The purpose of the current study was to evaluate the functional activity and storage viability (at 4 °C and 35 °C) of an immobilized as well as lyophilized multienzyme, viz., pectinase, cellulase, and amylase (PCA) that was produced by Bacillus subtilis NG105 under solid state fermentation (SSF) at 35 °C for 10 days using mosambi peel as a substrate. After SSF, the culture media was divided into two aliquots. From the first aliquot, the produced ME was extracted, precipitated, and further immobilized on calcium alginate beads (MEICA). In order to immobilize on mosambi peel matrix, the second aliquot was mixed with acetone and subsequently lyophilized (MELMP). Thus, ready MEICA and MELMP extracted 87.5 and 91.5% juice from mango pulp, respectively. In the reusability study, after 5 cycles, MEICA exhibited 23.8%, 24.4%, and 36.5% PCA activity, respectively. The PCA activity of MEICA and MELMP was examined after 60 days of storage at 4 °C. The result revealed that the PCA for MEICA declined from 100 to 66%, 58.2%, and 64.5%, respectively, while for MELMP, it dropped from 100 to 84.2%, 82.1%, and 69.7%, respectively. Further, after 60 days of storage, the reduction of total protein content (TPC) in free multienzyme (FME), MEICA, and MELMP was 92.2%, 91.5%, and 36.3% observed, respectively. In the localization study, the maximum levels of multienzyme activity were found in cell exudates. This study demonstrated that immobilizing of multienzyme through lyophilization on waste substrates like mosambi peel boosted its stability and shelf-life along with greatly reducing the cost of products.
Amylases and proteases are among the industrially most important enzymes for food processing, animal feed, brewing, starch processing, detergents, healthcare, leather processing, and biofuel production. In this study, we investigated the growth kinetics and statistically optimized the co-production of amylase and protease in a phylogenetically novel haloalkaliphilic actinomycete, Streptomyces lopnurensis KaM5 of seawater. The Plackett-Berman design using Minitab 14.0 software was employed to assess the impact of the nutritional factors, temperature, pH, and incubation time. Further, starch, yeast extract, NaCl concentrations, and incubation time were optimized by Box-Behnken design at their three levels. The Pareto charts, contour, surface plots, and individual factorial analysis expressed the variability and levels for the optimal enzyme production. ANOVA analysis admitted the statistical fitness and significance level among the variables. A two-fold increase in enzyme production was achieved by cost-effective co-production media. The study was further extended to growth kinetics associated with enzyme production. Specific growth rate (μ), maximal cell mass (Xmax), volumetric product formation (Pmax), rate of product formation (Qp), and generation time (g) were computed and analyzed. These parameters significantly improved when compared with the pre-optimized conditions, and the production economics of the enzyme was industrially viable. The initial studies on the characteristics of the enzymes suggested its ability to function under the combination of alkaline pH and high salt concentrations. The co-production of enzymes from extremophiles can be a potentially viable option for large-scale production and applications.
- MeSH
- acinární buňky cytologie enzymologie fyziologie MeSH
- amylasy metabolismus nedostatek MeSH
- chronická pankreatitida * diagnóza etiologie terapie MeSH
- diabetes mellitus MeSH
- dospělí MeSH
- exokrinní pankreatická insuficience * diagnóza etiologie terapie MeSH
- farmakoterapie MeSH
- klinický obraz nemoci MeSH
- komorbidita MeSH
- lidé MeSH
- pankreas exokrinní enzymologie metabolismus patofyziologie MeSH
- pankreatická lipasa metabolismus nedostatek MeSH
- pankreatin * aplikace a dávkování škodlivé účinky terapeutické užití MeSH
- prekurzory enzymů metabolismus nedostatek MeSH
- proteasy metabolismus nedostatek MeSH
- statistika jako téma MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Plagiodera versicolora (Laicharting, 1781) (Coleoptera: Chrysomelidae) is an important forest pest which damages many trees such as willow, poplar, and hazelnut. In order to find new microbes that can be utilized as a possible microbial control agent against this pest, we investigated the culturable bacterial flora of it and tested the isolated bacteria against P. versicolora larvae and adults. We were able to isolate nine bacteria from larvae and adults. The isolates were characterized using a combination of morphological, biochemical, and physiological methods. Additionally, we sequenced the partial sequence of the 16S rRNA gene to verify conventional identification results. Based on characterization studies, the isolates were identified as Staphylococcus sp. Pv1, Rahnella sp. Pv2, Rahnella sp. Pv3, Rahnella sp. Pv4, Rahnella sp. Pv5, Pantoea agglomerans Pv6, Staphylococcus sp. Pv7, Micrococcus luteus Pv8, and Rahnella sp. Pv9. The highest insecticidal activity against larvae and adults was obtained from M. luteus Pv8 with 50 and 40 % mortalities within 10 days after treatment, respectively. Extracellular enzyme activity of the bacterial isolates such as amylase, proteinase, lipase, cellulose, and chitinase was also determined. Consequently, our results show that M. luteus Pv8 might be a good candidate as a possible microbial control agent against P. versicolora and were discussed with respect to biocontrol potential of the bacterial isolates.
- MeSH
- amylasy metabolismus MeSH
- biologická ochrana MeSH
- brouci mikrobiologie MeSH
- celulosa metabolismus MeSH
- chitinasy metabolismus MeSH
- fenotyp MeSH
- larva mikrobiologie MeSH
- lipasa metabolismus MeSH
- Micrococcus luteus enzymologie izolace a purifikace patogenita MeSH
- mikrobiální testy citlivosti MeSH
- mikrobiota * MeSH
- Pantoea enzymologie izolace a purifikace patogenita MeSH
- proteasy metabolismus MeSH
- Rahnella enzymologie izolace a purifikace patogenita MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- Staphylococcus enzymologie izolace a purifikace patogenita MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Digestive processes and the effect of adipokinetic hormone (Pyrap-AKH) on the amount of nutrients (lipids, proteins, and carbohydrates), and on the activity of digestive enzymes (lipases, peptidases, and carbohydrases) were studied in the midgut of the firebug, Pyrrhocoris apterus. The analyses were performed on samples of anterior (AM), middle (MM) and posterior (PM) midgut parts. The results revealed that the digestion of lipids, carbohydrates and proteins take place in the acidic milieu. The Pyrap-AKH treatment increased significantly the level of lipids and proteins in the midgut, and also the level of triacylglycerols (TGs) predominantly in the AM, and the level of diacylglycerols (DGs) in the MM. The increase was not uniform for all present TG and DG species - those containing the linoleic fatty acid were predominant. No hormonal effect on lipase activity was recorded, while peptidase and glucosidase activity was increased in the MM and PM. All these facts indicate that the Pyrap-AKH probably stimulates digestion by more intensive food ingestion or turnover, and perhaps by the stimulation of metabolite absorption; the activation of digestive enzymes seems to be secondary or controlled by other mechanisms.
- MeSH
- amylasy metabolismus MeSH
- dietní proteiny metabolismus MeSH
- gastrointestinální trakt enzymologie MeSH
- Heteroptera metabolismus MeSH
- hmyzí hormony metabolismus MeSH
- kyselina pyrrolidonkarboxylová analogy a deriváty metabolismus MeSH
- lipasa metabolismus MeSH
- metabolismus lipidů MeSH
- metabolismus sacharidů MeSH
- oligopeptidy metabolismus MeSH
- proteasy metabolismus MeSH
- stravovací zvyklosti MeSH
- trávení MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: In barley, starch occurs in the form of granules with bimodal size distribution. Enzymatic hydrolysis of the starch granule is one of the most important reactions occurring during malting and mashing. Previous studies revealed the discrepancies in the assumption that barley varieties with better malting qualities should have a higher A/B (large/small starch granules) ratio. This led us to focus our attention on detailed analysis of two barley varieties, Jersey and Tolar, both with high malting quality but significantly differing in A/B (1.28 and 0.66, respectively), were chosen for more detailed analysis in the actual work. In this study, the capacity of gravitational field flow fractionation (GFFF) to monitor amylolysis of the starch granules was investigated. RESULTS: Isolated starch granules from these two barley cultivars were treated with amylases. The changes in starch granule size and bimodal distribution were studied by GFFF. Simultaneously, free sugars released during enzymatic digestion were observed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The changes in the fractogram and in the mass spectra reflect a correlation with the progress of enzymatic hydrolysis. CONCLUSION: The results show the interest in utilization of GFFF as a simple and cheap method for monitoring changes in the distribution of the starch granule size during amylolysis.
- MeSH
- amylasy metabolismus MeSH
- druhová specificita MeSH
- frakcionace tokem v poli metody MeSH
- gravitace MeSH
- hydrolýza MeSH
- ječmen (rod) chemie metabolismus MeSH
- jedlá semena chemie MeSH
- potravinářská technologie MeSH
- sacharidy biosyntéza MeSH
- škrob analýza metabolismus MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- velikost částic MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
The effect of long-term (30 days) exposure to PCZ (0.2, 50, and 500 microg l(-1)) on intestine-related biochemical markers in rainbow trout was investigated. Multiple biomarkers were measured, including digestive enzymes (proteolytic enzymes and amylase), antioxidant responses (TBARS, CP, SOD, CAT, GR and GPx) and energy metabolic parameters (RNA/DNA ratio, Na(+)-K(+)-ATPase). Exposure to 500 microg l(-1) PCZ led to significantly inhibited (p<0.01) proteolytic enzyme and amylase activity. Activities of the antioxidant enzymes SOD, CAT, and GPx gradually increased at lower PCZ concentrations (0.2 and 50 microg l(-1)). At the highest concentration (500 microg l(-1)), oxidative stress was apparent as significant higher (p<0.05) lipid peroxidation and protein carbonyls, associated with an inhibition of antioxidant enzymes activity. Moreover, energy metabolic parameters (RNA/DNA ratio, Na(+)-K(+)-ATPase) were significantly inhibited (p<0.01) in the intestines of fish exposed to 500 microg l(-1) PCZ, compared with controls. We suggest that long-term exposure to PCZ could result in several responses in intestine-related biochemical markers, which potentially could be used as indicators for monitoring residual PCZ present in the aquatic environment.
- MeSH
- amylasy metabolismus MeSH
- antioxidancia metabolismus MeSH
- biologické markery analýza MeSH
- chemické látky znečišťující vodu farmakologie MeSH
- karbonylace proteinů účinky léků MeSH
- Oncorhynchus mykiss metabolismus MeSH
- oxidační stres MeSH
- peroxidace lipidů účinky léků MeSH
- proteasy metabolismus MeSH
- střeva účinky léků metabolismus MeSH
- triazoly farmakologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
