Key Digestive Enzymes Research Articles

Exploring the Intestinal Microbiota and Metabolome Profiles Associated With Feed Efficiency in Pacific Abalone (Haliotis discus hannai).

Feed efficiency (FE) is critical to the economic and environmental benefits of aquaculture. Both the intestines and intestinal microbiota play a key role in energy acquisition and influence FE. In the current research, intestinal microbiota, metabolome, and key digestive enzyme activities were compared between abalones with high [Residual feed intake (RFI) = −0.029] and low FE (RFI = 0.022). The FE of group A were significantly higher than these of group B. There were significant differences in intestinal microbiota structures between high- and low-FE groups, while higher microbiota diversity was observed in the high-FE group. Differences in FE were also strongly correlated to variations in intestinal digestive enzyme activity that may be caused by Pseudoalteromonas and Cobetia. In addition, Saprospira, Rhodanobacteraceae, Llumatobacteraceae, and Gaiellales may potentially be utilized as biomarkers to distinguish high- from low-FE abalones. Significantly different microorganisms (uncultured beta proteobacterium, BD1_7_clade, and Lautropia) were found to be highly correlated to significantly different metabolites [DL-methionine sulfoxide Arg-Gln, L-pyroglutamic acid, dopamine, tyramine, phosphatidyl cholines (PC) (16:0/16:0), and indoleacetic acid] in the high- and low-FE groups, and intestinal trypsin activity also significantly differed between the two groups. We propose that interactions occur among intestinal microbiota, intestinal metabolites, and enzyme activity, which improve abalone FE by enhancing amino acid metabolism, immune response, and signal transduction pathways. The present study not only elucidates mechanisms of variations in abalone FE, but it also provides important basic knowledge for improving abalone FE by modulating intestinal microbiota.

Structure related α-glucosidase inhibitory activity and molecular docking analyses of phenolic compounds from Paeonia suffruticosa

High postprandial hyperglycaemia is an important determinant of the development and progression of type 2 diabetes. Thus, inhibition of key digestive enzymes such as α-amylase and α-glucosidase is considered an efficient approach to control glycaemic levels in diabetics. In search of α-amylase and α-glucosidase inhibitors, the root bark of Paeonia suffruticosa was screened for inhibitors, resulting in the isolation of eleven phenolic compounds (1–11). Their enzymes inhibitory activities and inhibition mechanism were investigated using an in vitro inhibition assay and molecular docking studies. Compounds 2, 5, 6, and 8–11 (IC50 between 290 and 431 µM) inhibited α-glucosidase more effectively than the reference compound acarbose (IC50 = 1463.0 ± 29.5 µM). However, the compounds (IC50 > 800 µM) were less effective against α-amylase than acarbose (IC50 = 16.6 ± 0.9 µM). Among them, compound 10 exhibited the highest α-glucosidase inhibitory effect with an IC50 value of 290.4 ± 9.6 µM. Compounds 2, 5, 9 10 and 11 were found to be competitive inhibitors, while compounds 6 and 8 were noncompetitive inhibitors of α-glucosidase. Computational analyses showed that the main binding forces between the compounds and the main residues were hydrogen bonds. The results suggest that these compounds have the potential to be developed as α-glucosidase inhibitors.

Rational design and development of a novel and highly specific near-infrared fluorogenic substrate for sensing and imaging of human pancreatic lipase in living systems

• A novel activatable near-infrared (NIR) fluorogenic substrate ( DDAO-ol ) for human pancreatic lipase (hPL) was devised. • DDAO-ol is found with the best combination of stability, specificity, sensitivity and reactivity towards hPL. • DDAO-ol offers a practical tool for high-throughput screening and characterizing hPL inhibitors. • DDAO-ol offers a powerful visualization tool for in situ sensing and imaging of PL in living cells, tissues and organs. Human pancreatic lipase (hPL), a crucial digestive enzyme responsible for the digestion of dietary lipids in humans, has been validated as a key target for treating obesity and as a potential biomarker for diagnosis of pancreatic diseases. This study aims to devise a novel and highly specific near-infrared (NIR) fluorogenic substrate for sensing and imaging of human pancreatic lipase (hPL) in biological systems. To achieve this aim, the oleate esters of various NIR fluorophores are rationally designed on the basis of the enzymatic properties and the preferred substrates of hPL. Following virtual screening and experimental validation, DDAO-ol is found with the best combination of stability, specificity, sensitivity and reactivity towards hPL. Under physiological conditions, DDAO-ol can be rapidly hydrolyzed by hPL to release the fluorophore DDAO , which emits strong NIR fluorescence signal around 660 nm. DDAO-ol offers a novel and practical tool to construct the assay for high-throughput screening and characterizing hPL inhibitors, which strongly facilitates the discovery of novel anti-obesity agents targeting hPL. Meanwhile, DDAO-ol is also a powerful visualization tool for sensing and imaging of mammalian pancreatic lipase (PL) in living systems (including living cells, tissues and organs) with low cytotoxicity and high imaging resolution. Collectively, this study reports a novel and highly specific NIR fluorogenic substrate for hPL, which offers a practical and reliable tool for deciphering the biological functions of hPL and the relevance of this key digestive enzyme to diseases.

Feeding Protocol Modulates the Digestive Process in Senegalese Sole (Solea senegalensis) Juveniles

Understanding the daily digestive function could lead to more efficient feeding practices in the farmed fish species. In this work, the effect of several daily feeding protocols was assessed on the daily expression of the genes related to the circadian system (arntl1, clock, per1, and cry1), appetite (cck), nutrients hydrolysis (try1, ctrb1, pga, atp4a, cel, and amy2a), peptides transport (slc15a1), pH, and activity of key digestive (trypsin, chymotrypsin, amylase, 4C- and 7C-like lipases) and absorptive (alkaline phosphatase) enzymes in the gastrointestinal tract (GIT) of Senegalese sole (Solea senegalensis) juveniles. Some of the tested feeding protocols demonstrated a significant influence on the entrainment of the peripheral oscillator. The synchrony of slc15a1 and some digestive proenzymes with per1 in some of the experimental treatments could underline the role of clock genes in the regulation of these biological functions. Unlike gene expression of the digestive proenzymes wherein feeding protocol occasionally affected the temporal organization, the influence of this factor was determinant on the daily average production and/or circadian pattern of their activities. Daily pH was maintained at approximately neutral levels in all the GIT segments, and no pepsin activity was detected. Fish fed a single diurnal meal showed the lowest activity for all the studied enzymes, except for amylase, suggesting an insufficient utilization of the ingested nutrients. Higher amylase activity in this group, on the other hand, implies the possible activation of alternative compensatory metabolic pathways. Overall, our results pointed to the suitability of higher feeding frequencies, especially during dark hours, for the aquaculture production of this species.

An Integrated Peptidomics and In Silico Approach to Identify Novel Anti-Diabetic Peptides in Parmigiano-Reggiano Cheese.

Simple SummaryDietary habits and sedentary lifestyle are widely established to be the major risk factors for the development of long-term chronic conditions, such as type-2 diabetes (T2D). In this framework, where consumer’s food choices are even more influenced by an ever-growing awareness on nutritional, environmental, and healthy aspects, the presence and identification of natural bioactive compounds are gaining increasing attention in the scientific community. Pharmacological treatment of T2D is based on the administration of molecules able to inhibit some key enzymes involved in the carbohydrate digestion and insulin secretion. The multiple side-effects of synthetic inhibitors led to an increased demand for natural food-derived anti-diabetic agents. The present work offers a new integrated approach for the identification and selection of new bioactive peptides, able to inhibit the key digestive enzymes implicated in the control of blood glucose level. The novelty lies in the development of a new, quick, and cost-effective integrated methodology supported by confirmed in vitro evidence. In fact, the present work successfully accomplished the identification of two selected candidates with a possible application in the diabetes management. Indeed, the functional and healthy role of Parmigiano-Reggiano cheese in human nutrition was assessed, highlighting its potential anti-diabetic properties.Inhibition of key metabolic enzymes linked to type-2-diabetes (T2D) by food-derived compounds is a preventive emerging strategy in the management of T2D. Here, the impact of Parmigiano-Reggiano (PR) cheese peptide fractions, at four different ripening times (12, 18, 24, and 30 months), on the enzymatic activity of α-glucosidase, α-amylase, and dipeptidyl peptidase-IV (DPP-IV) as well as on the formation of fluorescent advanced glycation end-products (fAGEs) was assessed. The PR peptide fractions were able to inhibit the selected enzymes and fAGEs formation. The 12-month-ripening PR sample was the most active against the three enzymes and fAGEs. Mass spectrometry analysis enabled the identification of 415 unique peptides, 54.9% of them common to the four PR samples. Forty-nine previously identified bioactive peptides were found, mostly characterized as angiotensin-converting enzyme-inhibitors. The application of an integrated approach that combined peptidomics, in silico analysis, and a structure–activity relationship led to an efficient selection of 6 peptides with potential DPP-IV and α-glucosidase inhibitory activities. Peptide APFPE was identified as a potent novel DPP-IV inhibitor (IC50 = 49.5 ± 0.5 μmol/L). In addition, the well-known anti-hypertensive tripeptide, IPP, was the only one able to inhibit the three digestive enzymes, highlighting its possible new and pivotal role in diabetes management.

Dual effects of quercetin on protein digestion and absorption in the digestive tract

Quercetin is a well-studied natural product with multiple pharmacological properties. In this study, we demonstrated that quercetin suppressed protein digestion in the intestinal fluid by inhibiting trypsin, a key digestive enzyme. However, we also observed a previously unknown property of quercetin: promoting the intestinal absorption of proteins. In addition, the promoted protein absorption was mediated by internalization of digested oligopeptides in the intestinal epithelia rather than increasing the intestinal paracellular permeability. Notably, four other flavonoids also achieved such enhanced intestinal absorption, suggesting that this effect was associated with the aglycone flavonol backbone, but not related to their inhibitory potencies against trypsin. This study demonstrates that quercetin exhibits dual effects on protein digestion and absorption: 1) suppressing protein digestion by inhibiting trypsin in the intestinal fluid; 2) promoting the intestinal absorption of oligopeptides in the intestinal villi cells.

Lactobacillus Sps in Reducing the Risk of Diabetes in High-Fat Diet-Induced Diabetic Mice by Modulating the Gut Microbiome and Inhibiting Key Digestive Enzymes Associated with Diabetes.

Simple SummaryType 2 diabetes (T2D) is increasingly spreading across the globe. The disease is linked to a disruption of gut microbiome. Probiotics are essential gut microbiota modulators proven to restore microbiota changes, thereby conferring health to its host. This study aimed to use probiotics (lactobacilli) and their metabolites as natural anti-diabetic therapy through the modulation of gut microbiota and inhibit diabetes-causing enzymes. Lactobacillus-treated high-fat diet mice showed lower blood glucose levels and body weight. Interestingly, our study also proved that the lactobacilli altered gut microbiota composition by suppressing opportunistic bacteria that are highly associated with metabolic diseases. Our findings substantiate the use of probiotics as natural anti-diabetic therapeutics.Obesity caused by a high-fat diet (HFD) affects gut microbiota linked to the risk of type-2 diabetes (T2D). This study evaluates live cells and ethanolic extract (SEL) of Lactobacillus sakei Probio65 and Lactobacillus plantarum Probio-093 as natural anti-diabetic compounds. In-vitro anti-diabetic effects were determined based on the inhibition of α-glucosidase and α-amylase enzymes. The SEL of Probio65 and Probio-093 significantly retarded α-glucosidase and α-amylase enzymes (p < 0.05). Live Probio65 and Probio-093 inhibited α-glucosidase and α-amylase, respectively (p < 0.05). In mice fed with a 45% kcal high-fat diet (HFD), the SEL and live cells of both strains reduced body weight significantly compared to HFD control (p < 0.05). Probio-093 also improved blood glucose level compared to control (p < 0.05). The gut microbiota modulatory effects of lactobacilli on HFD-induced diabetic mice were analyzed with qPCR method. The SEL and live cells of both strains reduced phyla Deferribacteres compared to HFD control (p < 0.05). The SEL and live cells of Probio-093 promoted more Actinobacteria (phyla), Bifidobacterium, and Prevotella (genus) compared to control (p < 0.05). Both strains exerted metabolic-modulatory effects, with strain Probio-093 showing more prominent alteration in gut microbiota, substantiating the role of probiotics in gut microbiome modulations and anti-diabetic effect. Both lactobacilli are potential candidates to lessen obesity-linked T2D.

Effects of the substitution of fishmeal with mealworm meal on enzymes, haemolymph and intestinal microbiota of the Pacific white shrimp

Shrimp farming is one profitable sector of aquaculture and the Pacific white shrimp (Litopenaeus vannamei) is the most cultivated species worldwide. Nutrition is considered as one of the greatest challenges in shrimp farming due to its dependence on fishmeal (FM) as an ingredient in the feed. The main goal of the present work was to analyse if the substitution of FM by yellow mealworm (Tenebrio molitor) meal (TM) influences digestive enzyme expression, haemolymph parameters or the intestinal microbiota of shrimp juveniles. Shrimps (average initial weight 4.42±0.02 g) were fed five experimental diets where FM protein was gradually substituted by TM protein (0, 25, 50, 75, and 100%). The shrimps were fed four times per day with an amount equivalent to approximately 6% of their biomass for 42 days. The total haemocyte count, protein concentration and prophenoloxidase activity in the haemolymph were not significantly altered (P&gt;0.05) when FM was substituted by TM. However, the agglutinating activity of shrimp serum determined against dog erythrocytes was higher in the shrimp group fed 25% TM. The absolute and specific activities of trypsin, chymotrypsin, lipase, α-amylase and the patterns of proteolytic activities were not affected by the dietary treatments. The shrimp gut bacterial microbiota profile was similar with the generaPseudoalteromonas, Rubritalea, Ruegeria, Tenacibaculum, andVibrio detected. These results show that TM is viable alternative for the replacement of FM, without any deleterious effects on expression of key digestive enzymes, gut microbiota or the immune system of the Pacific white shrimp.

Microplastics alter digestive enzyme activities in the marine bivalve, Mytilus galloprovincialis.

Microplastics are eaten by many invertebrates, particularly filter-feeding organisms like mussels. Since microplastics can be retained in the digestive system for extended periods, there is ample opportunity for them to interact with the functions of digestive enzymes. This study determined how the polymer type, size and concentration of ingested spherical microplastics affects the activities of seven key digestive enzymes in the digestive gland of Mytilus galloprovincialis, a common marine mussel. Polymer type significantly affected the activities of carbohydrase enzymes: polystyrene reduced amylase and xylanase activities, and increased cellulase activity. High concentrations of microplastics (5×104 microplastics L-1) caused a 2.5-fold increase in total protease activity. The activities of laminarinase, lipases and lipolytic esterases were unaffected by the polymer type, size or concentration of microplastics. Microplastics-induced changes to digestive enzyme activities can affect mussels' ability to acquire energy from food and reduce their energy reserves.

Characterization of digestive physiology in Pacific bluefin tuna Thunnus orientalis juveniles fed a raw fish feed and a commercial diet

This study examined the biochemical characteristics of key digestive enzymes in artificially produced Pacific bluefin tuna juveniles and revealed details of the digestive physiology of juveniles fed a commercial pellet diet and raw fish over long-term (225 days) and short-term (response to feeding) periods. pH and thermal profiles of key digestive enzymes of pepsin, trypsin, chymotrypsin, lipase and amylase were examined in juvenile bluefin tuna and the characteristics differed from those previously reported for larval and adult tuna. Long-term feeding (225 days) of the commercial diet markedly increased stomach and pyloric caeca size as well as storage/synthesis levels of the digestive enzymes. Highly increased gene expression levels of cholecystokinin (cck) and its receptor (cck1r) were also found in juveniles fed the commercial diet, suggesting cck might be involved in the gastrointestinal hypertrophy and digestion enhancement seen in the present study. In the short-term experiment (response to feeding), juveniles fed the commercial diet displayed faster gastric emptying, higher pH in the intestinal content and increased digestive enzymes storage/synthesis whereas the enzymes secretion was similar with those of juveniles fed raw fish. These indicate that juvenile bluefin tuna morphologically and physiologically adapted their digestive system to the artificial diet, however the adaptation was not sufficient to obtain a comparable performance as tuna fed raw fish feed. Biochemical characteristics of the digestive enzymes and the observed effects of artificial diet on digestive physiology could be good indicators to improve feeding strategies such as diet composition, feeding ration and frequency in juvenile tuna.

Ontogeny of Expression and Activity of Digestive Enzymes and Establishment of gh/igf1 Axis in the Omnivorous Fish Chelon labrosus.

Simple SummaryThick-lipped grey mullet (Chelon labrosus) feeds on the lowest trophic levels during adult stages, for which it is considered a viable candidate for an economically and environmentally sustainable aquaculture. Similar to most of marine fish species, C. labrosus produce a large number of eggs, leading to morphologically and anatomically larvae that are not completely mature and have to pass through substantial differentiation and development in their functional systems to acquire adult features. Therefore, the study of the development of digestive tract and of the growth regulation can provide useful information to adapt the feeding protocols and rearing conditions to the physiological requirements at each stage. This work aimed to evaluate the early ontogeny of key digestive enzymes and somatotropic factors at biochemical and/or transcriptional levels. Our results evidenced that maturation of the digestive system and acquisition of the adult mode of digestion occurs around 60 to 70 days post hatch (dph), when starch or other low-cost carbohydrate-based compounds could be used in formulated diets at increasing levels. Furthermore, our results implied an independent expression of the studied somatotropic genes during the first 40 dph and establishment of a functional growth hormone/insulin-like growth factor 1 axis from 50 dph onward.Thick-lipped grey mullet (Chelon labrosus) is a candidate for sustainable aquaculture due to its omnivorous/detritivorous feeding habit. This work aimed to evaluate its digestive and growth potentials from larval to early juvenile stages. To attain these objectives the activity of key digestive enzymes was measured from three until 90 days post hatch (dph). Expression of genes involved in digestion of proteins (try2, ctr, pga2, and atp4a), carbohydrates (amy2a), and lipids (cel and pla2g1b), together with two somatotropic factors (gh and igf1) were also quantified. No chymotrypsin or pepsin activities were detected. While specific activity of trypsin and lipase were high during the first 30 dph and declined afterward, amylase activity was low until 57 dph and increased significantly beyond that point. Expression of try2, ctr, amy2a, and cel increased continuously along development, and showed a peak at the end of metamorphosis. Expression of pla2g1b, pga2 and atp4a increased until the middle of metamorphosis and decreased afterwars. Most of these trends contrast the usual patterns in carnivorous species and highlight the transition from larvae, with high protein requirements, to post-larvae/juvenile stages, with omnivorous/detritivorous feeding preferences. Somatotropic genes, gh and igf1, showed approximately inverse expression patterns, suggesting the establishment of the Gh/Igf1 axis from 50 dph.

Evaluation of in vitro α-amylase and α-glucosidase inhibitory potential and hemolytic effect of phenolic enriched fractions of the aerial part of Salvia officinalis L.

BackgroundThe inhibition of α-amylase and α-glucosidase activities is one of the major practical strategies for the control of postprandial hyperglycemia. Salvia officinalis L. is known by its various bioactive compounds and its effective therapeutic properties towards illnesses including diabetes. ObjectiveThe present study aimed to evaluate the in vitro inhibitory effect of S. officinalis on key digestive enzymes linked to type 2 diabetes and to identify its hemolytic effect. MethodsHydro-methanol decoction extract and fractions of ethyl acetate and n-butanol were investigated for their in vitro α-amylase and α-glucosidase inhibitory activities, compared to acarbose as a standard. Furthermore, they were evaluated for their hemolytic effect. ResultsPhytochemical composition demonstrated the richness of S. officinalis in secondary metabolites. Extract and fractions inhibited the activity of both enzymes. They showed weak hemolytic activity. Quantitative estimation of total phenolic and flavonoids revealed that ethyl acetate fraction contained the highest amount of these compounds (450.51 ± 0.6 μg GAE/mg DE and 352.01 ± 0.78 μg CE/mg of DE, respectively). It showed the best antidiabetic activity tested both by α-amylase and α-glucosidase assays (IC50 = 46.52 ± 2.68 and 104.58 ± 0.06 μg/mL, respectively). Moreover, this fraction showed the least hemolytic effect (11.58 ± 0.1%). ConclusionsS. officinalis extract and fractions are promising sources of α-amylase and α-glucosidase inhibitors.

Inhibition of pancreatic lipase by environmental xenoestrogens.

Environmental xenoestrogens are the most accessible endocrine disrupting chemicals that have been reported with harmful effects on human health. Although the influences of xenoestrogens on the endocrine system have been extensively studied, it remains unclear whether these xenoestrogens can affect the digestive system in mammals. This study aimed to investigate the inhibitory effects and the underlying mechanism of six non-steroidal synthetic estrogens (including hexestrol, diethylstilbestrol, dienestrol, bisphenol A, bisphenol AF and bisphenol Z) on pancreatic lipase (PL), a key digestive enzyme responsible for lipid digestion and absorption in mammals. The results clearly demonstrated that hexestrol, diethylstilbestrol and dienestrol exhibited strong inhibition on PL, with the IC50 values of less than 1.0μM. Further investigations elucidated that these three synthetic estrogens functioned as mixed inhibitors of PL, with the Ki values of less than 1μM. Moreover, molecular dynamics simulations showed that diethylstilbestrol and its analogues might block the binding of substrate on PL via occupying the portal to the active site of PL and thereby inhibit the hydrolytic activity of this key enzyme. Collectively, these results suggested that diethylstilbestrol and its analogues were potent PL inhibitors, which might play a profound role in lipid absorption and weight gain in mammals.

Impact of starvation on digestive enzymes activities and plasma metabolites in Siberian sturgeon ( Acipenser baerii , Brandt, 1869)

To increase the current knowledge about the relationship between nutritional status and the digestive capacity of Siberian sturgeon (Acipenser baerii), we addressed the effect of starvation‐refeeding and macronutrient composition on growth parameters and key digestive enzyme activities in A. baerii. Acipenser baerii juveniles were fed four different diets for 3 weeks, then starved for 2 weeks and allowed to refed for 5 weeks with the same diets. Another group of fish were fed 10 weeks with the corresponding diets. Among 10‐week fed fish, high‐protein diets promoted higher body weight values, while the lowest specific growth rate was observed in fish fed a low‐protein, medium‐carbohydrate, high‐lipid diet (p .05) and α‐amylase activity in the intestine as well as pepsin in the stomach (p < .05). Our findings suggest that A. baerii maintains a high capacity to digest proteins and lipids after 2 weeks of starvation and that α‐amylase can be used as an indicator of the nutritional status in fish submitted to starvation‐refeeding cycles. Indeed, refeeding with high‐protein and CHO:L ratio diets after starvation could improve the growth rate of A. baerii in culture.

Expression Profile of the Digestive Enzymes of Manis javanica Reveals Its Adaptation to Diet Specialization

The expression of animal digestive enzymes reflects important dietary adaptations. The pangolin, also known as scaly anteater, is a specialized myrmecophage that consumes mainly ants and termites, but its digestive enzymes have not been fully investigated. Therefore, in this study, we used shotgun proteomic analysis to examine the protein components of the saliva and intestinal juice of a Sunda pangolin (Manis javanica) that died shortly after being rescued. The intestinal juice contained greater variety of digestive enzymes, including α-amylase, maltase-glucoamylase, α,α-trehalase, sucrase-isomaltase, pepsin A, trypsin, pancreatic endopeptidase E, carboxypeptidase A1, carboxypeptidase B, dipeptidyl-peptidase 4, and pancreatic triacylglycerol lipase. The digestive enzymes identified in the saliva were maltase-glucoamylase and trypsin, and chitinase which was also found in the intestinal juice. Compared with other animals, the Sunda pangolin has less intestinal protease diversity and lacks key digestive enzymes, such as chymotrypsin and pancreatic elastase. The expression profile of the digestive enzymes of the Sunda pangolin reveals animal’s adaptation to a diet consisting mainly of ants and termites. Our results will facilitate the preparation of artificial food for rescued pangolins and for those in captivity for conservation breeding efforts.

Inhibition of α-amylase, α-glucosidase and pancreatic lipase by phenolic compounds of Rumex maderensis (Madeira sorrel). Influence of simulated gastrointestinal digestion on hyperglycaemia-related damage linked with aldose reductase activity and protein glycation

In this work, we report the in vitro inhibitory potential of Rumex maderensis methanolic extracts (leaves, flowers, and stems) towards key digestive enzymes linked to type-2 diabetes and obesity (α-amylase, α-glucosidase, pancreatic lipase). The inhibitory activity towards aldose reductase and glycation of bovine serum albumin (BSA) is also reported; in these latter assays, the effect of simulated digestion on the bioactivities was evaluated. The inhibitory activities of R. maderensis extracts were statistically compared with the inhibition produced by reference compounds for each assay. The analysed extracts exhibited significant inhibitory activities, which decreased after the gastrointestinal digestion, possibly due to some loss of phenolics that took place during the digestion process. The most important results were observed during the BSA-glycation assay, in which the analysed extracts presented higher potency than a reference compound: aminoguanidine (AMG). This research is the first showing the potential anti-diabetic activity of R. maderensis.

Comparative transcriptome analysis reveals the expression and characterization of digestive enzyme genes in the hepatopancreas of the Chinese mitten crab

The Chinese mitten crab Eriocheir japonica sinensis is one of the most common aquaculture species cultivated in China. The crab is an omnivore, and its hepatopancreas absorbs and stores nutrients. The aim of this study was to elucidate the expressions of the digestive enzyme genes and determine their respective roles in regulating digestive capacity in E. j. sinensis. We sequenced the hepatopancreatic transcriptomes of crabs fed a meat diet (MD), a vegetarian diet (VD), or a mixed diet (MV) and compared the gene expression patterns of these three groups of crabs. A total of 305,887 unigenes were obtained, of which 8747, 10,963, and 8877 were significantly differentially expressed in the comparisons between the MD and MV, VD and MV, and MD and VD diets, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) database-based enrichment analysis revealed that the differentially expressed gene (DEG) responses in the hepatopancreases to the MD mainly involved the “pancreatic secretion,” “glutathione metabolism,” “sphingolipid metabolism,” “fatty acid metabolism,” and “glycerolipid metabolism pathways.” DEG responses to the VD based on KEGG analysis mainly involved the “galactose metabolism,” “starch and sucrose metabolism,” and “fructose and mannose metabolism” pathways. The key digestive enzymes, including trypsin, β-glucosidase, chitinase, and triacylglycerol lipase, were identified. Our results further our understanding of crustacean hepatopancreatic functions during food digestion and provide resources for further studies regarding the molecular basis of omnivorous diets in crustaceans.

Phenolic content, acute toxicity of Ajuga iva extracts and assessment of their antioxidant and carbohydrate digestive enzyme inhibitory effects

The management of blood glucose level is the hallmark in the treatment of type 2 diabetes, and this may be achieved through the inhibition of the digestive enzymes involved in carbohydrate metabolism. The aim of this work is the investigation of phenolic compounds content, antioxidant activity and for the first time, the in vitro anti-hyperglycemic potential of aerial part of Ajuga iva Schreber extracts through the inhibition of digestive enzymes (α-amylase and α-glucosidase), responsible of the digestion of poly and oligosaccharides. Test for total phenolic content showed that the methanol extract has the highest polyphenolic and flavonoid concentrations with (65.3 mg GAEs/g extract and 132.6 mg REs/g extract). The methanol extract has also exhibited the higher antioxidant activity compared to the aqueous extract in different tests with (IC50=0.187±0.016 mg/mL; 62.19±0.45 mg TE/g extract; 89.12±0.23 mg AAE/g extract) in DPPH, ABTS and FRAP tests, respectively. Ajuga iva extracts exhibited a remarkable inhibitory activity against key digestive enzymes linked to type 2 diabetes, with a more potent inhibitory effect against α-glucosidase and a considerable inhibition against α-amylase. The results indicated also that the tested extracts are non-toxic with an LD50 higher than 2 g/kg in female Swiss mice. These results suggested that the phenolic compounds in A. iva extracts maybe behind the antioxidant and antidiabetic activities. However, further investigations regarding the bioactive compounds and their mechanisms of action are required for developing new natural drugs against diabetes-related hyperglycemia.

Molecular basis of the digestive functionality in developing Persian sturgeon (Acipenser persicus) larvae: additional clues for its phylogenetic status

Persian sturgeon (Acipenser persicus) is a critically endangered species, mainly due to overexploitation for its caviar. The permanence of populations of this species in the Caspian Sea is fully dependent on restocking programs. Accordingly, it is considered as an interesting target for aquaculture for both restocking and commercial purposes. In addition, as a Chondrostei, it exhibits one of the slowest rates of molecular evolution among vertebrates and is propounded as an excellent candidate for phylogenetic analysis and evolutionary biology. In this study, the early ontogeny of some key digestive enzymes precursors was determined at molecular level, aiming to obtain basic knowledge on the acquisition of digestive capacity of this species, and at the same time, to advance in its phylogenetic status from the point of view of digestion. For this purpose, A. persicus cDNAs for β-actin (actb; used as an internal reference gene), bile-salt activated lipase (cel), trypsinogen 1 (try1), pepsinogen (pga), and gastric proton pump (atp4a) were amplified and cloned, and their subsequent expressions were measured by quantitative real-time PCR during the first 34days post hatch (dph). Two isoforms for pga and at least six for try1 were obtained in this study, probably due to the additional genome duplication which sturgeons suffered along evolution. Phylogenetic analysis of the deduced amino acids sequences from the studied genes demonstrated that this species has a close evolutionary distance to Holostei, coelacanths, and tetrapods, including amphibians, reptiles, birds and mammals. According to our results, expression of all the genes increased gradually over time and reached maximum levels around 18 dph. This pattern, which was comparable to length and weight data, could indicate that around 3 weeks after hatching, the digestive capacity of the Persian sturgeon changes from larval to juvenile mode.

Inhibitory Effect of Five Ganoderma Species (Agaricomycetes) against Key Digestive Enzymes Related to Type 2 Diabetes Mellitus.

Ganoderma mushrooms are widely used in clinical therapies and functional foods. The antidiabetic effect of Ganoderma has become a research hot spot in recent decades. To search for a superior antidiabetic Ganoderma extract, five common Ganoderma species (G. lucidum, G. sinense, G. tsugae, G. applanatum, and G. leucocontextum) were investigated. A total of 10 fractions, including a total triterpenes fraction and a crude polysaccharides fraction for each, were prepared for further assays. Activities of α-glucosidase and α-amylase are inhibited dominantly by triterpenes from all five Ganoderma species rather than the polysaccharides. G. lucidum triterpenes inhibits α-glucosidase and α-amylase most significantly with IC50 values of 10.02 ± 0.95 µg/mL and 31.82 ± 4.30 µg/mL. Even more, triterpenes content was positively correlated with anti-α-glucosidase and anti-α-amylase activities. Therefore, triterpenes were considered to be the active compounds in inhibiting α-glucosidase and α-amylase activity. It is hoped that the results will provide more systematic information for the application of Ganoderma in the functional food and traditional medicine industries in the future.