Monoterpene hydrocarbon biosynthesis by isolated leucoplasts of Citrofortunella mitis

The flowers of Narcissus poeticus are used for the isolation of valuable fragrance substances. So far, as the majority of these substances consist of volatile and sensitive to heat compounds, there is a need of developing effective methods for their recovery. In this study, freeze-dried N. poeticus inflorescences were extracted with pure supercritical CO2 (SFE-CO2) and its mixture with 5% co-solvent ethanol (EtOH) at 40 °C. Extract yields varied from 1.63% (12 MPa) to 3.12% (48 MPa, 5% EtOH). In total, 116 volatile compounds were identified by GC-TOF/MS in the extracts, which were divided into 20 different groups. Benzyl benzoate (9.44–10.22%), benzyl linoleate (1.72–2.17%) and benzyl alcohol (0.18–1.00%) were the major volatiles among aromatic compounds. The amount of the recovered benzyl benzoate in N. poeticus SFE-CO2 extracts varied from 58.98 ± 2.61 (24 MPa) to 91.52 ± 1.36 (48 MPa) mg/kg plant dry weight (pdw). α-Terpineol dominated among oxygenated monoterpenes (1.08–3.42%); its yield was from 9.25 ± 0.63 (12 MPa) to 29.88 ± 1.25 (48 MPa/EtOH) mg/kg pdw. Limonene was the major monoterpene hydrocarbon; (3E)-hexenol and heneicosanol dominated among alcohols and phenols; dihydroactinidiolide and 4,8,12,16-tetramethyl heptadecan-4-olide were the most abundant lactones; heptanal, nonanal, (2E,4E)-decadienal and octadecanal were the most abundant aldehydes. The most important prenol lipids were triterpenoid squalene, from 0.86 ± 0.10 (24 MPa) to 7.73 ± 0.18 (48 MPa/EtOH) mg/kg pdw and D-α-tocopherol, from 1.20 ± 0.04 (12 MPa) to 15.39 ± 0.31 (48 MPa/EtOH) mg/kg pdw. Aliphatic hydrocarbons (waxes) constituted the main part (41.47 to 54.93%) in the extracts; while in case of a 5% EtOH the percentage of alkanes was the lowest. The fraction of waxes may be removed for the separation of higher value fragrance materials. In general, the results obtained are promising for a wider application of SFE-CO2 for the recovery of fragrance substances from N. poeticus flowers.

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Algal Genes Encoding Enzymes for Photosynthesis and Hydrocarbon Biosynthesis as Candidates for Genetic Engineering

CYTOLOGIA ◽

10.1508/cytologia.83.7 ◽

2018 ◽

Vol 83 (1) ◽

pp. 7-17 ◽

Cited By ~ 2

Author(s):

Hidenobu Uchida ◽

Ko Kato ◽

Kensaku Suzuki ◽

Akiho Yokota ◽

Shigeyuki Kawano ◽

...

Keyword(s):

Genetic Engineering ◽

Genes Encoding ◽

Hydrocarbon Biosynthesis

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Active Multienzyme Assemblies for Long-Chain Olefinic Hydrocarbon Biosynthesis

Journal of Bacteriology ◽

10.1128/jb.00890-16 ◽

2017 ◽

Vol 199 (9) ◽

Cited By ~ 12

Author(s):

James K. Christenson ◽

Matthew R. Jensen ◽

Brandon R. Goblirsch ◽

Fatuma Mohamed ◽

Wei Zhang ◽

...

Keyword(s):

Active Sites ◽

Xanthomonas Campestris ◽

Fatty Acyl ◽

Biosynthetic Activity ◽

Content Type ◽

Large Structures ◽

Negative Stain ◽

Acyl Coenzyme A ◽

Hydrocarbon Biosynthesis ◽

Long Chain

ABSTRACT Bacteria from different phyla produce long-chain olefinic hydrocarbons derived from an OleA-catalyzed Claisen condensation of two fatty acyl coenzyme A (acyl-CoA) substrates, followed by reduction and oxygen elimination reactions catalyzed by the proteins OleB, OleC, and OleD. In this report, OleA, OleB, OleC, and OleD were individually purified as soluble proteins, and all were found to be essential for reconstituting hydrocarbon biosynthesis. Recombinant coexpression of tagged OleABCD proteins from Xanthomonas campestris in Escherichia coli and purification over His6 and FLAG columns resulted in OleA separating, while OleBCD purified together, irrespective of which of the four Ole proteins were tagged. Hydrocarbon biosynthetic activity of copurified OleBCD assemblies could be reconstituted by adding separately purified OleA. Immunoblots of nondenaturing gels using anti-OleC reacted with X. campestris crude protein lysate indicated the presence of a large protein assembly containing OleC in the native host. Negative-stain electron microscopy of recombinant OleBCD revealed distinct large structures with diameters primarily between 24 and 40 nm. Assembling OleB, OleC, and OleD into a complex may be important to maintain stereochemical integrity of intermediates, facilitate the movement of hydrophobic metabolites between enzyme active sites, and protect the cell against the highly reactive β-lactone intermediate produced by the OleC-catalyzed reaction. IMPORTANCE Bacteria biosynthesize hydrophobic molecules to maintain a membrane, store carbon, and for antibiotics that help them survive in their niche. The hydrophobic compounds are often synthesized by a multidomain protein or by large multienzyme assemblies. The present study reports on the discovery that long-chain olefinic hydrocarbons made by bacteria from different phyla are produced by multienzyme assemblies in X. campestris. The OleBCD multienzyme assemblies are thought to compartmentalize and sequester olefin biosynthesis from the rest of the cell. This system provides additional insights into how bacteria control specific biosynthetic pathways.

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Metabolic Engineering of Hydrocarbon Biosynthesis for Biofuel Production

Liquid, Gaseous and Solid Biofuels - Conversion Techniques ◽

10.5772/52050 ◽

2013 ◽

Cited By ~ 3

Author(s):

Anne M.

Keyword(s):

Metabolic Engineering ◽

Biofuel Production ◽

Hydrocarbon Biosynthesis

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The fatty acid elongase gene LmELO7 is required for hydrocarbon biosynthesis and cuticle permeability in the migratory locust, Locusta migratoria

Journal of Insect Physiology ◽

10.1016/j.jinsphys.2020.104052 ◽

2020 ◽

Vol 123 ◽

pp. 104052 ◽

Cited By ~ 1

Author(s):

Xiaoming Zhao ◽

Yang Yang ◽

Niu Niu ◽

Yiyan Zhao ◽

Weimin Liu ◽

...

Keyword(s):

Fatty Acid ◽

Locusta Migratoria ◽

Fatty Acid Elongase ◽

Migratory Locust ◽

Hydrocarbon Biosynthesis

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Volatile Constituents of Two Rare Subspecies of Thymus praecox

Natural Product Communications ◽

10.1177/1934578x1000500730 ◽

2010 ◽

Vol 5 (7) ◽

pp. 1934578X1000500 ◽

Cited By ~ 4

Author(s):

Danijela Vidic ◽

Sanja Ćavar ◽

Marija Edita Šolić ◽

Milka Maksimović

Keyword(s):

Essential Oil ◽

Monoterpene Hydrocarbon ◽

Oil Analysis ◽

Capillary Gc ◽

Headspace Volatiles ◽

Linalyl Acetate ◽

Volatile Constituents ◽

First Report ◽

Oxygenated Monoterpenes ◽

Ms Analysis

Hydrodistilled essential oil and the corresponding headspace volatiles of Bosnian wild growing Thymus praecox ssp. polytrichus (A. Kern. Ex Borbàs) Jalas and Thymus praecox ssp. skorpilii (Velen.) Jalas were subjected to capillary GC-MS analysis. This work presents a detailed essential oil analysis of these two rare Thymus subspecies from Bosnia, as well as the very first report on their headspace composition. Eighty-seven volatile constituents were identified in four samples. Two alcohols were the major constituents in the essential oil of T. praecox ssp. polytrichus, a monoterpene, linalool (13.9%), and a sesquiterpene, ( E)-nerolidol (10.4%), while linalyl acetate (36.7%) and linalool (22.7%) were the most abundant volatiles in the corresponding headspace sample. Oxygenated monoterpenes (57.5%) predominate in the essential oil of T. praecox ssp. skorpilii with linalyl acetate (28.7%) and linalool (14.4%) as the main representatives. Headspace sample of this subspecies also showed richness in linalyl acetate (52.4%), while the second most abundant compound was α-pinene (14.5%), a monoterpene hydrocarbon.

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Cloning, purification, crystallization and preliminary X-ray diffraction of the OleC protein fromStenotrophomonas maltophiliainvolved in head-to-head hydrocarbon biosynthesis

Acta Crystallographica Section F Structural Biology and Crystallization Communications ◽

10.1107/s1744309110031751 ◽

2010 ◽

Vol 66 (9) ◽

pp. 1108-1110 ◽

Cited By ~ 8

Author(s):

Janice A. Frias ◽

Brandon R. Goblirsch ◽

Lawrence P. Wackett ◽

Carrie M. Wilmot

Keyword(s):

Diffraction Data ◽

Unit Cell ◽

Biosynthetic Enzyme ◽

X Ray Diffraction ◽

Unit Cell Parameters ◽

Space Group ◽

X Ray ◽

Cell Parameters ◽

Hydrocarbon Biosynthesis

OleC, a biosynthetic enzyme involved in microbial hydrocarbon biosynthesis, has been crystallized. Synchrotron X-ray diffraction data have been collected to 3.4 Å resolution. The crystals belonged to space groupP3121 orP3221, with unit-cell parametersa=b= 98.8,c= 141.0 Å.

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