William Marsh Rice University

George Nelson Bennett

Ka-Yiu San

Ailen M Sanchez

Henry Lin

Irene Martinez

Hui Wu

Miles C Scotcher

San Ka-Yiu

Chandresh Thakker

Jiangfeng Zhu

Laurent Segueilha

Jonathan Silberg

Jian Wang

Thomas Bryce Causey

Matthew Sing Wong

Steven Wu

Xianpeng Cai

Ailen Sánchez

Ravishankar V Vadali

Joshua T Atkinson

Susan J Berrios-Rivera

Mary Lou Harrison

Cheryl Renee Dittrich

Ian J Campbell

Susana J Berrios-Rivera

Grant Balzer

Yipeng Wang

Ailen Genentech Inc Sanchez

Engineered protein electron carriers, microorganisms expressing the same, and methods detecting regulated electron flow are described.

Regulating electron flow using fragmented proteins

Microbes and methods used to convert renewable carbon sources such as glucose, sucrose, biomass hydrolysate, methanol or formate, to succinate-derived products, such as fumarate or malate, which are desirable products having many uses.

Microbial production of succinate derived products

The disclosure relates to a metabolic transistor in microbes such as bacteria and yeast where a competitive pathway is introduced to compete with a product pathway for available carbon so as to control the carbon flux in the microbe.

Metabolic transistor in bacteria

Method of cell culture, comprising adding a redox active compound with a redox potential of between &#x2212;0.116 to &#x2212;0.253 to a culture capable of forming hydrogen via a hydrogenase so that the redox potential is diverted from hydrogen to form a longer chain acids, e.g., butryic acid.

Long chain organic acid bioproduction

Methods of making C4+ hydrocarbon feedstocks using anaerobic microbes are described.

Making C4+ products in bacteria

Production of products by engineered bacteria is increased by regulating cellular respiration. Cellular respiration is controlled by reducing electron transfer enzyme activity. Some examples of electron transfer enzymes include NADH dehydrogenases, Succinate dehydrogenases, ubiquinone synthesis, cytochrome O, and cytochrome D. In one example, deletion of UbiCA prevents respiration. Respiration can the be controlled by addition of ubiquinone or expression of ubiCA.

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The disclosure relates to a metabolic transistor in bacteria where a competitive pathway is introduced to compete with a product pathway for available carbon so as to control the carbon flux in the bacteria.

Bacteria optimized to produce succinate and other feedstocks by growing on low cost carbon sources, such as sucrose.

Engineered bacteria produce succinate from sucrose

A method of increasing cellular NADPH levels by expressing one or more genes that encode an enzyme that causes the production of NADPH. The system is combined with other enzymes that require NADPH, thus improving the overall yield of the desired product.

Increasing NADPH-dependent products

Improved bacteria for making succinate and other 4 carbon dicarboxylates from the Krebs cycle have modifications to reduce acetate, lactate, EtOH and formate, as well as turn on the glyoxylate shunt, produce more NADH and overexpress In one embodiment, the bacteria are &#x394;adhE&#x394;ldhA&#x394;iclR&#x394;ack-pta plus PYCand NAD-dependant FDH.

Increasing bacterial succinate productivity

This invention is metabolically engineer bacterial strains that provide increased intracellular NADPH availability for the purpose of increasing the yield and productivity of NADPH-dependent compounds. In the invention, native NAD-dependent GAPDH is replaced with NADP-dependent GAPDH plus overexpressed NADK. Uses for the bacteria are also provided.

Native NAD-dependent GAPDH replaced with NADP-dependent GAPDH plus NADK

A new culture method for producing high levels of a metabolite, such as succinic acid uses oxygen rich culture without pH adjustment to increase the biomass, acclimation in under oxygen lean conditions having &#x3c;5% partial pressure of oxygen, and the production of high levels of succinate under oxygen deprived conditions. The method can be performed in a single reactor, and is amenable to efficient scale up.

Large scale microbial culture method

A method of producing biological products using bacteria with an inactivated ptsHI and wild type err and no added glucose transport activity and which consumes nearly all glucose in the media is described. The &#x394;ptsHI bacteria produce large quantities of recombinant protein without producing significant amounts of acetate. The bacteria grow well on standard LB broth without additional supplementation.

Reduced phosphotransferase system activity in bacteria

We have shown that the control of solventogenesis and sporulation can be genetically uncoupled in. In strain 824(pASspo), the absence of SpoIIE causes sporulation to be blocked at stage II. The cell remains in a vegetative state, and this allows solvent production to proceed for longer and for solvents to accumulate more rapidly and to a higher concentration. The characteristic drop in OD600 observed in wild type and control strains ofafter 48-72 hours as the cells transition from the solventogenic phase to sporulation is notably absent in the fermentations of 824(pASspo). Mutant S (wild type background, spoIIE disrupted), Mutant BS (Mutant B background, spoIIE disrupted), Mutant HS (Mutant H background, spoIIE disrupted) and Mutant bukS (buk- background, spoIIE disrupted) were generated to create stable solvent producing bacteria with complete inactivation of the SpoIIE protein. Similarity between the SpoIIE protein of, and otherspecies indicates that the techniques used incan be applied to other solvent producing

Blocking sporulation by inhibiting SpoIIE

Methods of increasing yields of succinate using aerobic culture methods and a multi-mutantstrain are provided. Also provided is a mutant strain ofthat produces high amounts of succinic acid.

Growing community of inventors

Houston, TX, United States of America

George Nelson Bennett