In the past there has been emphasis on the view that bioluminescent systems exhibit considerable diversity, both biologically and chemically [55]. While this certainly remains so, evidence is developing for interesting interand intra-phyletic similarities, especially in the structure of luciferins and the chemiluminescent reaction mechanisms. Bacteria. Although the reaction mechanism in bacteria is distinctly different from other well characterized systems, cross phyletic relationships between bacteria and other forms do occur. Luminous bacteria 'inhabit' other phyla; they are known to occur as symbionts in fish and other higher organisms [55), and thereby provide the host organism with a light source which may be used for one or more specific functions. The flashlight fish (Photohlepharon palpebratus), which possesses a pair of light organs situated just beneath the eyes, is a recently studied example [93]. (These fish are now on exhibit a t the Steinhart Aquarium in San Francisco.) Leiognathid fish, with internal luminous organs, have also been investigated [15,57, 581. The ability to luminesce occurs in several groups of marine bacteria, whose systematic relationships to each other and to nonluminous marine bacteria were described in detail by Reichelt and Baumann [ 105,1061. Luminous bacteria which are symbiotic fall within the genus Photobacteriurn; bacteria in the Beneckea genus have not been found in a symbiotic relationship and are generally isolated from seawater (K. Nealson and J. W. Hastings, unpublished). A more specific habitat for and function of the luminescence in the Beneckea species have not yet been uncovered. Insight concerning the function of the light emission is emerging from studies of the control of the synthesis of the luminescent system. In both Photobacterium and Beneckea species, luciferase is not synthesized constitutively [58, 951. A substance (termed autoinducer) synthesized by the cells themselves accumulates in the medium and results in the induction of luciferase synthesis [41]. By radioactive labeling, 5 peptides have been isolated which are specifically synthesized during the induction period [%I. From studies of mutants which require exogenous aldehyde for luminescence (see below), it was concluded that at least two genes are specifically involved in the in t.im biosynthesis of aldehyde factor for luminescence and that both genes are coordinately controlled with that for luciferase [29]. Luciferase synthesis is also subject to repression by glucose, reversible by cyclic

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1751-1097.1976.tb07282.x

Bioluminescence and chemiluminescence

Nature uses mechanochemical transduction processes to achieve diverse and vital functions, such as hearing, cellular adhesion and gating of ion channels. One fascinating example of biological mechanotransduction is the emission of light on mechanical stimulation. However, molecular-level transduction of force into luminescence in a synthetic system remains a challenge. Here, we show that bis(adamantyl)-1,2-dioxetane emits visible light when force is applied to a polymer chain or network in which this unit is incorporated. Bright-blue luminescence was observed on sonication of solutions of dioxetane-containing linear polymers and on the straining of polymer networks with dioxetane crosslinkers. Light is emitted from the adamantanone-excited state that forms on opening of the four-membered dioxetane ring. Increased sensitivity and colour tuning were achieved by energy transfer to suitable acceptors. High spatial and temporal resolutions highlight the potential to study the failure of polymeric materials in unprecedented detail.

Mechanically induced chemiluminescence from polymers incorporating a 1,2-dioxetane unit in the main chain

Chapter 225 – Rare-earth beta-diketonates

DNA probes and synthetic oligonucleotides in general present one of the key tools in modern molecular biology research and increasingly also in commercial applications. Along with the many applications that have been developed for and with DNA probes, faster and more sensitive detection methods are being developed. One of the most promising recent developments presents a method based on enzymatically triggered chemiluminescence. Details of this chemistry along with applications in molecular biology and immunology will be discussed and compared to conventional methods.

Applications of dioxetane chemiluminescent probes to molecular biology

We have synthesized and studied two 1,2-dioxetane-based chemiluminescent enzyme substrates: 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl- 1,2-dioxetane (AMPPD), and, 3-(2'-spiroadamantane)-4-methoxy-4-(3"-beta-D'-galactopyrano -yloxy)phenyl-1,2- dioxetane (AMPGD), which can be activated to chemiluminescence at 470 nm by alkaline phosphatase and beta-D-galactosidase, respectively. In addition, we observed that certain macromolecules enhance the luminescence of AMPPD. For example, the addition of 0.1% bovine serum albumin amplifies the luminescent signal and improves the detection limit for alkaline phosphatase by approximately one order of magnitude under certain conditions. This effect is due to the presence of a hydrophobic microenvironment provided by the enhancer which 'stabilizes' the dephosphorylated AMPPD emitter. Alkaline phosphatase catalysed chemiluminescence from AMPPD is constant for a prolonged period of time. Using AMPPD we were able to detect 0.01 attomole quantities of alkaline phosphatase immobilized on membrane supports and imaged on photographic film and, in solution, measured in a luminometer. AMPPD and AMPGD offer alternatives to colorimetric and fluorescent substrates for alkaline phosphatase and beta-D-galactosidase labels used in enzyme immunoassays. The simplicity and sensitivity of this chemiluminescent readout allowed the development of rapid clinical assays (e.g. beta-hCG, LH, TSH and others).

1,2-Dioxetanes: Novel chemiluminescent enzyme substrates. Applications to immunoassays

Adamantylideneadamantane-1,2-dioxetane. Chemiluminescence and decomposition kinetics of an unusually stable 1,2-dioxetane

Indirect chemiluminescence by 1,2-dioxetanes. Evaluation of triplet-singlet excitation efficiencies. Long range singlet-singlet energy transfer and an efficient triplet-singlet energy transfer

(1‐Adamantyl)‐α‐peroxylactone: Synthesis, Kinetics, and Luminescence

Unter Lichteinwirkung zerfallt Tetramethyl-1,2-dioxetan (1) in zwei Molekule Aceton mit einer Quantenausbeute Φ ≥ 1. Licht der Wellenlange λ ≤ 300 nm fuhrt in einer adiabatischen Photoreaktion mit hoher (quantitativer) Ausbeute zu Aceton im elektronisch angeregten Zustand, bestehend aus 77 % Triplett- und 23 % Singulett-Molekulen. Langerwelliges Licht vermindert die Gesamtausbeute an elektronisch angeregten Molekulen, begunstigt jedoch die Entstehung des Triplett-Zustands. Die sensibilisierte Zersetzung lauft nach einem Quanten-Kettenmechanismus ab und ist eine nahezu quantitative adiabatische Photoreaktion innerhalb des Triplett-Zustands. Der Nachweis der elektronisch angeregten Fragmente gelingt anhand ihrer Emission und durch chemische Titration. – Mit Hilfe der „Einzel-Photonen-Zahlung” ergibt sich die Lebensdauer einer moglichen Diradikal-Zwischenstufe als τ ≤ 10−10 s.



Photofragmentation of Tetramethyl-1,2-dioxetane. An Efficient Example of an Adiabatic Photoreaction



On irradition tetramethyl-1,2-dioxetane (1) cleaves into two molecules of acetone with a quantum yield of Φ ≥ 1. In an adiabatic photoreaction of high (quantitative) yield light of wave-lengths λ ≤ 300 nm produces electronically excited acetone, consisting of 77 % triplet and 23 % singlet state. Increasing of the wave-length of irradiation (λ ≤ 300 nm) diminishes the yield of excited states, but increases the share of acetone triplets. The sensitized cleavage follows a quantum-chain process and constitutes a nearly quantitative adiabatic photoreaction in the triplet manifold. The electronically excited fragments are detected by their emission and by chemical titration. – Application of „single-photon-counting” allows to limit the lifetime of a possible diradical intermediate to τ ≤ 10−10 s.

Photofragmentierung von Tetramethyl-1,2-dioxetan Ein effizientes Beispiel einer adiabatischen Photoreaktion

Aktivierte Chemilumineszenz von Tetramethyl-1,2-dioxetan; Ermittlung von Geschwindigkeitskonstanten und ein neuer Weg zur Bestimmung von Triplett/Singulett-Anregungsausbeuten bei der Thermolyse von 1,2-Dioxetanen

Hans-Christian Steinmetzer

Papers

Adamantylideneadamantane-1,2-dioxetane. Chemiluminescence and decomposition kinetics of an unusually stable 1,2-dioxetane

Indirect chemiluminescence by 1,2-dioxetanes. Evaluation of triplet-singlet excitation efficiencies. Long range singlet-singlet energy transfer and an efficient triplet-singlet energy transfer

(1‐Adamantyl)‐α‐peroxylactone: Synthesis, Kinetics, and Luminescence

Photofragmentierung von Tetramethyl-1,2-dioxetan Ein effizientes Beispiel einer adiabatischen Photoreaktion

Aktivierte Chemilumineszenz von Tetramethyl-1,2-dioxetan; Ermittlung von Geschwindigkeitskonstanten und ein neuer Weg zur Bestimmung von Triplett/Singulett-Anregungsausbeuten bei der Thermolyse von 1,2-Dioxetanen