNA analysis instrument.

We have adapted the techniques of DNA footprint analysis to an Applied Biosystems 3730 DNA Analyzer. The use of fluorescently labeled primers eliminates the need for radioactively labeled nucleotides, as well as slab gel electrophoresis, and takes advantage of commonly available automated fluorescent capillary electrophoresis instruments. With fluorescently labeled primers and dideoxynucleotide DNA sequencing, we have shown that the terminal base of each digested fragment may be accurately identified with a capillary-based instrument.