Meiothermus ruber thiolase – A new process stable enzyme for improved butanol synthesis

Reiße, Steven; Garbe, Daniel; Brück, Thomas

doi:10.1016/j.biochi.2014.03.013

Cited by 5 publications

(2 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Notably, the reaction cascade primarily focused on converting pyruvate to n -butanol utilizing seven enzyme activities, CoA and NADH as cofactors. In this approach, unbalanced concentrations of CoA-dependent intermediates and generated NAD + inhibit crucial enzyme activities, such as a crotonase similar 3-hydroxypropionyl-CoA dehydratase, thiolase, and β-hydroxybutyryl-CoA dehydrogenase, thereby limiting product yield (Engel et al, 1996 ; Sommer et al, 2013a ; Reisse et al, 2014 ). In order to sustain in vitro n -butanol biosynthesis in the presence of CoA and NAD + , therefore, required a time-dependent feeding of metabolic intermediates to overcome enzyme-specific feedback inhibition and temperature-dependent cofactor instability (Krutsakorn et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

In Vitro Bioconversion of Pyruvate to n-Butanol with Minimized Cofactor Utilization

Reiße

Haack

Garbe

et al. 2016

Front. Bioeng. Biotechnol.

Self Cite

View full text Add to dashboard Cite

Due to enhanced energy content and reduced hygroscopicity compared with ethanol, n-butanol is flagged as the next generation biofuel and platform chemical. In addition to conventional cellular systems, n-butanol bioproduction by enzyme cascades is gaining momentum due to simplified process control. In contrast to other bio-based alcohols like ethanol and isobutanol, cell-free n-butanol biosynthesis from the central metabolic intermediate pyruvate involves cofactors [NAD(P)H, CoA] and acetyl-CoA-dependent intermediates, which complicates redox and energy balancing of the reaction system. We have devised a biochemical process for cell-free n-butanol production that only involves three enzyme activities, thereby eliminating the need for acetyl-CoA. Instead, the process utilizes only NADH as the sole redox mediator. Central to this new process is the amino acid catalyzed enamine–aldol condensation, which transforms acetaldehyde directly into crotonaldehyde. Subsequently, crotonaldehyde is reduced to n-butanol applying a 2-enoate reductase and an alcohol dehydrogenase, respectively. In essence, we achieved conversion of the platform intermediate pyruvate to n-butanol utilizing a biocatalytic cascade comprising only three enzyme activities and NADH as reducing equivalent. With reference to previously reported cell-free n-butanol reaction cascades, we have eliminated five enzyme activities and the requirement of CoA as cofactor. Our proof-of-concept demonstrates that n-butanol was synthesized at neutral pH and 50°C. This integrated reaction concept allowed GC detection of all reaction intermediates and n-butanol production of 148 mg L−1 (2 mM), which compares well with other cell-free n-butanol production processes.

show abstract

Section: Introductionmentioning

confidence: 99%

In Vitro Bioconversion of Pyruvate to n-Butanol with Minimized Cofactor Utilization

Reiße

Haack

Garbe

et al. 2016

Front. Bioeng. Biotechnol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…silvanus, M. cerbereus, M. rufus e M. timidus e outras, que já foram isolados de solos e de fontes termais (CHEN et al, 2002b;CHUNG et al, 1997;PIRES et al, 2005;YU et al, 2014), e também são estudadas para aplicações biotecnológicas, principalmente M. ruber, que foi avaliada em relação à produção de butanol através de enzimas chaves como crotonases e tiolases, e em relação à degradação de penas de aves (REIßE; GARBE; BRÜCK, 2014BRÜCK, , 2015YAMAOKA et al, 2014) A frequência de tetranucleotídeos dos genomas funciona como uma assinatura de cada micro-organismo, de maneira que bactérias relacionadas filogeneticamente possuem frequências similares (DICK et al, 2009;PRIDE et al, 2003). A ANI é um método de hibridização DNA-DNA in silico que funciona de modo similar a frequência de tetranucleotídeos, onde valores iguais ou superiores a 95-96% normalmente indicam micro-organismos da mesma espécie (GORIS et al, 2007; KONSTANTINIDIS; Geitlerinema CCNMD22 possui 8 genes codificando glicosiltransferases e proteínas do operon rfb, relacionados com a EPS, além dos genes codificando wzA (2609076755), uma proteína de membrana que serve de canal para polissacarídeos (PEREIRA et al, 2009;REEVES et al, 1996).…”

Section: Recuperação De Genomasunclassified

Análise metagenômica da microbiota de solos de ambientes de mina de cobre

Castro¹

View full text Add to dashboard Cite

show abstract

Identification and optimization of a novel thermo- and solvent stable ketol-acid reductoisomerase for cell free isobutanol biosynthesis

2015

View full text Add to dashboard Cite

Meiothermus ruber thiolase – A new process stable enzyme for improved butanol synthesis

Cited by 5 publications

References 37 publications

In Vitro Bioconversion of Pyruvate to n-Butanol with Minimized Cofactor Utilization

In Vitro Bioconversion of Pyruvate to n-Butanol with Minimized Cofactor Utilization

Análise metagenômica da microbiota de solos de ambientes de mina de cobre

Identification and optimization of a novel thermo- and solvent stable ketol-acid reductoisomerase for cell free isobutanol biosynthesis

Contact Info

Product

Resources

About