Enzymes Amylase Research Articles

Production of Glucose Syrup Derived from Cassava Peel Starch Using Amylase Enzyme from Green Bean Seed Sprout Extract (Phaseolus radiatus L.)

The production of glucose syrup derived from cassava peels starch using amylase enzyme from green bean seed sprout extract (Phaseolus radiatus L.) has been carried out by isolating the enzyme from the green bean seed sprouts extract then purified by precipitation using ammonium sulphate with the saturation of 60% (w/v). Amylase enzyme activity was determined using the Nelson Somogyi method. The research results obtained the activity of the amylase enzyme in the deposition of 60% ammonium sulphate at 12.22 U/mL. Amylase enzyme was applied as a hydrolyzed starch from cassava peels with a weight ratio of raw material with water 10% (w / v) to produce glucose syrup with the addition of 0.25% enzyme; 0.5%; 0.75%; 1% (v / v). The glucose syrup obtained was then tested for viscosity, reducing sugars, water content, ash content, and organoleptics. The best glucose syrup analysis results from adding 1% amylase enzyme with a viscosity value of 238.71cP, water content of 12.45%, reduction sugar content of 33.81%, ash content of 0.49%, and dextrose equivalent value of 50.16%.

Production of alpha-amylase enzyme from grain sprouting

Alpha-amylase is considered to be one of the primary enzymes responsible for starch degradation in cereals. During grain germination, α-amylase initiates the conversion of starch into simple sugars to fuel embryo and coleoptile growth in the first few days of germination. The present study involves the estimation of Alpha amylase content in four different grains varieties, namely, wheat grain and barley up to 8 days after germination. It was observed that, alpha amylase content steadily increased, and found to be saturated after reaching the peak values. In all the samples, the peak values were observed in 7th day of germination. The enzymatic assay were done by estimation of maltose content by Spectrophotometry as starch degraded to maltose by alpha amylase enzyme.

Comparative Analysis of Enzymatic Activities and Transcriptional Profiles of Various Hepatic Enzymes between Male and Female Yellowfin Tuna (Thunnus albacares)

Yellowfin tuna (Thunnus albacares) is a valuable pelagic migratory fish with potential for aquaculture. Despite this, there is limited understanding of the biological and physiological characteristics of this species, particularly regarding sex differences in growth performance. The liver, a crucial organ for digestion and metabolism, plays a significant role in regulating fish growth. This study aimed to compare liver enzyme activities and transcriptome profiles between female and male yellowfin tuna to uncover the molecular mechanisms underlying difference between the sexes. The results revealed that female yellowfin tuna exhibited higher amylase and lipid metabolism enzyme activities, while male yellowfin tuna showed higher glucose-6-phosphate dehydrogenase and antioxidant enzyme activities. Additionally, through Illumina sequencing technology, the study generated 37.74 Gb of clean data and identified 36,482 unique genes (UniGenes) in the liver transcriptome. A total of 2542 differentially expressed genes were found, with enriched Gene Ontology terms and pathways related to metabolic processes, particularly lipid metabolism and transport. These findings suggest that female yellowfin tuna have superior digestive enzyme activities and lipid metabolism, while male yellowfin tuna excel in sugar metabolism, ATP production, and antioxidant defense. This study provides valuable insights into sex differences in yellowfin tuna and could aid in advancing full-cycle aquaculture practices for this species.

Production and Charaterization of Amylase Enzyme by Aspergillus Niger Isolated from Rhizospheric Soil Under Solid State Fermentation and Submerged Fermentation

Production and Charaterization of Amylase Enzyme by Aspergillus Niger Isolated from Rhizospheric Soil Under Solid State Fermentation and Submerged Fermentation

Changes in Digestive Enzyme Activities during Larval Development of Spotted Seatrout (Cynoscion nebulosus)

The spotted seatrout (Cynoscion nebulosus)—an important commercial species—has a high potential for aquaculture in the Gulf of Mexico. To optimize its feeding during larval rearing, this study aims to evaluate the primary gastric (pepsin), intestinal (leucine aminopeptidase and alkaline phosphatase), and pancreatic (alkaline protease, trypsin, chymotrypsin, amylase, and lipase) enzyme activities from hatching to day 30. A multivariate analysis identified three digestive enzyme development stages during the spotted seatrout larval transformation. The first stage occurred between 1 (mean ± standard error (SE) = 1.73 ± 0.14 millimeter (mm) standard length (SL)) and 3 (2.14 ± 0.07 mm SL) days after hatching (DAH); a period of digestive stability showed the highest activity in amylase and bile salt-dependent lipase. The second stage (from 4 (2.53 ± 0.09 mm SL) to 20 (10.92 ± 0.51 mm SL) DAH) was a period of digestive transition, during which leucine aminopeptidase, chymotrypsin, and alkaline proteases were identified as the predominant enzymes from 4 to 5 DAH. In the third stage—a period of digestive stability—pepsin was the major enzyme that occurred between 25 (16.51 ± 0.81 mm SL) and 30 (25.91 ± 0.82 mm SL) DAH. These results indicate that the spotted seatrout larvae have a digestive system adapted to lipids and carbohydrates at the onset of feeding, with an immediate transition to protein digestion when exogenous feeding begins. Additionally, the digestive system of the spotted seatrout may be considered mature at 25 DAH. Further research is needed to elucidate the mechanisms of digestive tract development in the spotted seatrout larvae.

301 Maxwell Lecture: Bacillus probiotics in nursery pigs: The effects of dose on gut health and performance

Abstract Weaning is one of the most stressful events in the life of a pig and critical for their health and development. To reduce the use of antibiotics and zinc oxide in nursery diets, many feed additives have been tested to demonstrate their efficacy in weaning pigs. Bacillus subtilis, a heat-resistant spore-forming bacteria, consumes oxygen in the digestive tract and can provide a favorable anaerobic environment for lactic acid-producing bacteria, resulting in their proliferation. It also produces several digestive enzymes, such as amylase, protease, and antioxidant enzymes. In addition, Bacillus species are known to alter the gut microbiome and increase the production of short-chain fatty acids in the gut. Therefore, Bacillus probiotics have been widely used in nursery diets to effectively improve growth performance and protect piglets from weaning stress and postweaning diarrhea. However, when the probiotics are supplemented in diets for weaning pigs, their efficacy is dependent on multiple factors, such as strains, the health condition of the animals, product composition, and the dose of the probiotics. There has been tremendous effort in establishing the optimal supplementation level of probiotics for nursery pigs to maximize the efficacy of their supplementation, but a high dose of probiotics could be detrimental to the gut health and microflora of nursery pigs, as it could increase the incidence of diarrhea and may have potential side effects in the immune system, antibiotic resistance, and the production of unfavorable metabolites in the gut. A study was conducted to demonstrate the effect of Bacillus probiotic supplementation for nursery pigs at the recommended and high doses (10 times greater bacterial counts than the recommended dose) compared with a control treatment with no probiotic supplementation. In the study, the Bacillus supplementation at the recommended dose did improve the early postweaning growth rate, increase the production of fecal short-chain fatty acids (acetate, propionate, and butyrate), and blood glucose concentrations, and alter the fecal microbiome. However, high-dosing probiotics in nursery diets did not show any beneficial effects that were shown in the pigs fed diets with Bacillus probiotics at the recommended dose. Bacillus-probiotic supplementation in nursery diets may enhance growth performance and benefit gut health, but a high dose of probiotics does not always lead to comparable positive effects for pigs. Therefore, it is critically important to test probiotics to optimize their efficacy and avoid unnecessary high-dosing in nursery diets, as it could be cost-ineffective and may not show benefits that can be observed with the recommended dose.

Formulation of Novel Microbial Consortia for Rapid Composting of Biodegradable Municipal Solid Waste: An Approach in the Circular Economy

Urbanization and rapid industrialization have led to the escalation of municipal solid waste generation and accumulation. Composting is widely recognized as a sustainable solution for solid waste management. However, its long-term investment is considered a disadvantage. The present research study discusses the rapid biotransformation of solid waste into valorized compost. Bacteria were isolated from soil, solid waste, and leachate samples from open dump sites. From the 18 different bacterial consortia created using potential isolates, the five most promising consortia were selected based on concurrent different enzyme production. These selected consortia were incorporated into typical compost bins with Municipal Solid Waste (MSW). Daily monitoring of enzymatic activity, pH, conductivity, bulk density, moisture, and temperature, along with other composting parameters, was conducted. The study’s results demonstrated that consortium No. 5, comprising Bacillus haynesii, Bacillus amyloliquefaciens, and Bacillus safensis, exhibited significant (p<0.05) enzyme activity of cellulase, amylase, lipase and proteinase enzymes during composting compared to the control and other treatment setups. Consortium No. 5 also facilitated rapid and successful composting, as evidenced by significant alterations of composting parameters by exhibiting a shorter average composting time, reducing it from 110±10 days to 20±3 days, showcasing the potential applicability of formulated bacterial consortium as a sustainable and greener approach to the global solid waste problem. The novelty of this study lies in the isolation of local bacterial strains from open dump sites soil, MSW, and MSW leachate samples, which were then utilized in the composting organic fraction of MSW, enhancing the potential for effective waste management.

Antioxidant Activity, Glycemic Response, and Functional Properties of Rice Cooked with Red Palm Oil.

High rice consumption levels accompanied by a lifestyle lacking in physical activity leads to obesity and diabetes due to the rice consumed generally has high digestibility and high glycemic index. Red palm oil (RPO) is a vegetable oil suggested to have the potential to reduce starch digestibility and increase the bioactive compounds of rice. This research aimed to find out the best cooking method to produce rice with a sensory quality similar to regular rice and to study the effect of the best cooking method on the glycemic response and physicochemical properties of rice. The results showed that RPO addition increased the antioxidant activities and total carotenoid levels of rice. The addition of RPO after cooking has better antioxidant activity and total carotenoid than before cooking. Adding 2% RPO before or after cooking produced rice with similar or better sensory quality than regular rice. Rice cooked with 2% RPO added before cooking had a lower glycemic response than regular rice, which was suggested to be caused by the increasing formation of the amylose lipid complex and the triglycerides that protected the starch from amylase enzyme. The formation of the amylose lipid complex and triglyceride layers protecting rice starch was confirmed by the new peaks of the FTIR spectra, the appearance of oil-coated starch morphology, and the changes in the proportion of C and O atoms. In conclusion, the addition of 2% RPO before the cooking process can be considered as a cooking method to produce rice for diabetic patients.

Integrative approach to the biochemical, and toxicological fingerprinting of Polygonum glabrum.: A computational and experimental synergy for a medicinal food plant

In this work, the biological and chemical fingerprints of methanol and DCM extracts from an important Polygonum species (Polygonum glabrum) was investigated. The biological fingerprint was assessed for its antioxidant, enzyme inhibitory, and cytotoxic effects. For the chemical characterization, total bioactive components, and individual secondary metabolites were determined using photometric, HPLC-PDA polyphenolic quantification, and UHPLC-MS methods. The main identified compounds were belonging to flavonoid and phenolic classes of phytochemicals. Methanol extracts contained the highest total phenolic (54.66 mg GAE/g extract) and flavonoid (27.85 mg GAE/g extract) content. The results of antioxidant assays showed that methanol extract displayed higher antioxidant (for DPPH, ABTS, FRAP, CUPRAC) activity compared to the DCM extract (which was most active for metal chelation assay). For the enzyme inhibition activity, the DCM extract was found to be active against cholinesterases, while the methanol extract exhibited the higher inhibition potential for tyrosinase, amylase, and glucosidase enzymes. Generally, the methanol extract exhibited higher biological activities which might be correlated with higher levels of the bioactive components. Moreover, three flavonoids (citrusin B, robinetin 3-rutinoside, and demethyltexasin) were selected for molecular docking studies against the tested enzymes. Furthermore, the utilization of Swiss ADME, pkCSM, and DFT computational analysis has enhanced our comprehension of the pharmacokinetic characteristics and electronic factors of the identified phytochemicals, hence advancing the possibility for drug discovery. To conclude, the results suggest that the P. glabrum extracts could be further studied as a potential candidate for developing new phyto-pharmaceuticals and functional ingredients.

Alpha-Amylase and Alpha-Glucosidase Inhibitory Potential of Overnight Soaked Aqueous Extract of Abelmoschus esculentus

Phytochemicals are secondary metabolites which act as antioxidants and possess various protective role.Alpha Amylase and Alpha Glucosidase enzymes catalyse hydrolysis of starch to simple sugars. Abelmoschus esculentus is a flowering plant of the Malvaceae family and is valued for its green seed pods. It naturally possesses anti-diabetic property, acts as a starch blocker as it prevents the absorption of starch in the body. Abelmoschus esculentus was cut vertically to open and added to 50ml of water. Soaked overnight and the aqueous extract was decanted and used for the phytochemical screening test followed by the evaluation of antioxidant and antidiabetic potential. The data were analysed statistically by a one-way analysis of variance (ANOVA) followed by Duncan’s multiple range test. Overnight soaked extract of Abelmoschus esculentus exhibited a significant antioxidant potential (IC50=280 µg/ml) and compared with standard vitamin C. Alpha amylase and alpha glucosidase inhibitory potential of Abelmoschus esculentus was analysed (IC50=380µg/ml and 320µg/ml respectively) and compared with standard acarbose. Though Abelmoschus esculentus is a common vegetable, its efficiency increases when soaking overnight.

Intensification of α-amylase activity using ultrasound and its application for food waste hydrolysis

The current research aims to study the ultrasound assisted intensification of activity of commonly applied amylase enzyme, under varying parameters like power, frequency, duty cycle, temperature and enzyme concentration. Two ultrasonic reactor configurations as bath and horn are used for comparing the efficacy of reactor configuration. The initial activity of native amylase was found as 5.77 U/g at optimum temperature of 35° C and pH 7. Under US bath treatment, α amylase attained maximum activity of 7.498 U/g at best treatment conditions of 120 W power, 30% duty cycle and 15 minutes of exposure time. In the case of US horn, application of 60 W power, 30% duty cycle, and 5 minutes of exposure increased the enzyme activity to 6.907 U/ g. Set of treatment approaches explored for obtaining highest yield of reducing sugars revealed the best approach of ultrasonic treatment of food waste followed by ultrasonic bath intensified amylase hydrolysis, yielding maximum amount of reducing sugars as 8.14 g/L in only 15 min. Overall the work elucidated the role of ultrasound in intensifying the enzyme activity as well as food waste hydrolysis maximizing the utilization of sustainable resource.

Isolation and Identification of Soil-Derived Amylase-Producing Bacteria

Soil is a vital ecosystem for various life forms, including bacteria. Bacteria are the main group of soil microbes, mostly because of their ability to produce various extracellular enzymes. Amylases are the enzymes that break down starch, which are essential in biotechnology industries such as food, fermentation, textiles, and paper manufacturing. The demand for amylase is often high since many enterprises do not have access to local supplies of the enzyme. The objective of the current investigation is to isolate and identify bacterial isolates that can produce the enzyme amylase from soil samples. Soil samples were collected from the unexplored Biligiriranga hills (11.9988° N, 77.1398° E) in south-western Karnataka Bacterial isolates were isolated from soil samples and were primarily screened on starch agar medium to determine the amylase production. Three out of fifteen bacterial isolates that produced amylase were selected based on the highest clear zone around the bacterial colony. These three bacterial isolates were identified both microscopically and biochemically. Gram staining demonstrated that the isolates are rod-shaped and gram-positive organisms. Biochemical studies indicated that the isolates were positive for the catalase and starch hydrolysis tests, whereas negative for the potassium hydroxide and methyl red tests. Besides, 16S rRNA gene amplification and sequencing confirmed the isolate as Bacillus subtilis. The 16S rRNA sequence of B. subtilis strain B has been deposited in the NCBI GenBank. The optimal growth conditions observed in B. Subtilis. The optimal growth conditions observed in B. Subtilis were 37° C temperature at pH 7 and incubation period for 37h. Thus, the isolation and identification of soil-derived amylase-producing bacteria are crucial for understanding their potential applications in various industrial processes, such as bioremediation, waste management, and the production of enzymes for food, pharmaceuticals, and biofuel industries.

Investigation of in vitro Biological Activity from Extracts of Ruellia tuberosa.

<b>Background and Objective:</b> <i>Ruellia tuberosa</i> is a common plant in the Mekong Delta and is widely used in many Vietnamese folk remedies. This study was conducted to investigate the potential use of roots, stems, leaves of <i>Ruellia tuberosa</i> as antioxidant, antimicrobial, α-amylase and α-glucosidase inhibitors. <b>Materials and Methods:</b> The extracts were tested for their ability to inhibit the enzymes α-amylase and α-glucosidase associated with diabetes. The antioxidant activities of the extracts were evaluated using 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) and 2,2-Azino-Bis-(3-Ethylbenzothiazoline-6-Sulfonic Acid) (ABTS), ferric reducing antioxidant power (FRAP), total antioxidant capacity (TAC) and reducing power (RP) assays. The antibacterial activity of extracts from <i>Ruellia tuberosa</i> was evaluated by the agar well diffusion method. <b>Results:</b> The root extract of <i>Ruellia tuberosa</i> has more polyphenols (32.49±0.72 mg GAE/g extract) and flavonoids (15.48±1.32 mg QE/g extract) than the other parts. Simultaneously, the root extract of <i>Ruellia tuberosa</i> has antioxidant activity (IC<sub>50</sub> values range from 117.67±2.82 to 569.20±7.68 μg/mL), inhibiting amylase (IC<sub>50</sub> = 266.72±10.58 μg/mL) and glucosidase (IC<sub>50</sub> = 147.13±3.58 μg/mL) enzymes more effectively than the other parts. Research results also show that extracts from <i>Ruellia tuberosa</i> are capable of inhibiting <i>Staphylococcus aureus</i>, <i>Escherichia coli</i> and <i>Pseudomonas aeruginosa</i> bacteria with minimum inhibitory concentrations ranging from 1280 to 10240 mg/mL. <b>Conclusion:</b> These results highlighted the potential using of <i>Ruellia tuberosa</i> extracts as natural antioxidant, antimicrobial, α-amylase and α-glucosidase inhibitors agents.

Kigelia africana (Lam.) Benth. fruit inhibits iron-induced lipid peroxidation and ?-amylase enzyme activity

Insufficiencies in insulin secretion and/or action cause diabetes mellitus (DM), a complex condition characterized by abnormal blood glucose levels. Diabetes is often treated by modifying either glucose metabolism, lipids metabolism, or both. A direct correlation has been found between high levels of lipid peroxide in individuals with diabetes and lower intracellular antioxidant activity. Therefore, this study was designed to investigate the effect of the aqueous extract of Kigelia africana fruit on the carbohydrate hydrolyzing enzyme, ?-amylase, and FeSO4-induced lipid peroxidation in the rat pancreas. The in vitro antioxidant potential, including assays for (1,1 diphenyl-2-picryl hydrazyl (DPPH), ferric reducing antioxidant property (FRAP), nitric oxide (NO), hydroxyl (OH) scavenging, and Fe2+ chelating abilities, of the fruit extract was also evaluated. The effect of the fruit extract on the ?-amylase enzyme, and FeSO4-induced lipid peroxidation in the rat pancreas was also evaluated. K. africana at a concentration of 0.5 mg/mL and 1 mg/mL scavenged DPPH radicals, with the lower concentration showing significant differences compared to the control. The K. africana fruit extract reduced ferric compounds significantly compared to the standard. The OH* radical scavenging results revealed that all concentrations of K. africana fruit scavenged OH* radicals without significant differences compared to the control. K. africana fruit also chelated iron significantly compared to the control. The fruit extract inhibited iron-induced lipid peroxidation in the rat pancreas and ?-amylase activity compared to the control. K. africana fruit displayed promising potential in inhibiting ?-amylase activity and associated lipid peroxidation in diabetics. Therefore, it holds strong promise for use in diabetes treatment, validating its traditional use for managing the condition.

Multigene phylogeny, bioactive properties, enzymatic and dye decolorization potential of selected marine fungi from brown algae and sponges of Mauritius

Marine fungi represent an important proportion of the microbial diversity in the oceans. They are attractive candidates for biotechnological purposes and industrial applications. Despite an increasing interest in mycology, marine fungi associated with sponges and algae have been poorly studied in Mauritius. The objectives of this study were to: 1) use multigene phylogenetic analyses to identify isolated marine fungi; 2) determine the differences in the antimicrobial and antioxidant properties of the fungal extracts; and 3) assess their enzyme activities and dye decolorization potential. Five fungal isolates viz Aspergillus chevalieri, Aspergillus iizukae, Aspergillus ochraceus, Exserohilum rostratum and Biatriospora sp. were identified based on phylogenetic analyses. There was no significant difference in the antimicrobial properties of the liquid and solid media extracts unlike the antioxidant properties (p < 0.05). The solid media extract of Aspergillus chevalieri (F2-SF) had a minimum inhibitory concentration of 0.156 mg/ml against Staphylococcus aureus while Aspergillus ochraceus (F25-SF) had a minimum inhibitory concentration of 0.313 and 2.5 mg/ml against Enterococcus faecalis and Salmonella typhi. The solid media extract of Biatriospora sp. (F34-SF) had a minimum inhibitory concentration of 0.195 and 1.563 mg/ml against Bacillus cereus, Escherichia coli and Enterobacter cloacae. An IC50 of 78.92 ± 4.71 μg/ml in the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging assay, ferric reducing antioxidant power (FRAP) value of 11.17 ± 0.20 mM Fe2+/g dry weight extract (DWE) and total phenolic content 360.35 ± 10.31 mg GAE/g DWE was obtained with the solid media extract of Aspergillus chevalieri (F2-SF). Aspergillus ochraceus (F25-SF) and Biatriospora sp. (F34-SF) solid media extracts showed lower IC50 values in the DPPH assay and higher total phenolic content as compared to the liquid media extracts. Aspergillus chevalieri was a good producer of the enzymes DNAse and lipase and had maximum percentage dye decolorization of 79.40 ± 17.72% on Congo red. An enzymatic index ≥ 2 was found for the DNAse and lipase and the maximum percentage dye decolorization of 87.18 ± 3.80% was observed with Aspergillus ochraceus on Methylene blue. Regarding Biatriospora sp., it was a moderate producer of the three enzymes amylase, DNAse and protease and had a maximum dye decolorization potential of 56.29 ± 6.51% on Crystal violet. This study demonstrates that Mauritian marine fungi possess good bioactive properties, enzymatic and dye decolorization potentials, that can potentially be considered for use in pharmaceutical and industrial applications.

Feeding rhythm of the zoea larvae of Scylla paramamosain: The dynamic feeding rhythm is not completely synchronized with photoperiod

The feeding rhythm is one of the key factors determining the success of artificial breeding of S. paramamosain. To understand the feeding rhythm of the different zoea larva developmental stages of S. paramamosain, the feeding rate, digestive enzyme activity, and expression of metabolism-related genes were investigated in the present study. The results showed that the S. paramamosain feeding rate has strong diurnal feeding rhythm, being significantly higher at 10:00–14:00 from stages ZI to ZIV. While the feeding rate peaked at 14:00 on Days 10 and 11, the peak shifted to 18:00 on Day 12. The activity of digestive enzymes amylase, pepsin and lipase decreased at night but increased in the daytime, showing a single-phase rhythm similar to that of the feeding rate, suggesting that the digestive enzyme activity was closely associated with the feeding rate during the larval development. Compared to pepsin and lipase, the activity of amylase was the most consistent with feeding rate. In particular, amylase activity peaked at 18:00 on Day 12. Due to its synchronicity with feeding activity, the activity of amylase could provide a potential reference for determining the best feeding time during zoea stages in S. paramamosain breeding. Moreover, the relative mRNA expression of metabolism-related genes SpCHH and SpFAS at most tested points was lower from 10:00 to 14:00, but higher at 18:00 to 6:00 of the next day. On the other hand, the expression patterns of SpHSL and SpTryp were converse to those of SpCHH and SpFAS. Our findings revealed that the S. paramamosain zoea has an obvious feeding rhythm, and the most suitable feeding time was 10:00–18:00 depending on different stages. The feeding rhythm is a critical aspect in aquaculture, influencing a series of physiological functions in aquatic animals. This study provides insights into the feeding rhythm during the zoea development of S. paramamosain, making a significant contribution to optimizing feeding strategy, improving aquafeed utilization, and reducing the impact of residual feed on water environment.

Hydrolysis profile of gadung (dioscorea hispida dennst) starch to glucose using alpha amylase enzyme

Indonesia is an agrarian Country that has abundant natural resources. It has potential to be used as industrial raw materials such as sugar reduction from starch. Gadung, which is abundant in Gandus and it can be monitored for its presence in the dry season, is one source of starch. The aims of this research is to obtain glucose with a high yield percentage, to know the optimum concentration of α-amylase enzyme to hydrolysis gadung starch into glucose, and to know the relationship between hydrolysis time and concentration the glucose formed by the addition of the enzyme variation. The implementation of the research is carried out in three stages, first is preparation of raw materials, hydrolysis of gadung starch into glucose enzymatically, and analysis of glucose level using the UV-Vis spectrophotometry with Nelson reaction. The free variables used are the time variation of hydrolysis 0, 10, 20, 30, 40, 50 minutes and alpha amylase enzyme 1 %, 2 %, 3 % b/v. The optimum hydrolysis results on this study were at 1 % b/v enzyme variation, 10 minutes hydrolysis time, and a temperature of 90 oC. The yield of glucose concentration is 255.35 ppm. From this reseach, it is known that the longer the starch hydrolysis takes place, the observed glucose tends to experience in fluctuative consentration caused by the increase of α- amylase enzyme variations.

Analysis of Amylase Activity in Bacteria Isolated from Hot Spring of Pentadio Resort

The hot spring of Pentadio Resort is one of the geothermal areas likely to be the place of bacteria or microorganisms that can produce amylolytic enzymes. This research aims to analyze bacteria in the Pentadio Resort hot springs that possess the potential to produce amylase enzymes, analyze the characteristics of bacterial isolates that produce amylase enzymes in Pentadio Resort hot springs and determine the types of bacteria that produce amylase enzymes in Pentadio Resort hot springs. The research employs a qualitative and descriptive method. The research involved several methods, including sample collection, media preparation, isolation and selection of thermophilic bacterial isolates, purification of thermophilic bacterial isolates, selection of isolates producing amylase enzymes, identification of selected isolates (motility test, biochemical test, gram staining), and molecular identification. The research yielded seven isolates, and isolate code A6 demonstrated the potential to produce amylase enzymes with a clear zone size of 23,26 mm, bacillus, gram positive, and positive motility. Subsequently, molecular identifidation was carried out in which, based on the results of phylogenetic tree reconstractiopisalate A6 was related to the bacteria Bacillus cereus strain MD 152, which belongs to the Bacillus cereus bacteria

Targeting intracellular nontuberculous mycobacteria and M. tuberculosis with a bactericidal enzymatic cocktail.

To address intracellular mycobacterial infections, we developed a cocktail of four enzymes that catalytically attack three layers of the mycobacterial envelope. This cocktail is delivered to macrophages, through a targeted liposome presented here as ENTX_001. Endolytix Cocktail 1 (EC1) leverages mycobacteriophage lysin enzymes LysA and LysB, while also including α-amylase and isoamylase for degradation of the mycobacterial envelope from outside of the cell. The LysA family of proteins from mycobacteriophages has been shown to cleave the peptidoglycan layer, whereas LysB is an esterase that hydrolyzes the linkage between arabinogalactan and mycolic acids of the mycomembrane. The challenge of gaining access to the substrates of LysA and LysB provided exogenously was addressed by adding amylase enzymes that degrade the extracellular capsule shown to be present in Mycobacterium tuberculosis. This enzybiotic approach avoids antimicrobial resistance, specific receptor-mediated binding, and intracellular DNA surveillance pathways that limit many bacteriophage applications. We show this cocktail of enzymes is bactericidal in vitro against both rapid- and slow-growing nontuberculous mycobacteria (NTM) as well as M. tuberculosis strains. The EC1 cocktail shows superior killing activity when compared to previously characterized LysB alone. EC1 is also powerfully synergistic with standard-of-care antibiotics. In addition to in vitro killing of NTM, ENTX_001 demonstrates the rescue of infected macrophages from necrotic death by Mycobacteroides abscessus and Mycobacterium avium. Here, we demonstrate shredding of mycobacterial cells by EC1 into cellular debris as a mechanism of bactericide.IMPORTANCEThe world needs entirely new forms of antibiotics as resistance to chemical antibiotics is a critical problem facing society. We addressed this need by developing a targeted enzyme therapy for a broad range of species and strains within mycobacteria and highly related genera including nontuberculous mycobacteria such as Mycobacteroides abscessus, Mycobacterium avium, Mycobacterium intracellulare, as well as Mycobacterium tuberculosis. One advantage of this approach is the ability to drive our lytic enzymes through encapsulation into macrophage-targeted liposomes resulting in attack of mycobacteria in the cells that harbor them where they hide from the adaptive immune system and grow. Furthermore, this approach shreds mycobacteria independent of cell physiology as the drug targets the mycobacterial envelope while sidestepping the host range limitations observed with phage therapy and resistance to chemical antibiotics.

Synthesis, biological activities, and molecular docking studies of triazolo[4,3-b]triazine derivatives as a novel class of α-glucosidase and α-amylase inhibitors.

In diabetes mellitus, amylase and glucosidase enzymes are the primary triggers. The main function of these enzymes is to break macromolecules into simple sugar units, which directly affect blood sugar levels by increasing blood permeability. To overcome this metabolic effect, there is a need for a potent and effective inhibitor capable of suppressing the enzymatic conversion of sugar macromolecules into their smaller units. Herein, we reported the discovery of a series of substituted triazolo[4,3-b][1,2,4]triazine derivatives as α-glucosidase and α-amylase inhibitors. All target compounds demonstrated significant inhibitory activities against α-glucosidase and α-amylase enzymes compared with acarbose as the positive control. The most potent compound 10k, 2-[(6-phenyl-[1,2,4]triazolo[4,3-b][1,2,4]triazin-3-yl)thio]-N-[4-(trifluoromethyl)phenyl]acetamide, demonstrated IC50 values of 31.87 and 24.64 nM against α-glucosidase and α-amylase enzymes, respectively. To study their mechanism of action, kinetic studies were also done, which determined the mode of inhibition of both enzymes. Molecular docking was used to confirm the binding interactions of the most active compounds.