Antitussive and immunomodulating activities of instant coffee arabinogalactan-protein

A low molecular mass arabinogalactan-protein (AGP) composed of galactose and arabinose with a low protein content, isolated from the instant coffee powder of Coffea arabica beans, has been tested on antitussive (in vivo) and immunomodulating (ex vivo) activities. The results of antitussive tests revealed a significant dose dependant cough-suppressive effect of coffee AGP. It was observed 30 or 60 min after AGP administration and its efficacy lasted during the entire experiment course. Immunological tests showed that AGP affected some mediators of immunocompetent cells of immune system as TNF-α, IFN-γ and IL-2 cytokines. It seems that coffee AGP is a good inductor of both pro-inflammatory cytokines TNF-α and IFN-γ, however, less potent in TNF-α induction in comparison with that of β-d-glucan. Evident induction of TNF-α, IL-2 and IFN-γ cytokines, pro-TH1 polarization supports our conclusion about bio-immunological efficacy of AGP with an emphasis on the cellular immunity.     

Implication of hydrogen peroxide in the mutagenicity of coffee

A cup of instant coffee (150 ml) of normal strength (15 mg/ml) was found to contain about 500 and 750 micrograms of hydrogen peroxide soon after its preparation at 37 degrees C and 80 degrees C, respectively, but the concentration of hydrogen peroxide in the coffee increased with time for up to 24 h after its preparation. Thus coffee contains a hydrogen peroxide generating system. As extracts of green coffee beans were found to have very low capacity to generate hydrogen peroxide, this generating system is produced by roasting coffee beans. Hydrogen peroxide itself was only weakly mutagenic to Salmonella typhimurium TA100, but in the presence of methylglyoxal, which is also present as a mutagenic component in coffee, hydrogen peroxide showed strong mutagenicity. Hydrogen peroxide and methylglyoxal seem to be responsible for most of the mutagenicity of instant coffee.     

Mutagenicity of instant coffee on cultured Chinese hamster lung cells

Coffee showed mutagenic activity in cultured Chinese hamster lung (CHL) cells as assessed by using diphtheria toxin resistance as a selective marker. Most of the mutagenicity was suppressed in the presence of sodium bisulfite. The contribution of methylglyoxal to the total mutagenicity of coffee was less than 3%.     

Biosynthesis of plant hormones during anaerobic digestion of instant coffee waste

A large amount of solid waste remains after the production of instant coffee. This waste has to be moved to dumps, where it poses a threat of environmental pollution. Treatment of this waste by anaerobic methanogenic thermophilic digestion produced, besides biogas, a digested slurry which was used as a growth medium for horticulture, and proved to be a suitable and economical substitute for peat moss. Biological tests with mung bean cuttings and Grevillea plantlets showed promotional effects on rooting of the slurry and its sieved fraction extract, washed with water (Capul). Green coffee beans, instant coffee waste, its anaerobically-digested slurry and Capul were extracted by various methods and the extracts were analyzed by TLC, HPLC and GC/MS. Examinations showed clearly the presence of IAA and IBA in free and bound forms in all the substrates. The values of free and bound IAA were calculated by use of an internal standard and GC/MS. The amount of conjugated IAA was found to be much higher than that of free IAA, in both the coffee beans and instant coffee waste (11.1 vs 2.7 nmol g^–1, respectively). In the digested slurry and Capul, however, most of the IAA was present as the free form and was approximately 23.5–33.0 nmol g^–1, which is almost ten times more than in the waste, and almost twice the total amount of IAA in coffee beans. It is postulated that the high levels of free IAA in the digested instant coffee waste are a result of catabolism of tryptophan by anaerobic bacteria.     

Structure of arabinogalactan oligosaccharides derived from arabinogalactan-protein of Coffea arabica instant coffee powder

Arabinogalactan-protein, previously isolated from instant coffee powder of Coffea arabica, has been subjected to partial mild acidic and enzymatic hydrolyses. Separation of obtained mixtures by size exclusion and HPLC chromatographies afforded series of oligosaccharides, structure of which were studied by NMR spectroscopy. Mild acidic hydrolysis afforded oligosaccharides without any αAraf substituent while after enzymatic hydrolysis αAraf was found in di-, tri-, and tetrasaccharides. In all cases αAraf was a terminal substituent linked separately to O3, O6, and to both, O3 and O6, of βGal residues. Identification of di-, tri-, and tetrasaccharides containing α-Araf enabled to distinguish in the ¹H NMR spectra αAraf signals linked to O6 and O3 of neighboring βGal unit. Composition of polymeric residues after enzymatic and mild acidic hydrolyses was also analyzed.     

Mutagenicity of instant coffee and its interaction with dimethylnitrosamine in the micronucleus test

The mutagenicity of instant coffee and its interaction with dimethylnitrosamine (DMN) were examined in mice using the micronucleus test. Although neither a single nor multiple administration of instant coffee by gavage induced a significant rise in micronucleated cells over the dose range tested (100-2500 mg/kg), there was a tendency for the number of micronucleated cells to increase in a dose-related fashion. When coffee was administered with DMN, the difference in the frequency of micronucleated cells was small in comparison to a single treatment with DMN alone, thus indicating a lack of synergism between coffee and DMN.     

A note on the nutritive value of dried instant coffee residue for broiler chickens and turkey poults

Diets containing progressively higher levels of instant coffee residue (ICR) (from 0 to 15% in 3% increments) were given for 11 days to 8-day-old chicks and turkey poults. A total collection of faeces was carried out for the last 4 days of the trial. Preliminary performance data indicated that liveweight gain (LWG), food intake (FI) and food conversion ratio (FCR) all deteriorated with progressively higher levels of ICR, although the decline was less evident in turkey poults. Values obtained for apparent metabolisable energy of ICR (fresh weight basis) were 2.36 and 9.94 MJ kg⁻¹ for broiler chicks and turkey poults, respectively, and although data for ether extract digestibility and nitrogen retention confirmed this species difference, there was no effect of species on acid detergent fibre digestibility. It was concluded that there is potential for the use of ICR in turkey diets providing problems of odour and texture can be overcome.     

Study of the causes of direct-acting mutagenicity in coffee and tea using the Ara test in Salmonella typhimurium

The mutagenic activities of 6 of the chemicals identified in coffee solutions were assayed with the Salmonella Ara test, under experimental conditions optimized for coffee mutagenicity. Caffeine was the only non-mutagenic compound. Among the other 5 chemicals, hydrogen peroxide was the strongest mutagen and chlorogenic acid the weakest; methylglyoxal, glyoxal and caffeic acid exhibited intermediate mutagenicities. The minimal mutagenic doses of these components correlated negatively with their relative concentrations in coffee. It was concluded that chlorogenic acid, caffeic acid, glyoxal and methylglyoxal cannot contribute alone to the mutagenicity of coffee in the Ara test, since their minimal mutagenic concentrations were much higher than their respective levels in the coffee samples assayed. By contrast, 40-60% of the mutagenic activity in coffee and also in tea could be attributed to their H2O2 contents. Catalase abolished more than 95% of the mutagenic activity of coffee, as detected by the Ara test. A similar sensitivity to catalase has been reported by other authors in relation to the coffee mutagenicity identified by the Salmonella His test. Nevertheless, the results presented in this paper suggest that the Ara forward and the His reverse mutation tests are sensitive to the mutagenicity of different constituents in coffee solutions. We propose that the His test, sensitive at high coffee doses, mainly recognizes the mutagenicity of methylglyoxal, whilst the Ara test, sensitive at low coffee doses, mainly detects the mutagenic activity of hydrogen peroxide. The data reported also suggest that the direct-acting mutagenicity(ies) detected by the Ara test in tea solutions is (are) based on similar, if not identical, mechanisms.     

A coronary vasoconstrictor substance is present in regular and “decaffeinated” forms of both percolated and instant coffee

Coffee contains a previously unrecognized vasoactive material with potent constrictor action on the coronary arteries of the heart when tested in two types of preparations. Contractions of beef coronary vessel strips developed in response to both regular and caffeine-reduced forms of coffee (0.02 – 1.0 ml/15 ml Krebs solution), and they were specifically blocked by the cholinergic antagonist atropine. A perfused slab of beef left ventricle also showed a vasoconstrictor response to coffee which was antagonized by atropine. In contrast, tea, in comparable volumes to those of coffee elicited only vascular relaxation, as did the methylxanthine caffeine. The possibility should be considered that the cholinomimetic substance described here is linked to the reported increased incidence of myocardial infarction in coffee but not in tea drinkers.     

A structural basis for the biosynthesis of the major chlorogenic acids found in coffee

Chlorogenic acids (CGAs) are a group of phenolic secondary metabolites produced by certain plant species and an important component of coffee (Coffea spp.). The CGAs have been implicated in biotic and abiotic stress responses, while the related shikimate esters are key intermediates for lignin biosynthesis. Here, two hydroxycinnamoyl-coenzyme A shikimate/quinate hydroxycinnamoyl transferases (HCT/HQT) from coffee were biochemically characterized. We show, to our knowledge for the first time, that in vitro, HCT is capable of synthesizing the 3,5-O-dicaffeoylquinic acid diester, a major constituent of the immature coffee grain. In order to further understand the substrate specificity and catalytic mechanism of the HCT/HQT, we performed structural and mutagenesis studies of HCT. The three-dimensional structure of a native HCT and a proteolytically stable lysine mutant enabled the identification of important residues involved in substrate specificity and catalysis. Site-directed mutagenesis confirmed the role of residues leucine-400 and phenylalanine-402 in substrate specificity and of histidine-153 and the valine-31 to proline-37 loop in catalysis. In addition, the histidine-154-asparagine mutant was observed to produce 4-fold more dichlorogenic acids compared with the native protein. These data provide, to our knowledge, the first structural characterization of a HCT and, in conjunction with the biochemical and mutagenesis studies presented here, delineate the underlying molecular-level determinants for substrate specificity and catalysis. This work has potential applications in fine-tuning the levels of shikimate and quinate esters (CGAs including dichlorogenic acids) in different plant species in order to generate reduced or elevated levels of the desired target compounds.     

Fluroxene mutagenicity

The commercially available volatile anesthetic fluroxene (2,2,2-trifluoroethyl vinyl ether) which contains the stabilizer N-phenyl-1-napthylamine, was tested for mutagenicity using four strains of S. typhimurium, TA1535, TA1537, TA98 and TA100, and one strain of E. coli, WP2. In addition, purified fluroxene; N-phenyl-1-napthylamine; trifluoroethanol, a major metabolite of fluoroxene; and urine from rats anesthetized with fluroxene were tested. Several procedures were utilized including exposure of bacteria to vapor in desiccators and in liquid suspension. Results indicate that fluroxene, but not its stabilizer, was mutagenic to strains TA1535, TA100 and WP2 only in liquid suspension and only in the presence of a rat-liver enzyme system. Trifluoroethanol and urine from fluroxene-treated rat were not mutagenic to any strain of bacteria. These findings indicate that fluroxene is a promutagen which requires preincubation before it is recognized. Further experiments were performed with enzymes prepared from mouse, hamster and human liver. Fluroxene was mutagenic only in the presence of enzymes prepared from Aroclor 1254 pretreated rodents. Since fluroxene was not mutagenic in the presence of enzymes prepared from three human livers, the significance of these findings to man are unclear.     

Biotechnological potential of coffee pulp and coffee husk for bioprocesses

Advances in industrial biotechnology offer potential opportunities for economic utilization of agro-industrial residues such as coffee pulp and coffee husk. Coffee pulp or husk is a fibrous mucilagenous material (sub-product) obtained during the processing of coffee cherries by wet or dry process, respectively. Coffee pulp/husk contains some amount of caffeine and tannins, which makes it toxic in nature, resulting the disposal problem. However, it is rich in organic nature, which makes it an ideal substrate for microbial processes for the production of value-added products. Several solutions and alternative uses of the coffee pulp and husk have been attempted. These include as fertilizers, livestock feed, compost, etc. However, these applications utilize only a fraction of available quantity and are not technically very efficient. Attempts have been made to detoxify it for improved application as feed, and to produce several products such as enzymes, organic acids, flavour and aroma compounds, and mushrooms, etc. from coffee pulp/husk. Solid state fermentation has been mostly employed for bioconversion processes. Factorial design experiments offer useful information for the process optimization. This paper reviews the developments on processes and products developed for the value-addition of coffee pulp/husk through the biotechnological means.     

Interaction with microsomal lipid as a major factor responsible for S9-mediated inhibition of 1,8-dinitropyrene mutagenicity

1,8-Dinitropyrene (1,8-DNP), present in polluted air, is a rodent carcinogen and a potent, direct-acting mutagen in salmonella typhimurium TA98. This mutagenicity is markedly reduced in the presence of mammalian hepatic S9 or microsomes. We demonstrate that at least a substantial part of this effect is attributable to non-enzymatic processes. The microsomal-dependent inhibition was unaffected by omission of an NADPH-generating system or when the cytochrome P-450 inhibitor, SKF-525A, or the cytochrome P-448 inhibitor, ellipticine, was incorporated in the metabolic activation system, suggesting that mixed function oxidases are not involved. Heat inactivation partially decreased the ability of induced S9 to reduce DNP mutagenicity. Substitution of S9 with a similar concentration of bovine serum albumin did not affect DNP activity. Thus non-specific binding to microsomal protein is not involved. However, when lipids, derived from uninduced microsomes, were added to incubations of DNP and S. typhimurium TA98, mutagenicity was decreased. Furthermore, substitution of microsomal lipids with a suspension of phosphatidylcholine (PC), a major lipid constituent of microsomes, affected DNP mutagenicity similarly. An increase in PC concentration resulted in a greater inhibitory effect. The reduction in DNP mutagenicity observed with microsomal lipids or with PC was less than that detected with uninduced S9, whilst the mutagenicity of 2-nitrofluorene was reduced to an approximately equal extent by lipids and S9. This phenomenon may be responsible for the S9-mediated detoxification of other mutagenic nitroaromatic compounds and may have important implications for mutagenicity testing.     

Germination of coffee seeds and its significance for coffee quality

Besides genotypic characteristics, the crucial factor that determines coffee quality is the mode of post-harvest treatment, i.e., the wet and dry processing. Up to now, the resulting characteristic flavour differences between these differentially processed coffees were attributed exclusively to differences in starting material. However, as these quality differences are still evident, even when identical coffee samples were processed by the two methods in parallel, the differences must be created by metabolic processes in the coffee beans themselves. Based on expression studies of the germination-specific isocitrate lyase and the resumption of cell cycle activity, monitored by the abundance of beta-tubulin, we evidence that germination is initiated in coffee seeds during the course of standard coffee post-harvest treatments. The extent and nature of the germination processes depend on the processing method. The coherence of metabolic events, substantial differences in the chemical composition of the coffee beans, and the generation of specific coffee qualities establishes the basis for a quite novel approach in coffee research.     

Fluroxene mutagenicity.

The commercially available volatile anesthetic fluroxene (2,2,2-trifluoroethyl vinyl ether) which contains the stabilizer N-phenyl-1-napthylamine, was tested for mutagenicity using four strains of S. typhimurium, TA1535, TA1537, TA98 and TA100, and one strain of E. coli, WP2. In addition, purified fluroxene; N-phenyl-1-napthylamine; trifluoroethanol, a major metabolite of fluoroxene; and urine from rats anesthetized with fluroxene were tested. Several procedures were utilized including exposure of bacteria to vapor in desiccators and in liquid suspension. Results indicate that fluroxene, but not its stabilizer, was mutagenic to strains TA1535, TA100 and WP2 only in liquid suspension and only in the presence of a rat-liver enzyme system. Trifluoroethanol and urine from fluroxene-treated rat were not mutagenic to any strain of bacteria. These findings indicate that fluroxene is a promutagen which requires preincubation before it is recognized. Further experiments were performed with enzymes prepared from mouse, hamster and human liver. Fluroxene was mutagenic only in the presence of enzymes prepared from Aroclor 1254 pretreated rodents. Since fluroxene was not mutagenic in the presence of enzymes prepared from three human livers, the significance of these findings to man are unclear.