Beta-glucuronidase Activity Research Articles

GsVAMP72, a novel Glycine soja R-SNARE protein, is involved in regulating plant salt tolerance and ABA sensitivity

Abiotic stress, especially high salinity, is a major threat to agricultural production. It has been well established that SNARE proteins sustain directed vesicle traffic to underpin plant growth and development, yet little is known about the role of SNARE protein in the capacity to withstand abiotic stress, especially in wild soybeans. Here we identified and characterized a GsCBRLK interacting protein, GsVAMP72, which is a putative vesicle-associated membrane protein in Glycine soja. GsVAMP72 protein has a longin domain at its N-terminus, belonging to R-SNARE family. Quantitative real-time (RT) PCR and beta-glucuronidase (GUS) activity assays revealed that the expression of GsVAMP72 was highly and rapidly induced by both high salt and ABA treatments. Overexpression of GsVAMP72 in Arabidopsis significantly reduced salt tolerance by modifying the ionic content and down-regulating expression of stress-responsive genes, including RD29A, COR47, KIN1, COR15A and RAB18. On the other hand, GsVAMP72 overexpression increased plant ABA sensitivity and altered the expression levels of ABA-responsive genes. Subcellular localization analysis showed that eGFP–GsVAMP72 fusion protein was observed on the plasma membrane-like and endosome-like structures but eGFP alone was distributing throughout the cytoplasm in Arabidopsis protoplasts and onion epidermal cells. GsVAMP72 promoter-controlled GUS activity was detected in both vegetative and reproductive organs, and was strongly induced by salt and ABA. In summary, we demonstrated that GsVAMP72 is a novel Glycine soja vesicle-associated membrane protein and is highly involved in regulating plant responses to salt and ABA stresses.

Detection and dynamics of fecal indicator bacteria in two Tyrrhenian lagoons (Sabaudia and Orbetello, central Italy)

1 - Coastal lagoons are particularly vulnerable to strong anthropogenic pressure. These environments can act as sink for allochthonous material, including harmful bacteria, with the ability to reduce their impact to adjacent coastal waters. 2 - This study investigates fecal pollution impairment in two Italian coastal lagoons and adjacent coastal waters. We utilised methodological approaches to gain fast and more precise information on the contamination and viability of fecal indicator bacteria. 3 - The analyses of total coliforms (TC) and Escherichia coli (E. coli) were performed in situ by enzyme assays. E. coli live/dead cells were enumerated by immunofluorescence technique (combined with propidium iodide) for the specific detection of enteropathogenic serotypes (12 EPEC and 2 EIEC). 4 - Overall TC enzyme activity showed a high degree of temporal variation whereas E. coli enzyme activity peaked in summer in both lagoons (beta-glucuronidase, 84 and 47 nmol MUFL-1h-1, Sabaudia and Orbetello respectively). Beta-glucuronidase activity showed a high correlation with viable cells counts of E. coli enteropathogenic strains (p<0.01). Significantly lower degree of contamination was observed in the adjacent coastal waters. 5 - Our findings describe these lagoons as very fragile systems which hydro-morphological features allow a long time preservation of microbial contaminants, including pathogenic serotypes. These findings imply potential consequences for the human health as well as the need for protective measures of these environments.

Regulation of AtSUS2 and AtSUS3 by glucose and the transcription factor LEC2 in different tissues and at different stages of Arabidopsis seed development

Sucrose synthase (SUS) is a key enzyme of carbon metabolism in heterotrophic tissues of plants. The Arabidopsis genome contains six SUS genes. Two members of this family, namely AtSUS2 (At5g49190) and AtSUS3 (At4g02280) are strongly and differentially expressed in Arabidopsis seed. Expression analysis was carried out using SUS:promoter-GUS fusion lines in a wild-type genetic background or in a mutant carrying a lesion in the transcription factor LEAFY COTYLEDON 2 (LEC2; At1g28300). The accumulation patterns of mRNA, protein, and SUS activity were altered in the lec2 mutant during seed development 9-18 days after flowering. This indicates that LEC2 acts epistatically on the expression of AtSUS2 and AtSUS3. It appears that LEC2 is required for cotyledon-specific expression of both SUS genes but it is not responsible for expression in the radicle tip during embryo development. The AtSUS2 promoter was induced in planta by feeding of glucose but less so by sucrose and trehalose. Non-phosphorylable glucose analogs such as 3-O-methyl-glucose and 2-deoxyglucose also caused an induction, suggesting that sugar signaling proceeds by a hexokinase-independent pathway, possibly involving hexose sensing. Analysis of transgenic lines carrying of truncated versions of the AtSUS2:promoter fused to Beta-glucuronidase activity revealed an internal 421 bp region that was responsible for expression in seeds. Bioinformatic sequence analysis revealed regulatory cis-elements putatively responsible for the spatio-temporal pattern of AtSUS2 expression such as the SEF3 (aaccca) and W-box (ttgact) motifs. These findings are discussed in relation to the roles played by AtSUS2, AtSUS3 and LEC2 in the biosynthesis of seed storage products in Arabidopsis.

Mucopolisacaridosis tipo VII: ¿causa infrecuente de hidrops fetal?

We report the case of a pregnant woman who was diagnosed with hydrops fetalis at 17 weeks in two consecutive pregnancies. Once an immune origin was ruled out, the association with lysosomal storage diseases was investigated during the second pregnancy. This showed decreased activity of beta-glucuronidase in cultured cells and a glycosaminoglycan profile in amniotic fluid, indicative of Sly disease or mucopolysaccharidosis type VII which is inherited in an autosomal recessive pattern.

Variation in GUS activity in vegetatively propagated Hevea brasiliensis transgenic plants

Hevea brasiliensis transgenic plants are regenerated from transgenic callus lines by somatic embryogenesis. Somatic embryogenesis is not yet available for commercial propagation of Hevea clones, which requires conventional grafting of buds on rootstock seedlings (budding). The stability of transgene expression in budded plants is therefore necessary for further development of genetic engineering in rubber trees. Transgene expression was assessed by fluorimetric beta-glucuronidase (GUS) activity in fully developed leaves of in vitro plants from transgenic lines and their sub-lines obtained by budding. A large variation in GUS activity was found in self-rooted in vitro plants of five transgenic lines, and the absence of activity in one line suggested transgene silencing. Beyond confirming transmissibility of the reporter gene by budding and long-term expression, a quantification of GUS activity revealed that greater variability existed in budded plants compared to self-rooted mother in vitro plants for three transgenic lines. Although somatic embryogenesis provided more stable GUS activity, budding remained an efficient way of propagating transgenic plants but transgene expression in budded plants should be verified for functional analysis and further development.

Functional analysis of the HS185 regulatory element in the rice HSP70 promoter

A series of HSP70 promoter deletion constructs was established. Analysis of beta-glucuronidase activities from the promoter deletion constructs in transient expression assays identified a cis-element, located from -493 to -308 bp upstream of the ATG start site. This element was designated as HS185 and has a crucial role in HSP70 promoter activity. HS185 has some characteristics of a miniature inverted-repeat transposable element (MITE), such as terminal inverted repeats (TIRs) (GGTCCCACA) and a putative target site duplication. There are 362 copies of homologous sequences of HS185 in the rice genome, which are preferentially distributed to non-coding regions. Based on these sequence features, we propose that HS185 is an uncharacterized rice MITE, possibly derived from the rice transposon Mutator-like element VIII family. Further transient expression assays showed that HS185 inhibited the enhancer activity of the cauliflower mosaic virus 35S promoter. These results demonstrate that not only is HS185 necessary for HSP70 promoter activity, but it also has a functional role as an insulator. This study explored new regulatory functions of non-coding repeat sequences in rice.

Impact of beta-glucan on the faecal microbiota of polypectomized patients: A pilot study

Beta-glucans are polysaccharides present in the cell walls of higher plants, in the seeds of some cereals, and certain yeasts and fungi also produce them. It is suggested that they exhibit, among many other health benefits, protective effects against carcinogenesis in the colon, but there is not enough human data to support this. The aim of the study was to determine the effect of barley-derived beta-glucan in the gut microbiota of polypectomized patients. Subjects were randomly assigned to consume 125 g of bread per day with beta-glucan (3 g/d), or without (placebo group), for 3 months. Thirty-three polypectomized men and women (mean age 57.6 years) were recruited into the study, but only 20 completed. Subjects did not consume any probiotics, prebiotics or antibiotics 2 months prior the intervention, or during the study. Stool samples were collected at baseline, on days 30 and 90 of intervention, as well as 2 weeks after the intervention, for enumeration of total aerobes and anaerobes, coliforms, E. coli, enterococci, Bacteroides spp., Clostridium perfringens, bifidobacteria, lactobacilli and Candida spp. Faecal bacterial enzyme activity (beta-glucuronidase and beta-glucosidase), pH, faecal moisture and the concentration of volatile fatty acids in the faeces were measured. Gastrointestinal symptoms were also recorded. Overall, no significant differences were observed in bacterial viable counts between the two feeding groups. Group specific analysis for β-glucan group revealed significantly decreased total coliform counts on the 30th day of the trial compared to the baseline ( p = 0.041). Clostridium perfringens concentration increased without reaching statistical significance, on the 30th day, while it decreased significantly on the 90th day of the intervention compared to the 30th day ( p = 0.016). An increase was noted in the molar ratio of acetate on the 90th day of the trial compared to placebo ( p = 0.018). The molar ratio of butyrate presented a trend to increase on the 30th day, which decreased ( p = 0.013) on the 90th day and then increase 2 weeks after the intervention ( p = 0.017) compared to placebo. A decrease was recorded in the β-glucan group in the bloating and abdominal pain score after the 30th day of the intervention (Day 30–37) compared to placebo. During β-glucan administration we did not observe any changes on beta-glucuronidase or beta-glucosidase activity, faecal pH, or on faecal moisture.

Alteration of rat skin acid and alkaline phosphatase and beta-glucuronidase activity after topical application of fluocinolone acetonide cream.

Alteration of rat skin acid and alkaline phosphatase and beta-glucuronidase activity after topical application of fluocinolone acetonide cream.

Some aspects of the inhibitory activity of hypolaetin-8-glucoside in acute inflammation.

Hypolaetin-8-glucoside (H-8-G) has been examined for its mode of action in several models of acute inflammation. Its anti-inflammatory activity in carrageenan-induced inflammation of the rat hind-paw is not affected either by adrenalectomy or by phentolamine given with propranolol. H-8-G and its aglycone, hypolaetin, did not antagonize the actions of histamine, 5-hydroxytryptamine (5-HT), bradykinin or prostaglandin E2 (PGE2) on various smooth muscle preparations in-vitro, but protected erythrocytes from heat-induced lysis. The glycoside was more potent than troxerutin on capillary permeability increased by histamine and exerted inhibitory effects on protein exudation, leucocyte migration and beta-glucuronidase activity in the carrageenan air pouch, thereby showing some difference from indomethacin. These results are discussed in relation to the features of non-steroidal anti-inflammatory drugs (NSAID) and flavonoid anti-inflammatory actions.

Histostaining for Tissue Expression Pattern of Promoter-driven GUS Activity in Arabidopsis

Promoter-driven GUS (beta-glucuronidase) activity is the most commonly used technique for tissue-specific expression patterns in Arabidopsis. In this procedure, GUS enzyme converts 5-bromo-4-chloro-3-indolyl glucuronide (X-Gluc) to a blue product. The staining is very sensitive. Processed samples can be examined under dissecting microscope or Differential Interference Contrast (Nomaski) microscope for bright blue color over cleared transparent background. Note this assay does not provide accurate information to subcellular levels., 启动子驱动的GUS（β-葡糖醛酸糖苷酶）活性是拟南芥中组织特异性表达模式最常用的技术。 在该程序中，GUS酶将5-溴-4-氯-3-吲哚基葡糖苷酸（X-Gluc）转化为蓝色产物。 染色非常敏感。 处理的样品可以在解剖显微镜或微分干涉对比（Nomaski）显微镜下在透明背景上检查明亮的蓝色。 注意这个测定不提供准确的信息亚细胞水平。

Effects of Probiotic Lactobacillus paracasei-enriched Artichokes on Constipated Patients

To determine whether the consumption of artichokes enriched with a probiotic Lactobacillus paracasei strain affects fecal microbiota composition, fecal enzyme activity, and short-chain fatty acids production and symptom profile in patients suffering from constipation. Constipation is a common gastrointestinal disorder often related to the food diet. The beneficial effects of probiotics and prebiotics on human health are under investigation. Moreover, recent studies assessed the suitability of some vegetables, particularly olives and artichokes, to vehicle probiotic strains into the gastrointestinal tract. For 15 days, 8 volunteers (3M/5F age 40+/-14 y) integrated their normal diet with artichokes (180 gr) enriched with 20 billions of L. paracasei LMGP22043. Faecal samples were subjected to microbiologic and biochemical analyses. Besides, investigations on symptom profile of the volunteers and stool consistency were carried out by using a validated questionnaire (Gastrointestinal Symptom Rating Scale) and the Bristol stool form chart. The gut of all volunteers resulted to be colonized by the probiotic strain after 15 days feeding. No significant differences in the microbiological counts throughout the experimental period were registered, whereas a significant increase of butyric and valeric acids with a concomitant decrease of lactic acid was registered. At the same time, the fecal beta-glucuronidase activity was significantly reduced. Finally, the analysis of symptom profile indicated a marked reduction in abdominal distension and feeling of incomplete evacuation. These preliminary data suggest that novel approaches for treating constipation can come through ingestion of probiotic vegetable products that, acting as symbiotics, can ameliorate this common disorder.

GhMPK7, a novel multiple stress-responsive cotton group C MAPK gene, has a role in broad spectrum disease resistance and plant development

Mitogen-activated protein kinase (MAPK) cascades play a pivotal role in environmental responses and developmental processes in plants. Previous researches mainly focus on the MAPKs in groups A and B, and little is known on group C. In this study, we isolated and characterized GhMPK7, which is a novel gene from cotton belonging to the group C MAPK. RNA blot analysis indicated that GhMPK7 transcript was induced by pathogen infection and multiple defense-related signal molecules. Transgenic Nicotina benthamiana overexpressing GhMPK7 displayed significant resistance to fungus Colletotrichum nicotianae and virus PVY, and the transcript levels of SA pathway genes were more rapidly and strongly induced. Furthermore, the transgenic N. benthamiana showed reduced ROS-mediated injuries by upregulating expression of oxidative stress-related genes. Interestingly, the transgenic plants germinated earlier and grew faster in comparison to wild-type plants. beta-glucuronidase activity driven by the GhMPK7 promoter was detected in the apical meristem at the vegetative stage, and it was enhanced by treatments with signal molecules and phytohormones. These results suggest that GhMPK7 might play an important role in SA-regulated broad-spectrum resistance to pathogen infection, and that it is also involved in regulation of plant growth and development.

Bioconversion of glycyrrhizinic acid in liquorice into 18-β-glycyrrhetinic acid by Aspergillus parasiticus Speare BGB

A filamentous fungi strain, Aspergillus parasiticus Speare BGB, producing beta-glucuronidase was screened to transform glycyrrhizinic acid (GL) in liquorice into 18-beta-glycyrrhetinic acid (GA). Under the following cultivate conditions in shake flask, 1% GL (purity 30%), medium capacity 40% of flask, the initial pH value at 4.5, cultivate temperature of 32 degrees C, inoculum size of 5% and culturing time for 96 h the bioconversion ratio of GL into GA could reach 95%. A variety of parameters of submerged state fermentation, including the growth characteristics of A. parasiticus Speare BGB, the change amount of GL and GA, and the activity of beta-glucuronidase, were monitored simultaneously. GA was separated and purified by macroporous resin, silica gel column chromatography followed by recrystalization with the final purity over 98%. Purified product was identified as GA by the infrared absorption spectrum, molecular weight, and nuclear magnetic resonance. This study provided a new and efficient approach of obtaining GA by microbial transformation.

Identification of Valproic Acid Glucuronide Hydrolase As a Key Enzyme for the Interaction of Valproic Acid with Carbapenem Antibiotics

Plasma levels of valproic acid (VPA) are decreased by concomitant use with carbapenem antibiotics, such as panipenem (PAPM). One of the plausible mechanisms of this interaction is the inhibition of VPA glucuronide (VPA-G) hydrolysis by carbapenems in the liver. To elucidate this interaction mechanism, we purified VPA-G hydrolase from human liver cytosol, in which the hydrolytic activity was mainly located. After chromatographic purification, the VPA-G hydrolase was identified as acylpeptide hydrolase (APEH). APEH-depleted cytosol, prepared by an immunodepletion method, completely lacked the hydrolytic activity. These results demonstrate that APEH is a single enzyme involved in PAPM-sensitive VPA-G hydrolysis in cytosol. In addition, the hydrolytic activity of recombinant human APEH was inhibited by PAPM and the inhibition profile by typical esterase inhibitors (diisopropyl fluorophosphate, 5,5'-dithiobis(2-nitrobenzoic acid), p-chloromercuribenzoic acid, and d-saccharic acid 1,4-lactone) was similar to that of human liver cytosol. Cytosolic VPA-G hydrolase activity was slightly inhibited by cholinesterase and carboxylesterase inhibitors. beta-Glucuronidase activity remained in APEH-depleted cytosol, whereas VPA-G hydrolase activity was completely abolished. Thus, either cholinesterase, carboxylesterase, or beta-glucuronidase in cytosol would not be involved in VPA-G hydrolysis. Taken together, APEH plays a major role in the PAPM-sensitive VPA-G hydrolysis in the liver. These findings suggest that APEH could be a key enzyme for the drug interaction of VPA with carbapenems via VPA-G hydrolysis.

Proteomic Analysis of Quorum Sensing-Dependent Proteins in Burkholderia glumae

Burkholderia glumae, the causal agent of bacterial rice grain rot, utilizes quorum sensing (QS) systems that rely on N-octanoyl homoserine lactone (synthesized by TofI) and its cognate receptor TofR to activate toxoflavin biosynthesis genes and an IclR-type transcriptional regulator gene, qsmR. Since QS is essential for B. glumae pathogenicity, we analyzed the QS-dependent proteome by 2-dimensional gel electrophoresis. A total of 79 proteins, including previously known QS-dependent proteins, were differentially expressed between the wild-type BGR1 and the tofI mutant BGS2 strains. Among this set, 59 proteins were found in the extracellular fraction, and 20 were cytoplasmic. Thirty-four proteins, including lipase and proteases, were secreted through the type II secretion system (T2SS). Real-time RT-PCR analysis showed that the corresponding genes of the 49 extracellular and 13 intracellular proteins are regulated by QS at the transcriptional level. The T2SS, encoded by 12 general secretion pathway (gsp) genes with 3 independent transcriptional units, was controlled by QS. beta-Glucuronidase activity analysis of gsp::Tn3-gusA gene fusions and electrophoretic mobility shift assays revealed that the expression of gsp genes is directly regulated by QsmR. T2SS-defective mutants exhibited reduced virulence, indicating that the T2SS-dependent extracellular proteins play important roles in B. glumae virulence.

Preventive effect of caffeic acid on lysosomal dysfunction in isoproterenol‐induced myocardial infarcted rats

We evaluated the preventive effect of caffeic acid (CA) on lysosomal enzymes in isoproterenol (ISO)-treated myocardial infarcted rats. Male albino Wistar rats were pretreated with CA (15 mg/kg) daily for a period of 10 days. After the pretreatment period, ISO (100 mg/kg) was subcutaneously injected to rats twice at an interval of 24 h. The activity of serum creatine kinase-MB and lactate dehydrogenase was increased significantly (P < 0.05) in ISO-induced myocardial infarcted rats. The levels of plasma thiobarbituric acid reactive substances and lipid hydroperoxides were significantly (P < 0.05) increased, and the level of plasma-reduced glutathione was significantly (P < 0.05) decreased in ISO-induced myocardial infarcted rats. The activities of lysosomal enzymes (beta-glucuronidase, beta-N-acetylglucosaminidase, beta-galactosidase, cathepsin-B and cathepsin-D) were increased significantly (P < 0.05) in the serum and heart of ISO-induced myocardial infarcted rats. ISO induction also resulted in decreased stability of membranes, which was reflected by lowered activities of beta-glucuronidase and cathepsin-D in different fractions except cytosol. Pretreatment with CA (15 mg/kg) to ISO-treated rats significantly (P < 0.05) prevented the changes in the activities of cardiac marker enzymes, the levels of lipid peroxidation products, reduced glutathione and the activities of lysosomal enzymes in the serum, heart, and subcellular fractions. Oral treatment with CA (15 mg/kg) to normal control rats did not show any significant effect. Thus, the results of our study showed that CA prevented the lysosomal membrane damage against ISO-induced myocardial infarction. The observed effects of CA are due to membrane-stabilizing, antilipo peroxidative, and antioxidant effects.

Main airborne Ascomycota spores: characterization by culture, spore morphology, ribosomal DNA sequences and enzymatic analysis

The aim of this work was to identify the main allergy-related Ascomycetes fungal spores present in the atmosphere of Porto, using different and complementary techniques. The atmospheric sampling, performed in the atmosphere of Porto (Portugal) from August 2006 to July 2008, indicated Cladosporium, Penicillium, Aspergillus and Alternaria as the main fungal spore taxa. Alternaria and Cladosporium peaks were registered during summer. Aspergillus and Penicillium highest values were registered from late winter to early spring. Additionally, the Andersen sampler allowed the culture and isolation of the collected viable spores subsequently used for different identification approaches. The internal-transcribed spacer region of the nuclear ribosomal repeat unit sequences of airborne Ascomycetes fungi isolates revealed 11 taxonomically related fungal species. Among the identified taxa, Penicillum and Aspergillus presented the highest diversity, while only one species of Cladosporium and Alternaria, respectively, were identified. All selected fungal spore taxa possessed phosphatase, esterase, leucine arylamidase and beta-glucosidase enzymatic activity, while none had lipase, cystine arylamidase, trypsin or beta-glucuronidase activity. The association between the spore cell wall morphology, DNA-based techniques and enzymatic activity approaches allowed a more reliable identification procedure of the airborne Ascomycota fungal spores.

Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 Alleviated Symptoms of Salmonella Infection, as Determined in Wistar Rats Challenged with Salmonella enterica Serovar Typhimurium

Wistar rats were administered daily with Lactobacillus plantarum 423 and Enterococcus mundtii ST4SA through intragastric gavage (1 x 10(8) cfu of each strain and a combination of the two strains). Sterile saline was used as placebo. After 7 days, the animals were challenged by infection with 2 x 10(8) CFU Salmonella enterica serovar Typhimurium. After 1 day of treatment with L. plantarum 423 and E. mundtii ST4SA, the feed and water intake, and body weight of the rats increased. The faecal moisture content and beta-glucuronidase activity remained more-or-less constant after 2 days of treatment with E. mundtii ST4SA, L. plantarum 423 and a combination of the two strains. Reduced levels of endotoxin were recorded in blood samples taken from rats that received L. plantarum 423 and E. mundtii ST4SA. Although both strains alleviated symptoms of S. enterica serovar Typhimurium infection, L. plantarum 423 administered as a single culture proved more effective than E. mundtii ST4SA. Less promising results were recorded when L. plantarum 423 was administered in combination with E. mundtii ST4SA. This suggests that L. plantarum 423 is more effective than E. mundtii and should be the preferred probiotic to alleviate symptoms of S. enterica serovar Typhimurium infection.

Bioinformatics Characterization of Potential New Beta-Glucuronidase from Streptococcus equi subsp. zooepidemicus

Recently, the gene coding for a new beta-glucuronidase enzyme has been identified and cloned from Streptococcus equi subsp. zooepidemicus. This is another report of a beta-glucuronidase gene cloned from bacterial species. The ORF Finder analysis of a sequenced DNA (EMBL, AJ890474) revealed a presence of 1,785 bp large ORF potentially coding for a 594 aa protein. Three protein families in (Pfam) domains were identified using the Conserved Domain Database (CDD) analysis: Pfam 02836, glycosyl hydrolases family 2, triose phosphate isomerase (TIM) barrel domain; Pfam 02837, glycosyl hydrolases family 2, sugar binding domain; and Pfam 00703, glycosyl hydrolases family 2, immunoglobulin-like beta-sandwich domain. To gain more insight into the enzymatic activity, the domains were used to generate a bootstrapped unrooted distance tree using ClustalX. The calculated distances for two domains, TIM barrel domain, and sugar-binding domain were comparable and exhibited similarity pattern based on function and thus being in accordance with recently published works confirming beta-glucuronidase activity of the enzyme. The calculated distances and the tree arrangement in the case of centrally positioned immonoglobulin-like beta-sandwich domain were somewhat higher when compared to other two domains but clustering with other beta-glucuronidases was rather clear. Nine proteins, including beta-glucuronidases, beta-galactosidase, and mannosidase were selected for multiple alignment and subsequent distance tree creation.

β-glucuronidase and β-glucosidase activity of Lactobacillus and Enterococcus Isolated from Human Feces

The domination of microorganisms characterized by excessive activity of the so-called fecal enzymes may be one of the reasons of the large intestine cancers. These enzymes are mainly those that belong to the hydrolase and reductase classes and their excessive activity may lead to disorders in the functioning of the digestive tract. The aim of tise research was to determine the activity of beta-glucuronidase and beta-glucosidase of Lactobacillus and Enterococcus strains isolated from the feces of healthy children, aged 1 and 8, and adults, aged 30 and 80. The analysis included 10 strains isolated from the feces of individuals in each of the age groups. beta-glucuronidase activity in the case of the isolates from children, depending on the strain, equaled from about 0.15 mM/h/mg of protein to 0.26 mM/h/mg of protein and was lower, respectively, by 52.35% and 57.81%, than the beta-glucosidase activity. Simultaneously, the activity of the Lactobacillus enzymes from children was 2.4 times higher, and in case of the isolates obtained from adults they were 4.6 and 2.7 times higher than the activity of the Entercoccus enzymes. The highest beta-glucuronidase activity was observed in Lactobacillus isolates coming from an 80-year-old subject. The differences between the activity of Enterococcus beta-glucuronidase isolated from the feces of 1 and 8 year old children were statistically insignificant. On the other hand, in the case of the subjects aged 30 and 8 the isolates were characterized by activity lower by, respectively. 48% and 37% than the isolates coming from children. The highest beta-glucosidase activity was discovered in the case of Lactobacillus and Enterococcus coming from children, which was higher by 32% than the activity of the isolates from adult persons. Therefore, it was determined that the activity of beta-glucuronidase of Lactobacillus strains isolated from feces from people aged 80 was the highest, and the isolates of the examined microorganisms coming from children were characterized by the highest beta-glucosidase activity.