The Aldehyde Content of Luminous Bacteria and of an “Aldehydeless” Dark Mutant

Abstract

Fatty aldehydes, present in the luminescent cells of Photobacterium phosphoreum and Achromobacterfischeri, and td a very slight extent in the cells of a visually dark, "aldehydeless" mutant of the latter species, were extracted, purified, and oxidized to the corresponding acids. The acids were analyzed by mass spectrometry. The results, in Cell-free extracts of lumitous bacteria that produced an NADH-activated luminescence were reported in 1953 (1). Evidence soon followed that the system required FMN (2) and, in addition, a factor "KCF" obtained from kidney cortex (3), subsequently identified as hexadecanal (4). This aldehyde could be replaced by any one in a series of saturated, straightchain aldehydes having chain lengths from C7 to C16 with varying degrees of effectiveness (5). Until now, however, no evidence has been reported that such aldehydes are specifically required for luminescence of the bacterial system in vivo. In fact, the specific requirement of an aldehyde is questionable on the grounds that a terminal nitrite group can satisfactorily replace the terminal aldehyde group (6).Recent evidence has shown that the above type of aldehydes of chain lengths from C9 to C14 are oxidized to the corresponding acids by the bacterial luminescent system in vitro (7-9), with a quantum yield for each aldehyde amounting to 0.17 0.01 photons per aldehyde molecule reacted (7). Suggestive evidence that aldehydes function in luminescence also in vivo has been available in the fact that certain dark mutants, which seem identical to the parent strain except for failing to emit visible luminescence, respond practically at once by emitting a bright glow of light when a suitable aldehyde is added to the dark cells (10). Moreover, an aldehydogenic compotond in extracts of Photobacterium phosphoreum has been described (11).The present study demonstrates the presence of saturated fatty aldehydes in luminescent bacterial cells of Achromo- During extraction and purification of aldehydes from the bacteria, assays were made, to monitor the procedure at each step, on the basis of the amount of light emitted with a bacterial luciferase preparation from A. fischeri. To a specimen containing aldehyde for assay, a combination of the luciferase preparation, FMN, and NADH was added, and the total luminescence was measured under conditions described (see explanation of Fig. 2, p. 2088, ref. 7).Preparation of Bacterial Cells. Each species Was cultivated on the surface of hard agar, consisting of 30 g of Difco BactoNutrient Agar Dehydrated per liter of 3% (w/v) NaCl, plus 10 ml of glycerol, plus 5 g of powdered CaCO3, autoclaved, and aseptically poured into sterile aluminum baking pans with sheet aluminum covers, or into sterile fiberglass-type pans with covers, to a depth of about 0.5 inch (1.3 cm) (the deeper than usual medium favors better growth and luminescence).A total surface area of 1.3 m2 -was inoculated for each extraction. The growth of A. fischeri was harvested after incubation for 24 hr at 250, and that of P. phosp...

Evidence for tetradecanal as the natural aldehyde in bacterial bioluminescence.

“…Although the results presented are for B. harveyi, similar observations in vivo indicate that the conclusions are also applicable for Photobacterium fischeri (8) and P. leiognathi. The small quantities of aldehyde that could be isolated from two species of luminous bacteria were mostly of chain lengths 12, 14, and 16 carbons, in relative abundance of 5, 63, and 30%, respectively, in one species and 36, 32, and 20% in the other (14). It is also interesting that with luciferase isolated from all three Photobacterium species the rate of the reaction is much greater with tetradecanal than with aldehydes of other chain lengths, especially the naturally occurring even-numbered ones (ref.…”

“…The in vitro reaction involves the mixed function oxidation of FMNH2 and a long-chain aliphatic aldehyde to give the corresponding acid and light emission. Pure Beneckea harveyi luciferase is not highly specific with regard to aldehyde chain length (8)(9)(10)(11)(12)(13)(14) carbons), even though. some distinctions can be made, especially with luciferases from different bacterial species (3,4).…”

Induction of biochemical active constituents of Jojoba (Simmondsia chinensis (Link) Schneider) callus affected by hormones

“…Aristolene with the following biological activity: has a smell that attracts predators and wolves (Scrivner et al, 1984), is an antibiotic bacterium, Bacillus, and is used for throat congestion (Zour), it is anti-inflammatory, the cradle of the nerves (Tian-Shung et al, 2004), pain reliever and has luminous bacteria (Shimomura et al, 1974); 4. 7-Hexadecenal with the following biological activity: used for treating patients with diabetes, by increasing the secretion of insulin in the body (Turgumbayeva et al, 2015), it is found in the liver of sheep and thus attractive to wolves and used in hunting wild animals (Scrivner et al, 1984), has Luminous Bacteria "Aldehydeless" Dark Mutant (Shimomura et al, 1974); 5. Tricosene which has kairomone effect (Ananthakrishnan et al, 1991).…”

“…These major active constituents are divided into five fractions: 1. 1-Hexadecanol which has the following biological activity: a composite medium used in the synthesis of other compounds (Turgumbayeva et al, 2015), has flavors and smells like composite simmondsin (Scrivner et al, 1984) and has luminous bacteria (Shimomura et al, 1974); 2. nonadecatriene compound acting as pheromone that attracts male insects (Nilsson, 2009); 3. Aristolene with the following biological activity: has a smell that attracts predators and wolves (Scrivner et al, 1984), is an antibiotic bacterium, Bacillus, and is used for throat congestion (Zour), it is anti-inflammatory, the cradle of the nerves (Tian-Shung et al, 2004), pain reliever and has luminous bacteria (Shimomura et al, 1974); 4.…”

“…1-Hexadecanol which has the following biological activity: a composite medium used in the synthesis of other compounds (Turgumbayeva et al, 2015), has flavors and smells like composite simmondsin (Scrivner et al, 1984) and has luminous bacteria (Shimomura et al, 1974); 2. nonadecatriene compound acting as pheromone that attracts male insects (Nilsson, 2009); 3. Aristolene with the following biological activity: has a smell that attracts predators and wolves (Scrivner et al, 1984), is an antibiotic bacterium, Bacillus, and is used for throat congestion (Zour), it is anti-inflammatory, the cradle of the nerves (Tian-Shung et al, 2004), pain reliever and has luminous bacteria (Shimomura et al, 1974); 4. 7-Hexadecenal with the following biological activity: used for treating patients with diabetes, by increasing the secretion of insulin in the body (Turgumbayeva et al, 2015), it is found in the liver of sheep and thus attractive to wolves and used in hunting wild animals (Scrivner et al, 1984), has Luminous Bacteria "Aldehydeless" Dark Mutant (Shimomura et al, 1974); 5.…”