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This research aimed to characterize the enzymatic activity potency of 10 isolates of Streptomyces, to test their antifungal activity against C. gloeosporioides, a red chili plant pathogen, and to test the ability of selected isolate in inhibiting C. gloeosporioides colonization on red chili seed germination. The enzyme activities tested were amylase, cellulase, protease, and chitinase. The antifungal activity test of the isolate against C. gloeosporioides used the dual culture method. The ability of selected isolates to inhibit pathogen colonization on seed germination was tested in a growth chamber experiment. The results showed that 8 isolates indicated amylase activity, while 4 isolates indicated cellulase and protease activity and 2 isolates (LB and ST20) indicated positive chitinase activity. Identified Streptomyces (S. prunicolor ST20 and S. andamanensis LB) were able to inhibit the fungus growth through non-volatile metabolites (7.9 and 50.2%) which was higher than through volatile metabolites (12.6 and 19%). Inhibition test of the pathogen colonization on the seed germination at 2 days after seed germination, all of the seed germination medium surfaces of control treatment were colonized by the fungus, while those on the treatments of Streptomyces inoculation had not been colonized. However, at 6 days after seed germination, the treated medium surface was colonized by the fungus at a lower intensity than the control treatment, whereas around the roots of ST20 inoculation had lower fungus spores than those of LB inoculation. In the future, we need to examine the fungus control by Streptomyces spp on seedling development and plant growth. [ABSTRACT FROM AUTHOR]

Thraustochytrids is a family of microalga which lives in the marine ecosystem and mangrove habitats. Indonesia with a mangrove area of 3.49 million hectares, is a country with the largest mangrove habitats in the world. With such potency, the Indonesian mangrove has the largest Thraustochytrids habitat and it is hoped that the potency will attract Indonesian scientists to explore and develop a bio industry based on thraustochytrids. Thraustochytrids have potent biotechnology as a source of hydrolytic enzymes, carotenoids, squalene, biodiesel, and polyunsaturated fatty acids (PUFA), specifically docosahexaenoic acid (DHA). Among those biotechnology products, DHA is the most promising product to be commercialized. According to a number of studies, DHA is critical throughout all life stages from the need for fetal development, the prevention of preterm birth, and the prevention of cardiovascular disease to the improvements in the cognitive function and the eye health of adults and the elderly. A recent study showed that n-3 PUFA indirectly or directly modulated the gut microbiota which acts as a prebiotic. However, cost production for producing dry thraustochytrids is relatively expensive. One of the ways to reduce those costs in production is by using low-cost substrates. Recently, it is already possible to produce thraustochytrid on a commercial scale using low-cost substrate palm oil mill effluent (POME), so cost production of dry thraustochytrids became economically feasible. Indonesia, as the number one crude palm oil (CPO) producer of 44,5 million tons has the great opportunity to develop an industry base DHA. [ABSTRACT FROM AUTHOR]

The purpose of the research was to study the features of the influence of microbiological preparations based on associative nitrogen-fixing bacteria on the enzymatic activity of the rhizosphere and the elements of spring barley productivity. The research was carried out as part of a vegetation experiment. Soil samples were taken in three terms, which corresponded to the phases: tillering, budding and heading. In the experiment, microbiological preparations flavobacterin (Flavobacterium sp. strain 30) were considered; mizorin (Arthrobacter mysorens, strain 7), rhizoagrin. (Agrobacterium radiobacter strain 204). Enzymes belonging to the classes of oxidoreductases (catalase, peroxidase, and polyphenol oxidase) and hydrolases (urease, phosphatase, and invertase) have been studied. The enzymatic activity of the barley rhizosphere was evaluated against the background of the application of mineral fertilizers in combination with microbiological preparations and without them. During the experiment, it was noted that the activity of the studied enzymes depended on the phase of plant development and the type of microbial preparation. The most significant increases in ear productivity elements were found with the combined use of mineral fertilizers and microbiological preparations. But even without a mineral background, inoculation with microbiological preparations gives a significant increase compared to the control variant. [ABSTRACT FROM AUTHOR]

Sustainability is influenced by various aspects especially in the field of agriculture. Agricultural sustainability includes meeting the food resource demands but among the disturbing factors is pathogen infestation. More innovative technologies are needed to overcome with the sustainability of centralized agricultural systems developing technologies which not adversely impact the environment. Environmental sustainability problems with the usage of fertilisers, agricultural machinery, irrigation water, and pesticides are driving this trend. As a result, a new method is required that integrates biological and ecological processes into food production while reducing the usage of harmful agents. Agricultural sustainability indicates an increasing awareness of the benefits of ecological and agronomic natural management, modification, and design, as well as a focus on genotype enhancement using a number of current biological techniques. Among the pathogens in agriculture is Glomerella cingulate, a plant pathogenic fungus that cause anthracnose disease that infects a variety of plants. The fungus secretes cutinase, a degradative and hydrolytic enzyme that degrade cutin in order to traverse the cuticular barrier of the infected plant. In order to do genetic manipulation and study the regulatory for the production of this enzyme, protoplast is needed as a tool in genetics and molecular research and in transformation of DNA. In this study, a protocol for isolation of protoplast from G. cingulata was established including the effects of mycelial age, concentration of enzymes and duration of lytic digestion on the yield of protoplasts. The highest yield of protoplast (7.2x107 protoplasts/ml) was achieved from a 3-day mycelial culture which was treated with 10 mM dithiothreitol and incubated with 4 mg/ml lysing enzyme for 3 hours at 30˚C. The frequencies of regeneration of the isolated protoplast are at 0.4% to 0.6% on a complete medium compared to 0.03% on Potato Dextrose Yeast Extract Agar. The use of protoplast isolation is significant for a variety of genetic modification applications [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the effect of feed pellet containing three leaves mixtures as tannin sources (Acacia mangium Willd, Swietenia mahagoni, and Artocarpus heterophyllus) on rumen fermentation parameters in vitro, especially on ruminal enzymes activity and gas production kinetics. Pelleted leaves were applied to substitute the use of concentrate in the diet (grass:concentrate, 60:40) with three substitution levels equal to tannin concentration at 0%, 1%, and 2% based on dry matter (DM). All dietary treatments were used as a substrate for in vitro fermentation by Menke and Steingass gas production method at 48 hours of incubation time. Fermentation medium and gas production after incubation were used to determine rumen hydrolytic enzymes activity and gas production kinetics. Data were analysed by one-way analysis of variance (ANOVA) and continued by Duncan's Multiple Range Test (DMRT). The application of pelleted leaves mixture as a tannin source of up to 2% could reduce the activity of amylase, CMC-ase, and protease significantly (P<0.05). β-glucosidase enzyme activity was decreased significantly at 2% tannin. Gas production from soluble fractions (a) was decreased significantly (P<0.05) but gas production from insoluble fraction (b), the gas production rate (c), and the amount of (a+b) fraction was not affected (P>0.05). The substitution of concentrate with pelleted leaves as tannin source could decrease in vitro rumen hydrolytic enzymes activity and the kinetics of gas production in terms of insoluble fraction. [ABSTRACT FROM AUTHOR]

Endophytic communities provide benefits to their host plants such as protecting plants against herbivores, insects, or pathogens, as well as being able to stimulate plant growth. The association of endophytic bacteria with plants can affect plant productivity directly or indirectly. One of the contributions of endophytic bacteria is as a producer of extracellular enzymes. Mangrove areas are the main ecosystems that support important life in coastal and marine areas. Many benefits are obtained from mangrove plants. One of them is microorganisms associated with mangrove plants. Enzymes from microorganisms are more beneficial and their production is faster than enzymes derived from plants and animals. Endophytic bacteria associated with a mangrove species, Rhizophora apiculata, are a source of hydrolytic enzymes such as proteases and cellulases which are produced for their ecological contribution and also to support the growth of their host plants. The aims of this research were isolate endophytic bacteria from the R. apiculata and screening qualitatively the proteolytic and cellulolytic activities of the endophytic bacterial isolates. The isolates were screened for protease activity on skimmed milk agar (1%) and cellulase activity on Carboxymethyl Cellulose (CMC) agar. In this study, 18 isolates of endophytic bacteria were isolated from the leaves of the mangrove plant R. apiculata and 9 isolates had proteolytic activity and 13 isolates had cellulolytic activity or could produce cellulase. Endophytic bacteria R. apiculata which has the ability to produce proteases and cellulases qualitatively can be explored further to be developed as a source of hydrolytic enzymes from endophytic bacteria. This works contributes to our knowledge of the endophytic bacterial as source of enzymes present in mangrove plants. [ABSTRACT FROM AUTHOR]

Potential sources of thermostable enzymes are thermophilic bacteria. In the research, nucleotide base sequences of 16S rRNA gene from thermophilic bacteria have been identified. The bacteria showed activity of hydrolytic enzyme exo-type on inulin substrate. The bacteria were isolated from 'Batu Bajanjang' hot spring, Solok (West Sumatra) that labeled as BB3 isolate. Gene of 16S rRNA fragment was isolated using PCR technique with BactF1 and UniB1 primers. The nucleotide base sequences of 16S rRNA gene were determined using the dideoxy-Sanger method. Analysis of the gene fragment (1348 bp) from BB3 isolate indicated as Fictibacillus gelatini or Bacillus gelatini species. Activity of extracellular enzyme indicated exo-type action based on analysis of the inulin hydrolysis end products using Thin Layer Chromatography (TLC). So, the enzyme is the potential to produce high fructose syrup from inulin. [ABSTRACT FROM AUTHOR]

Rapid global economy and population increase play a significant role in food waste (FW) production. Improper food waste management is a considerable challenge for the entire world since FW pollutes the environment and risks human and animal health. However, transforming food waste into value-added chemicals is an area of intensive research since they contain organic compounds. To this effect, we investigate the possibility of enhancing the food waste hydrolysis using microorganisms. In this research, FW was first blended and diluted at different concentration ratios (Fw: water) of 2:1,1:1,1:2 and 1:3. After a simple solid-liquid separation, the liquid was hydrolysed by fungi rich in hydrolytic enzymes at their log phases. The moisture content of FW was 15.68 %. This study aims to produce glucose from food waste using the three types of hydrolytic microorganisms: Aspergillus oryzae, aspergillus aculeatus and Candida rugosa. Moreover, we discussed the best ratio variation of microbes towards food waste hydrolysis. This study analysed glucose levels using the DNS method by measuring the absorbance through a spectrophotometer at 540 nm. The results showed that the highest glucose concentration of 16.5 g/l was obtained after 48 hours by mixing all microorganisms using a ratio of 2:1 (FW 2:1 M). The highest glucose (g/l) concentration from a ratio of 2:1 obtained from FW hydrolysis by Candida rugosa, Aspergillus oryzae and Aspergillus aculeatus after 48 hours were 13.05,14.31,15.9, respectively. Therefore, the current finding indicates that hydrolytic microorganisms could save the pretreatment cost of food waste and greatly enhance food waste degradation. [ABSTRACT FROM AUTHOR]

Xylanases are one of the industrially important hydrolases with potent applications. The development of an economically feasible process for the production of low-cost xylanase is a challenging task. Computer-aided simulation of such process resolve bottlenecks thereby predicting the economic feasibility. In this present study, a model process for the production of xylanase from a mesophilic bacterium Bacillus subtilis have been designed using the Intelligen Inc., SuperPro Designer v9.5. The model consists both upstream and downstream processes for the production of xylanase. Four different scaleup strategies were employed to identify the most economically feasible combination that could be exercised in the production process. Based on the model constructed, the case involving PBA chromatography along with high-pressure homogenizer was found to yield a higher percentage of xylanase (99%) and found to be the most economically feasible process among other cases. [ABSTRACT FROM AUTHOR]

Studies of the effect of microflora on soil fertility and crop yields have formed the direction of ecologized agriculture. It is based on the creation of drugs based on microorganisms of different with different life strategies. Biologically active preparations with the direction of action to improve the level of soil fertility, as a result of the work of effective microorganisms, are designated as EM preparations. Action of biologically active preparation «Baikal EM-1» on indices of enzymatic activity of soils and level of accumulation of microbial toxicosis is studied. The general goal of the work is to identify changes that occur in the soil under the influence of a complex of bacteria with different life strategies that are part of the drug. Experiments were carried out in greenhouse conditions when growing early-ripe pepper varieties adapted to the conditions of Siberia. The level of microbial toxicosis of soils of different compositions was assessed using the phytotesting method. Control of soil fertility level is performed by analysis of activity of hydrolytic enzymes of invertase and phosphatase. The possibility of using the preparation Baikal EM 1 to improve biochemical indices of artificial soils is described. [ABSTRACT FROM AUTHOR]

The aim of the study is to evaluate the enzymatic activities of Candida albicans and Candida parapasilosis isolated from bovine mastitis. Two hundred and fifty milk samples were isolated from cows with mastitis in Basrah province were collected and examined using rapid diagnostic tests. Candida Spp were identified as proportion of 116/250 (46.4%). Based on conventional method and ID - Yst card system Vitek 2, C. albicans was the predominant 60/116 (51.7%), followed by C.parapasilosis as 15/116 (12.9%). Some randomly chosen isolates of C. albicans and C. parapasilosis were used for detection of their enzymatic activities by API ZYM test. In case of C. albicans (n=30) the isolates produced n-acetylo-B-glucosyloaminidase strongly when the percentage was 30/30 (100%), esterase and valine arylamidase (C4) 29/30 (96.6%), leucine arylamidase 28/30 (93.3%), esterase lipase (C8) and naphthol-AS-BI- phosphohydrolase 25/30 (83.3%), α-glucosidase 23(76.6%), β-glucosidase 22 (73.3%), α-mannosidase 4 (13.3%), with a lowest activity of cystine arylamidase 1(3.3%). Regarding C. parapasilosis (n=10) the isolates showed a highest activity of leucine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase and α-glucosidase which was 10/10 (100%) for each, followed by esterase lipase (C8) 9/10 (90%), valine arylamidase and alkaline phosphatase 8/10 (80%) for both and esterase (C4) 7/10 (70%). [ABSTRACT FROM AUTHOR]

Indonesia, one of the Asian countries, has many kinds of fermented food which coverage area are from Sumatera to Papua. Fermented food product is processed food through fermentation process, where certain microorganisms play an important role during the process. Growth of undesirable wild microorganisms may cause fermentation failure and results in undesirable products. Many researches have been done to explore fermented foods from several areas such as dadih and tempoyak (Sumatera); ikan peda, petis, oncom, tape, tempe and kecap (Jawa); brem, urutan, bebontot, brengkes (Bali); sour horse milk (NTB); sei (NTT); and bekasang (Sulawesi). From these products, the usage of many kinds of microorganisms are also explored. Most of them are of a group known as lactic acid bacteria (LAB), which has been used as starter culture, as well as probiotic. By using LAB as starter culture, some benefit such as acceleration in the process and ensure quality and safety of the products, were shown. During fermentation, LAB produce lactic acid and other metabolites that respectively lower the pH and limit the growth of pathogenic microorganisms. The bacteria also release hydrolytic enzymes (lipases and proteases), which are able to break down macromolecules, such as lipids and proteins, resulting in the production of precursors for specific aroma. The discussion in this paper is focused on the role of LAB on safety and quality of fermented food, especially urutan (Balinese fermented or dried sausage) and sour horse milk. [ABSTRACT FROM AUTHOR]

Lipase is an important class of industrial hydrolytic enzyme that is produced mainly by bacteria. The goal of this study was to screen and determine the optimum conditions for lipase production by a thermo-halophilic bacterium, named Pria Laot Sabang 80 (PLS 80), isolated from an underwater hot spring. Screening of lipase was conducted on agar media supplemented with olive oil and rodhamine B. PLS 80 exhibited lipase activity based on the formation of orange red fluorescence area around the colonies observed under UV light. The growth curve and lipase activity of the isolate was studied at various temperature, incubation time, pH and salt concentrations. The growth curve was measured by dry cell biomass and lipase activity using spectrophotometry method by p-nitrophenyl palmitate as the substrate. The isolate showed the highest biomass production after 16 hours of fermentation at 70 °C, pH 9 and with an addition of 3 M NaCl. The highest activity of lipase was 24.3 U/mL. This study shows that PLS 80 produced a thermo-halostable alkaline lipase. [ABSTRACT FROM AUTHOR]

Corncob is solid waste generated from the corn processing industry. Corncobs are composed of cellulose, hemicellulose, and lignin. Hemicellulose in corncobs is the second largest component after cellulose. The high content of hemicellulose, especially xylan in corn led to their potential use as raw material for xylose. Xylose can be used as substrate for xylitol production. Corncob xylose can be obtained through the chemical or enzymatical hydrolysis of xylan. Enzymatic hydrolysis of xylan produces more specific product without by-products such as furfural. Before starting the enzymatic hydrolysis, pre-treatment is necessary to degrade lignin in the cell wall so that the the access of enzyme to the materials are increased. This research aims to determine the most appropriate type of pre-treatment to maximize xylose production resulting from the enzymatic hydrolysis process using xylanase. The descriptive method use in this research and followed by t-Test with the level confidenceof 95%. The experimental variations are heat treatment in autoclave, ozonolysis, soaking in a ammonia solution, ozonolysis followed by heating, and soaking the material in ammonia followed by heattreatment. The results showed that pre-treatment is effective to increasethe accesibility of enzyme. Heat treatment as well as soaking in ammonia followed by heat treatment were the most efficient proccesses to produce xylose with the condition of enzymatic hydrolysis of 52 h, at 50oC. [ABSTRACT FROM AUTHOR]

A process for maltodextrin production from avocado seed starch was reported in this study. Response surface methodology was used to investigate the effects of three independent variables for hydrolysis of the starch using a commercial food-grade α-amylase, Termamyl SC. These variables included enzyme concentration (0.05 – 0.15% starch), pH (5.0 – 6.0) and hydrolysis time (1.0 – 3.0 h), while the temperature fixed at 95°C. The result showed that the optimum conditions were using enzyme concentration at 0.12%, pH at 5.5 and 2.75 h of the incubation time. Under the optimum conditions, the recovered starch yield was 79.8% and the maltodextrin powder had 15.8 of dextrose equivalent. [ABSTRACT FROM AUTHOR]

Industry is constantly forced to look for new ways to make production processes environmentally safe and products more competitive. In this regard, microbiological enzyme preparations increasingly replace conventional chemical catalysts in a number of industrial processes. Such preparations, in addition to environmental friendliness and high activity, have a number of advantages over enzyme preparations of vegetable and animal origin. With the aim of creating new recombinant lipase, we analyzed data on the activity, specificity and structure of this enzyme, as well as on its coding genes, and searched for suitable producer in microorganism collections. The target gene of lipase from microorganisms was isolated and cloned by integration into the plasmid vectors of the PET system (Novagen). Expression of cloned gene was carried out in E. coli cells (Bl-21, C-41). The optimal time for induction of biosynthesis of recombinant enzyme was determined, high levels of target enzyme in cell culture was achieved - up to 40% of the total protein volume. Isolation and purification of recombinant enzyme was performed by Ni-affinity chromatography, using Ni-NTA agarose (Qiagen, USA). The activity of the obtained enzymes was determined by standard biochemical methods. The activity indices of the obtained recombinant lipase are 12 times higher than the activity of the native enzyme. A method for purifying wastewater using the obtained recombinant lipase was developed by the authors. The maximum yield of fatty acids and their salts as a result of hydrolysis by lipase is observed when the enzyme is introduced in an amount of 10 U/g of fat after 4 days of hydrolysis of waste water from the fat and oil enterprise at a temperature of 40±2 °C in a stationary mode. After 24 hours of hydrolysis, the concentration of fat in the wastewater is reduced to the maximum permissible concentration. [ABSTRACT FROM AUTHOR]

The pancreas of cattle is one of the most important by-products produced in the meat processing industry. Meat processing plants of medium capacity per year produce at least 40 tons of pancreas of cattle. The pancreas is a valuable raw material for the biotechnology, because it has high contents of hydrolytic enzymes. Among them, the most important enzymes are ribonuclease, proteases, amylase and lipase. The most perspective is the implementation of low-waste technology, involving the isolation of the maximum number of enzymes in a single technological process. The purpose of this work is to select the conditions for the isolation of pancreatic lipase from the pancreas, which is widely used in the production of detergent powders, the food industry, and the production of biodiesel. During the course of the investigations, a sequence of pancreas treatment was established. According to this sequence, during the first stage, amylase is isolated by treating the pancrease with aqueous-salt solutions. In the second stage, lipase is isolated by treating the solid pancreas residue with organic solvents. The conditions for isolating lipase from the pancreas, namely the type of organic extractant, the temperature of the process, the concentrating of the lipase solution by ultrafiltration, and precipitation are selected. As a result of the investigations, a sample of lipase with an enzymatic activity of at least 2000 U / g was obtained, which was successfully tested for the hydrolysis of fat-containing wastes of the meat processing industry. [ABSTRACT FROM AUTHOR]

This study aimed to characterize the probiotic properties of lactic acid bacteria from the naturally fermented milk of Indonesia, namely dangke and dadih. Fifty-one representative lactic acid bacteria belonging to the species Lactobacillus Plantarum, Lactococcus lactis subsp. lactis and Enterococcus faecium were evaluated in vitro for potential probiotic properties based on their bile salt resistance, low pH tolerance, antimicrobial activity, antibiotic susceptibility and adherence to Caco-2 colon cancer cells. In addition, bacteriocin related gene (plantaricin A), bile salt hydrolase (bsh) and mannose-specific adhesin (msa) genes in the genome of lactobacilli were also examined. None of the dangke isolates, which belonged to the species L. lactis subsp. lactis tolerated low pH. However, eight of the isolates were able to grow in the presence of bile salts. It was observed that L. plantarum strain S1.30 and SL2.7 from dadih tolerated low pH, survived bile salt concentrations and were resistant to vancomycin. Furthermore, these strains also contained bacteriocin regulating gene (plantaricin A) and msa and bsh genes in their genome. However, only the strain S1.30 exhibited optimal antimicrobial activity against the selected pathogens and was able to adhere to Caco-2 cells by up to 82.24±0.14%. Antagonistic activity of L. lactis subsp. lactis from dadih and dangke was not detected. However, 73.94±1.26% adherence to Caco-2 cells was demonstrated by L. lactis subsp. lactis strain SL3.34 sourced from dangke. These results suggest that Lactobacillus plantarum strain S1.30 associated with dadih fulfilled the in vitro probiotic criteria and could be exploited for further in vivo evaluation. In addition, dadih was an effective probiotic carrier compared to dangke. [ABSTRACT FROM AUTHOR]

It is known that the use of lignocellulosic hydrolysates reduces the production cost of biofuel such as biobutanol and bioethanol. But for the most successful application of the hydrolysates for the biofuel production, it is necessary to apply an inexpensive and effective detoxification method and to use of cost-effective growth factors. In the present study, we evaluated the use of an acid hydrolysate of spruce and an enzymatic hydrolysate of miscanthus cellulose for the biobutanol production. To remove inhibitors from the hydrolysates, we applied the traditional physicochemical method with overliming and the biodetoxification method based on the use of the specially adapted activated sludge. Calcium hydroxide (150 g/L) was used for the neutralization. The biological method of detoxification of lignocellulosic hydrolysates was carried out under non-sterile conditions at room temperature by the specially adapted activated sludge of the urban wastewater treatment plants. The acetone-butanol fermentation was carried out by a strain of bacteria Clostridium acetobutylicum ATCC 824. The treatment by overliming removed 84-85 % and 83-86% of 5-hydroxymethylfurfural (5-HMF) and furfural from the hydrolysates respectively. Using the method of biodetoxification the content of furfural decreased by 98% and concentration of 5-HMF -- by 97-99%. In the present study as an inexpensive source of growth substances for the fermentation of the hydrolysates it has been suggested to use decantate of the brewer's spent grain. The obtained results showed that the brewer's spent grain can be used in the biofuel production as efficiently as the synthetic growth substances. [ABSTRACT FROM AUTHOR]

This study focused on the comparison among the usage of immobilized porcine pancreas lipase (PPL) on different hydrotalcite carriers (uncalcined and calcined hydrotalcite - like compound Mg/Al) and free lipase as the catalysts to hydrolyze of roselle (Hibiscus sabdariffa L.) seed oil. The reaction conditions were investigated including the ratio of oil to buffer, ratio of enzyme to substrate, the temperature of the hydrolysis, pH. The calcined hydrotalcite showed a higher lipase immobilization yield and a better reusability than the uncalcined hydrotalcite (87.15% and 86.78%, respectively). [ABSTRACT FROM AUTHOR]

In this study, the antioxidant capacity of peptide fractions isolated from the Tra Catfish (Pangasius hypophthalmus) by-product-derived proteolysate using ultrafiltration centrifugal devices with 5 distinct molecular-weight cutoffs (MWCOs) of 1 kDa, 3 kDa, 5 kDa, 10 kDa, and 30 kDa was investigated. Firstly, the chemical composition of the Tra Catfish by-products was analyzed. The result showed that the Tra Catfish by-products contained 58.5% moisture, 33.9% crude protein, 50.1% crude lipid and 15.8% ash (on dry weight basis). Secondly, the effects of hydrolysis time, enzyme content on the antioxidant potential of the proteolysate were studied using DPPH• (2,2-diphenyl-1-picrylhydrazyl) radical scavenging method (DPPH• SM) and FRAP (Ferric Reducing Antioxidant Potential) method. Alcalase® 2.4 L FG was used for hydrolysis. The result of antioxidant activity of the hydrolysate showed that the 50% DPPH• inhibition concentration (IC50) of the hydrolysate reached about 6775 µg/mL which was 1645-fold higher than that of vitamin C and 17-fold higher than that of BHT (ButylatedHydroxytoluene) with the degree of hydrolysis (DH) of the hydrolysate of 14.6% when hydrolysis time was 5 hours, enzyme/substrate (E/S) ratio was 30 U/g protein, hydrolysis temperature was 55°C, and pH was 7.5. The antioxidant potential of hydrolysate using FRAP method reached about 52.12 µM Trolox equivalent which was 53-fold and 18-fold lower than those of vitamin C and BHT, respectively, when the hydrolysis time was 5 h, enzyme/substrate ratio was 30 U/g protein, temperature was 50°C, and pH level was 8. Next, the proteolysate was further fractionated using MWCOs of 1 kDa, 3 kDa, 5 kDa, 10 kDa, and 30 kDa and the peptide fractions were investigated for their antioxidant activity. The result showed that the <1 kDa fraction showed strongest antioxidant activity with the IC50 of 1313.31 ± 50.65 µg/mL and FRAP value of 906.90 ± 44.32 µM Trolox equivalent. The second strongest fraction was 1-3 kDa with the IC50 and FRAP value of 2897.85 ± 128.38 µg/mL and 517.7 ± 21.08 µM Trolox equivalent, respectively. The fractions of 3-5 kDa, 5-10 kDa were the third and fourth strongest, respectively. The 10-30 kDa fraction showed weakest antioxidant capacity with the IC50 of 8012.88 ± 280.68 µg/mL and FRAP value of 92.92 ± 0.61 µM Trolox equivalent. The antioxidant peptides derived from Tra Catfish by-products proteolysate showed the potential to be used as natural antioxidant components in pharmaceutical and food industry. [ABSTRACT FROM AUTHOR]

Glycosaminoglycans (GAGs) are natural biocompounds which join to construct cartilage tissuses, it can be extracted from cartilage of sharks, pigs, cows, chickens, etc. GAGs contain a Chondroitin sulfate (CS) content which is a supplement of functional food used for preventing and supporting treatment of arthritis and eye diseases. Therefore, the GAGs extraction from byproducts of the industry of cattle and poultry slaughter to identify the CS content by papain enzyme is necessary. In this study, the optimal hydrolysis conditions were obtained by response surface methodology (RSM). The independent variables were coded as: pH (x1), enzyme concentration (x2), incubation temperature (x3) and hydrolysis time (x4). The results of the analysis of variance (ANOVA) shown that the variables actively affected GAGs content. The optimal conditions of hydrolysis were derived at pH of 7.1, ratio of enzyme per substances of 0.62% w/wpo, temperature of 65oC and hydrolysis time of 230 minutes, GAGs content reached 14.3% of the dry matter of raw material. Analyzes by HPLC revealed that 56.17% of the dry preparations of GAGs were CS compound, were equivalent to 8.11% of the dry matter of chicken keel cartilage. Molecular weight of the dry preparations GAGs was 259.6 kDa. The dry preparations included the contents of moisture 12.2%, protein 8.42%, lipid 0%, ash 10.03% and extracted GAGs 69.35%. [ABSTRACT FROM AUTHOR]

This work is aimed at studying of the kinetics of the cellulose hydrolytic hydrogenation process. In the course of the study, the methodology of the process in a high-pressure reactor under subcritical water conditions in the presence of the optimal synthesised catalyst, 3% Ru based on hypercrosslinked polystyrene MN 270 (Purolite Ltd, UK) (3% Ru/MN 270) was developed. The optimal parameters of the cellulose hydrolytic hydrogenation process were selected: the temperature - 205 °C, the hydrogen partial pressure - 6 MPa, reaction time - 60 min, Ru/cellulose ratio (mmol Ru in the composite per 1 g of cellulose) - 0.042/1, ruthenium content in the composite - 3% (by weight). Under these conditions, cellulose conversion was 64.0%, sorbitol selectivity was 43.5%. The mechanism of the cellulose conversion in subcritical water in the presence of 3%Ru/MN 270 catalyst was proposed. Besides, an adequate kinetic model of the cellulose conversion process was selected basing on the proposed mechanism. The kinetic parameters for the selected model were calculated: activation energy = 45 kJ/mol, the pre-exponential factor K0 = 8.92· 106, the reaction rate constant K = 0.102713 (mol/mol)·s-1. [ABSTRACT FROM AUTHOR]

Paddy straw is one of the most abundant lignocellulose wastes and has potential as a feedstock for sugar production. In this study, dilute acid pretreatment and enzymatic hydrolysis are the process that selected for the production of sugar. The paddy straw was pretreated with 1% (v/v) of sulfuric acid at 95 °C for 60 minutes followed by enzymatic hydrolysis. Optimization of enzymatic hydrolysis is desirable to achieve high yield of sugar from solid residues with high cellulose content. The optimization has been carried out by using Central composite design (CCD) to analyze the effect of pH, temperature and enzyme dosing for the enzymatic hydrolysis. For both the process, the concentration of sugar was analyzed by using Dinitrosalicylic acid (DNS) reagent with the aid of the standard glucose curve. The results showed the highest sugar yield from dilute acid pre-treatment was 1.14 g/L. For the optimum condition for enzymatic hydrolysis was at pH 5, 50 °C and 0.10 mL of enzyme and this produced 4.55 g/L of fermentable sugar. [ABSTRACT FROM AUTHOR]

In recent years, the input of antibiotics into soils has sharply increased. The environmental behavior of antibiotics is not well known and the precise environmental risk assessment is not practical. Long-term studies have shown the efficiency of the diagnostics and monitoring of soils with biochemical methods, particularly with the help of the indices of soil enzymatic activity. We studied the impact antibiotics (benzylpenicillin, tilosin and nystatin) at different concentrations (100, 500 mg/kg) on hydrolase (phosphatase and invertase) and oxidoreductase (catalase and dehydrogenase) activities in model experiments. The applied doses of antibiotics had definite suppressing effects on the soil enzymatic activity (20-70% of the control). Among the studied enzymes belonging to oxidoreductases and hydrolases, catalase and phosphatase had the highest and the lowest resistance to the antibiotics, respectively. The effect of antibiotics on enzymatic activity of soil lasted for a long time. The studied parameters were not completely recovered in 120 days. [ABSTRACT FROM AUTHOR]

In this work, the ability of commercial Trichoderma reesei cellulases preparation, Celluclast® or in combination with Accellerase®BG β-glucosidase to hydrolyse pretreated oil palm empty fruit bunch (OPEFB) was evaluated. Celluclast® alone hydrolyzed OPEFB to produce 2.41¯0.44 mg glucose per gram OPEFB. However, the production of glucose was significantly improved with supplementation of Accellerase®BG (8.12¯0.93 mg/g). This result suggested that the endoglucanases and exoglucanases in Celluclast® and β-glucosidase in Accellerase®BG able to work synergistically to increase the production of glucose from OPEFB. In addition, the production of xylose was also improved by 30% when the enzyme mixture was used. The result suggested that the mixture of Celluclast® with Accellerase®BG work synergistically to improve the production of sugars by removing the inhibition by cellobiose for complete cellulose hydrolysis. The production of glucose and xylose from OPEFB wastes showed the potential of this biomass as the source of renewable energy and fine chemicals production in Malaysia. [ABSTRACT FROM AUTHOR]

Edible bird's nest protein hydrolysates (EBN) were prepared via enzymatic hydrolysis to investigate its antioxidant activity. Two types of enzyme (alcalase and papain) were used in this study and EBN had been hydrolysed with different hydrolysis time (30, 60, 90 and 120 min). Antioxidant activities in EBN protein hydrolysate were measured using DPPH, ABTS+ and Reducing Power Assay. From this study, increased hydrolysis time from 30 min to 120 min contributed to higher DH, as shown by alcalase (40.59%) and papain (24.94%). For antioxidant assay, EBN hydrolysed with papain showed higher scavenging activity and reducing power ability compared to alcalase. The highest antioxidant activity for papain was at 120 min hydrolysis time with ABTS (54.245%), DPPH (49.78%) and Reducing Power (0.0680). Meanwhile for alcalase, the highest antioxidant activity was at 30 min hydrolysis time. Even though scavenging activity for EBN protein hydrolysates were high, the reducing power ability was quite low as compared to BHT and ascorbic Acid. This study showed that EBN protein hydrolysate with alcalase and papain treatments potentially exhibit high antioxidant activity which have not been reported before. [ABSTRACT FROM AUTHOR]

Cutinase is an important biocatalyst for various industrial applications. This enzyme which has dual functionality comparable to esterases and lipases, is efficient in the hydrolysis of soluble esters and emulsified triacylglycerols. Naturally-occurring enzymes usually have disadvantages when applied in non-natural catalysis due to Glomerella cingulata cutinase enzyme thermostability. It is postulated that by increasing the rigidity at certain amino acid positions showing high mobility based on the three-dimensional structure of G. cingulata cutinase, the improvement in thermostability will be achieved. The amino acid N82 of G. cingulata cutinase was selected based on its high B-factor value determined via the B-FITTER program. Megaprimer PCR was employed to introduce mutations at the chosen site by randomization using NNK degenerate primers. About 300 transformants were selected for screening of positive cutinase variants. The N82_V14 cutinase variant was observed to be more thermostable at an almost 2-fold increase when exposed at 50°C for 1 hr as compared to the wild-type enzyme. This study may provide valuable information regarding thermal stability of cutinases denaturation at high temperatures. [ABSTRACT FROM AUTHOR]

β-amylase (E.C 3.2.1.2) is an enzyme commonly found in plants and bacteria. The enzyme is an exo-acting carbohydrolase which hydrolyzes α-1.4-glucosidic linkages of starch, removing maltose units from the non-reducing end of the polysaccharide chain, producing β-maltose and β-limit dextrin as the final product. β-amylase is widely distributed in the higher plants such as sweet potato. Besides the use in starch hydrolysis, starch-converting enzymes are also used in a number of other industrial applications, such as laundry and porcelain detergents or as anti-stalling agents in baking. This enzyme was extracted from Dioscorea hispida Dennst in 0.05 M acetate buffer pH 4.8 and followed by ammonium sulfate fractionation at cold temperature (10°C). Ammonium sulfate fractionation was shared into fraction of 0-60%, 60-70%, 70-80% and 80-100%. The fraction containing high of specific activity (determined by Somogyi-Nelson and Lowry methods) was futher purified by dialysis. Fraction with high enzyme activity of β-amylase were fraction 60-70% and 70-80%, with specific activity of Dioscorea hispida Dennst were 1.32 and 1.55 mg sugar.mg protein-1.minute-1, whereas specific activity of crude extract enzyme was 0.21 mg sugar.mg protein-1.minute-1. After purified with dialysis, fraction with high enzyme activity of β-amylase were fraction of 60-70% and 70-80%, with specific activity of Dioscorea hispida Dennst was 2.72 and 4.24 mg sugar.mg protein-1.minute-1. The purified Dioscorea hispida Dennst β-amylase from dialysis showed increasing in spesific activity the crude enzyme as much as 24 folds. The characterization of enzyme showed that Dioscorea hispida Dennst derived enzyme had optimum pH of 5.5 and temperature of 70°C. The kinetic parameters of purified Dioscorea hispida Dennst β-amylase showed that the KMapp, Vmaxapp value and Hill constant were 0.0211 mg/ml, 9.63 mg sugar.minute-1 and 1.34, respectively. [ABSTRACT FROM AUTHOR]

Genomic data mining was carried out and revealed a total of seventeen ß-glucosidases in filamentous fungi Aspergillus niger. Two of them belonged to glycoside hydrolase family 1 (GH1) while the rest belonged to genes in family 3 (GH3). These proteins were then named according to the nomenclature as proposed by the International Union of Biochemistry (IUB), starting from the lowest pI and glycoside hydrolase family. Their properties were predicted using various bionformatic tools showing the presence of domains for signal peptide and active sites. Interestingly, one particular domain, PA14 (protective antigen) was present in four of the enzymes, predicted to be involved in carbohydrate binding. A phylogenetic tree grouped the two glycoside hydrolase families with GH1 and GH3 related organisms. This study showed that the various domains present in these ß-glucosidases are postulated to be crucial for the survival of this fungus, as supported by other analysis. [ABSTRACT FROM AUTHOR]

Edible bird nest (EBN) is a dried glutinous secretion from the salivary glands of several different swiftlet species. It is widely consumed as a health food due to its high beneficial effects to human health and has been considered to be one of the most precious food items by the Chinese for thousands of years. The aim of this study was to evaluate the effect of enzymatic hydrolysis on protein solubility (μg/g), the degree of hydrolysis (DH%), and peptide content (μg/g) of edible bird's nest hydrolysate. Initial protein solubility of boiled EBN was 25.5mg/g EBN. Treating the solubilized EBN with pancreatin 4NF for 1.0 - 1.5hours increased EBN protein solubility to 163.9mg/g and produced hydrolysate with DH% of 86.5% and 109.5mg/g peptide. EBN hydrolyzed with alcalase for 1.5 hours produced hydrolysate with protein solubility of 86.7mg/g, 82.7 DH% and had 104.1mg/g peptide content. [ABSTRACT FROM AUTHOR]

Fish protein hydrolysate (FPH) was prepared from freshwater catfish (Clarias batrachus) by using Alcalase® 2.4L and Papain. The effect of hydrolysis time (30, 60, 120, 180 min) with enzyme concentration of 1% (v/w substrate); pH = 8.0, 7.0 was studied to determine the degree of hydrolysis (DH), peptide content, proximate composition and amino acid profile. Results showed that the highest DH of Alcalase and Papain FPH were 58.79% and 53.48% after 180 min at 55°C incubation respectively. The peptide content of both FPH increased as hydrolysis time increases. FPH showed higher crude protein content and lower fat, moisture and ash content compared to raw catfish. The major amino acids of both hydrolysates were Glu, Lys and Asp. Content of essential amino acids of Alcalase and Papain hydrolysates were 44.05% and 43.31% respectively. [ABSTRACT FROM AUTHOR]

Two preparation methods of crude extracts of psychrophilic yeast Glaciozyma antarctica were compared in order to obtain a good recovery of intracellular proteins. Extraction with mechanical procedures using sonication was found to be more effective for obtaining good yield compare to alkaline treatment method. The procedure is simple, rapid, and produce better yield. A total of 52 proteins were identified by combining both extraction methods. Most of the proteins identified in this study involves in the metabolic process including glycolysis pathway, pentose phosphate pathway, pyruyate decarboxylation and also urea cyle. Several chaperons were identified including probable cpr1- cyclophilin (peptidylprolyl isomerase), macrolide-binding protein fkbp12 and heat shock proteins which were postulate to accelerate proper protein folding. Characteristic of the fundamental cellular processes inferred from the expressedproteome highlight the evolutionary and functional complexity existing in this domain of life. [ABSTRACT FROM AUTHOR]

Soil samples were collected from different farming fields of the Training Farm ( Aleksandras Stulginskis University): extensive (abandoned barley Hordeum vulgaris L. field, meadow and fodder galega (Galega orientalis Lam.) stand), conventional, and organic (winter wheat Triticum aestivum L. and and legumes mixtures) rotation crops in the middle of vegetation, June of 2007-2009. The objective of this study was to determine the soil bioactivity responses (saccharase and urease activity) on different land management systems and main recourses of ecological factors such as soil genesis type, content of soil organic carbon (SOC), total nitrogen (N), and C/N ratio. The trial was comprised of eight experimental plots (three land management systems and two crops groups: legumes and gramineous). Hydrolases activity correlated stronger with SOC (r = 0.7) or ratio C/N (r = 0.6) than that with total nitrogen content (r = 0.5-0.6) Significantly higher mean value of saccharase (27.00 and 12.6 mg CG g-1) and urease (5.78 and 4.16 mg NH4+-N g-1 24 h-1) were observed in oatvetch-barley of conventional farming and winter wheat-oat-pea-barley rotation fields of Organic Farm respectively. In most of the studies, it was observed that enzymes behave differently with different SOC content. Without mineral fertilizers application resulted in the least activity of urease (1.13 mg NH4+-N g-1) and saccharase (8.40 mg CG g-1 24 h-1). A significant increase of hydrolases activity has been observed in conventional (144- 413% of urease and 49-50% of saccharase) and organic (219-269% of urease and 78-221% of saccharase) farming as compared. [ABSTRACT FROM AUTHOR]