scholarly journals Exploring cellulolytic microorganisms from coffee industry by-products and their enzyme properties

2021 ◽  
Vol 924 (1) ◽  
pp. 012075
Author(s):  
A A Brahmanti ◽  
E Martati ◽  
A K Wardani
Keyword(s):  
Enzyme Activity ◽  
Specific Activity ◽  
Cellulase Activity ◽  
Cellulase Production ◽  
Zone Formation ◽  
Cellulolytic Microorganisms ◽  
Activity Measurements ◽  
Congo Red Staining ◽  
Immense Potential ◽  
By Products

Abstract Cellulolytic microorganism has immense potential due to their cellulase production, enzyme complexity and widespread habitat of life. This study was conducted to obtain microbial cellulase with vast industrial applicability from the coffee industry by-product in East Java, Indonesia. Fifty-four isolates with significant clear zone formation were obtained by Congo red staining in CMC agar plates. Eighteen bacteria, two yeasts and two moulds with high cellulolytic index were subjected to protein content determination as well as reducing sugar analysis in various conditions such as pH, temperature, addition of metal ions, surfactant and inhibitor agent. The specific activity measurements of all the crude enzymes result in the highest value of cellulase activity produced by isolate C12 which was 0.401 ± 0.018 U/mg. This enzyme activity was known to be optimum at 50°C and pH 9. It was also stimulated by K+, Na+, Mg2+, Fe3+, and SDS. However, the enzyme activity was inhibited by EDTA at 10 mM concentration. The use of coffee industry by-products as the source of cellulolytic microorganisms offers a promising approach for its various types of indigenous microorganisms and their unique property of cellulase produced that is useful for industrial application.

scholarly journals Response surface methodology-artificial neural network based optimization and strain improvement of cellulase production by Streptomyces sp.

2020 ◽  
Vol 36 (4) ◽  
Author(s):  
Ega Soujanya Lakshmi ◽  
Manda Rama Narasinga Rao ◽  
Muddada Sudhamani
Keyword(s):  
Neural Network ◽  
Artificial Neural Network ◽  
Response Surface Methodology ◽  
Response Surface ◽  
Wild Strain ◽  
Cellulase Activity ◽  
Cellulase Production ◽  
Optimal Conditions ◽  
Zone Formation ◽  
Artificial Neural

ABSTRACT Thirty seven different colonies were isolated from decomposing logs of textile industries. From among these, a thermotolerant, grampositive, filamentous soil bacteria Streptomyces durhamensis vs15 was selected and screened for cellulase production. The strain showed clear zone formation on CMC agar plate after Gram’s iodine staining.  Streptomyces durhamensis vs15 was further confirmed for cellulase production by estimating the reducing sugars through dinitrosalicylic acid (DNS) method. The activity was enhanced by sequential mutagenesis using three mutagens of ultraviolet irradiation (UV), N methyl-N’-nitro-N-nitrosoguanidine (NTG) and Ethyl methane sulphonate (EMS). After mutagenesis, the cellulase activity of GC23 (mutant) was improved to 1.86 fold compared to the wild strain (vs15). Optimal conditions for the production of cellulase by the GC 23 strain were evaluated using Response Surface Methodology (RSM) and Artificial Neural Network (ANN). Effect of pH, temperature, duration of incubation, , and substrate concentration on cellulase production were evaluated. Optimal conditions for the production of cellulase enzyme using Carboxy Methyl Cellulase as a substrate are 55 oC of temperature, pH of 5.0 and incubation for 40 h. The cellulase activity of the mutant Streptomyces durhamensis GC23 was further optimised to 2 fold of the activity of the wild type by RSM and ANN.  

Moving into Nanotechnology Roles to Mimic and Boost Enzyme Activity

2018 ◽  
pp. 421-440
Author(s):  
Maria Laura Soriano
Keyword(s):  
Enzyme Activity ◽  
Catalytic Activity ◽  
Enzyme Immobilization ◽  
Mesoporous Materials ◽  
Specific Activity ◽  
Artificial Enzymes ◽  
Catalytic Nanoparticles ◽  
Solid Liquid ◽  
By Products ◽  
Insight Into

A new tendency toward the design of artificial enzymes based on nanostructures (nanodots, nanofibers, mesoporous materials) has emerged. On one hand, nanotechnology bestows self-catalytic nanoparticles with a specific activity to achieve efficient reactions with low number of by-products. On other hand, the nanoparticles may behave as nanometric scaffolds for hosting enzymes, promoting their catalytic activity and stability. In this case, enzyme immobilization requires the preservation of the catalytic activity by preventing enzyme unfolding and avoiding its aggregation. These approaches render many other advantages like hosting/storing enzymes in nanotechnological solid, liquid, and gel-like media. This chapter focuses on the most up-to-date approaches to manipulate or mimic enzyme activity based on nanotechnology, and offers examples of their applications in the most promising fields. It also gives new insight into the creation of reusable nanotechnological tools for enzyme storage.

scholarly journals Screening Cellulolytic Bacteria from the Digestive Tract Snail (Achatina fulica) and Test the Ability of Cellulase Activity

2016 ◽  
Vol 8 (3) ◽  
pp. 386
Author(s):  
Wijanarka Wijanarka ◽  
Endang Kusdiyantini ◽  
Sarjana Parman
Keyword(s):  
Enzyme Activity ◽  
Digestive Tract ◽  
Generation Time ◽  
Cellulase Activity ◽  
Biology Education ◽  
Activity Measurement ◽  
Bacterial Isolates ◽  
Achatina Fulica ◽  
Cellulolytic Bacteria ◽  
Activity Measurements

<p>On the research of enzyme production levels observed cellulase produced by bacteria in the digestive tract of the isolation of the Snail (<em>Achatina fulica</em>). Isolation of bacteria based on the ability of bacteria to grow on CMC media. The purpose of this study was to determine cellulase activity by cellulolytic bacteria. Some bacterial isolates were identified as cellulolytic bacteria, they were KE-B1, KE-B2, KE-B3, KE-B4, KE-B5, and KE-B6. Isolates KE-B6 was the best isolates. Furthermore KE-B6 isolates were grown on media production to determine the pattern of growth and enzyme activity. Measurement of cell growth was conducted by inoculating starter aged 22 hours at CMC production of liquid medium. Cellulase enzyme activity measurements was performed by the DNS method. The results showed that the highest activity by new isolate bacteria KE-B6 and its value of the activity of 0.4539 U/mL, growth rate (µ) 0.377/hour and generation time (g) 1.84 hour. This research expected cellulase of producing bacteria were easy, inexpensive and efficient. This enzyme can be used as an enzyme biolytic once expected to replace expensive commercial enzyme. The biotylic enzyme can be applied to strains improvement (protoplast fusion).</p><p><strong>How to Cite</strong></p><p>Wijanarka, W., Kusdiyantini, E. &amp; Parman, S. (2016). Screening Cellulolytic Bacteria from the Digestive Tract Snail (<em>Achatina fulica</em>) and Test the Ability of Cellulase Activity. <em>Biosaintifika: Journal of Biology &amp; Biology Education</em>, 8(3), 386-392. </p>

scholarly journals Cellulase activity of bacteria isolated from water of mangrove ecosystem in Aceh Province

DEPIK ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 243-250
Author(s):  
Irma Dewiyanti ◽  
Darmawi Darmawi ◽  
Zainal Abidin Muchlisin ◽  
Teuku Zahrial Helmi ◽  
Iko Imelda Arisa ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Specific Activity ◽  
Methyl Cellulose ◽  
Cellulase Activity ◽  
Mangrove Ecosystem ◽  
Microbiology Laboratory ◽  
Cellulolytic Bacteria ◽  
Cellulase Enzymes ◽  
Cellulase Enzyme ◽  
Aceh Province

Cellulolytic bacteria that produce cellulase enzymes play an essential role in degrading cellulose in their habitat. The presence of cellulolytic bacteria strongly supports the fertility and productivity in mangrove waters. The objectives of the study are to analyze the activity of cellulase enzyme qualitatively through the cellulolytic index and quantitatively through the activity and specific activity of the cellulase enzyme from bacteria isolated from the water of mangrove ecosystems in Aceh Province. The qualitative experiment of enzyme activity was carried out at the Microbiology laboratory SKIPM Aceh, and a quantitative experiment of enzyme activity was conducted at the Microbiology Laboratory, Biology Department, IPB. Isolation of cellulolytic bacteria isolated from mangrove water used Carboxy Methyl Cellulose (1% CMC) selective media and carried out by spread plate method. The ability of bacteria to produce cellulase was tested qualitatively using the spot technique, this test was carried out using 1% Congo Red. Furthermore, the quantitative testing of cellulase enzymes activity adopted the DNS spectrophotometric method. The specific activity of the cellulase enzyme can be determined by using the Lowry method. There were 21 isolates that had a clear zone and had the ability to produce cellulase enzymes from 49 isolates that were successfully purified. The highest cellulolytic index (CI) produced using BAM421 isolate with the value of 5.50 was included in the high category, followed by BAM326 and BAM132 isolates, with values of 1.55 and 1.05 were categorized into the medium category. The other isolates were in the low cellulolytic index category. The isolate with the highest CI value was further tested using the quantitative enzyme activity test. The highest cellulase enzyme activity of BAM421 occurred at 24hr (0.0029 U/ml). The highest specific cellulase activity of BAM421 was at 24hr with the value of 0.210 U/mg. The result concluded that the qualitative test showed CI values can be categorized into low, medium, and high. Moreover, the value of the quantitative assay described that the cellulase enzyme and the specific enzyme activities of the bacteria were low in the study area.Keywords:Cellulolytic indexQuantitative testMangrove watersCellulase enzymeMicroorganismTRANSLATE with x EnglishArabicHebrewPolishBulgarianHindiPortugueseCatalanHmong DawRomanianChinese SimplifiedHungarianRussianChinese TraditionalIndonesianSlovakCzechItalianSlovenianDanishJapaneseSpanishDutchKlingonSwedishEnglishKoreanThaiEstonianLatvianTurkishFinnishLithuanianUkrainianFrenchMalayUrduGermanMalteseVietnameseGreekNorwegianWelshHaitian CreolePersian //  TRANSLATE with COPY THE URL BELOW Back EMBED THE SNIPPET BELOW IN YOUR SITE Enable collaborative features and customize widget: Bing Webmaster PortalBack//

Production and Optimization of Cellulase Activity of Thermomonospora Viridis Isolated From Rice Straw

2021 ◽  
Vol 50 (2) ◽  
pp. 395-404
Author(s):  
Faozia Faleha Sadida ◽  
Ma Manchur
Keyword(s):  
Enzyme Activity ◽  
Metal Ions ◽  
Rice Straw ◽  
Cellulase Activity ◽  
Nitrogen Sources ◽  
Cellulase Production ◽  
Tetraacetic Acid ◽  
Carbon And Nitrogen Sources ◽  
Carbon And Nitrogen ◽  
Optimum Ph

A highly cellulolytic actinomycete SR1 was locally isolated from rice straw and provisionally identified as Thermomonospora viridis. Optimum pH, temperature, carbon and nitrogen sources for its cellulase production were 6.5, 35°C, Carboxymethyl cellulase (CMC) and yeast extract, respectively whereas those of cellulase activity were 7.5, 40°C, CMC and peptone respectively. The effects of various metal ions and different reductant and inhibitors on its cellulase activity were investigated. Univalent Ag+ was found to decrease the enzyme activity whereas increased by bivalent Mg2+. Ethylene diamine tetraacetic acid (EDTA) caused remarkable decrease of cellulase activity but β-Mercaptoethanol stimulated its cellulase activity. Bangladesh J. Bot. 50(2): 395-404, 2021 (June)

IMPROVED PROCEDURES FOR PREPARATION AND CHARACTERIZATION OF MYROTHECIUM CELLULASE: PART 2. PURIFICATION PROCEDURES

1963 ◽  
Vol 41 (1) ◽  
pp. 671-696
Author(s):  
D. R. Whitaker ◽  
K. R. Hanson ◽  
P. K. Datta
Keyword(s):  
Ammonium Sulphate ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Cellulase Activity ◽  
Starch Gel Electrophoresis ◽  
Polymethacrylic Acid ◽  
Citrate Buffer ◽  
Starch Gel ◽  
By Products

Two methods are described for purification of the cellulase of Myrothecium verrucaria from concentrated culture filtrates. The steps of method I are (1) fractionation with ammonium sulphate, (2) elution through Sephadex G25, (3) elution through Sephadex G75, (4) precipitation with polymethacrylic acid, and (5) elution through Amberlite CG50 with citrate buffer containing a gradient of urea concentration. The steps of method II are precipitation by saturated ammonium sulphate, (2) and (4) as above, elution through DEAE-cellulose with phosphate buffer containing 7 M urea, followed by (2) and (3) as above. The two methods gave final products with identical specific activities toward carboxymethyl cellulose; the increase in specific activity was approximately 12-fold. Starch-gel electrophoresis at pH 6.8 showed no major indications of heterogeneity. The purified enzyme was unstable but could be stored frozen in dilute salt solution.Enzyme passed through DEAE-cellulose without an inhibitor of cellulase activity was contaminated by DEAE-substituted oligoglucosides and subject to proteolysis. Urea could be replaced by cellobiose as an inhibitor but the latter gave enzyme contaminated by products of transfer reactions.Binding of urea by the resin is shown to influence significantly the resolution attainable in chromatographic fractionations on Amberlite CG-50 with buffers containing gradients of urea concentration.Procedures for dialysis and desalting of cellulases and a compact reaction vessel for pH stats are described.

IMPROVED PROCEDURES FOR PREPARATION AND CHARACTERIZATION OF MYROTHECIUM CELLULASE: PART 2. PURIFICATION PROCEDURES

1963 ◽  
Vol 41 (3) ◽  
pp. 671-696 ◽  
Author(s):  
D. R. Whitaker ◽  
K. R. Hanson ◽  
P. K. Datta
Keyword(s):  
Ammonium Sulphate ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Cellulase Activity ◽  
Starch Gel Electrophoresis ◽  
Polymethacrylic Acid ◽  
Citrate Buffer ◽  
Starch Gel ◽  
By Products

Two methods are described for purification of the cellulase of Myrothecium verrucaria from concentrated culture filtrates. The steps of method I are (1) fractionation with ammonium sulphate, (2) elution through Sephadex G25, (3) elution through Sephadex G75, (4) precipitation with polymethacrylic acid, and (5) elution through Amberlite CG50 with citrate buffer containing a gradient of urea concentration. The steps of method II are precipitation by saturated ammonium sulphate, (2) and (4) as above, elution through DEAE-cellulose with phosphate buffer containing 7 M urea, followed by (2) and (3) as above. The two methods gave final products with identical specific activities toward carboxymethyl cellulose; the increase in specific activity was approximately 12-fold. Starch-gel electrophoresis at pH 6.8 showed no major indications of heterogeneity. The purified enzyme was unstable but could be stored frozen in dilute salt solution.Enzyme passed through DEAE-cellulose without an inhibitor of cellulase activity was contaminated by DEAE-substituted oligoglucosides and subject to proteolysis. Urea could be replaced by cellobiose as an inhibitor but the latter gave enzyme contaminated by products of transfer reactions.Binding of urea by the resin is shown to influence significantly the resolution attainable in chromatographic fractionations on Amberlite CG-50 with buffers containing gradients of urea concentration.Procedures for dialysis and desalting of cellulases and a compact reaction vessel for pH stats are described.

scholarly journals Enhanced production of cellulase by Escherichia coli engineered with UV-mutated cellulase gene from Aspergillus niger UVMT-I

BioResources ◽  
2019 ◽  
Vol 14 (4) ◽  
pp. 9054-9063
Author(s):  
Qurat-ul-ain Daud ◽  
Zara Hamid ◽  
Tayyaba Sadiq ◽  
Asif Abbas ◽  
Shahzad Hussain Shah ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Specific Activity ◽  
Cellulase Activity ◽  
Pcr Amplification ◽  
Type A ◽  
Cellulase Production ◽  
Cellulase Gene ◽  
Wild Type ◽  
E Coli ◽  
Enhanced Production

Enhanced cellulase production was studied with ultraviolet mutagenesis and the mutated cellulase gene in E. coli DH5α was cloned for production under controlled conditions. Aspergillus niger inoculum was exposed to UV radiation for different time intervals. The UV exposure of 10 min to A. niger yielded 330 μmol/min/mg specific activity. The mRNA of mutant A. niger yielding maximum enzyme activity was isolated and used for the synthesis of cDNA. The cDNA prepared from mRNA was used for the PCR amplification of mutated cellulase gene with primers designed on the basis of a cellulase gene database from A. niger. The amplified cellulase gene was cloned into E. coli DH5α followed by expression in E. coli BL21. The cellulase activity by wild type A. niger, A. niger UVMT-I, and recombinant E. coli was compared by analysis of variance test. The specific activity of cellulase by recombinant E. coli was maximum (441 μmol/min/mg), followed by A. niger UVMT-I (330 μmol/min/mg) and wild type A. niger (96 μmol/min/mg).

A New Method for Plasminogen Standardization

1968 ◽  
Vol 20 (03/04) ◽  
pp. 548-554
Author(s):  
J Gajewski ◽  
G Markus
Keyword(s):  
Proteolytic Activity ◽  
Specific Activity ◽  
Molar Ratio ◽  
Sharp Transition ◽  
Equivalence Point ◽  
Constant Amount ◽  
Residual Level ◽  
Activity Measurements ◽  
Complete Saturation ◽  
Human Plasminogen

SummaryA method for the standardization of human plasminogen is proposed, based on the stoichiometric interaction between plasminogen and streptokinase, resulting in inhibition of proteolytic activity. Activation of a constant amount of plasminogen with increasing amounts of streptokinase yields linearly decreasing activities, as a function of streptokinase, with a sharp transition to a constant residual level. The point of transition corresponds to complete saturation of plasmin with streptokinase in a 1:1 molar ratio, and is therefore a measure of the amount of plasminogen present initially, in terms of streptokinase equivalents. The equivalence point is independent of the kind of protein substrate used, buffer, pH, length of digestion and, within limits, temperature. The method, therefore, is not subject to the variations commonly encountered in the usual determination based on specific activity measurements.

scholarly journals Study of Extraction and Enzymatic Properties of Cell-Envelope Proteinases from a Novel Wild Lactobacillus plantarum LP69

Catalysts ◽  
2018 ◽  
Vol 8 (8) ◽  
pp. 325 ◽  
Author(s):  
He Chen ◽  
Jie Huang ◽  
Binyun Cao ◽  
Li Chen ◽  
Na Song ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Lactobacillus Plantarum ◽  
Industrial Development ◽  
Extraction Efficiency ◽  
Specific Activity ◽  
Cell Envelope ◽  
Serine Proteinase ◽  
Kinetic Studies ◽  
Predicted Values ◽  
Hydrolyze Whey Protein

Lactobacilli cell-envelope proteinases (CEPs) have been widely used in the development of new streams of blockbuster nutraceuticals because of numerous biopharmaceutical potentials; thus, the development of viable methods for CEP extraction and the improvement of extraction efficiency will promote their full-scale application. In this study, CEP from a novel wild Lactobacillus plantarum LP69 was released from cells by incubating in calcium-free buffer. The extraction conditions of CEP were optimized by response surface methodology with the enzyme activity and specific activity as the detective marker. The optimal extraction conditions were: time of 80 min, temperature of 39 °C and buffer pH of 6.5. Under these conditions, enzyme activity and specific activity were (23.94 ± 0.86) U/mL and (1.37 ± 0.03) U/mg, respectively, which were well matched with the predicted values (22.12 U/mL and 1.36 U/mg). Optimal activity of the crude CEP occurred at pH 8.0 and 40 °C. It is a metallopeptidase, activated by Ca2+, inhibited by Zn2+ and ethylene-diamine-tetra-acetic acid, and a serine proteinase which is inhibited by phenylmethylsulfonyl fluoride. Kinetic studies showed that CEP from LP69 could hydrolyze whey protein, lactoglobulin and casein. Our study improves the extraction efficiency of CEPs from LP69, providing the reference for their industrial development.

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