Addition of formate dehydrogenase increases the production of renewable alkane from an engineered metabolic pathway

Jaroensuk, Juthamas; Intasian, Pattarawan; Kiattisewee, Cholpisit; Munkajohnpon, Pobthum; Chunthaboon, Paweenapon; Buttranon, Supacha; Trisrivirat, Duangthip; Wongnate, Thanyaporn; Maenpuen, Somchart; Tinikul, Ruchanok; Chaiyen, Pimchai

doi:10.1074/jbc.ra119.008246

Cited by 19 publications

(17 citation statements)

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“…It was shown that addition of formate dehydrogenase (FDH) from Xanthobacter sp. 91 could increase intracellular NADH/NAD + ratio in cells which can increase higher level of reduced Fd and increase yield of alkane production ~50% (23). Here, we further explored why the alkane product was limited at ~50% by further exploring whether this cell can generate other types of by-products.…”

Section: Production Of Fatty Alcohols By E Coli Cells Containing Adomentioning

confidence: 98%

“…pET17b, pCDFduet-1 and pRSFDuet-1 vectors were applied for construction. The same plasmid sets as the previous study [23] were used to investigate alcohol and alkane production by PmADO in whole-cell catalysts (pET17b-acr1, pCDFDuet-1-fd-fnr-pmado, pRSFDuet-1-trxA-trxB and pRSFDuet-1-fdh). To investigate further about the ability of PmADO for fatty alcohol production, pET17bmmcar and pCDFDuet-1-pmado were used in whole-cell bio-catalysis assays.…”

Section: Chemicals and Reagentsmentioning

confidence: 99%

“…Alternatively, if the engineered microbe contains a gene encoding an aldehyde-deformylating oxygenase (ADO), end product of the pathway is alkane/alkene instead of fatty alcohol [22,23]. It has previously been reported that ADO can also convert fatty aldehydes to fatty alcohols with one-carbon less (C n-1 ) as a side reaction in very low yields [24].…”

Section: Introductionmentioning

confidence: 99%

“…Aldehyde-deformylating oxygenase (ADO) from cyanobacteria Prochlorococcus marinus (PmADO) is a non-heme diiron enzyme catalyzing the O 2 -dependent production of one carbon less (C n-1 ) alkanes/alkenes from aldehydes [25]. This enzyme has been attractive for applications in metabolic engineering to produce hydrocarbon [22,23,26]. ADO belongs to the ferritin-like di-iron proteins superfamily [27,28], in which both iron atoms are coordinated with two histidine and four glutamate residues [25,[29][30][31].…”

Section: Introductionmentioning

confidence: 99%

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Unusual Aldehyde Reductase Activity for Production of Full-length Fatty Alcohol by Cyanobacterial Aldehyde Deformylating Oxygenase  

Buttranon

Intasian

Treesukkasem

et al. 2020

Preprint

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Background: Aldehyde-deformylating oxygenase (ADO) is a non-heme di-iron enzyme that catalyzes deformylation of aldehydes to generate alkanes/alkenes. In this study, we report for the first time that under anaerobic or limited oxygen conditions, Prochlorococcus marinus (PmADO) can generate full-length fatty alcohols from fatty aldehydes without eliminating a carbon unit. Results: Unlike the native activity of ADO which requires electrons from the Fd/FNR electron transfer complex, the aldehyde reduction activity of ADO requires only NADPH. Our results demonstrated that yield of alcohol products can be affected by oxygen concentration and type of aldehyde. Under O2-scant conditions (10-15%), yields of octanol and dodecanol were around 40-60% and could be increased up to 80% under strict anaerobic conditions (>0.0004%). Unexpectedly, Fe2+ cofactor is not involved in the aldehyde reductase activity of PmADO because yields of alcohols obtained from holo- and apo-enzymes were similar under anaerobic conditions. The direct hydride transfer activity of PmADO is highly specific to substrates; NADPH not NADH can be used as a reductant to reduce medium-chain fatty aldehydes (C6-C10) with decanal as the most preferred substrate (the highest kcat/Km value with 98% bioconversion yield). Molecular dynamics (MD) simulations was used to identify a binding site of NADPH which is located close to the aldehyde binding site. In the metabolic engineered cells containing PmADO, dual activities of alkane and alcohol production could be detected. Conclusion: The findings reported herein highlight a new activity of PmADO which may be applied as a biocatalyst for industrial synthesis of fatty alcohols in the future.

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Section: Production Of Fatty Alcohols By E Coli Cells Containing Adomentioning

confidence: 98%

Section: Chemicals and Reagentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Unusual Aldehyde Reductase Activity for Production of Full-length Fatty Alcohol by Cyanobacterial Aldehyde Deformylating Oxygenase  

Buttranon

Intasian

Treesukkasem

et al. 2020

Preprint

Self Cite

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“…The placement of rare codons with an mRNA region can promote stability in addition to ribosomal stalling allowing extra time for folding of problematic peptide regions [ 157 ]. These studies purchased synthetic genes from commercial manufacturers including GenScript USA Inc. [ 132 , 158 , 159 ], Invitrogen [ 96 , 160 ], Sloning BioTechnology GmbH, and Synbio Technologies [ 47 ] that carry out codon optimisation on their products as a default service. The Graphical Codon Usage Analyser [ 41 ] and the Genescript Rare Codon Analysis Tool [ 72 ] were used for in-house codon analysis [ 6 , 40 ].…”

Section: Role Of Systems Biology In Addressing the Challenges Around The Formation Of Inclusion Bodiesmentioning

confidence: 99%

Recombinant expression of insoluble enzymes in Escherichia coli: a systematic review of experimental design and its manufacturing implications

2021

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Recombinant enzyme expression in Escherichia coli is one of the most popular methods to produce bulk concentrations of protein product. However, this method is often limited by the inadvertent formation of inclusion bodies. Our analysis systematically reviews literature from 2010 to 2021 and details the methods and strategies researchers have utilized for expression of difficult to express (DtE), industrially relevant recombinant enzymes in E. coli expression strains. Our review identifies an absence of a coherent strategy with disparate practices being used to promote solubility. We discuss the potential to approach recombinant expression systematically, with the aid of modern bioinformatics, modelling, and ‘omics’ based systems-level analysis techniques to provide a structured, holistic approach. Our analysis also identifies potential gaps in the methods used to report metadata in publications and the impact on the reproducibility and growth of the research in this field.

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Detection of cellular metabolites by redox enzymatic cascades

Tinikul

Trisrivirat

Chinantuya

et al. 2022

Biotechnology Journal

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Detection of cellular metabolites that are disease biomarkers is important for human healthcare monitoring and assessing prognosis and therapeutic response. Accurate and rapid detection of microbial metabolites and pathway intermediates is also crucial for the process optimization required for development of bioconversion methods using metabolically engineered cells. Various redox enzymes can generate electrons that can be employed in enzyme‐based biosensors and in the detection of cellular metabolites. These reactions can directly transform target compounds into various readout signals. By incorporating engineered enzymes into enzymatic cascades, the readout signals can be improved in terms of accuracy and sensitivity. This review critically discusses selected redox enzymatic and chemoenzymatic cascades currently employed for detection of human‐ and microbe‐related cellular metabolites including, amino acids, d‐glucose, inorganic ions (pyrophosphate, phosphate, and sulfate), nitro‐ and halogenated phenols, NAD(P)H, fatty acids, fatty aldehyde, alkane, short chain acids, and cellular metabolites.

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Addition of formate dehydrogenase increases the production of renewable alkane from an engineered metabolic pathway

Cited by 19 publications

References 53 publications

Unusual Aldehyde Reductase Activity for Production of Full-length Fatty Alcohol by Cyanobacterial Aldehyde Deformylating Oxygenase

Unusual Aldehyde Reductase Activity for Production of Full-length Fatty Alcohol by Cyanobacterial Aldehyde Deformylating Oxygenase

Recombinant expression of insoluble enzymes in Escherichia coli: a systematic review of experimental design and its manufacturing implications

Detection of cellular metabolites by redox enzymatic cascades

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Addition of formate dehydrogenase increases the production of renewable alkane from an engineered metabolic pathway

Cited by 19 publications

References 53 publications

Unusual Aldehyde Reductase Activity for Production of Full-length Fatty Alcohol by Cyanobacterial Aldehyde Deformylating Oxygenase&nbsp;&nbsp;

Unusual Aldehyde Reductase Activity for Production of Full-length Fatty Alcohol by Cyanobacterial Aldehyde Deformylating Oxygenase&nbsp;&nbsp;

Recombinant expression of insoluble enzymes in Escherichia coli: a systematic review of experimental design and its manufacturing implications

Detection of cellular metabolites by redox enzymatic cascades

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Unusual Aldehyde Reductase Activity for Production of Full-length Fatty Alcohol by Cyanobacterial Aldehyde Deformylating Oxygenase

Unusual Aldehyde Reductase Activity for Production of Full-length Fatty Alcohol by Cyanobacterial Aldehyde Deformylating Oxygenase