Chromophore distribution and ultraviolet visual pigment in the compound eyes of the Japanese fireflies Luciola cruciata and L. lateralis (Coleoptera, Lampyridae)

Based on electroretinogram measurements, fireflies are reported to possess two spectral-sensitivity peaks: one in the near-ultraviolet and the other in the green region. This suggests that at least two kinds of visual pigment should be present (Lall and Lloyd 1989). Analyses by high pressure liquid chromatography revealed that there are two kinds of chromophores, i.e., retinal (A1) and 3-hydroxyretinal (A3), in the compound eye of Japanese fireflies of the genus Luciola (Gleadall et␣al. 1989). Through selective light-adaptation experiments and in vitro analyses by high pressure liquid chromatography, it is demonstrated that the A1 is the chromophore of an ultraviolet-sensitive visual pigment and the A3 is the chromophore of a green-sensitive pigment. The absolute amounts of A1 and A3 were, respectively, 32.1 ± 3.9 and 111.1 ± 35.2 pmol in male, and 26.9 ± 6.9 and 63.4 ± 18.2 pmol in female (mean ± SD per animal). These results indicate that the amount of green-sensitive visual pigment in males is significantly higher than in females. A difference in the distribution of these chromophores is demonstrated in the eyes of female (but not male) fireflies: in the dorsal region of the female eye, the A1:A3 ratio is higher than that for the rest of the eye. Accepted: 28 February 1998     

Studies with general acyl-CoA dehydrogenase from pig kidney. Inactivation by a novel type of "suicide" inhibitor, 3,4-pentadienoyl-CoA

3,4-Pentadienoyl-CoA, an allenic substrate analog, is a potent inhibitor of the flavoprotein pig-kidney general acyl-CoA dehydrogenase. The analog reacts very rapidly (k = 2.4 X 10(3) min-1) with the native oxidized enzyme to form a covalent flavin adduct probably involving the isoalloxazine position N-5. This species is inactive, but activity may be regained by two pathways. The allenic thioester can be displaced (k = 0.3 min-1) by a large excess of octanoyl-CoA substrate upon reversal of covalent adduct formation. Alternatively, the enzyme inactivator adduct slowly decomposes (t1/2 = 75 min) to form the strongly thermodynamically favoured 2,4-diene and catalytically active, oxidized enzyme. During this latter process 15-20% of the activity is irreversibly lost probably due to covalent modification of the protein. These data suggest that 3,4-pentadienoyl-CoA should be considered a suicide substrate of the acyl-CoA dehydrogenase. The mechanism of the reactions, and in particular the 3,4----2,4 tautomerization, are consistent with a catalytic sequence initiated by abstraction of an alpha-hydrogen as a proton.     

Design of Peptides Using α,β-Dehydro-residues: Synthesis,Crystal Structure and Molecular Conformation of N-Boc-L-Val-ΔPhe–ΔPhe-L-Ala-OCH3

To obtain general rules of peptide design using α,β-dehydro-residues, a sequence with two consecutive ΔPhe-residues, Boc-L -Val-ΔPhe–ΔPhe- L -Ala-OCH₃, was synthesized by azlactone method in solution phase. The peptide was crystallized from its solution in an acetone/water mixture (70:30) in space group P6₁ with a=b=14.912(3) Å, c= 25.548(5) Å, V=4912.0(6) ų. The structure was determined by direct methods and refined by a full matrix least-squares procedure to an R value of 0.079 for 2891 observed [I?3σ(I)] reflections. The backbone torsion angles ?₁=?54(1)°, ψ₁= 129(1)°, ω₁=?177(1)°, ?₂ =57(1)°, ψ₂=15(1)°, ω₂ =?170(1)°, ?₃=80(1)°, ψ₃ =7(2)°, ω₃=?177(1)°, ?₄ =?108(1)° and ψ^T₄=?34 (1)° suggest that the peptide adopts a folded conformation with two overlapping β-turns of types II and III′. These turns are stabilized by two intramolecular hydrogen bonds between the CO of the Boc group and the NH of ΔPhe³ and the CO of Val¹ and the NH of Ala⁴. The torsion angles of ΔPhe² and ΔPhe³ side chains are similar and indicate that the two ΔPhe residues are essentially planar. The folded molecules form head-to- tail intermolecular hydrogen bonds giving rise to continuous helical columns which run parallel to the c-axis. This structure established the formation of two β-turns of types II and III′ respectively for sequences containing two consecutive ΔPhe residues at (i+2) and (i+3) positions with a branched β-carbon residue at one end of the tetrapeptide.     

Optical activity of polypeptides in the infrared. Predicted CD of the amide I and amide II bands.

The Miyazawa-Blout-Krimm (M-B-K) treatment of polypeptide absorption in the infrared is extended to the calculation of circular dichroism (CD), linear dichroism, and oriented CD for the amide I and amide II transitions. Matrix methods are applied to the α helix and β structures using measured values for the strengths and directions of the transition dipole moments and empirical values from M-B-K for the coupling constants. Relatively small aggregates, a 36-residue helix, and 8-chain × 4-residue β sheets, are large enough to show calculated absorption agreeing with M-B-K results, which are based on infinite lattices. In all cases the predicted CD is an approximately conservative couple. The strongest CD should appear in the α helix, Δε/ε ?± 10^?3 for both transitions. The amide II transition should show moderate CD couples in both β structures, Δε/ε ? (+2 to ?1) × 10^?4. The amide I transitions in β structures should show weak CD couples, Δε/ε = (+3 to ?2) × 10^?5, except that the negative branch in the antiparallel structure may be detectable (Δε/ε ? ?2 × 10^?4) because absorption is very low at its wavelength peak. CD on oriented samples should be enhanced over the unoriented cases, giving values as large as Δε/ε = 3 × 10^?3 because particular directions of observation allow the light to avoid much of the absorption in the sample. If all three structures are considered as helices, then the larger distance of the transition dipoles from the axis in the α helix, and the orientations of the transitions in the different structures, are the factors that, in terms of our previous theoretical work [Snir and Schellman (1973) J. Phys. Chem. 77 , 1653] satisfactorily explain the calculated results. Simple dipole–dipole interaction is calculated to make a substantial contribution to the coupling between groups.     

pH-dependent structural conformations of B-phycoerythrin from Porphyridium cruentum

B-phycoerythrin from the red alga Porphyridium?cruentum was crystallized using the technique of capillary counter-diffusion. Crystals belonging to the space group R3 with almost identical unit cell constants and diffracting to 1.85 and 1.70?? were obtained at pH values of 5 and 8, respectively. The most important difference between structures is the presence of the residue His88α in two different conformations at pH?8. This residue is placed next to the chromophore phycoerythrobilin PEB82α and the new conformation results in the relocation of the hydrogen-bond network and hydration around PEB82α, which probably contributes to the observed pH dependence of the optical spectrum associated with this chromophore. Comparison with the structures of B-phycoerythrin from other red algae shows differences in the conformation of the A-ring of the chromophore PEB139α. This conformational difference in B-phycoerythrin from P.?cruentum enables the formation of several hydrogen bonds that connect PEB139α with the chromophore PEB158β at the (αβ)(3) hexamer association interface. The possible influence of these structural differences on the optical spectrum and the ability of the protein to perform energy transfer are discussed, with the two pH-dependent conformations of His88α and PEB82α being proposed as representing critical structural features that are correlated with the pH dependence of the optical spectrum and transient optical states during energy transfer. DATABASE: Structural data have been deposited in the Protein Data Bank under accession numbers 3V58 and 3V57. STRUCTURED DIGITAL ABSTRACT: B-phycoerythrin beta?and?B-phycoerythrin alpha?bind?by?x-ray crystallography?(View interaction).     

Why 11-cis-Retinal?

The C₂₀ diterpenoid compound retinal is the chromophore of thevisual pigments the rhodopsins, and the pigments present inHalobacterium halobium, namely, bacteriorhodopsin (proton pump),halorhodopsin (chloride pump), and the sensory rhodopsins (phototaxisreceptor). In all cases, they are bound covalently to the receptorprotein by a protonated Schiff base. However, in rhodopsins,the retinal is the 11-cis isomer, whereas in H. halobium pigmentsit is the all-trans isomer. Why did Nature choose retinal asthe chromophore, and why 11-cis in some cases and all-transin other cases? Also why is the chromophore a protonated Schiffbase? These points are addressed after giving an outline ofthe current status of the various photoreceptor pigments     

An Alternative Pathway Mediates the Mouse and Human Cone Visual Cycle

One of the fundamental mysteries of the human visual system is the continuous function of cone photoreceptors in bright daylight. As visual pigment is destroyed, or bleached, by light [1], cones require its rapid regeneration, which in turn involves rapid recycling of the pigment's chromophore. The canonical visual cycle for rod and cone pigments involves recycling of their chromophore from all-trans retinol to 11-cis retinal in the pigment epithelium, adjacent to photoreceptors [2]. However, shortcomings of this pathway indicate the function of a second, cone-specific, mechanism for chromophore recycling [3]. Indeed, biochemical [3], [4], [5], [6] and [7] and physiological [8] studies on lower species have described a cone-specific visual cycle in addition to the long-known pigment epithelium pathway. Two important questions remain, however: what is the role of this pathway in the function of mammalian cones, and is it present in higher mammals, including humans? Here, we show that mouse, primate, and human neural retinas promote pigment regeneration and dark adaptation selectively in cones, but not in rods. This pathway supports rapid dark adaptation of mammalian cones and extends their dynamic range in background light independently of the pigment epithelium. This pigment-regeneration mechanism is essential for our daytime vision and appears to be evolutionarily conserved.     

Irreversible inactivation of mammalian Δ5-3β-hydroxysteroid dehydrogenases by 5, 10-secosteroids. Enzymatic oxidation of allenic alcohols to the corresponding allenic ketones

Novel 4,5-allenic 3β-hydroxy-5,10-secosteroids have been synthesized by sodium borohydride reduction of the corresponding conjugated allenic 3-oxo-5, 10-secosteroids. The secosteroid allenic alcohols are substrates for bovine adrenal and human placental Δ⁵-3β-hydroxysteroid dehydrogenases, and the resulting electrophilic conjugated allenic ketones are shown to inactivate these dehydrogenases in a time-dependent manner. Inactivated enzyme did not recover activity after filtration through Sephadex G-25. In contrast, the secosteroid allenic alcohols were not oxidized at C-3 by the bacterial 3β(and 17β)-hydroxysteroid dehydrogenase from $\text{P. testosteroni}$, nor did the corresponding allenic ketones inactivate this enzyme when incubated directly.     

Synthetic molecules designed as potential inhibitors in ecdysone biosynthesis

For fundamental studies as well as applied research, we have set up a programme for the synthesis of irreversible and selective inhibitors of ecdysone biosynthesis. Suicide substrate type inhibitors have been synthesized in order to react in the last steps of ecdysone biosynthesis on C-22 and C-25 hydroxylases which are cytochrome P-450-dependent monooxygenases. The most active inhibitors are formed of a steroid nucleus on which a short side chain, with an acetylenic or an allenic function closely linked to an hydroxyl group, has been grafted. The exact form of the steroid nucleus (as in cholesterol, 7-dehydrocholesterol, 3-dehydrocholesterol or a molecule with a saturated B cycle) did not appear essential for the activity of the chemicals. These molecules injectedin vivo in larvae ofLocusta migratoria induced some morphological modifications of the adults and a very slight delay in metamorphosis.

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Résumé Dans le but d'inhiber la biosynthèse de l'ecdysone au cours du développement chez l'Insecte, plus de 50 molécules ont été synthétisées selon la conception des substrats-suicides. Deux enzymes des dernières étapes de la biosynthèse de l'ecdysone, parfaitement connues, les C-22 et C-25 hydroxylases, qui sont des monooxygénases à cytochrome P-450, ont été choisies pour cibles. A partir d'une molécule de base présentant à la fois un bon pouvoir inhibiteur et des caractéristiques d'activité de type substrat-suicide, c'està-dire irréversibilité de l'action, spécificité de l'inhibition et sélectivité sur l'enzyme visé, de nombreuses modifications ont été introduites. Elles ont concernés notamment la longueur de la cha?ne latérale, la nature et la position du groupement inhibiteur, la position du ou des groupements hydroxyles et la nature du noyau stéro?de. Des divers résultats obtenus, il est possible de conclure que les meilleurs inhibiteurs, qui diminuent significativement la production d'ecdysone des glandes prothoraciques incubéesin vitro, sont composés d'une cha?ne latérale courte à groupement inhibiteur alcyne ou allénique dans le voisinage d'un groupement hydroxyle et d'un noyau stéro?de qui peut-être, indifféremment, de type cholestérol, 7-déshydrocholestérol, 3-déshydrocholestérol ou porter un cycle B saturé. On peut noter que les noyaux qui se rapprochent le plus de celui des ecdystéro?des diminuent ou empêchentl' activité. Malheureusement, jusqu'à présent, ces molécules injectéesin vivo chez des larves deLocusta migratoria n'ont qu'une action réduite: elles ne produisent, dans les meilleurs cas, que de rares retards de métamorphose et quelques modifications morphologiques chez les adultes.

     

The role of mutants in the search for the photoreceptor for phototropism in higher plants

Early attempts to identify the chromophore of the photoreceptor for phototropism are reviewed. Carotenoids and flavins were the principal candidates, but studies with grass coleoptiles devoid of carotenoids suggest that at least in these organs carotenoids are most unlikely to play that role. The status of characterization of a gene for a putative photoreceptor protein is also reviewed. As the action spectrum for phototropism resembles the absorption spectrum of a flavoprotein, flavoproteins are attractive candidates at present, especially since the CRY1 photoreceptor in Arabidopsis thaliana that mediates blue light-dependent hypocotyl growth suppression has flavin adenine dinucleotide as one of its two chromophores. As the second chromophore appears to be pterin, pterins should not be ruled out as candidate chromophores for the photoreceptor for phototropism.     

Preparation of η-1,3-dienyl, η-1,2-dienyl and η-cyclobutenone complexes via reactions of transition-metal carbonyl containing anions with allenic electrophiles

The preparation and reaction chemistry of 1,3- and 1,2-diene and related complexes derived from metal carbonyl containing anions and allenic electrophiles are addressed. The preparation of some CpFe(CO)₂ η¹-diene complexes and their conversion into CpFe(CO) η³-diene complexes is presented followed by reactions of CpMo(CO)₃⁻, CpW(CO)₃⁻ and CpMo(CO)₂PR₃⁻ anions with allenic electrophiles which produce metal complexed cyclobutenones (via CO and alkene insertions from the initially formed product) and 1,2-diene complexes, respectively. Lastly, the reactions of PPh₃(CO)₃Co⁻ anions with allenic electrophiles are outlined which result in several different coordination geometries depending on the reaction conditions used.     

Configurational effects in antibody–antigen interactions studied by microcalorimetry

In this paper we study the binding of two monoclonal antibodies, E3 and E8, to cytochrome c using high-sensitivity isothermal titration calorimetry. We combine the calorimetric results with empirical calculations which relate changes in heat capacity to changes in entropy which arise from the hydrophobic effect. The change in heat capacity for binding E3 is ?350 ± 60 cal K^?1 mol^?1 while for E8 it is ?165 ± 40 cal K^?1 mol^?1. This result indicates that the hydrophobic effect makes a much larger contribution for E3 than for E8. Since the total entropy change at 25°C is very similar for both antibodies, it follows that the configurational entropy cost for binding E3 is much larger than for binding E8 (?77 ± 15 vs. ?34 ± 11 cal K^?1 mol^?1). These results illustrate a case of entropy compensation in which the cost of restricting conformational degrees of freedom is to a large extent compensated by solvent release. We also show that the thermodynamic data can be used to make estimates of the surface area changes that occur upon binding. The results of the present study are consistent with previous hydrogen–deuterium exchange data, detected using 2D NMR, on the two antibody–antigen interactions. The NMR study indicated that protection from exchange is limited to the binding epitope for E8, but extends beyond the epitope for E3. These results were interpreted as suggesting that a larger surface area was buried on cytochrome c upon binding to E3 than to E8, and that larger changes in configurational entropy occur upon binding of E3 than E8. These findings are confirmed by the present study using isothermal titration calorimetry. © 1995 Wiley-Liss, Inc.     

Phototactic behaviour of Eucryptorrhynchus scrobiculatus and E. brandti (Coleoptera: Curculionidae) adults*

Eucryptorrhynchus scrobiculatus and E. brandti (Coleoptera: Curculionidae) are destructive weevils on Ailanthus altissima in China. This study examined phototactic behaviour of E. scrobiculatus and E. brandti in response to eight light-emitting diodes (LEDs) in the laboratory and field. Effects of gender, starvation, and light and dark experience on the phototactic behaviour of the insects were evaluated. The results demonstrated that, the two species of weevil were phototactic insects and most sensitive to violet light (400–405?nm), followed by blue–violet light (420–430?nm). They were less sensitive to red (655–660?nm), white (6000–6500?k), blue–green (470–480?nm), yellow (590–595?nm), blue (450–455?nm), and green (515–530?nm) light. In the light intensity range of 200–1000?lux, the light intensity had no significant effect on the phototatic behaviour of E. scrobiculatus and E. brandti. Phototactic behaviour of the insects was affected by gender. The phototaxis indices of the two species of weevil increased with starvation, reaching a plateau after 2 or 3?d of starvation. The phototaxis indices of E. scrobiculatus and E. brandti were significantly affected by various wavelengths of light following exposure to the light for 3?h or in different dark experience time. In the mark-release-recapture test, the number of E. scrobiculatus and E. brandti adults trapped by violet (400–405?nm) light traps is the largest. The information provided here provides a basis for survey and control of E. scrobiculatus and E. brandti.     

Haem oxygenase: A functionally diverse enzyme of photosynthetic organisms and its role in phytochrome chromophore biosynthesis,cellular signalling and defence mechanisms

Haem oxygenase (HO) is a universal enzyme that catalyses stereospecific cleavage of haem to BV IX α and liberates Fe⁺² ion and CO as by‐product. Beside haem degradation, it has important functions in plants that include cellular defence, stomatal regulation, iron mobilization, phytochrome chromophore synthesis, and lateral root formation. Phytochromes are an extended family of photoreceptors with a molecular mass of 250 kDa and occur as a dimer made up of 2 equivalent subunits of 125 kDa each. Each subunit is made of two components: the chromophore, a light‐capturing pigment molecule and the apoprotein. Biosynthesis of phytochrome (phy) chromophore includes the oxidative splitting of haem to biliverdin IX by an enzyme HO, which is the decisive step in the biosynthesis. In photosynthetic organisms, BVα is reduced to 3Z PΦB by a ferredoxin‐dependent PΦB synthase that finally isomerised to PΦB. The synthesized PΦB assembles with the phytochrome apoprotein in the cytoplasm to generate holophytochrome. Thus, necessary for photomorphogenesis in plants, which has confirmed from the genetic studies, conducted on Arabidopsis thaliana and pea. Besides the phytochrome chromophore synthesis, the review also emphasises on the current advances conducted in plant HO implying its developmental and defensive role.     

Aggregation‐Induced Emission of 1,8‐Naphthalimide?Casein Micelle: Investigation by Synchronous Spectrographic Method

A novel 1,8‐naphthalimide probe 1 , bearing two acetic‐acid moieties was synthesized. The acetic‐acid groups, docked into the sub‐domains of casein micelle and bound with tryptophan residues, and the 1,8‐naphthalimide chromophore adsorbed on the surface of casein micelle, forming a supermolecule, 1 ? casein micelle, which exhibited the aggregation‐induced synchronous emission (AISE) characters. The effect of pH on the intensity of supermolecule was investigated, and the result indicated that the emission enhancement was mainly due to the 1,8‐naphthalimide chromophore aggregated onto the casein micelle. Based on AISE, a novel casein quantification method was developed, which exhibited a good linear range of 0.05–10.0 μg ml^?1 and 0.07–9.5 μg ml^?1 with the detection limits of 2.8 and 3.0 ng ml^?1. The effects of metal ions and pH on the system of 1 ? casein micelle were investigated. The proposed method was applied to determine casein in milk samples, and the results were in good agreement with the result of the Biuret method.     

The Synthesis of Pyrazol0[3,4-c]Pyridine C-Nucleosides

Abstract

The synthesis of two members of the pyrazolo[3,4-c]pyridine C-nucleoside family is accomplished utilizing the 1.3-dipolar cycloaddition reaction of a diazo-ribofuranosyl derivative and an appropriate allenic diester.     

Adaptation to different climates results in divergent phenotypic plasticity of wing size and shape in an invasive drosophilid

The phenotypic plasticity of wing size and wing shape of Zaprionus indianus was investigated in relation to growth temperature (17°C to 31°C) in two natural populations living under different climates, equatorial and subtropical. The two populations were clearly distinguished not only by their wing size (the populations from the colder climate being bigger in size), but also by the shape of the response curves to growth temperature i.e., their reaction norms. In this respect, the temperature at which the size of the wing was maximum was about 3°C higher in the equatorial population. Such a difference in size plasticity is already found in two other nonclosely related species, might be a general evolutionary pattern in drosophilids. Wing shape was investigated by calculating an ellipse included into the wing blade, then by considering the ratio of the two axes, and also by analysing the angular position of 10 wing-vein landmarks. For an overall shape index (ratio of the two axes of the ellipse), a regular and almost linear increase was observed with increasing temperature i.e., a more round shape at high temperatures. Wing shape was also analysed by considering the variations of the various angles according to temperature. A diversity of response curves was observed, revealing either a monotonous increase or decrease with increasing temperature, and sometimes a bell shape curve. An interesting conclusion is that, in most cases, a significant difference was observed between the two populations, and the difference was more pronounced at low temperatures. These angular variations are difficult to interpret in an evolutionary context. More comparative studies should be undertaken before reaching some general conclusions.     

Photo-induced DNA cleavage reaction characteristics of propargylic sulfones possessing anthraquinone chromophore

DNA cleavage potency of propargylic sulfones possessing anthraquinone chromophore 1 under UV-irradiation was evaluated in comparison with the dark reaction. 1 showed inefficient DNA cleavage activity, while having considerably strong DNA binding ability. This result is accounted for by spatial conditions that the activated alkylating allenic site of intercalated 1 could not effectively approach to DNA bases, most probably guanine moiety, and thereby led to insufficient DNA strand cleavage. In contrast, the DNA cleavage activity of 1 was notably enhanced upon UV-irradiation (lambda(ex)=365 nm) followed by incubation. Under UV-irradiation, further DNA cleavage were occurred primary at 5'-G of GG steps within DNA. A DNA cleavage mechanism for 1, by which photo-induced one-electron oxidation of 5'-G of GG steps may occur along with ordinary alkylation, has been proposed.     

A pilot study to assess the effect of acute exercise on brain glutathione

The brain is highly susceptible to oxidative stress due to its high metabolic demand. Increased oxidative stress and depletion of glutathione (GSH) are observed with aging and many neurological diseases. Exercise training has the potential to reduce oxidative stress in the brain. In this study, nine healthy sedentary males (aged 25?±?4 years) undertook a bout of continuous moderate intensity exercise and a high-intensity interval (HII) exercise bout on separate days. GSH concentration in the anterior cingulate was assessed by magnetic resonance spectroscopy (MRS) in four participants, before and after exercise. This was a pilot study to evaluate the ability of the MRS method to detect exercise-induced changes in brain GSH in humans for the first time. MRS is a non-invasive method based on nuclear magnetic resonance, which enables the quantification of metabolites, such as GSH, in the human brain in vivo. To add context to brain GSH data, other markers of oxidative stress were also assessed in the periphery (in blood) at three time points [pre-, immediately post-, and post (～1?hour)-exercise]. Moderate exercise caused a significant decrease in brain GSH from 2.12?±?0.64?mM/kg to 1.26?±?0.36?mM/kg (p?=?.04). Blood GSH levels increased immediately post-HII exercise, 580?±?101?µM to 692?±?102 µM (n?=?9, p?=?.006). The findings from this study show that brain GSH is altered in response to acute moderate exercise, suggesting that exercise may stimulate an adaptive response in the brain. Due to the challenges in MRS methodology, this pilot study should be followed up with a larger exercise intervention trial.     

Simple ITC method for activity and inhibition studies on human salivary α-amylase

Isothermal titration calorimetry (ITC) has an increasing significance in enzyme kinetic studies owing to its general applicability and sensitivity. In the present work, we aimed at developing a simple ITC-based screening procedure for the measurement of human salivary α-amylase (HSA) activity. Reaction of two substrates was studied with three independent methods (ITC, HPLC and spectrophotometry). ITC experiments were made using free and chromophore-containing maltooligomers of different length as substrates. Detailed studies revealed that maltoheptaose or longer oligomers could model properly starch and the presence of aromatic chromophore group did not affect the K_M values considerably. It is the first time, when ITC was used to investigate of HSA-catalysed hydrolysis of different substrates (2-chloro-4-nitrophenyl-4-O-α-D-galactopyranosyl-maltoside, maltoheptaose and starch) in the presence of acarbose inhibitor. All measured IC₅₀ values are in micromolar range (0.9, 18.6 and 29.0?μM, respectively) and increased in parallel with the degree of polymerisation of substrates.