In the last years, biotechnological production of platform chemicals for fuel components has become a major focus interest. Although ligno-cellulosic material is considered as suitable feedstock, almost inevitable pretreatment this recalcitrant may interfere with subsequent fermentation steps. study, fungus Ustilago maydis was used to produce itaconic acid chemical synthesis potential biofuels such 3-methyltetrahydrofuran. No studies, however, have investigated how biomass precisely...

Abstract An immense number of chemical reactions are carried out simultaneously in living cells. Nature’s optimization approach encompasses the assembly cascades and to embed them finely tuned metabolic networks. With vast progress field biocatalysis, man‐made cascades, especially redox have reached a degree complexity that needs tools for improved control optimization. Combined strategies from engineering synthetic biology lead establishment artificial pathways with minimized interference...

Abstract Background Terpenes are remarkably diverse natural structures, which can be formed via two different pathways leading to common intermediates. Among those, sesquiterpenes represent a variety of industrially relevant products. One important produced product is β-farnesene as precursor for jet fuel additive. So far, microbial terpene production has been mostly limited known hosts, only able grow on heterotrophic substrates. Results In this paper, we the first time describe by...

Abstract We describe the development of a kinetic model for simulation and optimization an in vivo redox cascade E. coli which combination alcohol dehydrogenase, enoate reductase, Baeyer–Villiger monooxygenase is used synthesis lactones. The was to estimate concentrations active enzyme sequential biotransformations identify bottlenecks together with their reasons how overcome them. estimated adapted Michaelis–Menten parameters from vitro experiments isolated enzymes these values simulate...

Abstract Synthetic enzyme cascades in living cells often lack efficiency owing to the formation of byproducts by endogenous enzymes or toxicity cascade intermediates. Highly reactive aldehyde species can trigger a metabolic stress response, and this leads undesired side reactions decreased yields. Owing background Escherichia coli ( E. ), aldehydes may be irreversibly oxidized carboxylic acids reduced corresponding alcohols. Herein, we applied an approach equilibrate concentration vivo. We...

Abstract This paper describes the stabilization of flavin‐dependent monooxygenases under reaction conditions, using an engineered formulation additives (the natural cofactors NADPH and FAD, superoxide dismutase catalase as catalytic antioxidants). way, a 10 3 ‐ to 4 ‐fold increase half‐life was reached without resource‐intensive directed evolution or structure‐dependent protein engineering methods. The stabilized enzymes are highly valued for their synthetic potential in biotechnology...

Abstract Terpenoids have an impressive structural diversity and provide valuable substances for a variety of industrial applications. Among terpenes, the sesquiterpenes (C 15 ) are largest subclass with bioactivities ranging from aroma to health promotion. In this article, we show gram‐scale production sesquiterpene α‐humulene in final aqueous concentrations 2 g L −1 recombinant strain Cupriavidus necator pKR‐hum fed‐batch mode on fructose as carbon source n‐dodecane extracting organic phase...

ABSTRACT This paper describes the development of a biocatalytic process on multi‐dozen gram scale for synthesis precursor to Nylon‐9, specialty polyamide. Such materials are growing in demand, but their corresponding monomers often difficult synthesize, giving rise approaches. Here, we implemented cyclopentadecanone monooxygenase as an Escherichia coli whole‐cell biocatalyst defined medium, together with substrate feeding–product removal concept, and optimized downstream processing (DSP). A...

This paper describes the measurement and analysis of in vivo activity stability cyclohexanone monooxygenase from Acinetobacter sp. NCIMB 9871 (CHMO), a model Baeyer-Villiger monooxygenase, recombinant host Escherichia coli. enzyme was often described as poorly stable vitro, has recently been found to deactivate rapidly absence its essential cofactors anti-oxidants. Its scarcely studied, so far. Under conditions common for overexpression CHMO we investigated ability support these properties...

Abstract A synthetic cascade for the transformation of primary alcohols into polyhydroxylated compounds in Escherichia coli , through situ preparation cytotoxic aldehyde intermediates and subsequent aldolase‐mediated C−C bond formation, has been investigated. An enzymatic toolbox consisting alcohol dehydrogenase AlkJ from Pseudomonas putida dihydroxyacetone‐/hydroxyacetone‐accepting aldolase variant Fsa1‐A129S was applied. Pathway optimization performed at genetic process levels. Three...

The objective of the study was to develop a strategy for identification new vitamin B12-producing species and characterize their production capability using fast sensitive LC-MS/MS method developed in this study.Searching homologues bluB/cobT2 fusion gene known be responsible active B12 form P. freudenreichii shown successful strains. analysis identified strains via showed ability Terrabacter sp. DSM102553, Yimella lutea DSM19828 Calidifontibacter indicus DSM22967 produce B12. Further...

Abstract Review: [217 refs.