Isolation from tobacco extracts

His-tagged GUS-fusion proteins have been produced and isolated from tobacco chloroplasts. His-tagged proteins have also been extracted by foam frachonahon (Crofcheck et al., 2003,2004) or by a modihed intein expression system (Morassutti et al., 2002). 

Isol. from tobacco (Nicotiana tabacum), hop oil (Humulus lupulus), clover (Trifolium repens), tea, tomatoes, various fruits and other vegetable sources. Industrial solvent, particularly for coating systems, used in the manuf. of resins and in lubricating oil refining. Reagent for sepn. of amines by glc. Used for extraction separation of Au. Liq. Part misc. H2O. dj 0.805. Fp —85.9°. Bp 79.6°. 1.3814. Forms constant-boiling mixt. with H2O, B.p. 73.4°, contg. 11.3% H2O. 

Intact tobacco plants were exposed to 0.60-0.70 yl/1 ozone for 1 hr mitochondria isolated from visibly injured tissue demonstrated an inhibition of oxidative phosphorylation in conjunction with an increase in respiration (6). However, when detached tobacco leaves were fumigated with 1.0 yl/1 ozone for 1-5 hr, the mitochondria extracted from the tissue prior to s3rmptom development exhibited reduced oxygen uptake and reduced oxidative phosphorylation (7 ). In an experiment of similar design when ozone was bubbled through a solution of isolated mitochondria, both respiration and oxidative phosphorylation were reduced (7 ). 

Solanesol was isolated from waste tobacco by extracting with hexane, treating with KOH, and purifying by chromatography on alumina with hexane/diethyl ether, 9 1. It is also obtainable from potato leaves. 

Supercritical fluid extraction processes are particularly appropriate for the separation and isolation of biochemicals where thermal decomposition, chemical modification, and physiologically-active solvents are undesirable. Examples of these bioseparations include the extraction of oils from seeds using carbon dioxide (1), of nicotine from tobacco using carbon dioxide-water mixtures (2), and of caffeine from coffee beans again using carbon dioxide-water mixtures (3). 

The most common use of scC02 is in the extraction of caffeine from coffee or tea, nicotine from tobacco, and essential oils from plants. The isolation of products is simple, with the evaporation of the solvent with no residue. Another important application is in supercritical fluid chromatography (SFC). 

The increased resistance of some primitive and wild strains of cotton to bollworms and tobacco budworms has been attributed to certain terpenoids. Some of these were isolated from flower bud extracts and have been identified as hemigossypolone (64), heliocide H2 (65), heliocide H3 (66), heliocide H, (67) and heliocide M (68). 

Phytuberin has also been found32 in tobacco leaves infiltrated with Pseudomonas Solanacearum. In addition phytuberin was isolated from Pseudomonas Solanacearum U-7. The methylene chloride extract from the callus contained phytuberin (55) and phytuberol (56). Both (55) and (56) were absent from healthy tissues. They were identified33 as stress compounds in leaves ofNocotianatabacum CV SamsunNN. sylvestris. 

In an important experiments Nirenberg and Matthaei, in 1961, isolated ribosomes from E. coli and mixed them with crude extracts of soluble materials, also from E. coli cells. The extracts included tRNA molecules and aminocyl-tRNA synthases. The 20 amino acids, ATP, and an ATP-generating system (PEP + pyruvate kinase) were added. Nirenberg showed that under such conditions protein was synthesized by ribosomes in response to the presence of added RNA. For example, RNA from tobacco mosaic virus (Chapter... 

In 1968, Schlotzhauer and Schmeltz (3465) noted that hexane extractables from tobacco constituted about 6% of the original tobacco weight, whereas the extracted tobacco residue constituted about 94% of the original tobacco weight. Pyrolysis of the hexane extractables and the extracted tobacco residne indicated that, of the total B[a]P determined in these two pyrolysates, abont 60% was found in the hexane extractables pyrolysate (6% of the original tobacco) and 40% in the pyrolysate of the extracted tobacco residue (94% of the original tobacco). Schlotzhaner and Schmeltz (3466) also demonstrated by pyrolysis of individual components isolated from the hexane extract that the following were precursors of the PAHs in the pyrolysates n-dotriacontane, stearic acid and linoleic acid, phytol, sqnalene, and P-sitosterol. 

Kaneko, H. and K. Ijichi The aroma of cigar tobacco. Part 1. Isolation of 2-hydroxy-2,6,6-trimethylcyclohexy-lidene-1-acetic acid lactone (dihydroactinidiolide) from ether extract of cigar leaves Agr. Biol. Chem. Japan 32 (1968) 1337-1340. 

Noma, M., N. Kawashima, and M. Noguchi Isolation and characterization of hydroxyvaleric acid derivatives from tobacco leaves and the hydrolysate of tobacco extract J. Biol. Chem. 41 (1977) 913-915. 

Hallard et al. (208) used SSS isolated from a transgenic tobacco cell line containing the Str gene for an assay of secologanin in plant material. Incubation with the crude enzyme, the plant extract, and tryptamine results... 

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