Amylase Enzyme Activity Research Articles

Phytochemical evaluation and pharmacognostic standardization of Syzygium palghatense endemic to Western Ghats

BackgroundThe plants belonging to the genus Syzygium has received much attention since ancient times due to its multidimensional uses for mankind. These plants in general, reported to contain a wide array of phytomolecules with a broad spectrum of biological activities that include antihypoglycemic, antioxidant, anti-inflammatory, anticarcinogenic, and antihypertensive properties. The pharmacological potential of S. palghatense which is endemic to Western Ghats has not yet been investigated. The present study was intended to examine the potential benefits of the leaves and bark of S. palghatense.ResultsMorpho-anatomical studies of S. palghatense provided useful details for the identification. The pharmacological characterization of the plants shows significant results in their studied parameters. The total phenolic and flavonoid contents were higher in methanolic extracts of leaves and bark of S. palghatense. Methanolic extract of the leaves and bark have superior antioxidant capacity when compared with the chloroform, ethyl acetate, and distilled water extracts. A significant result in alpha amylase and alpha glucosidase enzyme activity was exhibited when antidiabetic study was performed.ConclusionThe current study showed that the methanolic extract of S. palghatense leaves and bark have significant pharmacognostic properties and potential antioxidant and antidiabetic compounds. Phytochemical studies portray the presence of several biologically active secondary metabolites, including essential oil, alkaloid, glycosides, carbohydrate, tannins, reducing sugar, phenolics, and saponins, which may be the reason for its biological properties. As compared to bark, leaves of S. palghatense possess significant activity toward all the biological properties tested.

Influence of Oral Administration of Bacillus subtilis on Growth Performance and Physiological Responses in Litopenaeus vannamei Cultured in Low Saline Waters.

The present study examined the effect of Bacillus subtilis on growth performance, digestive enzyme activity, and immune response of juvenile white-leg shrimp (Litopenaeus vannamei) in 100 m2 lined ponds. Control (without B. subtilis) and experimental (with B. subtilis) diets were fed at 10% of animal body weight, four times daily for 6weeks in duplicate ponds. Significantly increased final weight, weight gain, specific growth rate, average daily growth, and survival rate were observed in the B. subtilis treated group (P < 0.05). Significantly higher (P < 0.05) digestive enzyme activities of protease, lipase, amylase, and cellulase were found in the B. subtilis supplemented diet fed group. At the end of trial, a challenge study, using Vibrio alginolyticus, found lower (15.00 ± 2.88%) cumulative mortality in the B. subtilis treated group compared to control (73.33 ± 3.33%). Activities of immune and antioxidant enzymes (phenoloxidase, SOD, and catalase) were significantly higher (P < 0.05) in the B. subtilis-incorporated diet fed group. Interestingly, histopathology of the hepatopancreas and intestine revealed that the B. subtilis-supplemented group showed no deformity. Overall, the study found inclusion of B. subtilis in P. vannamei diet improves growth, physiometabolic activities, and immunity, indicating B. subtilis as a potential probiotic for shrimp feed.

Combined effect of Spirulina and ferrous fumarate on growth parameters, pigmentation, digestive enzyme activity, antioxidant enzyme activity and fatty acids composition of Yellow River carp (Cyprinus carpio)

This study was conducted to investigate the effects of Spirulina and ferrous fumarate supplementations into diets on growth parameters, pigmentation, digestive enzyme activity, antioxidant enzyme activity and fatty acids composition of Yellow River carp. Three experimental groups included control, Spirulina (diet 1), mixture of Spirulina and ferrous fumarate treatments (diet 2) in 3 replications in 9 aquariums. Fifteen fish in each aquarium were fed twice a day for six weeks. Compared to the control group, the Yellow River carp fed with the Spirulina and ferrous fumarate had a higher specific growth rate (P < 0.05). The total carotenoids content in Yellow River carp skin and scale was significantly higher for fish fed diet with Spirulina and ferrous fumarate relative to all other diets (P < 0.05). The color parameters of L *, a *, b*, Hue (Hab°) and Chroma (ch*) were calculated for all treatments. Obtain findings revealed that the Spirulina and ferrous fumarate treatment could provide the desired skin color exactly of Yellow River carp. The Spirulina and ferrous fumarate supplementation significantly elevated intestinal digestive capacity by increasing amylase, lipase, and trypsin enzyme activities (P < 0.05). The results obtained indicated that Spirulina and ferrous fumarate supplementation could improve intestinal health through enhancing intestinal digestive enzyme activity and reducing tissue peroxidation. In addition, the Spirulina and ferrous fumarate has a satisfactory nutritional value of improving the fatty acid profile. Therefore, the use of Spirulina and ferrous fumarate as feed additive could be practiced in the aquaculture species.

Physiological Characteristics and Comparative Secretome Analysis of Morchella importuna Grown on Glucose, Rice Straw, Sawdust, Wheat Grain, and MIX Substrates.

Morels (Morchella sp.) are economically important edible macro-fungi, which can grow on various synthetic or semi-synthetic media. However, the complex nutritional metabolism and requirements of these fungi remain ill-defined. This study, based on the plant biomass commonly used in the artificial cultivation of morels, assessed and compared the growth characteristics and extracellular enzymes of Morchella importuna cultivated on glucose, rice straw, sawdust, wheat grain, and a mixture of equal proportions of the three latter plant substrates (MIX). M. importuna could grow on all five tested media but displayed significant variations in mycelial growth rate, biomass, and sclerotium yield on the different media. The most suitable medium for M. importuna was wheat and wheat-containing medium, followed by glucose, while rice straw and sawdust were the least suitable. A total of 268 secretory proteins were identified by liquid chromatography coupled with tandem mass spectrometry detection. Functional classification and label-free comparative analysis of these proteins revealed that carbohydrate-active enzyme (CAZYme) proteins were the predominant component of the secretome of M. importuna, followed by protease, peptidase, and other proteins. The abundances of CAZYme proteins differed among the tested media, ranging from 64% on glucose to 88% on rice straw. The CAZYme classes of glycoside hydrolases and carbohydrate-binding module were enriched in the five secretomes. Furthermore, the enzyme activities of CMCase, lignase, amylase, xylase, pNPCase, and pNPGase were detected during the continuous culture of M. importuna in MIX medium, and the relative expression of the corresponding genes were detected by quantitative real-time PCR. The combined data of growth potential, secretome, extracellular enzyme activity, and gene expression on different substrates inferred that M. importuna was weak in lignocellulose degradation but a good starch decomposer. Specifically, in terms of the degradation of cellulose, the ability to degrade cellulose into oligosaccharides was weaker compared with further degradation into monosaccharides, and this might be the speed-limiting step of cellulose utilization in M. importuna. In addition, M. importuna had a strong ability to decompose various hemicellulose glycosidic bonds, especially α- and β-galactosidase. Only a very few lignin-degradation-related proteins were detected, and these were in low abundance, consistent with the presence of weak lignin degradation ability. Furthermore, the presence of lipase and chitinase implied that M. importuna was capable of decomposition of its own mycelia in vitro. The study provides key data that facilitates a further understanding of the complex nutritional metabolism of M. importuna.

AKTIVITAS ENZIM HIDROLASE PADA PENAPISAN ISOLAT ACTINOMYCETES TOLERAN SALINITAS

Actinomycetes is one of the microorganisms which have the ability of enzymes hydrolases such as cellulose, amylase, protease, and manannase. Manufacturer of enzymes of the hydrolase can be used as a candidate probiotic. A candidate probiotic should have a good resistance in different environmental conditions including pH and salinity. This research is conducted to filter the candidate’s probiotics from bacteria Actinomycetes that tolerance salinity and has the activity of the enzyme hydrolase. Testing the activity of the enzyme was preformed qualitatively on the variation of pH 4, 7, and 9.8. Theability of Atinomycetes produce enzymes hydrolases is indicated by the colony on media containing the substrate.From the test results, cellulase enzyme activity with the largest enzymatic index value is 3.5 AF2 isolates pH 9.8. Amylase enzyme activity with the largest enzymatic index value is 3.66 RH1 isolates pH 7. Protease enzyme activity with the largest enzyme index value is 0.66, AF2 isolates at pH 7. Mananase enzyme activity with the largest enzymatic index value is 0.85, namely AF2 isolates pH 4. In testing the effect of salinity test isolates were able to grow at variations in the concentration of NaCl 0, 3, and 6%.

The effect probiotics from the types of nuts in feeding to the for bacterial populations, enzyme activity and feed FCR for milkfish (Chanos chanos)

Nuts contain oligosaccharides which can function as prebiotics. Prebiotics are fibers that the body cannot digest and become food for probiotics. This study aimed to analyze the bacterial population, enzyme activity and feed conversion ratio of milkfish after being given various prebiotics from legumes in the feed. Milkfish measuring 4.13 ± 2.75 g are reared in a glass aquarium measuring 50 × 45 × 45 cm filled with water with a salinity of 15-20 ppt. Fish are maintained at a density of 15 individuals/aquarium for 60 days and are fed 5% body weight/day. The study was designed in a completely randomized design with 5 prebiotic treatments of soybeans, peanuts, green beans, kidney beans, and control. The results showed that the treatment had a significant effect (p <0.05) on the bacterial population, as well as the protease and amylase enzyme activity, but not on FCR. The highest bacterial population was red bean prebiotics (2,082 × 103 CFU/mL), followed by green beans (2,001 × 103 CFU/mL), soybeans (972 × 103 CFU/mL). The highest activity (0.243 u/mL) and amylase enzyme (0.745 u/mL) was the prebiotic kidney beans, followed by soybeans (0.236 u/mL) amylase enzyme (0.506 u/mL), peanuts, protease enzymes (0.129 u/mL) amylase enzyme (0.554 u/mL) and green bean protease enzyme (0.156 u/mL) amylase enzyme (0.543 u/mL). Meanwhile, the FCR ranges from 1.43 ±.0.85 to 2.87 ±.1.70. It can be concluded that the best type of prebiotic is red beans.

Change in malate dehydrogenase and alpha amylase activities in Rubus fruticosus and Valeriana jatamansi treated granary weevil, Sitophilus granarius.

Poor storage conditions provide favorable environment to stored grain pests for their growth. The bio-pesticides are the best alternatives to synthetic pesticides. Present study was conducted to compare toxicity of Rubus fruticosus and Valeriana jatamansi against granary weevil, Sitophilus granarius and subsequent changes in enzyme activity responsible for grain damage. In current research 5 g of R. fruticosus fruit and V. jatamansi rhizome powders were tested separately against S. granarius, in 50 g wheat whole grains for seven days in comparison with the control. The enzymatic activity of malate dehydrogenase and α-amylase was observed in the cellular extracts of S. granarius. The insects were crushed and homogenized in phosphate-buffer solution and centrifuged at 10000 rpm for 5 minutes. For the enzymatic measurement supernatant was tested; the spectrophotometer was adjusted at 340 nm. The reagents were mixed and incubated at 25 °C for five minutes. The cuvettes were placed in the experimental and reference sites of spectrophotometer and recorded the change in absorbance for 3-4 minutes. There was 5.60% and 14.92% reduction in the activity of malate dehydrogenase in R. fruticosus and V. jatamansi, treated insects, respectively. The alpha amylase enzyme activity was 6.82% reduced and 63.63% increase in R. fruticosus and V. jatamansi, treated insects, respectively. Present study addresses that both plant powders are effective against granary weevil by altering enzyme activities so both the plant powders can be used as bio-pesticides against the stored grains pests.

Ethanol and Sugar Production from Sorghum (Sorghum bicolor) Starch at Varying Acidity

Sorghum (Sorghum bicolor), in addition to producing foodstuffs, feed, industrial use, is also potential for ethanol production. Sorghum is also a potential developed because it has a complete adaptation power in marginal land and other non-productive lands. The use of sorghum starch for ethanol production is a renewable fuel alternative and friendly for the environment. The aim of the research was to study the fermentation of sorghum starch into simple sugars and ethanol at varying acidity (pH). The study is experimental research with two treatment factors: pH (4; 6; 7; 8; 9; 10) and sugar levels. Data were collected and analyzed with simple and multiple linear regression analyses. The result showed that the pH treatment applied to influence the enzyme fermentation of sorghum starch. The conclusion of the study is the optimum pH of amylase enzyme activity in fermenting sorghum starch into glucose is pH 8. The relationship between the amount of alcohol, sugar, and pH is very strong at 95.1%. Multiple regression equations to explain the relationship of the three variables that the amount of alcohol (Y) produced in sorghum starch fermentation can be estimated additive by the amount of sugar formed (X1) and pH (X2).

Characteristic of Biomaterial from Fermentation Product and its Effectto Enzyme Activity of &lt;i&gt;Clarias&lt;/i&gt; sp.

The physical characteristics of fermented products from restaurant waste food with BIOMS probiotic at a dose of 8% which was incubated for 7 days were investigated in order to study effect of fermentation to the physical characteristic of biomaterial and its effect to enzyme activity of Clarias sp. Fermented products are added to the feed with doses of 0, 10%, 20%, 30% and 40% and physical tests include breaking time, sinking rate test, durability test, and stability test. Furthermore, the feed was given to catfish for 40 days and the enzyme activity (amylase and protease) was analyzed in its digestive tract at the end of the study. It is showed that the addition of fermented products decreased the physical quality of the feed. The best physical quality was found in the control feed with an average value of breaking time of 22.98 hours, an average value of sinking rate of 0.53 cm/s, a durability test of 97.23 %, and a stability test value of 93.53% (10 minutes), 88.8% (30 minutes), and 76.26% (60 minutes). For feed products with doses of fermented products of 10%, 20%, 30%, and 40%, the values ​​of breaking time and sinking rate, durability and stability were lower compared to that of control feed products. Meanwhile, the enzyme activity in the digestive tract of catfish shows that giving a dose of 20% fermented products in the feed resulted in the best composition of amylase and protease enzyme activity of 9.021 units/mL and 275 units/mL. These results indicated that improvements are needed in the fermentation process to produce better physical quality and suitable for catfish feed.

Nutritional evaluation of some Indian tree pods for livestock feeding

The objective of this study was to evaluate promising tree pods for feeding to the livestock particularly for the small ruminants. Out of eight tested tree pods, seven tree pods, i.e. White siris (Albizia procera), Siris (Albizia lebbeck), White kheri (Acacia senegal), Babul (Acacia arabica), Khejri (Prosopis cineraria), Vilayati babul (Prosopis juliflora) and Sajna (Moringa oleifera) were collected from semiarid region of Rajasthan while one tree pods e.g., Jungle jalebi (Enterolobium timoba) was collected from Dehradun, Uttrakhand, India. Most of the tree pods were rich in CP content. On an average, OM, CP, EE, NDF, ADF and cellulose content of these tree pods were found to be 91.1, 16.7, 2.5, 43.3, 34.7 and 25.4% on DM basis, respectively. Rumen protozoal number decreased due to inclusion of Enterolobium timoba tree pods in the incubation media. The TVFA and propionate production were higher for Acacia Senegal, Acacia arabica tree pods followed by Moringa oleifera tree pods while ammonia nitrogen concentration was lower due to inclusion of Enterolobium timoba tree pods in the incubation media. All the tested tree pods had no effect on xylanase, β-glucosidase and amylase enzyme activity. However, specific activity of carboxymethyl cellulase enzyme reduced due to addition of Enterolobium timoba tree pods in the incubation medium. Highest IVDMD was observed for Acacia arabica tree pods followed by Acacia senegal and Moringa oleifera tree pods. The results indicated that Acacia arabica, Acacia senegal and Moringa oleifera are good tree pods for feeding to the animals.

Development of a novel multi-strain wheat Qu with high enzyme activities for Huangjiu fermentation.

Wheat Qu has long been used as a fermentation starter to produce Huangjiu. Wheat Qu quality depends on its microbial community structure and the hydrolytic enzymes generated by the micro-organisms. Strain YF1 and YF2 were successfully screened as they exhibited high acidic protease (231.9 ± 1.4U g-1 ) and cellulase (7.1 ± 0.6U g-1 ) activities. Based on a morphological and sequence analysis of the internal transcribed spacer (ITS) gene, YF1 and YF2 were identified as Rhizopus oryzae and Aspergillus niger, respectively. Cooked wheat Qu was produced using mixed fungal starter fermentations with Aspergillus oryzae SU-16, YF1, and YF2. For Qu-making, the optimized conditions for fermentation time, water content, and inoculum size were 47.8h, 69.4%, and 6.1%, respectively. Under these conditions, compared with single-strain cooked wheat Qu, enzyme activities of amylase, acidic protease, and cellulase increased by 27.4%, 657.1%, and 1276.2%, respectively. Short peptides and free amino acids contents increased by 19.6% and 131.8%, respectively. This wheat Qu was used for Huangjiu brewing, and the alcohol content increased by approximately 14.6% because of the increased starch hydrolysis efficiency mainly attributed to its high enzyme activity. Using mixed fungal strains as starter cultures may be an efficient strategy to improve wheat Qu quality, with great potential for application in industrial Huangjiu production. © 2021 Society of Chemical Industry.

Biological Control of Phytopathogenic Fungi by Kluyveromyces marxianus and Torulaspora delbrueckii Isolated from Iraqi Date Vinegar

Yeasts are distributed in all environments and have been reported as potential biocontrol agents against various phytopathogenic fungi. To investigate their enzymatic and biological activities, 32 yeasts were isolated from 15 date vinegar samples. Evaluation of the antagonistic activities of isolated yeasts against the plant pathogens Fusarium oxysporium, Sclerotinia sclerotiorum, and Macrophomina phaseolina indicated that there are two yeasts had the highest inhibitory effect against plant pathogens, these yeasts identified as Kluyveromyces marxianus and Torulaspora delbrueckii using traditional and molecular methods. These yeast isolates were tested for fungal cell wall degrading enzymes (in vitro), and results indicated that the yeasts had strong protease and amylase enzyme activity and moderate chitinase and cellulase enzyme activity. The antagonistic activities of each yeast were evaluated using a dual culture technique. The results showed that K. marxianus inhibited the mycelial growth of F. oxysporium, S. sclerotiorum, and M. phaseolina by 70.5, 57.5, and 75.5%, respectively, whereas T. delbrueckii inhibited mycelial growth of F. oxysporum, S. sclerotiorum, and M. phaseolina by 55.3%, 66.2%, and 31.11%, respectively. The biofilm production assay indicated that the tested yeast could form biofilms as a mechanism of antagonistic activity against phytopathogenic fungi.

Chemical composition and in vitro antiobesity and in vivo anti-hyperlipidemic effects of Ceratotheca sesamoides, Jatropha tanjorensis, Mucuna flagellipes, Pterocarpus mildbraedii and Piper guineense

Ethnopharmacological relevanceCeratotheca sesamoides, Jatropha tanjorensis, Mucuna flagellipes, Pterocarpus mildbraedii and Piper guineense are all medicinal plants found in West Africa and used traditionally for the treatment of various ailments such as; hypertension, arteriosclerosis, anemia, malaria and flatulence. Aim of the studyThe purpose of this study is to ascertain the in vitro anti-obesity and in vivo anti-hyperlipidemic activity of extracts and fractions of selected plants. Materials and methodsThe ethanolic extract of the aforementioned five plants were tested in vitro for their activity against porcine pancreatic lipase and alpha-amylase at a concentration of 1mg/ml. Thereafter, the IC50 values of the plants active against alpha-amylase were determined. The ethanolic extract of P. mildbraedii and M. flagellipes were fractionated using n-hexane, n-butanol, ethyl acetate and water, thus the four fractions each obtained were used for further in vitro and in vivo studies. Thirty-five male rats were divided into seven groups out of which groups 2–7 were made hyperlipidemic using poloxamer 407 (1.0 g/kg body weight) intraperitoneally. Groups 1, 2 and 3 were the normal, hyperlipidemic and standard drug control groups respectively, whereas groups 4–7 were the test groups. ResultsC. sesamoides, M. flagellipes and P. mildbraedii inhibited alpha-amylase activity, while M. flagellipes and P. mildbraedii had activity against pancreatic lipase. The n-hexane fraction of P. mildbraedii and M. flagellipes inhibited both lipase and amylase enzyme activities among the four fractions of each plant tested. The GCMS analysis of n-hexane fractions revealed the presence of two major compounds in each plant which were 9-octadecenoic acid and hexadecanoic acid in P. mildbraedii, and hexadecanoic acid and 9,12-octadecadienoic acid in M. flagellipes. The in vivo studies revealed that the n-hexane fraction of P. mildbraedii and M. flagellipes significantly decreased (p < 0.05) total triglycerides, total cholesterol, and low density lipoprotein cholesterols when compared to the hyperlipidemic control rats. ConclusionsP. mildbraedii and M. flagellipes have the potency to act as antiobesity and anti-hyperlipidemic agents.

Effect of fermented rice bran concentration using Lactobacillus sp in artificial feed on growth performance and enzyme activity of tilapia Oreochromis niloticus (Linnaeus, 1758)

This study aims to determine the effect of the dose of fermented rice bran using Lactobacillus sp. on growth performance and enzyme activity in tilapia. This research was conducted in Sidenreng Rappang Regency. The test animal used was tilapia larvae measuring 1.17 g. The maintenance container is a tarp pool with a size of 1 m x 1 m x 1 m filled with fresh water as much as 85 L. The study was designed using a completely randomized design consisting of four treatments and three replications, thus there were 12 experimental units. The treatment was determined based on the dose of fermented rice bran using Lactobacillus sp. which differ, namely 0, 10, 15 and 20%. The test feed was given to tilapia larvae, which were stocked with 20 fish per container for 60 days of rearing. The specific growth rate data of tilapia were analysed using analysis of variance (ANOVA) and enzyme activity was analysed descriptively based on the viability of tilapia. The results showed that tilapia fed with 20% fermented rice bran had a significant effect on the specific growth rate, protease and amylase enzyme activity. The optimal dosage of fermented rice bran using Lactobacillus sp. is 15-20%.

Literature study of production dry cellulase from Trichoderma reesei, Aspergillus niger, and Bacillus subtilis

Cellulase is an enzyme that can degrade cellulose through a catalytic process. The enzyme that works synergistically to release sugar (glucose). This review aims to determine cellulase enzymes’ activation from three different microorganisms, Aspergillus niger, Bacillus subtilis, and Trichoderma reesei. Enzymes’ activation using various kinds of lignocellulosic substrates and the effect of adding carrier agents on the freeze-drying process. The cellulase enzyme from Trichoderma reesei has an activity of 3.4 IU / mL. This enzyme is produced by pre-treatment 1% H2O2 in lignocellulosic media. Aspergillus niger has a cellulase enzyme activity of 0.229 IU / mL using a sugarcane bagasse substrate that has been pre-treated by shiitake mushrooms. The cellulase enzyme from Bacillus subtilis has an activity of 0.907 IU / mL, with the addition of CMC levels of 5% as an inducer. Cellulase enzymes in liquid form are susceptible to denaturation during storage. The production of cellulase enzymes in solid form is expected to maintain the stability of the resulting cellulase enzyme activity longer. The addition of non-reducing sugar as a carrier agent in the freeze-drying process is reported to preserve the biological activity. The addition of sucrose and trehalose, each with a concentration of 300 mM, maintained the amylase enzyme activity up to 90%. The addition of sucrose with an optimum concentration of 60 mM protects both total protein stability and good lysozyme activity. It is suspected that a carrier agent with an optimal concentration can maintain protein stability in cellulase enzyme activity in this review.

Effect of Rhamnolipids on Pathogenicity Characteristics of Microorganisms in Organic Compost

Background: Rhamnolipids are hydrophilic glycolipids, often classified as biosurfactants. They are produced by different bacterial species. Rhamnolipids are extensively studied in biological research because of their interesting features like antimicrobial, antifungal, and antiviral activities. Materials and Methods: To study the antimicrobial effect of rhamnolipid, we conducted a crosssectional study on the eight different pathogenic bacterial strains from November 2019 to June 2020. These bacterial strains were isolated from the organic compost. Both disk diffusion and broth microdilution methods were used to evaluate the inhibitory effects of rhamnolipids on these pathogenic bacteria. Also, protease and amylase enzyme activities were evaluated in these eight bacterial isolates. Results: For Bacillus stearothermophilus, within the area of 31.5 mm, no growth was observed, hence proving the inhibitory effect of rhamnolipid. After calculating the minimum inhibitory (MIC) and minimum lethal concentrations (MLC) for each bacterial strain, it has been found that the studied bacteria were more susceptible to rhamnolipids than most of the antibiotics. Strains were also quantified for their enzymatic activity of proteases and amylases. The bacterial strains of Bacillus stearothermophilus, Brevibacillus brevis, Bacillus licheniformis, and Bordetella petrii showed maximum protease activity. Whereas Escherichia coli, Enterococcus faecalis, Enterobacter aerogenes, and Pseudomonas aeruginosa showed high amylase activity. Conclusion: Rhamnolipids can be used as a potential antimicrobial agent for treatment of infections.

Metagenomic-based Approach for the Analysis of Yeast Diversity Associated with Amylase Production in Lai (Durio kutejensis)

This study reported the application of a next generation sequencing (NGS) analysis of yeast diversity in native Indonesian fruit, Durio kutejensis, collected from Borneo, Central Kalimantan. The analysis was designed to observe the microbial consortium associated with solid state fermentation (SSF) for amylase production. Together with the additional data from culture-dependent analysis, we observed the morphological features, molecular characteristics, and amylase concentration produced by each isolate. We performed Solid State Fermentation (SSF) for amylase production and the enzyme activity was then determined using UV-Vis spectrophotometer at 540 nm. Result obtained from metagenomic approach consist of 4 group that fungal species included in the Ascomycota identified as Botryosphaeria dothidea (1.35%), Lasiodiplodia crassispora (17.62%), Aureobasidium pullulans (55.02%), Paraphoma chrysanthemicola (11.38%), Preussia funiculate (1.90%), Sporormiella intermedia (0.82%), Myrothecium gramineum (1.35%), Fusarium oxysporum (6.24%), Fusarium proliferatum (3.25%) and Phialemoniopsis curvata (1.08%). The results of isolation using culturable medium in the form of YMA obtained 40 yeast isolates. A total of 40 representative isolates from durian fruit were screened, two positive amylase isolates based on clear zones formed were DU 4.2 (Candida sorboxylosa) and DU4.22 (Cyberlindnera fabianii) isolates with amylolytic index of DU 4.2 isolates at 0.24 and DU 4.22 at 0.72 with an incubation time of 48 h. The highest amylase enzyme activity was found in isolate DU 4.2 of 31.21 U / mL.

Isolation, molecular identification and enzyme activity of amylase producing thermophilic bacteria from hot springs

In this era, enzyme exploration in Indonesia continues to increase, and the amylase enzyme of about 30% of the total production is used worldwide. The great needs for amylase enzyme and its uses in several industrial and health fields lead to the search for potential natural resources in its production, including hot springs, which are very promising because of their thermophilic and thermostable properties. The aim of this study was to isolate, determine the activity and molecular identification of amylase-producing thermophilic bacteria from natural hot springs Dolok Tinggi Raja, Simalungun, North Sumatra. This study includes measurement of enzyme activity using the DNS method and molecular identification. The results were obtained from 20 isolated isolates, 6 isolates had amylase activity with the highest activity, namely the UTMTR VAR A10 isolate with an amylase enzyme activity of 0.2733 Unit/mL. Based on the molecular identification using 16SrRNA gene amplification based on the BLAST program and phylogenetic analysis with the MEGA X program, it was shown that the UTMTR VAR A10 isolate is a bacterium Bacillus licheniformis strain UTMTR VAR A10. Further research is needed to purify the enzyme in such a way that it can be applied to the industrial world.

The potential of amylase enzyme activity against bacteria isolated from several lakes in East Java, Indonesia

Abstract. Nisa IK, Prabaningtyas S, Lukiati B, Saptawati RT, Rodiansyah A. 2021. The potential of amylase enzyme activity against bacteria isolated from several lakes in East Java, Indonesia. Biodiversitas 21: 42-49. Indonesia is one country that has water resources having an abundance of microbial diversity, but not explored massively. This study aims to measure the amylase activity quantitatively from 53 amylolytic bacterial isolates from Ranu Pani, Ranu Regulo, Ranu Grati, and Ngebel Lake; also it identifies the isolate with the highest amylase enzyme activity. The amylase enzyme activity test calculates with DNS (Dinitrosalycylic acid) method, molecular identification of the highest bacterial isolate is based on the 16S rRNA gene. Its relationship is determined through the phylogenetic tree with the Neighbor-Joining (NJ) method. The results showed that the fifty-three bacterial isolates have amylase activity about 0.000-0.016 units/mL. The KN bacterial isolate from Ranu Ngebel was the highest amylase activity, producing enzyme around 0.016 units/mL, while isolate G20 from Ranu Grati was the lowest, reaching about 0.0001 Unit/mL. Based on the morphological and molecular identification, the KN bacterial isolate is classified as the Bacillus cereus group with 99.4-100% sequence similarity, closely related to Bacillus paramycoides (NR 157734.1).

Determination of the optimal priming interval of rumen fluids used as inocula for the in vitro digestibility trials through radial enzyme diffusion method

Context Determination of the neutral detergent fibre digestibility is one of the important parameters to consider when formulating diets. However, the in vitro determination shows low repeatability because of the source of rumen-fluid inoculum. Priming of the rumen fluid inocula, obtained through an oesophageal probe, has been proposed to overcome this issue. Aim The objective of the study was to investigate the evolution of the microbial enzymatic activities of different rumen fluids during a priming procedure, to establish the fermentation interval that minimises the differences among rumen-fluid degradative potentials. Methods Three farms for each type of diet were involved in the study. Rumen fluids were obtained from dry and lactating cows fed the following four diet types: 100% hay or a diet with 80:20 forage:concentrate ratio (F:C) as dry-cow diets, and ad libitum hay and concentrate, or a total mixed ration (both at 60:40 F:C) as lactating-cow diets. On each farm, rumen fluid was collected from three Holstein cows by using an oesophageal probe, and mixed. Two aliquots of each rumen fluid mix were added to the medium containing the same priming substrate in an in vitro batch-fermentation system. During the incubation, the fermentation fluids were sampled in duplicate at 0-, 1-, 2-, 4-, 8-, 24- and 48-h intervals. Enzymatic activities of amylase, cellulase and xylanase were determined by radial enzyme diffusion method. Key results Initial enzymatic activities were quite variable and increased with an increasing incubation time. By 24 h, amylase showed similar values among high-concentrate diet fermentation fluids, and a lower data dispersion in comparison to the other intervals; cellulase was characterised by similar values in all the fermentation fluids derived from diets including concentrates, and xylanase showed similar activity in the fermentation fluids derived from high-concentrate diets. Development of the enzymatic activity of the fermentation fluids derived from the 100% hay diet differed from the others. Conclusions A 24-h priming procedure was needed to stabilise and equalise the enzymatic activity of the rumen fluid from cows fed high-concentrate diets. This was not observed in rumen fluid from cows fed hay-based diets. Implications The 24-h-primed rumen fluid can be used to increase the repeatability of neutral detergent fibre digestibility determination.