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Carob liqueur is an alcoholic drink (minimum 15% v/v of ethanol and 100 g/L of sugar) typical for the Mediterranean countries. In the current work, carob macerate produced by maceration of carob pods in hydroalcoholic base at different maceration conditions was characterized for the first time based on its aroma compounds/profile, physicochemical parameters, and chromatic characteristics. The results confirm the migration process of bioactive compounds, aroma compounds, and sugars flowing from the carob pod to the hydroalcoholic base. Changes in ethanol concentration modify the physical properties of the solvent and influence the phenolic and aroma compounds extraction, color, and acidity of the obtained samples. The higher content of phenolic compounds was determinate in the samples obtained in the darkness. The amounts of phenols were in the range of some red fruit liqueurs or walnut liqueurs, and sugars (mostly sucrose) ranging between 96 and 107 g/L. Twenty‐six (out of total 94) aroma compounds were detected in all samples, of which 17 esters, 3 alcohols, 4 ketones, and 2 acids. Low molecular weight ethyl esters, ethyl hexanoate, ethyl 2‐methyl propanoate, ethyl octanoate, ethyl benzoate, ethyl butanoate, and ethyl cinnamate, were the most abundant. Carob pod maceration in 50% v/v hydroalcoholic base (1:5 solid to liquid ratio) in darkness at room temperature during 8 weeks can be recommended as optimal maceration conditions for production of the aromatic carob macerate with functional properties., The results confirm migration process such as: bioactive compounds, aroma compounds, and sugars flows from the carob pod to the alcoholic base. Changes in ethanol concentration modify the physical properties of the solvent and influence the phenolic and aroma compounds extraction, colour, and acidity of the obtained samples. The higher content of phenolic compounds was determinate in the samples obtained in the darkness. The amounts of phenols were in the range of some red fruit liqueurs or walnut liqueurs.

During the ageing period wine spirits are changing their color, chemical composition and sensory characteristics. These changes should be simply monitored. The aim of this study was to develop partial least squares regression (PLS) models for higher alcohols and phenols in wine spirits as well as to show the feasibility of the NIR spectroscopy combined with chemometric tools to distinguish wine spirits and brandies with different ageing degree. To get the reference values, the usual methods for the analysis of spirits drinks were used. Ethanol, esters, acids, methanol and higher alcohols were studied. Wine spirits and brandies phenol composition was determined by liquid chromatography. Principal component analysis (PCA) was used to classify the wine spirits and brandies according to their phenolic and higher alcohols composition. Moreover, the Partial least squares regression (PLS regression) was used to calibrate and predict expected contents of higher alcohols and phenols in the wine spirits. Success of the classification of samples by ageing based on individual alcohols was 93.8 %, while success of the classification based on individual phenols raised to 100 %. This efficiency of the prediction was evaluated by use of linear discriminator analysis (LDA).

Research background. During the current SARS-CoV2 pandemic, as well as earlier SARS and MERS epidemics, it has been observed that COVID19-positive women on average tend to have milder symptoms and lower fatality rates than men. There is a number of differences between the sexes known to contribute to different immune responses and severity of the disease, one being the effect of estrogen via estrogen receptor signalling. We wondered if estrogen might also affect the SARS-CoV2 more directly, perhaps by binding to the surface glycoprotein (S protein), thus possibly reducing its infectivity. Experimental approach. To assess whether there is a possibility for estrogen binding on the SARS-CoV2 S protein, we used BLAST and HHpred to compare protein sequences of S protein and human estrogen receptor β, while 3D structures of a potential estrogen binding site and an active site of estrogen receptor β were visualized and compared using PyMOL. Results and conclusions. By comparing the sequence of SARS-CoV2 S protein with the human estrogen receptor β, we identified a potential estrogen binding site on S protein and further determined that it also shares notable similarities with the active site of ER β when observed in 3D structure of their respective proteins. As a control, SARS-CoV2 S protein was compared with the human androgen receptor, and no such similarities were found. The potential estrogen binding site is part of coronavirus S2 superfamily domain, which is involved in host-virus membrane fusion during infection and appears to be conserved throughout the Coronaviridae family. Novelty and scientific contribution: This preliminary communication shows that SARS-CoV2 S protein features a potential estrogen binding site. Hopefully, this will prompt a more comprehensive study on the possibilities of estrogen binding on the S protein and the effect this might confer on the virus infectivity. © 2021, University of Zagreb. All rights reserved.

In Croatia, as part of the Mediterranean region where carob is grown, only carob liquor is produced, mostly as a domestic product. In contrast, pure spirit production by alcoholic fermentation and distillation of carob is very rare and the compounds responsible for the characteristic bouquet of carob spirit are unknown. Therefore, the aim of this study was to evaluate the potential of carob as a raw material (source of sugars, nutrients and yeast inhibitors) for the production of carob spirit using commercial yeast strains. Moreover, a preliminary characterization of the volatiles in the obtained distillates was considered. Fermentations were carried out in carob mash with or without added nutrients by five Saccharomyces cerevisiae strains at three different temperatures (20°C, 30°C and room temperature). The obtained wine was subjected to fractional distillation in a copper still to produce carob spirit with an alcohol content of 40 % vol. Analysis of sugars and fermentation products was performed by high performance liquid chromatography. Gas chromatography and gas chromatography coupled with MS detection was used to analyse the volatile components of the carob wine and brandy. Carob flour and S. cerevisiae strains can be efficiently used for the fermentation process to produce carob wine with ethanol content ranging from 46.4 to 50.5 gL-1 and corresponding yield coefficients ranging from 0.45 to 0.49 gg-1. More than ninety compounds detected in carob spirit, namely ethyl 2-methyl butanoate, ethyl 2-methyl propanoate, ethyl cinnamate, ethyl hexanoate, beta-ionone, ethyl butanoate and ethyl octanoate, contribute significantly to the fruity and floral bouquet of carob spirit. Thus, by using the tested yeast strains, a novel brandy rich in volatiles can be obtained. Carob brandy can be 82 an additional product in the carob processing chain and new potential, especially for small carob growers.

Velika količina lignoceluloznog otpada, kojeg čine uglavnom celuloza, hemiceluloza i lignin, nastaje u šumarstvu, poljoprivredi i prehrambenoj industriji. Obzirom na dostupne količine ovih jeftinih i obnovljivih sirovina u Republici Hrvatskoj, važno je razmotriti mogućnosti njihove primjene u proizvodnji bioetanola i raznih drugih biotehnoloških proizvoda. Lignocelulozni hidrolizati su složene smjese heksoza i pentoza te drugih spojeva od kojih neki mogu djelovati kao inhibitori fermentacije. U ovom radu dan je pregled kvasaca koji se koriste i/ili istražuju za proizvodnju etanola iz lignoceluloznih hidrolizata.

Yeasts have an important role in the biotechnological production of ethanol, as well as in the production of fermented food and beverages. Mycotoxins, secondary metabolites of molds, can contaminate a large number of foods due to their ubiquity and therefore present great risk to human health. Other than their toxicological effects on humans, mycotoxins can have negative effects on various properties of yeasts and consequently limit their use in industry. Therefore, the aim of this work was to determine the effects of aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) on metabolic activity and biomass growth of selected yeasts S. cerevisiae, S. uvarum, C. utilis and K. marxianus. The presence of fermentation products was determined with high-performance liquid chromatography after 24h of incubation. In the early stage, all mycotoxin-exposed yeasts had lower metabolic activity as well as biomass growth when compared to control. However, concentrations of metabolic products and biomass reached the control levels by the end of the process, with the exception of C. utilis exposed to zearalenone (ZEA). Adaptive response to exposed mycotoxins suggests that certain yeasts could be used to control mycotoxin concentrations in the production of fermented food and beverages.

Zbog bolje učinkovitosti i sigurnosti potreba za enantiomerno čistim kiralnim spojevima u stalnom je porastu, pogotovo u farmaceutskoj industriji. Da bi se te potrebe zadovoljile potrebne su nove, učinkovitije i jeftinije ali i ekološki prihvatljive metode njihove priprave. U skladu s tim veliku je pažnju privukla biokataliza gdje se kao biokatalizatori koriste mikroorganizmi ili enzimi izolirani iz njih. Posljednjih godina znanstvena zajednica se okrenula istraživanju upotrebe svježeg, netaknutog biljnog materijala kao biokatalizatora. Upotreba biljnog materijala ne zahtijeva dodatak skupih kofaktora budući da se različiti enzimski sustavi i potrebni kofaktori, te njihov regeneracijski sustav već nalaze u biljnom tkivu. Ovisno o upotrijebljenoj biljci (voću, povrću, začinima), pojedini enantiomeri se mogu dobiti u visokom prinosu i s visokim enantiomernim viškom, što je kritični faktor za razvoj biološki aktivnih spojeva i njihovu bioaktivnost. U ovom radu dan je pregled istraživanja potencijalne upotrebe različitih biljnih vrsta i njihovih dijelova kao biokatalizatora u stereoselektivnim transformacijama pogodnih spojeva s ciljem dobivanja novih enentiomerno čistih spojeva. Primjena biljnih biokatalitičkih sustava u skladu je s principima zelene kemije i predstavlja značajnu alternativu sintetskim kemijskim postupcima.

Poljoprivreda, šumarstvo i prehrambena industrija izvori su velike količine lignocelulozne biomase, koja može poslužiti kao lako dostupna i jeftina obnovljiva sirovina za dobivanje različitih bioproizvoda. Jedan od takvih proizvoda je i bioetanol. Ovaj rad daje pregled bakterija koje se koriste i/ili istražuju za proizvodnju bioetanola iz lignoceluloznih sirovina. U navedenim istraživanjima proizvodnje bioetanola pomoću bakterija primijenjuju se različiti pristupi kako bi se povećala ekološka i ekonomska efikasnost procesa. Pored uobičajenih bioprocesa, koji se provode u više faza i uz pomoć monokulture, razvijaju se i visokointegrirani (konsolidirani) bioprocesi uz primjenu mikrobnih kokultura., Agriculture, forestry and food industry are sources of large quantities of lignocellulosic biomass, which can be used as an easily accessible and cheap renewable raw material for production of different bioproducts. One of these bioproducts is also bioethanol. This paper provides an overview of bacteria used and/or investigated for bioethanol production from lignocellulose-containing feedstocks. In the research of bioethanol production using bacteria, various approaches are applied in order to increase ecological and economic efficiency of bioprocess. In addition to conventional multi-stage bioprocesses that are carried out using monoculture, highly integrated (consolidated) bioprocesses and applications of microbial cocultures are also in developing stage.

In this paper the effect of aflatoxin B1, ochratoxin A and zearalenon on morphology, growth parameters and metabolic activity of yeasts Saccharomyces cerevisiae, Saccharomyces uvarum, Candida utilis and Kluyveromyces marxianus was determined. The results showed that the three mycotoxins affected the morphology of all these yeasts, primarily the cell diameter, but not their final cell count. Fourier transform infrared spectroscopy showed that the yeast membranes bound the mycotoxins, C. utilis in particular. The cell membranes of most yeasts underwent denaturation, except S. uvarum exposed to ochratoxin A and zearalenone. In the early stage of fermentation, all mycotoxin-exposed yeasts had lower metabolic activity and biomass growth than controls, but fermentation products and biomass concentrations reached the control levels by the end of the fermentation, except for C. utilis exposed to 20 μg/mL of zearalenone. The adaptive response to mycotoxins suggests that certain yeasts could be used to control mycotoxin concentrations in the production of fermented food and beverages.

Organic farming preserves biodiversity and organic products can be the source of many microbial species. The species diversity in organically grown wheat, spelt and rye was investigated in order to find strains suitable for sourdough fermentation. Colonies representing various morphological appearances were isolated and catalase-negative colonies were identified by mass spectrometer Microflex LT ™ MALDI-TOF. The fermentation products (lactic, acetic, formic and phenyllactic acid) were determined by high performance liquid chromatography, while the antifungal activity was determined using an overlay agar method. Wheat flours showed less microbial biodiversity than the rye and spelt flours. The most common genera in the tested flour were Lactobacillus, Pediococcus and Enterococcus. Isolated Lactobacillus farciminis, Pediococcus pentosaceus, Leuconostoc citreum and Lactobacillus brevis showed the best acidification activity. Lactobacillus brevis, Pediococcus pentosaceus, Weissella cibaria and Lactobacillus farciminis showed significant antifungal activity against A. niger 357 and Penicillium sp. 505. The prefermented medium of Lactobacillus farciminis were characterized by high content of lactic and phenyllactic acid.

The efficiency of gas phase plasma at atmospheric pressure by using an electrical discharge in gas argon on the inactivation of microorganisms was examined. The gas phase plasma was applied to suspensions of pure cultures Escherichia coli 3014, Staphylococcus aureus 3048, Salmonella sp. 3064, Listeria monocytogenes ATCC 23074 and Bacillus cereus 30. The experiments were planned and performed according to a statistical experimental design, specifically central composite design, which considered three independent variables: volume (2, 3 and 4 mL), gas flow (0.75, 1 and 1.25 L/min) and treatment time (3, 4 and 5 min). Two studied parameters, volume and treatment time, substantially affected the inactivation. For plasma treatment, the inactivation can be attributed to UV radiation and plasma reactive oxygen species (ROS). It was found that Gram-negative bacteria were more susceptible to the plasma treatment than Gram-positive bacteria, and that the susceptibility of Gram-positive bacteria was remarkably species-dependent. Complete inactivation of Escherichia coli, Salmonella sp., and Listeria monocytogenes was achieved when optimal combination of parameters was applied.

Certain species of lactic acid bacteria (LAB), as well as other microorganisms, can bind metal ions to their cells surface or transport and store them inside the cell. Due to this fact, over the past few years interactions of metal ions with LAB have been intensively investigated in order to develop the usage of these bacteria in new biotechnology processes in addition to their health and probiotic aspects. Preliminary studies in model aqueous solutions yielded LAB with high absorption potential for toxic and essential metal ions, which can be used for improving food safety and quality. This paper provides an overview of results obtained by LAB application in toxic metal ions removing from drinking water, food and human body, as well as production of functional foods and nutraceutics. The biosorption abilities of LAB towards metal ions are emphasized. The binding mechanisms, as well as the parameters influencing the passive and active uptake are analyzed.

Western diet is frequently low in essential metal ions. A common method for preventing metal ion deficiency is pharmacological supplementation, especially in a highly available form such as metalo-protein complexes. Accordingly, in this work, specific lactic acid bacteria were assessed for their ability to bind copper ions. Significant amounts of copper ions were bound, and the binding potential was found to be strain specific. Differences among the strains were evaluated with the Langmuir model for biosorption. Binding of copper was a fast process, strongly influenced by ionic strength, pH and biomass concentration. During the process, copper ions significantly reduced cell viability. Discharge of copper ions in a simulated gastrointestinal tract was examined ; 85-90% of copper ions bound to LAB were discharged in the gastrointestinal model system.

S-adenosyl-l-methionine (SAM) is an important molecule in the cellular metabolism of mammals. In this study, we examined several of the physiological characteristics of a SAM-accumulating strain of the yeast Scheffersomyces stipitis (M12), including SAM production, ergosterol content, and ethanol tolerance. S. stipitis M12 accumulated up to 52.48 mg SAM/g dry cell weight. Proteome analyses showed that the disruption of C-24 methylation in ergosterol biosynthesis, a step mediated by C-24 sterol methyltransferase (Erg6p), results in greater SAM accumulation by S. stipitis M12 compared to the wild-type strain. A comparative proteome-wide analysis identified 25 proteins that were differentially expressed by S. stipitis M12. These proteins are involved in ribosome biogenesis, translation, the stress response, ubiquitin-dependent catabolic processes, the cell cycle, ethanol tolerance, posttranslational modification, peroxisomal membrane stability, epigenetic regulation, the actin cytoskeleton and cell morphology, iron and copper homeostasis, cell signaling, and energy metabolism.

Aims The aim of this study was to investigate and compare two nonthermal techniques in the inactivation of moulds. Methods and Results High power ultrasound (20 kHz) and nonthermal gas phase plasma treatments were studied in the inactivation of selected moulds. Aspergillus spp. and Penicillium spp. were chosen as the most common mould present in or on food. Experimental design was introduced to establish and optimize working variables. For high power ultrasound, the greatest reduction of moulds (indicated by the total removal of viable cells) was obtained after ultrasound treatments at 60°C (thermosonication) for 6 and 9 min (power applied, 20–39 W). For plasma treatment, the greatest inactivation of moulds was observed for the longest treatment time (5 min) and lowest sample volume (2 ml), (AP12, AP13, PP12 and PP13). Conclusions The great amount of applied energy required for achieving a partial log reduction in viable cells is the limiting factor for using high-power ultrasound. However, both treatment methods could be combined in the future to produce beneficial outcomes. Significance and impact of the study This study deals with nonthermal food processing techniques and the results and findings present in this study are the root for further prospective studies. The food industry is looking for nonthermal methods that will enable food preservation, reduce deterioration of food compounds and structure and prolong food shelf life.

Trigonopsis variabilisd-amino acid oxidase accounts for 35% ofEscherichia coliprotein when addedd-methionine suppresses the toxic activity of the recombinant product. PermeabilizedE. colicells are reusable and stabilized enzyme preparations. The purified oxidase lacks the microheterogeneity of the natural enzyme. Oriented immobilization of a chimeric oxidase maintains 80% of the original activity in microparticle-bound enzymes.

Lactic acid bacteria (LAB) as starter culture in food industry must be suitable for large-scale industrial production and possess the ability to survive in unfavorable processes and storage conditions. Approaches taken to address these problems include the selection of stressresistant strains. In food industry, LAB are often exposed to metal ions induced stress. The interactions between LAB and metal ions are very poorly investigated. Because of that, the influence of non- toxic, toxic and antioxidant metal ions (Zn, Cu, and Mn) on growth, acid production, metal ions binding capacity of wild and adapted species of Leuconostoc mesenteroides L3, Lactobacillus brevis L62 and Lactobacillus plantarum L73 were investigated. The proteomic approach was applied to clarify how the LAB cells, especially the adapted ones, protect themselves and tolerate high concentrations of toxic metal ions. Results have shown that Zn and Mn addition into MRS medium in the investigated concentrations did not have effect on the bacterial growth and acid production, while copper ions were highly toxic, especially in static conditions. Leuc. mesenteroides L3 was the most efficient in Zn binding processes among the chosen LAB species, while L. plantarum L73 accumulated the highest concentration of Mn. L. brevis L62 was the most copper resistant species. Adaptation had a positive effect on growth and acid production of all species in the presence of copper. However, the adapted species incorporated less metal ions than the wild species. The exception was adapted L. brevis L62 that accumulated high concentration of copper ions in static conditions. The obtained results showed that L. brevis L62 is highly tolerant to copper ions, which allows its use as starter culture in fermentative processes in media with high concentration of copper ions.

Bread has been an essential part of human nutrition and culture for thousands of years. Every step of it’s production, from careful raw material selection to dough preparation, processing and baking, could be considered as art that ensure unlimited abundance of color, taste and aroma. Diversity of bakery products is result of the diversities in tradition, culture and geographic origin. All around the world alluring and aromatic bakery products always contain sourdough. Sourdough is a complex microbial ecosystem that is composed of lactic acid bacteria and yeasts. The use of lactic acid bacteria in the bread production has long tradition, from spontaneous fermentations (leavening the dough only), to the development and use of defi ned and functional starter cultures. This review covers the sourdough technology, with major emphasis on production of wheat bread with enhanced nutritional value, prolonged shelf life, better taste and aroma., Kruh je od pamtivijeka neizostavni dio ljudske prehrane i kulture. Svaki korak u njegovoj proizvodnji, od pažljivog izbora sirovina do pripreme i obrade tijesta te procesa pečenja, predstavlja dio umjetnosti koja u konačnici osigurava neograničeno bogatstvo boje, okusa i arome kruha. Raznolikost pekarskih proizvoda rezultat je različitosti tradicije, kulture te geografskog podrijetla. Međutim, san svakog pekara, primamljivi i mirisni pekarski proizvodi diljem svijeta, uvijek sadržavaju kiselo tijesto. Kiselo tijesto je kompleksni mikrobni ekosustav kojeg čine bakterije mliječne kiseline i kvasci. Upotreba bakterija mliječne kiseline u pripremi kruha ima dugu tradiciju, od spontanih fermentacija gdje je njihova uloga bila samo dizanje tijesta, preko defi niranih starter kultura, do razvoja i primjene funkcionalnih starter kultura. U ovom radu dat je prikaz tehnologije proizvodnje kiselog tijesta s naglaskom na proizvodnju pšeničnog kruha povećane nutritivne vrijednosti i trajnosti, bogatijeg okusa i arome.

Yeast biomass as well as biomass enriched with trace minerals have been demonstrated to be useful in improving animal health and growth performance. In this work, process for the production of Saccharomyces cerevisiae biomass enriched with manganese, a microelement with antioxidant properties in the form of high bioavailable Mn-protein complex, has been studied. The influences of media composition, Mn2+ concentration and Mn salt were investigated in shaken cultures. The change of biomass and ethanol yields was not observed in molasses media with addition of Mn, while in sucrose media the decrease was observed at Mn2+ concentrations higher than 0.8 mM (added as sulphate) and 0.2 mM (added as chloride). It was established that aeration mode (anaerobic, shaken flask or aerated culture) influences amount and dynamic of Mn incorporation into the yeast biomass, and that this incorporation was S. cerevisiae strain dependent. The Fourier transform infrared (FTIR) spectrophotometry on blank and Mn loaded biomass suggested that carboxyl groups, N-H groups of secondary amide, and sulfonate groups are involved in mechanism of manganese binding.

The production of Saccharomyces cerevisiae cells enriched with copper and the effects of adding copper ions to different media on yeast cell growth and ethanol production were studied. In the media Cu(2+) concentrations of up to 0.094 mM had no effect on alcoholic fermentation, whereas higher Cu(2+) concentrations markedly decreased yeast cell growth rate and ethanol production. Under static conditions, the maximum amounts of copper uptake (i.e., 1.16 mg/g, 1.2 mg/g and 0.81 mg/g dry matter yeast biomass for glucose, sucrose and molasses media, respectively) were obtained after 8 h of fermentation, whereas under dynamic conditions smaller amounts of copper uptake (i.e., 0.98 mg/g, 1.02 mg/g and 0.7 mg/g dry matter yeast biomass for glucose, sucrose and molasses media, respectively) were obtained after 6 h of fermentation.

Procedures for the production of Saccharomyces cerevisiae biomass enriched with iron and the effects of the iron ions addition into the molasses medium on the yeast growth and the production of ethanol were studied. The growth of the yeast S. cerevisiae and the ethanol production in media with different concentrations of Fe were monitored in the batch process under semiaerobic and anaerobic conditions. The highest biomass concentration and ethanol production were achieved in the medium with 0.6-0.8 g l-1 of Fe under both (semiaerobic and anaerobic) conditions. Kinetics of the iron ions accumulation in yeast cells during 24 h of growth in the batch process under semiaerobic and anaerobic conditions were monitored. In anaerobic conditions the maximum uptake (10 mg g-1 d.m. yeast biomass) was obtained after 12 h of fermentation, while in semiaerobic conditions a four times lower uptake (2.5 mg g-1 d.m. yeast biomass) was obtained after 16 h of fermentation.

One of the most important antioxidants in virgin olive oil (VOO) are phenolic compounds. The amount of phenolic substances present in VOO, as well as the fatty acid composition, is related to the health beneficial effects that make VOO a very valuable and appreciated commodity. The level of these substances is a very important parameter in the evaluation of VOO quality since phenols are strictly related both to the oil’s resistance to oxidation because of their antioxidative properties. The aim of the present study was to define parameters for the characterization, polyphenolic composition and related antioxidative properties of monovarietal olive oil from cv. Drobnica grown in Dalmatia (Croatia). We also examined the antioxidant activity, total antioxidant capacity and oxidative stability of Drobnica’s oil. Besides several qualitative parameters (free acidity, peroxide value and UV spectrophotometric indices), the analyses of major (fatty acids) and minor components (phenolic fraction and their antioxidant power evaluated by ABTS●+ and electrochemical test) were also carried out. Radical scavenging activity of phenolic extracts was performed by ABTS+ and DPPH methods. An amperometric flow injection method (FIA) was used for the evaluation of the antioxidant power (AOP) of the phenolic extracts of the oil samples. The evaluation of correlation coefficients between parameters DPPH, ABTS, AOP and certain fatty acids and phenolic groups were performed. Olive oil from Drobnica showed high proportion of oleic and palmitic acid. Stable 18:1/18:2 ratio leads to conclusion that this oil has pretty high oxidative stability. Moderate positive correlation between DPPH and linoleic acid was detected, as well as moderate positive correlation between DPPH and the ratio of oleic and linoleic acids. The positive correlation was detected between DPPH and secoiridoides derivates.