Tannase Enzyme Research Articles

Tamarindus indica seed induced tannase production from Aspergillus niger

Introduction and Aim: Tannase an inducible extracellular microbial enzyme is extensively produced by different fermentation techniques and finds wide applications in food, pharmaceutical and textile industries. The present investigation focused on tannase production from Aspergillus niger through submerged (SmF) and solid-substrate fermentation (SSF) using different media. The enzyme was also induced using tamarind seeds. Methods and Materials: Aspergillus niger was grown in various submerged media such as Tannase screening broth (TSB), Czapek Dox broth (CDB), Sabouraud’s Dextrose broth (SDB) and Potato Dextrose broth (PDB). Among solid substrates wheat bran, ground nut cake, coconut cake and tamarind seed powder used individually and in mixed form. Results: In SmF, the highest tannase activity of 713.6 U/ml was found in TSB, followed by CDB (590.6 U/ml), SDB (465.6 U/ml) and PDB (424.8 U/ml). In all media highest activity was on 4th day while in SDB on 6th day of incubation. Tannase production was also carried out in CDB supplemented with 2% processed tamarind seeds to test for its induction, where the highest (936 U/ml) activity was in raw seeds than in processed seeds. SSF in mixed substrates showed highest activity of 1708.6 U/gds followed by 1278.0, 986.5, 826.3 and 723.6 U/gds in tamarind seeds, wheat bran, ground nut cake and coconut cake, respectively. Conclusion: Tannase enzyme is found to be induced by raw tamarind seed supplemented submerged medium than with processed seeds. In SSF with tamarind seed showed the highest tannase activity indicating that the tannic acid present in raw seeds serve to induce Aspergillus niger for increased production of tannase enzyme.

Estimation of Gallic Acid in Triphala Using Enzymatic Hydrolysis

A well-known Ayurvedic formulation, Triphala has numerous health benefits like appetite stimulation, controls diabetes, reduces cholesterol, relieves stress, alleviates inflammation and treats a variety of bacterial and fungal infections along with digestive problems. The phytoconstituents flavonoids, tannins, polyphenols and many chemical compounds are responsible for its claimed health benefits. One of the very significant compounds is gallic acid, which is reported to possess hepatoprotective, anti-inflammatory, antineoplastic and antioxidant properties that are beneficial in treatment of several diseases. In the present work using gallic acid as a marker, an attempt has been made to extract gallic acid from tannins present in Triphala by enzymatic hydrolysis and subsequent estimation of total gallic acid content by high performance thin layer chromatography (HPTLC). Rhizopus oryzae fungus is cultured for the production of enzyme tannase, which converts tannins into gallic acid. The extracted gallic acid is estimated in the biocatalyzed sample at optimal process parameters by high performance thin layer chromatography (HPTLC) method. The gallic acid content after treatment with enzyme was found to be doubled 0.296 ± 0.018 mg/ml, Rf value 0.37 at 254nm which was 0.197 ± 0.008 mg/ml and Rf value was 0.37 prior to enzymatic treatment. Gallic acid in Triphala is present in free form as well as the constituent of tannins. There is a remarkable yield of gallic acid from tannins using the enzyme Tannase in the conversion process. Application of such advanced technology increases the gallic acid yield from response surfaces in herbal and ayurvedic products containing gallic acid. The extraction condition of phenolic compounds can be optimised using enzyme hydrolysis. The effective factors including the solid to solvent ratio, enzyme concentration, particle size and extraction time can be optimised. Keywords: Triphala, Gallic acid, Tannin, Rhizopus oryzae, Tannase, HPTLC

Pestalotiopsis mangiferae isolated from cocoa leaves and concomitant tannase and gallic acid production

The enzyme tannase is of great industrial and biotechnological importance for the hydrolysis of vegetable tannins, reducing their undesirable effects and generating products for a wide range of processes. Thus, the search for new microorganisms that permit more stable tannase production is of considerable importance. A strain of P. mangiferae isolated from cocoa leaves was selected and investigated for its capacity to produce tannase enzymes and gallic acid through submerged fermentation. The assessment of the variables affecting tannase production by P. mangiferae showed that tannic acid, ammonium nitrate and temperature were the most significant (8.4 U/mL). The variables were analyzed using Response Surface Methodology - RSM (Box-Behnken design), with the best conditions for tannase production being: 1.9% carbon source, 1% nitrogen source and temperature of 23 °C. Tannase activity doubled (16.9 U/mL) after the optimization process when compared to the initial fermentation. A pH of 7.0 was optimal for the tannase and it presented stability above 80% with pH between 4.0 and 7.0 after 2h of incubation. The optimal temperature was 30 °C and activity remained at above 80% at 40–60 °C after 1 h. Production of gallic acid was achieved with 1% tannic acid (0.9 mg/mL) and P. mangiferae had not used up the gallic acid produced by tannic acid hydrolysis after 144 h of fermentation. A 5% tannic acid concentration was the best for gallic acid production (1.6 mg/mL). These results demonstrate P. mangiferae’s potential for tannase and gallic acid production for biotechnological applications.

Effects of enzyme treatment on volatile and non-volatile compounds in dried green tea leaves.

Green tea contains polyphenols, mainly four catechins, including (-)-epigallocatechin gallate, (-)-epicatechin gallate, (-)-epigallocatechin, and (-)-epicatechin. Enzyme tannase is known to hydrolyze gallated catechins into non-gallated catechins and gallic acid (GA). In this study, dried green tea leaves were treated with tannase to determine changes of volatile and non-volatile compounds by the hydrolysis. The results indicated that (-)-epigallocatechin, (-)-epicatechin, and GA increased, while (-)-epigallocatechin gallate and (-)-epicatechin gallate decreased after the treatment. The GA level increased in the treated samples, which increased titratable acidity significantly, while the pH became lower. Furthermore, the antioxidant activity of the tannase-treated tea leaves increased. The level of glycosidically bound aromas decreased with the concomitant increase of corresponding volatile compounds, while some alcohols derived from fatty acids decreased significantly after the treatment. These results suggest that tannase-treatment influences both volatile and non-volatile compounds in dried green tea leaves.

Structural diversity and substrate preferences of three tannase enzymes encoded by the anaerobic bacterium Clostridium butyricum

Tannins are secondary metabolites that are enriched in the bark, roots, and knots in trees and are known to hinder microbial attack. The biological degradation of water-soluble gallotannins, such as tannic acid, is initiated by tannase enzymes (EC 3.1.1.20), which are esterases able to liberate gallic acid from aromatic-sugar complexes. However, only few tannases have previously been studied in detail. Here, for the first time, we biochemically and structurally characterize three tannases from a single organism, the anaerobic bacterium Clostridium butyricum, which inhabits both soil and gut environments. The enzymes were named CbTan1-3, and we show that each one exhibits a unique substrate preference on a range of galloyl ester model substrates; CbTan1 and 3 demonstrated preference toward galloyl esters linked to glucose, while CbTan2 was more promiscuous. All enzymes were also active on oak bark extractives. Furthermore, we solved the crystal structure of CbTan2 and produced homology models for CbTan1 and 3. In each structure, the catalytic triad and gallate-binding regions in the core domain were found in very similar positions in the active site compared with other bacterial tannases, suggesting a similar mechanism of action among these enzymes, though large inserts in each enzyme showcase overall structural diversity. In conclusion, the varied structural features and substrate specificities of the C. butyricum tannases indicate that they have different biological roles and could further be used in development of new valorization strategies for renewable plant biomass.

Characterization of Apple Juice Clarified by Tannase from Serratia marcescens IMBL5 Produced using Agro-industrial Waste Materials

In the present study, clarification of apple juice with tannase from S. marcescens IMBL5 produced using various agro-waste materials was carried out. Sugarcane bagasse was found to be the most suitable source for the augmented production of tannase enzyme by response surface methodology with the temperature at 40 °C, pH 4.5 and the incubation period of 96 hrs. The enzyme was quantified and partially purified through protein precipitation. The partially purified tannase with gelatin clarified about 62% of the apple juice in 3 hr of incubation at room temperature and it was gently increased with the incubation period. The detannification was characterized by estimating tannin content of the clarified juice. The amount of total reducing sugar in the juice was increased almost 50 % after 5 hours of incubation period. FTIR spectrum of the clarified juice revealed that the conformational changes that occurred in the functional groups. The spectrum absorptions between 500 and 1700 cm-1 mainly reflected the C=O stretch of the pectins and acids and C–O modes of the carbohydrates that correspond to the absorption zones of the sugars. The HPLC analysis of the clarified apple juice indicate the presence of phenolic compounds and sugar derivatives such as gallic acid, catechin, caffeic acid, epicatechin, glucose and sucrose which confirms the quality and clarity of the apple juice using the tannase enzyme.

Dynamic Modeling of Tannase Production from Bacillus cereus: A Framework Simulation based on Fed Batch Strategy

Tannase enzyme is a metabolite of great interest in the industry. Typical examples of its use can be found in wines and beer production, beverage and juice fruits clarification and leather production. However, tannase production on an industrial scale is limited to the operation batch mode. For this reason, its production is low and severe limitations take the place of being carried on a large scale. To improve production, this research proposes an operation strategy based on Fed-batch mode. The kinetic constants were taken from the literature to simulate trends obtained through a Feed-batch mode of operation. One of the most important findings of this research focuses on increasing tannase production with found values of 0.380 U/g. The latter indicates that tannase production could be almost twice the concentration obtained with the traditional batch mode (0.1900 U/g). Results obtained in this research may be promising for the enzyme production industry. Using computational techniques, it is possible to identify an improvement without investing in excessive experimentation and resources.

Cross‐linked tannase‐carbon nanotubes composite in elevating antioxidative potential of green tea extract

Multi-walled carbon nanotubes (MWCNT)-tannase composite was investigated as an immobilized biocatalyst on the basis of its facile preparation, low cost, and excellent aqueous dispersibility. Cross-linked tannase enzymes, obtained in the presence of glutaraldehyde, were composited with MWCNT via physical adsorption. Multiple techniques were applied to investigate, and corroborate the successful adsorption of cross-linked tannase onto the MWCNT structure. Green tea infusion extract post-treatment using the composite preparation showed elevated radical scavenging activities relative to the control. Green tea infusion extract exhibited a markedly reduced EC50 value on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals following its treatment with the enzyme composite, which represents 20%-34% enhancement in its free radical scavenging capacity. Stoichiometry and number of reduced DPPH were determined and compared. The antioxidative potential of a widely consumed, health-beneficial green tea is elevated by the treatment with MWCNT-tannase composite. PRACTICAL APPLICATIONS: Cross-linked tannase enzymes were composited with pristine multi-walled carbon nanotubes via simple physical adsorption. The composite presents key advantages such as low specific volume compared to other well-known immobilization media, inert, facile enzyme composition, and ease of recovery for repeated use. The work demonstrated carbon nanotube prosthetic utility in the biotransformation of food-based health commodity sought after for its nutritional benefits. The approach is of both industrial- and agricultural importance, and is a promising and viable strategy to obtain a natural, functional food supplement for the multi-billion dollar well-being and health-related industries.

Diverse methanogens, bacteria and tannase genes in the feces of the endangered volcano rabbit (Romerolagus diazi).

BackgroundThe volcano rabbit is the smallest lagomorph in Mexico, it is monotypic and endemic to the Trans-Mexican Volcanic Belt. It is classified as endangered by Mexican legislation and as critically endangered by the IUCN, in the Red List. Romerolagus diazi consumes large amounts of grasses, seedlings, shrubs, and trees. Pines and oaks contain tannins that can be toxic to the organisms which consume them. The volcano rabbit microbiota may be rich in bacteria capable of degrading fiber and phenolic compounds.MethodsWe obtained the fecal microbiome of three adults and one young rabbit collected in Coajomulco, Morelos, Mexico. Taxonomic assignments and gene annotation revealed the possible roles of different bacteria in the rabbit gut. We searched for sequences encoding tannase enzymes and enzymes associated with digestion of plant fibers such as cellulose and hemicellulose.ResultsThe most representative phyla within the Bacteria domain were: Proteobacteria, Firmicutes and Actinobacteria for the young rabbit sample (S1) and adult rabbit sample (S2), which was the only sample not confirmed by sequencing to correspond to the volcano rabbit. Firmicutes, Actinobacteria and Cyanobacteria were found in adult rabbit samples S3 and S4. The most abundant phylum within the Archaea domain was Euryarchaeota. The most abundant genera of the Bacteria domain were Lachnoclostridium (Firmicutes) and Acinetobacter (Proteobacteria), while Methanosarcina predominated from the Archaea. In addition, the potential functions of metagenomic sequences were identified, which include carbohydrate and amino acid metabolism. We obtained genes encoding enzymes for plant fiber degradation such as endo 1,4 β-xylanases, arabinofuranosidases, endoglucanases and β-glucosidases. We also found 18 bacterial tannase sequences.

Evaluation of Activity of Tannases Produced by Isolated from Syrian Woody Soils Trichoderma citrinoviride and T. brevicompactum on Tannins Degradation

This study was conducted during the period 2019–2020 to identify Trichoderma spp. isolated from woody soils and assessing their efficacy for biodegradation of tannins through tannase enzyme activity produced. Results obtained confirmed the presence of two species; Trichoderma citrinoviride, which was isolated from Maysalon area near Damascus, and characterized by yellowish green colony with dense growth of spores at the center of the colony, and Trichoderma brevicompactum isolated from the Balluran area near Lattakia characterized by yellow colonies with concentric rings. Trichoderma citrinoviride had higher biodegradation activity, measured by degrading different tannins concentrations (2, 4, 6%) collected from Queircus coccifera from Bmelka area in Tartous in liquid medium and led to 85, 87 and 90% degradation, for the three concentrations, respectively, following 12 days incubation. The activity of the produced tannase was measured to be 37.9 units/mg. Whereas, the decomposition rate of the three tannin concentrations by Trichoderma brevicompactum reached 67, 80 , 89%, respectively, again after 12 days of incubation, with enzyme activity measured to be 35.2 units/mg. Keywords: Trichoderma, tannins, tannase, biodegradation, enzyme activit

Kinetic Studies of Aspergillus ficuum Tannase Enzyme and its Application in Detannification of Grape Juice

This investigation was carried out to study the characterization and kinetics of the partially purified Aspergillus ficuum tannase enzyme for commercial scale application in the clarification and detannification of juices like grape. pH, temperature and optimum substrate concentration were determined for the tannase enzyme activity from Aspergillus ficuum. Also, the Crude and partial purified enzyme were tested for detannification grape juice quality characteristics. Results evidenced that it was found that the optimum partial purified tannase parameters were: temperature 50 oC, PH 5.5 and substrate concentration 1.6% as a tannic acidwith 6.155 specific activities of partial purified tannase enzyme with 225.46% relative activity higher than its control. The tannase enzyme showed a higher temperature and lower pH optima than the soluble enzyme and showed 81.61% and 66.36 % tannin removal from grape juice by crude enzyme and partial purified enzyme treatment, respectively at 37 oC, which has not been reported earlier. Also, maximum velocity of reaction (Vmax) and Michaeli’s constant (Km) values were 0.72 and 1.45 for partially purified tannase enzyme activity, respectively. Overall the quality of grape juice was better in partial pure tannase treated juice compared with untreated and crude enzyme treated juice. The present study exposed thatthis enzyme could demonstrate a valuable tool for a number of biotechnological applications. Also, the application of this enzyme on juice clarification also states that it can be employed in fruit juice processing for removal of tannin efficiently.

Optimized production of tannase and gallic acid from fruit seeds by solid state fermentation

Purpose: To investigate the possibility for gallic acid production from different tannin-rich fruit seeds using Aspergillus oryzae via solid-state fermentation.
 Methods: Fruit seeds of apple, guava, tamarind, black plum and watermelon were analyzed to estimate the synthesis of an enzyme tannase and its product gallic acid. Various physicochemical parameters were optimized to increase the gallic acid yield. Gallic acid was extracted by Soxhlet apparatus and identified by Fourier-transform infrared spectroscopy (FTIR). It was quantitatively determined by high performance liquid chromatography (HPLC).
 Results: Amongst the various substrates tested, black plum seeds gave the highest activity of 34.40 U/g for tannase and 16.66 mg/g for gallic acid under optimized physicochemical conditions, i.e., 1:3 substrate: moisture ratio, 30 °C, 96 h incubation period and pH 5.5. Addition of carbon source had a negative effect on production while ammonium sulphate (0.2 %) as nitrogen source increased the yield of both products. The gallic acid produced was 98.5 % pure, compared to the standard.
 Conclusion: Production of tannase and gallic acid via solid-state fermentation conditions has been optimized in vitro. The optimized conditions can be utilized on a commercial scale for economically viable production of gallic acid.

In-vitro biotransformation of tea using tannase produced by Enterobacter cloacae 41.

Tannase is a widely used enzyme that improves the quality of tea by facilitating the release of water-soluble polyphenolic compounds, as well as reduces the formation of tea creams. The microbial tannase enzymes are often employed for tea biotransformation by hydrolyses esters of phenolic acids, including the gallated polyphenols found in blacks teas. The study was focused to investigate the tannase enzyme mediated biotransformation of black tea such as CTC-(Crush, tear, curl) & Kangra orthodox which are commonly used by the south Indian peoples. HPLC spectral analysis revealed that tannase treatment on tea cream formation (CTC & Kangra orthodox tea) allows the hydrolysis of the EGC, GA, ECG, and EGCG. A significant reduction in the formation of tea cream and increased antioxidant activity has been observed in the CTC (1.62 fold) and Kangra orthodox (1.55 fold). The results revealed that tannase treatment helps to improve the quality of black tea infusions with respect to cream formation, the intensity of colour, and sensory characteristics of tea. The result of this study indicates that E. cloacae 41 produced tannase can be used to improve the quality of both tea samples.

Клонирование, выделение и изучение свойств новой рекомбинантной танназы Aspergillus niger

A new recombinant Aspergillus niger tannase (tannin acyl hydrolase) produced by the Penicillium verruculosum fungus has been studied. A strain with a high level of extracellular tannase (TAN2) secretion (80% of the total extracellular protein) was obtained by cloning the tan2 gene (PDB Acc. No: MT828303) into the recipient strain. The tannase enzyme preparation degraded tannins in black tea extracts. TAN2 was isolated in homogeneous form using chromatographic methods; the enzyme had a high activity against gallotannin (53 U/mg) and less activity against propyl gallate (4.7 U/mg). Homogeneous TAN2 showed temperature and pH optima of 45 °C and 3.5, respectively. At a temperature of 50 °C, TAN2 retained above 80% activity for 3 h; at 60 °C, it retained about 75% of its activity for 90 min; at 70 °C, the enzyme was completely inactivated within 10 min. Tannase was characterized by a high tolerance to NaCl, the activity against gallotannin exceeded 50% of the initial value in solutions with a salt concentration of up to 5 M. The tannase activity was stimulated by Ca2+, Mg2+, Zn2+, Mn2+, Cu2+, Cd2+, Pb2+ by 3--64% and inhibited by 4-65% in the presence of Co2+, Fe3+ and Fe2+ ions. tannase, tannins, Aspergillus niger, Penicillium verruculosum This work was supported by the Ministry of Science and Higher Education of the Russian Federation.

Response Surface Optimization and Impact of Immobilized Enzymes Naringianse and Tannase on the Quality Parameters of Citrus maxima Juice

Pomelo has been reported as a rich source of flavanone glycoside with antioxidants and exhibits favorable health effects such as antimicrobial, anti-inflammatory, antiatherogenic, antitumor, and anti-clotting activity. Despite all the beneficial health impacts of Citrus maxima, it still has lower commercial value because of its juice's bitterness due to the presence of naringin and tannic acid. Therefore, an attempt has been made for the cost-effective and economic debittering process using naringinase and tannase enzymes. The 17 experiments were planned according to RSM, BBD to analyze the effect of independent variables with three levels of each, i.e., Enzyme ratio ((Naringinase: tannase) (100:0, 50:50, 0:100)), incubation temperature (30°C, 40°C, 50°C) and incubation time (2, 3, 4 hrs) on physicochemical quality of Citrus maxima Juice. The study's result indicated that independent variables affected the responses (pH, TSS, TA, Naringin content, Tannin content, TPC, and Vitamin C content). Optimization was done using Design Expert 10.0.1 software to debitter and clarify citrus maxima juice by immobilized enzymes. The optimum values were found to be 54.55, 50°C, and 4 hrs. The values for pH, TSS, TA, Naringin content, Tannin content, TPC and Vitamin C content were found to be 3.17, 6.256 °Brix, 0.885 %citric acid , 220.549 μg/ml ,0.311mg/ml, 1256.721 mg GAE/L,30.309 mg/100ml respectively. From the study, it could be concluded that the maximum debittering and clarification of citrus maxima juice could be done under processing conditions, i.e., enzyme ratio 50:50, incubation temperature 50°C and incubation time 4 hrs.

Combined isoflavones biotransformation increases the bioactive and antioxidant capacity of soymilk.

Isoflavones are phenolic secondary metabolites mainly occurring in soy and soybean products. Compared to glycoside forms, isoflavone aglycones present higher biological activities. This study evaluated the potential of microbial and enzymatic treatments in biotransformed isoflavones in their biologically active forms in soymilk. Seven different cultures of lactic acid bacteria and bifidobacteria associated with the action of immobilized tannase enzyme were screened for isoflavone glycoside biotransformation ability. The biotransformed soymilk samples were characterized regarding isoflavone profile, total phenolic content, and in vitro antioxidant activities. All bacterial strains showed a good growth capacity in soymilk matrix and produced β-glucosidase enzyme, which hydrolyzed isoflavone glycosides into aglycones in soymilk after 24h of fermentation. The microbial fermentation followed by tannase reaction (FT processes) resulted in the highest increase of bioactive aglycones (10.3- to 13.1-fold for daidzein, 10.4- to 12.3-fold for genistein, and 3.8- to 4.7-fold for glycitein), compared to control soymilk. Further, FT processes enhanced the total phenolic content (53-70%) and antioxidant activity by ORAC (69-102%) and FRAP (49-71%) assays of the soymilk matrix. Therefore, the combination of microbial fermentation and tannase treatment is a promising strategy to obtain a fermented soy product rich in bioactive isoflavones with greater health-promoting potential. KEY POINTS: • Bacterial cultures and tannase enzyme displayed isoflavone deglycosylation activity. • The addition of tannase following the fermentation maximized the isoflavone conversion. • Increased isoflavone aglycones contributed to the improved antioxidant activity of soymilk.

Contributions of protein microenvironment in tannase industrial applicability: An in-silico comparative study of pathogenic and non-pathogenic bacterial tannase

Tannase is an inducible industrially important enzyme, produced by several microorganisms. A large number of bacteria have reported as tannase producers; however, some of them are pathogenic in nature. Therefore, it is quite uncertain whether the application of these tannase enzymes from such pathogenic bacteria is suitable for industries and human welfare. Till date, there is no clear evidence regarding which group of bacteria (non-pathogenic or pathogenic) is better suited for their application in the edge of industries with particular reference to the food industry. The present study is following the findings of the above queries. In this study, a large number of tannase protein sequences have been retrieved from the databases, including both non-pathogenic and pathogenic bacterial species. Physiochemical and evolutionary properties of those sequences have been evaluated. Results have shown that non-pathogenic bacterial tannase possesses a high number of acidic and basic amino acid residues as compared to their pathogenic counterparts. The acidic and basic amino acid residues of tannase provide unique microenvironment to it. In the other hand, the numbers of disorder forming residues are higher in tannase sequences of pathogenic bacteria. The study of tannase microenvironment leads in the formation of salt bridges, which finally favoring the stability and proper functioning of tannase. This is the first report of such observation on tannase enzyme using in silico approach. Study of the microenvironment concept will be helpful in protein engineering.

Immobilization and characterization of tannase from Penicillium rolfsii CCMB 714 and its efficiency in apple juice clarification

In the present study a newly tannase enzyme from Penicillium rolfsii CCMB 714 was immobilized on calcium alginate and its efficiency in clarifying juice was analyzed. The efficiency of immobilization was 99.5% and the enzyme maintained an activity above 50% after 6 cycles of reuse. Immobilization of tannase enzyme changed the optimum temperature from 33 to 44 °C, and the optimum pH from 4.0 to 4.3. The immobilized tannase enzyme showed greater thermal and pH stability when compared to the free form, maintaining 80% of its activity at 50 °C and more than 50% of residual activity at all pH tested for up to 16 h. The immobilized enzyme was also resistant to almost all additives used (cations, SDS and EDTA). When tested in apple juice, the immobilized tannase efficiently clarified the juice showing the reduction of more than 70% in turbidity. Such results showing that the P. rolfsii CCMB 714 immobilized tannase was suitable to clarify apple juice and have a potential for future beverage industrial application mainly in juice processing.

Comparative analysis of enzymatic activity of tannase in non-conventional yeasts to produce ellagic acid

Abstract Non-conventional yeasts represent an alternative for microorganisms that are capable to produce the enzyme Tannase (EC 3.1.1.20), which has many applications is the food and pharmaceutical industry as it stands out for its ability to produce ellagic acid out of the enzymatic hydrolysis of ellagitannins. Ellagic acid has been studied in the medical area for its action against various diseases, as is the case of cancer or heart problems. The investigation was conducted to evaluate seven non-conventional yeasts - Debaryomyces hansenii PYC ISA 1510, Debaryomyces hansenii PYC 2968, Candida parapsilosis, Candida utilis, Pichia pastoris, Pichia kluyveri, Issatchenkia terricola - as producers of the enzyme tannase. This was done in mediums of fermentation with tannic acid (1-3%) and YPD without tannic acid, to determine the activity of the enzyme by two spectrophotometric methods, the first being with rhodamine and the second with methyl gallate, to detect the presence of gallic acid as a product of the hydrolysis of tannic acid. The main results showed that the evaluated non-conventional yeasts all presented activity with the Tannase enzyme, obtaining better detection results with the rhodamine technique for the stability of the formed complex, which gives more precision of the obtained data.

A newly developed tannase enzyme from Aspergillus terreus versus commercial tannase in the diet of lactating Damascus goats fed diet containing pomegranate peel

Inclusion of tannase enzyme in tannin-rich diets of lactating animals can be explored as a strategy to overcome the negative effects of tannins on feed utilization and lactational performance. The present study aimed to compare the effect of replacing commercially available exogenous tannase with a newly developed tannase, produced from Aspergillus terreus, on feed utilization and lactational performance of lactating Damascus goats fed tannin-rich pomegranate peel based diet. Thirty lactating Damascus goats were randomly stratified into 3 treatments and fed pomegranate peel-based basal diet at 10% (containing 7.13% tannins DM basis) without exogenous tannase as a control treatment or with addition of 500 IU of new tannase/kg feed daily, or 500 IU of commercial tannase/kg feed daily for 9 weeks. Tannases decreased feed intake and increased diet digestibility; however, new tannase increased (P<0.05) diet digestibility relative to commercial tannase. Tannase treatments increased (P<0.05) blood total protein and glucose concentrations. New tannase improved (P<0.05) milk production and feed efficiency, without affecting milk composition. Moreover, new tannase reduced (P<0.05) total unsaturated fatty acids (UFA) and increased (P<0.05) poly UFA and total conjugated linoleic acid concentrations, while the commercial tannase increased (P<0.05) total SFA and atherogenic index. Tannases reduced (P<0.05) milk total essential amino acids (EAA), total non-EAA, total conditionally EAA, and total branched amino acids. It is concluded that tannases enhanced the utilization of tannin-rich diet by lactating goats, with better results for the new tannase produced from A. terreus compared with the commercial tannase.