Enzyme Source Research Articles

Edible Flowers: A New Source of Minerals

Abstract: Edible flowers have become very popular in recent years as a potent and natural source of various important minerals, vitamins, flavonoids, enzymes and aromatic compounds. Various anti-bacterial, anti-viral, anti-fungal & anti-inflammatory substances have been found in various edible flowers e.g. Rose, Marigold, Lilac, Chrysanthemum, Chinese hibiscus, Roselle, Tulip, Magnolia, Water lily etc. These flowers are almost calorie-free, high in antioxidants, have inhibitory effects against cancer, obesity and inflammation, prevent neuronal degradation, help to diminish blood pressure, demonstrate gastro-protective and hepato-protective effects. Edible flowers show antioxidant activities, inhibitive activities on free radicals and ROSscavenging properties due to presence of various phenolic compounds e.g. Quercetin, Rutin, Gallic acid etc. Pollen, nectar and petals are rich source of proteins, carbohydrates, lipids. The colour of flowers is characterized by various carotenoids, anthocyanins etc. Harvesting of edible flowers should be done in cool climate during day time in fully bloomed condition to retain quality and shelf-life. Edible flowers are generally used in salads, soups, beverages, and herbal medicines across the world. People with conditions like hay fever, allergies or asthma must be careful in consuming these flowers. However, in upcoming years, there are immense possibilities of a rapid increase in the consumption of edible flowers when cultivated organically.

Biochemical Characteristics of Laccases and Their Practical Application in the Removal of Xenobiotics from Water

The rapid growth of the human population in recent decades has resulted in the intensive development of various industries, the development of urban agglomerations and increased production of medicines for animals and humans, plant protection products and fertilizers on an unprecedented scale. Intensive agriculture, expanding urban areas and newly established industrial plants release huge amounts of pollutants into the environment, which, in nature, are very slowly degraded or not decomposed, which leads to their accumulation in water and terrestrial ecosystems. Researchers are scouring extremely contaminated environments to identify organisms that have the ability to degrade resistant xenobiotics, such as PAHs, some pharmaceuticals, plasticizers and dyes. These organisms are a potential source of enzymes that could be used in the bioremediation of industrial and municipal wastewater. Great hopes are pinned on oxidoreductases, including laccase, called by some a green biocatalyst because the end product of the oxidation of a wide range of substrates by this enzyme is water and other compounds, most often including dimers, trimers and polymers. Laccase immobilization techniques and their use in systems together with adsorption or separation have found application in the enzymatic bioremediation of wastewater.

Hybrid De Novo Whole-Genome Assembly, Annotation, and Identification of Secondary Metabolite Gene Clusters in the Ex-Type Strain of Chrysosporium keratinophilum.

Chrysosporium is a polyphyletic genus belonging (mostly) to different families of the order Onygenales (Eurotiomycetes, Ascomycota). Certain species, such as Chrysosporium keratinophilum, are pathogenic for animals, including humans, but are also a source of proteolytic enzymes (mainly keratinases) potentially useful in bioremediation. However, only a few studies have been published regarding bioactive compounds, of which the production is mostly unpredictable due to the absence of high-quality genomic sequences. During the development of our study, the genome of the ex-type strain of Chrysosporium keratinophilum, CBS 104.66, was sequenced and assembled using a hybrid method. The results showed a high-quality genome of 25.4 Mbp in size spread across 25 contigs, with an N50 of 2.0 Mb, 34,824 coding sequences, 8002 protein sequences, 166 tRNAs, and 24 rRNAs. The functional annotation of the predicted proteins was performed using InterProScan, and the KEGG pathway mapping using BlastKOALA. The results identified a total of 3529 protein families and 856 superfamilies, which were classified into six levels and 23 KEGG categories. Subsequently, using DIAMOND, we identified 83 pathogen-host interactions (PHI) and 421 carbohydrate-active enzymes (CAZymes). Finally, the analysis using AntiSMASH showed that this strain has a total of 27 biosynthesis gene clusters (BGCs), suggesting that it has a great potential to produce a wide variety of secondary metabolites. This genomic information provides new knowledge that allows for a deeper understanding of the biology of C. keratinophilum, and offers valuable new information for further investigations of the Chrysosporium species and the order Onygenales.

Valorization of orange peels exploiting fungal solid-state and lacto-fermentation.

Orange peels can serve as a cost-effective raw material for the production of lactic acid. Indeed, given their high concentration of carbohydrates and low content of lignin, they represent an important source of fermentable sugars, recoverable after a hydrolytic step. In the present article, the fermented solid, obtained after 5 days of Aspergillus awamori growth, was used as the only source of enzymes, mainly composed of xylanase (40.6IU g-1 of dried washed orange peels) and exo-polygalacturonase (16.3IU g-1 of dried washed orange peels) activities. After the hydrolysis, the highest concentration of reducing sugars (24.4g L-1 ) was achieved with 20% fermented and 80% non-fermented orange peels. The hydrolysate was fermented with three lactic acid bacteria strains (Lacticaseibacillus casei 2246 and 2240 and Lacticaseibacillus rhamnosus 1019) which demonstrated good growth ability. The yeast extract supplementation increased the lactic acid production rate and yield. Overall, L. casei 2246 produced the highest concentration of lactic acid in mono-culture. To the best of our knowledge this is the first study exploiting orange peels as low-cost raw material for the production of lactic acid avoiding the employment of commercial enzymes. The enzymes necessary for the hydrolyses were directly produced during A. awamori fermentation and the reducing sugars obtained were fermented for lactic acid production. Despite this preliminary work carried out to study the feasibility of this approach, the concentrations of reducing sugars and lactic acid produced were encouraging, leaving open the possibility of other studies for the optimization of the strategy proposed here. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Mechanistic Investigation of the Time-Dependent Aldehyde Oxidase Inhibitor Hydralazine.

The anti-hypertensive agent hydralazine is a time-dependent inhibitor of the cytosolic drug-metabolizing enzyme aldehyde oxidase (AO). Glutathione (GSH) was found to suppress the inhibition of AO by hydralazine in multiple enzyme sources (human liver and kidney cytosol, human liver S9, rat liver S9, and recombinant human AO) and with different AO substrates (zoniporide, O6 -benzylguanine, and dantrolene). Hydralazine-induced AO inactivation was unaffected when GSH was added to the incubation mixture after pre-incubation of hydralazine with AO (rather than during the pre-incubation), suggesting that GSH traps a hydralazine reactive intermediate prior to enzyme inactivation. Consistent with previous reports of 1-phthalazylmercapturic acid formation when hydralazine was incubated with N-acetylcysteine, we detected a metabolite producing an MS/MS spectrum consistent with a 1-phthalazyl-GSH conjugate. O6 -Benzylguanine, an AO substrate, did not protect against hydralazine-induced AO inactivation, implying that hydralazine does not compete with O6 -benzylguanine for binding to the AO active site. Catalase also failed to protect AO from hydralazine-induced inactivation, suggesting that hydrogen peroxide is not involved. However, an allosteric AO inhibitor (thioridazine) offered some protection, indicating a catalytic role for AO in the bioactivation of hydralazine. AO inhibition by phthalazine (a substrate and inhibitor of AO and a metabolite of hydralazine) was unaffected by the presence of GSH. GSH also prevented hydralazine from inhibiting the nitro-reduction of dantrolene by AO. Furthermore, the GSH-hydralazine combination stimulated dantrolene reduction. Phthalazine inhibited only oxidation reactions, not reduction of dantrolene. Together, these results support the hypothesis that hydralazine is converted to a reactive intermediate that inactivates AO. SIGNIFICANCE STATEMENT: These studies suggest that a reactive intermediate of hydralazine plays a primary role in the mechanism of aldehyde oxidase (AO) inactivation. Inactivation was attenuated by glutathione and unaffected by catalase. Phthalazine (hydralazine metabolite) inhibited AO regardless of the presence of glutathione; however, phthalazine inhibited only oxidation reactions, while hydralazine inhibited both oxidation and reduction reactions. This report advances our mechanistic understanding of hydralazine as an AO inhibitor and provides information to facilitate appropriate use of hydralazine when probing AO metabolism.

Mining of Thermostable Alpha-amylase Gene from Geothermal Springs using a Metagenomics Approach

The geothermal springs are said to contain the greatest diversity of undiscovered microorganisms, making them the best source for enzymes with economic significance. The untapped microbial diversity living in the geothermal springs can be mined for novel genes, bioactive substances, and industrially significant biocatalysts using the metagenomics technique. Metagenome was extracted from soil samples of various geothermal springs of India. Metagenome was screened for various carbohydrate degrading enzymes (amylase, cellulase, xylanase, amylopullulanase) using degenerate primers-based Polymerase chain reaction amplifications. Further amplicons were cloned, sequenced and analysis of data was done using various bioinformatics tools, e.g., Blast analysis, Protparam and phre2 program. We have isolated numerous enzymes, including cellulase, amylase, amylopullulanase, and xylanase, from diverse geothermal spring in different parts of India using sequence and function-based metagenomics. In this study, we describe the metagenomics-based isolation of a thermostable amylase from the geothermal spring of Odisha. The amylase gene (1503 bp) was amplified using the metagenome as a template using degenerate primers and cloned into the linearized T vector. The putative gene was likely to encode a protein of 469 amino acids with a molecular weight of 53895.05 Da with pI-7.78. Sequence analysis showed its maximum identity of 98.95% with Bacillus licheniformis alpha-amylase gene. Homology modeling of the amylase protein was done using the phyre2 program, which shows it belongs to the (trans) glycosidase superfamily and contains the catalytic TIM alpha/beta-barrel fold. Hence, we can conclude that geothermal springs are hotspots for the mining of industrially robust biocatalysts.

Safety evaluation of the food enzyme phytepsin from Cynara cardunculus L.

The food enzyme phytepsin (EC3.4.23.40) is extracted from the pistils of the cardoon (Cynara cardunculus L.) by different manufacturers represented by the Dirección General de Salud Pública, Gobierno de Canarias, España. It is intended to be used in milk processing for cheese production. As no concerns arose from the source of the food enzyme, from its manufacture, and based on a history of safe use and consumption, the Panel considered that toxicological data and the estimation of dietary exposure were not required. A search for the similarity of the amino acid sequences of the food enzyme to known allergens was made and no matches were found. The Panel considered that allergic reactions to this phytepsin cannot be excluded in individuals allergic to cardoon. However, the likelihood of allergic reactions to the phytepsin from C. cardunculus L. is expected not to exceed the likelihood of allergic reactions to cardoon. As the prevalence of allergic reactions to cardoon is low, also the likelihood of such reactions to occur to the food enzyme is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

Enhancement of phycocyanobilin biosynthesis in Escherichia coli by strengthening the supply of precursor and artificially self-assembly complex

Phycocyanobilin (PCB) is widely used in healthcare, food processing, and cosmetics. Escherichia coli is the common engineered bacterium used to produce PCB. However, it still suffers from low production level, precursor deficiency, and low catalytic efficiency. In this study, a highly efficient PCB-producing strain was created. First, chassis strains and enzyme sources were screened, and copy numbers were optimized, affording a PCB titer of 9.1 mg/L. Most importantly, the rate-limiting steps of the PCB biosynthetic pathway were determined, and the supply of precursors necessary for PCB synthesis was increased from endogenous sources, affording a titer of 21.4 mg/L. Then, the key enzymes for PCB synthesis, HO1 and PcyA, were assembled into a multi-enzyme complex using the short peptide tag RIAD-RIDD, and 23.5 mg/L of PCB was obtained. Finally, the basic conditions for PCB fermentation were initially determined in 250 mL shake flasks and a 5-L bioreactor to obtain higher titers of PCB. The final titer of PCB reached 147.0 mg/L, which is the highest reported titer of PCB so far. This research provided the foundation for the industrial production of PCB and its derivatives.

Amanita sp. from subtropical region of Saudi Arabia as a source of chitinase enzyme and its antifungal activity

Mushroom products have been used as a biotechnological tool for many applications. Particularly, thermostable chitinase plays a vital role in biowaste management and biological control. In the present investigation, Amanita sp. was recorded in the subtropical region of Saudi Arabia, therefore, it was utilized for chitinase production using substrates chitin and dead fungal mycelia (DFM). Compared with the DFM, chitin was more suitable for chitinase activity at different temperatures and pH. Amanita sp. produced chitinase up to 70 °C, but the optimum was 50 °C. The chitinase activity was 4.98, 3.5, and 0.9 U.mg-1 with the use of chitin, while it was 4.6, 3.1, and 0.6 U.mg-1 with the use of DFM at 50, 60, and 70 °C, respectively. Chitinase activity was stable up to 60 °C, then it began to decrease at 70 °C. The chitinase activity was better at pH 4 and 5 than pH 8 and 9. The antifungal effect of the produced chitinase at 50 °C was more effective than at 60 °C. For instance, the Alternaria alternata colony radius was 3.50 cm and 2.26 cm at 50 °C while it was 4.35 cm and 4.13 cm at 60 °C when using DFM and chitin, respectively.

Marine-derived fungi as biocatalysts.

Marine microorganisms account for over 90% of ocean biomass and their diversity is believed to be the result of their ability to adapt to extreme conditions of the marine environment. Biotransformations are used to produce a wide range of high-added value materials, and marine-derived fungi have proven to be a source of new enzymes, even for activities not previously discovered. This review focuses on biotransformations by fungi from marine environments, including bioremediation, from the standpoint of the chemical structure of the substrate, and covers up to September 2022.

Salmon oyster mushroom: growth characteristics and protease production on a laboratory scale

Pleurotus have an easy cultivation and are excellent sources of proteolytic enzymes. The aimed was evaluate the mycelial growth and extracellular protease activity of Pleurotus ostreatoroseus. The mushroom was grown on potato dextrose agar (BDA) with yeast extract (YE) 0.5% (w/v). The radial mycelial growth velocity (RCV) and mycelial density occurred on different media containing 0.5% (w/v) YE: Potato Dextrose agar (BDA), GYP agar (glucose, yeast extract and peptone), malt extract agar (MEA), purple yam extract agar (EIR), torn yam extract agar (CEIN), aria extract agar (EA). Proteolytic activity was determined by the gelose block method. In mycelial growth (VCM) is used: cupuaçu exocarp (CC), açaí (Sac), pineapple (CsAb) and sawdust (SER), supplemented with rice bran (FA) or wheat bran (FT) were used. Proteases were extracted in sterile distilled water under 180 rpm agitation at 30 °C. Significant RCV was observed in MEA+YE (12.32 ± 0.10 mm/day). However, GYP agar showed strong mycelial density. Significant substrate mixture with strong mycelial density occurred on Sac+FA (0.61 ± 0.09 cm/day). The tested media were efficient in the production of extracellular proteases by this mushroom.

Improving chestnut physicochemical properties through fermentation – Development of chestnut Amazake

The increased awareness of population regarding the impact of consumption habits is leading to interest in new, innovative, diversified and health promoting foods. In this work, two new amazake fermented products were developed with chestnut (Castanea sativa Mill.), using rice or chestnut koji as source of glycolytic enzymes. The analysis of the amazakes evolution showed improvements in chestnuts physicochemical characteristics. The fermented products presented higher values of soluble protein, sugars, starches, antioxidant capacity, and similar values of ascorbic acid for chestnut koji amazake. The adhesiveness increased, which is related to the enhanced concentrations of sugars and starches. The evolution into less structured products was observed in the firmness followed by a consistent decrease of the viscoelastic moduli.The developed chestnut amazakes can represent a suitable alternative to traditional amazake, creating an opportunity for valorisation of chestnut industrial by-products, as new, tasty, and nutritive fermented products with potential functional characteristics.

Is the Phytotoxic Effect of Digestive Fluid of Nepenthes x ventrata on Tomato Related to Reactive Oxygen Species?

The digestive fluid of pitcher plants is a rich source of enzymes and secondary metabolites, but its impact on higher plant growth and development remains unknown. The aim of the study was to determine the phytotoxicity of the digestive fluid of the pitcher plant (Nepenthes x ventrata) on the germination of tomato (Solanum lycopersicum L.) seeds, elongation growth and cell viability of roots of tomato seedlings. The digestive fluid was collected from pitchers before feeding and four days after feeding; the pH and electrical conductivity of the fluid were determined. Undiluted and 50% fluids were used in the study. An inhibition of germination of tomato seeds, by around 30% and 55%, was respectively observed in 50% and 100% digestive fluids collected before and after feeding. Digestive fluid did not affect the root growth of tomato seedlings; a slight (6%) inhibition was only observed after the application of 100% digestive fluid from an unfed trap. The roots of the tomato seedlings treated with undiluted fluid were characterized by reduced cell viability. Reactive oxygen species (H2O2 and O2•-) were mainly localized in the root apex regardless of the used phytotoxic cocktail, and did not differ in comparison to control plants.

Biochemical and Kinetic Study for the Partial Purified Thioredoxin Reductase from Healthy Human Serum

The research included the separation of thioredoxin reductase (TrxR) from human blood serum using different biochemical techniques. Two proteinous peaks had been isolated by gel filtration using sephadex)G-50) and three proteinous peaks isolated from sephadex (G-100) that produced by ammonium sulphate precipitation (65%). The approximately molecular weight of the isolated protein as a source of enzyme using gel filtration chromatography (G-100) was (118059 + 925) Dalton.
 The results showed two peaks of enzyme when using ion exchange-type DEAE-cellulose and when using electrophoresis technique type SDS-PAGE indicated the presence of two bands from peak of separation column type sephadex G-100 and one band of each peak, resulting from the ion exchange process with molecular weights of 58231 +490 Dalton.
 The results showed that the optimum conditions of purified enzyme from human serum was at (200 µg/ml) of protein as a source of the enzyme using (0.4 mol/l) citric acid-Na2HPO4 buffer solution at pH (7) act for (25) minutes at (40°C). Using Line Weaver-Burk plot, the values of maximum velocity (Vmax) and Michaelis constant (Km) were found to be (0.182 µmol/ min) and (0.013 mol/l) respectively using 5,5-- dithio bis (2-nitrobenzoic acid) (DNTB) as a substrate. Finally, this was, also, involved the study of the effect of some chemicals and drugs on the enzyme activity. The results showed that gundinium - HCl is uncompetitive inhibitor, while bromoacetic acid is noncompetitive inhibitor and zinc sulphate is a competitive inhibitor for the enzyme at different concentrations of inhibitors, but dexathamazone was an activator to the enzyme.

Dynamics and thermodynamics of bimolecular reactions at heterogeneous biological interfaces.

The behaviors of biomolecules at heterogeneous biological aqueous-nonaqueous interfaces markedly differ from those in bulk water. We found that various biomolecular reactions, including protein unfolding, protein-ligan binding, chlorophyll demetallation, redox reactions, and crystallization, are accelerated by a factor of 103 or higher at the interface of micron-sized water droplets (microdroplets) (PNAS, 2015; QRevPhys, 2015 and 2017). The FRET analysis revealed that molecules are highly localized at the nanoscale regime near the water surface, which is in part responsible for the observed accelerated dynamics (JPCB, 2020). Besides, the thermodynamics of biomolecular reactions are also altered near the water surface. We found that molecules are in highly ordered states near the aqueous-nonaqueous interface (Sci Adv, 2020, Sci Rep, 2020). This minimized entropy lowers entropic barriers so that thermodynamically unfavorable reactions such as phosphorylation and RNA formation can proceed without any enzyme or ATP energy source at room temperature (PNAS 2017, 2018). Moreover, the charge exchange between water molecules and biomolecules spontaneously occurs at the water interface, leading to the oxidation of water molecules and the reduction of biomolecules in the Krebs cycle (JACS, 2019; Nat Commun, 2018; PNAS, 2018, 2019; Chem Sci, 2019). Vibrational Stark spectroscopy coupled with stimulated Raman excited fluorescence microscopy showed the presence of a strong intrinsic interfacial electric field formed at the surface of the microdroplet (JPCL, 2020). This strong electric field, combined with the partial desolvation effect, can be responsible for the unusual behaviors of biomolecules at the aqueous-nonaqueous interface (Annu Rev Phys Chem, 2020). These findings suggest that the dynamics and thermodynamics of biomolecular reactions occurring in cells with various aqueous-nonaqueous interfaces and compartments may significantly differ from those in bulk water.

황칠나무 추출물의 고초균 발효물로 제조된 가쓰오부시 단백가수분해물의 Lactobacillus plantarum 발효를 통한 고농도 GABA 생산

To develop a multi-functional ingredient, the bioconversion of katsuobushi protein was optimized using Bacillus subtilis HA and Lactobacillus plantarum KS2020. The Dendropanax morbiferus extract (DME) culture with protease activity (102 unit/mL) was prepared by B. subtilis with 2% glucose and 1% skim milk through one day of alkaline fermentation. Katsuobushi protein was effectively hydrolyzed by the DME culture at 60°C for 3 hours, resulting in a tyrosine content of 156.85 mg%. Subsequently, a second lactic acid fermentation was carried out with 10% monosodium glutamate (MSG) using L. plantarum KS2020 to produce higher levels of GABA. Following co-cultivation for three days, DME exhibited a pH of 8.3 (0% acidity). After seven days, the viable cell count of L. plantarum increased to 9.33 CFU/mL, but viable Bacillus cells were not detected. Taken together, a multi-functional ingredient with enriched GABA, peptides, probiotics, and umami flavor was developed through lactic acid fermentation using hydrolyzed katsuobushi protein. These results indicate that katsuobushi protein could be used as a byproduct to produce a palatable protein hydrolysate using alkaline-fermented DME culture as a proteolytic enzyme source.

Fish By-Products: A Source of Enzymes to Generate Circular Bioactive Hydrolysates.

Fish viscera are usually discarded as waste, causing environmental problems, or as low-value by-products. This study describes a self-sufficient and zero waste approach to obtain enzymes and protein hydrolysates from fish by-products. Firstly, recovery steps of viscera enzymatic extract were applied, and the resulting raw extract was stable at a pH range of 8-9 and at temperatures between 40 and 50 °C. The application of the extracted enzymes and alcalase on fish by-products hydrolysis was also determined. The selected conditions for the enzymatic hydrolysis were 10% (E/S) for 6 h using viscera enzymatic extract and 3% (E/S) for 2 h using alcalase. Fish protein hydrolysates (FPH) proved to have a notable antioxidant capacity with similar activity, ~11 mg ascorbic acid/g dry extract (ABTS assay) and ~150 mg Trolox/g dry extract (ORAC assay). FPH were also able to inhibit the angiotensin-converting enzyme, however, alcalase hydrolysates revealed a higher antihypertensive potential, IC50 of 101 µg of protein/mL. In general, FPH obtained by both enzymes systems maintained these bioactivities after the passage throughout a simulated gastrointestinal tract. The hydrolysates also displayed important technological properties, namely oil absorption capacity (~1 g oil/g sample) and emulsifying property (~40%). Therefore, it will be conceivable to use fish by-products based on a circular economy approach to generate added value compounds for animal and human nutrition.

Differences in Carbon Sequestration Ability of Diverse Tartary Buckwheat Genotypes in Barren Soil Caused by Microbial Action.

Planting plants to increase soil carbon input has been widely used to achieve carbon neutrality goals. Tartary buckwheat not only has good barren tolerance but is also rich in nutrients and very suitable for planting in barren areas. However, the effects of different genotypes of Tartary buckwheat roots and rhizosphere microorganisms on soil carbon input are still unclear. In this study, ozone sterilization was used to distinguish the sources of soil organic acids and C-transforming enzymes, and the contribution of root and rhizosphere microorganisms to soil carbon storage during the growth period of two genotypes of tartary buckwheat was studied separately to screen suitable varieties. Through the analysis of the experimental results, the conclusions are as follows: (1) The roots of Diqing tartary buckwheat have stronger carbon sequestration ability in a barren environment than Heifeng, and the microorganisms in Diqing tartary buckwheat soil will also increase soil carbon input. Therefore, Diqing tartary buckwheat is more suitable for carbon sequestration than Heifeng tartary buckwheat in barren soil areas. (2) In the absence of microorganisms, the rhizosphere soil of tartary buckwheat can regulate the storage of soil organic carbon by secreting extracellular enzymes and organic acids. (3) The structural equation model showed that to promote carbon sequestration, Heifeng tartary buckwheat needed to inhibit microbial action when planted in the barren area of Loess Plateau, while Diqing tartary buckwheat needed to use microbial-promoting agents. Adaptive strategies should focus more on cultivar selection to retain carbon in soil and to assure the tolerance of fineness in the future.

The Genues Trichoderma: Ecologicological Diversity, Taxonomic Classification and Its Agricultural, Human Health, Industrial and Environmental Applications

The genus Trichoderma is a diverse group of free-living fungi in the family Hypocreaceae (class Ascomycetes), that are living at different ecosystems in a wide range of climatic zones and that can be found all over the globe. The common ecological nich of the genus Trichoderma includes Natural soils, Decaying wood, Agricultural Habitats, Endophytes and mushroom related substrates. Trichoderma classified base on their morphological and cultural characteristics, conidiophore structure as well as the size of conidia. Now adays, molecularmethods including DNA fingerprinting and sequence analysis of multiple genes (ITS1- 5.8 rDNA-ITS2 and genes encoding translation-elongation factor 1-alpha (tef1) as well as some specific molecular databases have been used for taxonomic classification of this fungi. Trichodermahave been used agriculture biological control of plant diseases and as plant growth promoter, in industry as sources of cellulolytic and hemicellulolytic enzymes used in different industries, in human health producer of secondary metabolites that have clinical importance and in enviromt they are as a potential biodegrader of toxic compounds and soil bioremediation.Therefore, this review article presented the ecological diversiy, taxonomic classification and application of genus Trichoderma.

17β-Hydroxysteroid dehydrogenases types 1 and 2: Enzymatic assays based on radiometric and mass-spectrometric detection.

The 17β-hydroxysteroid dehydrogenase type 1 (HSD17B1) has a key role in estrogen biosynthesis as it catalyzes the reduction of estrone to the most potent estrogen, estradiol. Estradiol has a high affinity for estrogen receptors and thus stimulates their transactivation, which leads to cell proliferation and numerous other effects. HSD17B2 catalyzes the oxidation of estradiol to the less potent estrone, thereby decreasing estrogen receptor activation, which results in reduction of estrogen-associated effects. HSD17B1 and HSD17B2 overexpressing E.coli homogenates or recombinant enzymes can be used for screening and development of drugs against various pathologies such as cancer, endometriosis or osteoporosis. Here we describe the preparation of HSD17B1 and HSD17B2 bacterial homogenates and purified recombinant HSD17B1 protein as enzyme sources as well as enzymatic assays based on radiometric and mass-spectrometric detection for enzyme characterization.