Gene Cloning of an alcohol dehydrogenase from thermophilic alkane-degrading Bacillus thermoleovorans B23

Kato, Tomohisa; Miyanaga, A.; Haruki, Mitsuru; Imanaka, Tadayuki; Morikawa, Masaaki; Kanaya, Shigenori

doi:10.1016/s1389-1723(01)80122-5

Cited by 7 publications

(2 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thermoanaerobacter ethanolicus MDR multiple substrates ABY93890 [108] ADH1 Pyrococcus furiosus DSM 3638 SDR low promiscuity AAC25556 [109] BtADH Bacillus thermoleovorans B23 MDR low promiscuity AB040809 [110] ApADH Aeropyrum pernix K1 MDR multiple substrates APE2239 [111] LbADH Lactobacillus brevis ATCC 14869 SDR multiple substrates Q84EX5 [112] ReADH Rhodococcus erythropolis MDR multiple substrates AAN73270 [113] Kacr1 Kluyveromyces aestuarii SDR low promiscuity BAD01116 [114,115] Cscr1…”

Section: Sadhmentioning

confidence: 99%

Stairway to Stereoisomers: Engineering Short‐ and Medium‐Chain Ketoreductases To Produce Chiral Alcohols

Shanbhag

2023

ChemBioChem

View full text Add to dashboard Cite

The short-and medium-chain dehydrogenase/reductase superfamilies are responsible for most chiral alcohol production in laboratories and industries. In nature, they participate in diverse roles such as detoxification, housekeeping, secondary metabolite production, and catalysis of several chemicals with commercial and environmental significance. As a result, they are used in industries to create biopolymers, active pharmaceutical intermediates (APIs), and are also used as components of modular enzymes like polyketide synthases for fabricating bioactive molecules. Consequently, random, semi-rational and rational engineering have helped transform these enzymes into product-oriented efficient catalysts. The rise of newer synthetic chemicals and their enantiopure counterparts has proved challenging, and engineering them has been the subject of numerous studies. However, they are frequently limited to the synthesis of a single chiral alcohol. The study attempts to defragment and describe hotspots of engineering short-and medium-chain dehydrogenases/reductases for the production of chiral synthons.

show abstract

Section: Sadhmentioning

confidence: 99%

Stairway to Stereoisomers: Engineering Short‐ and Medium‐Chain Ketoreductases To Produce Chiral Alcohols

Shanbhag

2023

ChemBioChem

View full text Add to dashboard Cite

show abstract

“…The alcohols are further oxidized by alcohol and aldehyde dehydrogenases to fatty acids, which readily enter the β-oxidation pathway 7,21 . A zinc-independent alcohol dehydrogenase from the thermophillic bacterium Geobacillus thermoleovorans B23 oxidizes medium-chain alkanols into their respective alkanals, using NAD + as a cofactor 10 . Aldehyde dehydrogenase from the same bacterium is able to catalyze the NAD + -dependent final step in the medium-chain oxidation 11 .…”

Section: Introductionmentioning

confidence: 99%

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

Brinkman

Schipper

Bongaerts

et al. 2012

JoVE

View full text Add to dashboard Cite

This work puts forward a toolkit that enables the conversion of alkanes by Escherichia coli and presents a proof of principle of its applicability. The toolkit consists of multiple standard interchangeable parts (BioBricks)(9) addressing the conversion of alkanes, regulation of gene expression and survival in toxic hydrocarbon-rich environments. A three-step pathway for alkane degradation was implemented in E. coli to enable the conversion of medium- and long-chain alkanes to their respective alkanols, alkanals and ultimately alkanoic-acids. The latter were metabolized via the native β-oxidation pathway. To facilitate the oxidation of medium-chain alkanes (C5-C13) and cycloalkanes (C5-C8), four genes (alkB2, rubA3, rubA4and rubB) of the alkane hydroxylase system from Gordonia sp. TF6(8,21) were transformed into E. coli. For the conversion of long-chain alkanes (C15-C36), theladA gene from Geobacillus thermodenitrificans was implemented. For the required further steps of the degradation process, ADH and ALDH (originating from G. thermodenitrificans) were introduced(10,11). The activity was measured by resting cell assays. For each oxidative step, enzyme activity was observed. To optimize the process efficiency, the expression was only induced under low glucose conditions: a substrate-regulated promoter, pCaiF, was used. pCaiF is present in E. coli K12 and regulates the expression of the genes involved in the degradation of non-glucose carbon sources. The last part of the toolkit - targeting survival - was implemented using solvent tolerance genes, PhPFDα and β, both from Pyrococcus horikoshii OT3. Organic solvents can induce cell stress and decreased survivability by negatively affecting protein folding. As chaperones, PhPFDα and β improve the protein folding process e.g. under the presence of alkanes. The expression of these genes led to an improved hydrocarbon tolerance shown by an increased growth rate (up to 50%) in the presences of 10% n-hexane in the culture medium were observed. Summarizing, the results indicate that the toolkit enables E. coli to convert and tolerate hydrocarbons in aqueous environments. As such, it represents an initial step towards a sustainable solution for oil-remediation using a synthetic biology approach.

show abstract

Cloning and expression of three ladA-type alkane monooxygenase genes from an extremely thermophilic alkane-degrading bacterium Geobacillus thermoleovorans B23

2014

Self Cite

View full text Add to dashboard Cite

1An extremely thermophilic bacterium, Geobacillus thermoleovorans B23, is capable of 2 degrading a broad range of alkanes (with carbon chain lengths ranging between C11 and 3 C32) at 70 °C. Whole--genome sequence analysis revealed that unlike most alkane--degrading 4 bacteria, strain B23 does not possess an alkB--type alkane monooxygenase gene. Instead, it 5 possesses a cluster of three ladA--type genes, ladAα B23 , ladAβ B23 , and ladB B23 , on its 6 chromosome, whose protein products share significant amino acid sequence identities, 49.8, 734.4, and 22.7%, respectively with that of ladA alkane monooxygenase gene found on a 8 plasmid of Geobacillus thermodetrificans NG 80--2. Each of the three genes, ladAα B23 , 9 ladAβ B23 , and ladB B23 , was heterologously expressed individually in an alkB1 deletion mutant 10 strain, Pseudomonas fluorescens KOB2Δ1. It was found that all three genes were functional 11 in P. fluorescens KOB2Δ1, and partially restored alkane degradation activity. In this study, 12we suggest that G. thermoleovorans B23 utilizes multiple LadA--type alkane 13 monooxygenases for the degradation of a broad range of alkanes. 14 15

show abstract

Gene Cloning of an alcohol dehydrogenase from thermophilic alkane-degrading Bacillus thermoleovorans B23

Cited by 7 publications

References 12 publications

Stairway to Stereoisomers: Engineering Short‐ and Medium‐Chain Ketoreductases To Produce Chiral Alcohols

Stairway to Stereoisomers: Engineering Short‐ and Medium‐Chain Ketoreductases To Produce Chiral Alcohols

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

Cloning and expression of three ladA-type alkane monooxygenase genes from an extremely thermophilic alkane-degrading bacterium Geobacillus thermoleovorans B23

Contact Info

Product

Resources

About