scholarly journals A set of simple methods for detection and extraction of laminarinase

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ananthamurthy Koteshwara ◽  
Nancy V. Philip ◽  
Jesil Mathew Aranjani ◽  
Raghu Chandrashekhar Hariharapura ◽  
Subrahmanyam Volety Mallikarjuna
Keyword(s):  
Ammonium Sulfate ◽  
Ammonium Sulphate ◽  
Gold Standard ◽  
Direct Detection ◽  
Low Cost ◽  
Direct Analysis ◽  
Reducing Sugar ◽  
Ammonium Sulfate Precipitation ◽  
Simple Method ◽  
Functional Step

AbstractA carefully designed ammonium sulfate precipitation will simplify extraction of proteins and is considered to be a gold standard among various precipitation methods. Therefore, optimization of ammonium sulfate precipitation can be an important functional step in protein purification. The presence of high amounts of ammonium sulphate precludes direct detection of many enzymatically active proteins including reducing sugar assays (e.g. Nelson-Somogyi, Reissig and 3,5-dinitrosalicylic acid methods) for assessing carbohydrases (e.g. laminarinase (β (1–3)-glucanohydrolase), cellulases and chitinases). In this study, a simple method was developed using laminarin infused agarose plate for the direct analysis of the ammonium sulphate precipitates from Streptomyces rimosus AFM-1. The developed method is simple and convenient that can give accurate results even in presence of ammonium sulfate in the crude precipitates. Laminarin is a translucent substrate requiring the use of a stain to visualize the zones of hydrolysis in a plate assay. A very low-cost and locally available fluorescent optical fabric brightener Tinopal CBS-X has been used as a stain to detect the zones of hydrolysis. We also report simple methods to prepare colloidal chitin and cell free supernatant in this manuscript.

Brewer’s spent grain as substrate for enzyme and reducing sugar production using Penicillium sp. HC1

2021 ◽  
Author(s):  
Marcela Bernal-Ruiz ◽  
Alejandro Correa-Lozano ◽  
Laura Gomez-Sánchez ◽  
Balkys Quevedo-Hidalgo ◽  
Lilia Carolina Rojas-Pérez ◽  
...  
Keyword(s):  
Ammonium Sulfate ◽  
Reducing Sugars ◽  
Low Cost ◽  
Xylanase Activity ◽  
Reducing Sugar ◽  
Endoglucanase Activity ◽  
Brewer’S Spent Grain ◽  
Penicillium Sp ◽  
Brewing Process ◽  
Spent Grain

Brewer’s spent grain (BSG) is the main solid waste from the brewing process. It is recognized as a valuable resource for biobased industries because of its composition, high availability, and low cost. The objective of this study was to employ BSG as a substrate to produce the enzymes endoglucanase, cellobiohydrolase, β-glucosidase, and xylanase, as well as reducing sugars using Penicillium sp. HC1. For enzyme production, we evaluated BSG submerged fermentation at different concentrations (1%, 3%, and 5%, w/v) and two sources of nitrogen (yeast extract and ammonium sulfate) on different days (6, 10, and 12) in a 100 mL Erlenmeyer flask. The highest enzyme activity was obtained after 10 days. The enzyme extract obtained using 3% BSG (w/v) and 5 g L-1 of ammonium sulfate showed the highest xylanase activity (25013 ± 1075 U L-1). Using BSG 5% (w/v) without nitrogen supplementation, the endoglucanase activity was 909.7±14.2 U L-1 while underthe same conditions but using BSG 3% (w/v), the β-glucosidase and cellobiohydrolase activity was 3268.6 ±229.9 U L-1 and 103.15±8.1 U L-1, respectively. Maximum reducing sugar concentrations using an enzyme dosage of 1000 U g-1 of xylanase were: 2.7 g L-1 xylose, 1.7 g L-1 arabinose, and 3.3 g L-1 glucose after 6 h of hydrolysis. Result s demonstrated it is possible to produce enzymes and reducing sugars using Penicillium sp. HC1 and BSG as substrate and BSG grinding only as pretreatment. 

scholarly journals Distributed Acoustic Sensing Using Chirped-Pulse Phase-Sensitive OTDR Technology

Sensors ◽  
2019 ◽  
Vol 19 (20) ◽  
pp. 4368 ◽  
Author(s):  
María R. Fernández-Ruiz ◽  
Luis Costa ◽  
Hugo F. Martins
Keyword(s):  
Direct Detection ◽  
Low Cost ◽  
High Sensitivity ◽  
Coherent Detection ◽  
Simple Method ◽  
Acoustic Sensing ◽  
Chirped Pulse ◽  
Distributed Sensor ◽  
Phase Sensitive ◽  
Distributed Acoustic Sensing

In 2016, a novel interrogation technique for phase-sensitive (Φ)OTDR was mathematically formalized and experimentally demonstrated, based on the use of a chirped-pulse as a probe, in an otherwise direct-detection-based standard setup: chirped-pulse (CP-)ΦOTDR. Despite its short lifetime, this methodology has now become a reference for distributed acoustic sensing (DAS) due to its valuable advantages with respect to conventional (i.e., coherent-detection or frequency sweeping-based) interrogation strategies. Presenting intrinsic immunity to fading points and using direct detection, CP-ΦOTDR presents reliable high sensitivity measurements while keeping the cost and complexity of the setup bounded. Numerous technique analyses and contributions to study/improve its performance have been recently published, leading to a solid, highly competitive and extraordinarily simple method for distributed fibre sensing. The interesting sensing features achieved in these last years CP-ΦOTDR have motivated the use of this technology in diverse applications, such as seismology or civil engineering (monitoring of pipelines, train rails, etc.). Besides, new areas of application of this distributed sensor have been explored, based on distributed chemical (refractive index) and temperature-based transducer sensors. In this review, the principle of operation of CP-ΦOTDR is revisited, highlighting the particular performance characteristics of the technique and offering a comparison with alternative distributed sensing methods (with focus on coherent-detection-based ΦOTDR). The sensor is also characterized for operation in up to 100 km with a low cost-setup, showing performances close to the attainable limits for a given set of signal parameters [≈tens-hundreds of pe/sqrt(Hz)]. The areas of application of this sensing technology employed so far are briefly outlined in order to frame the technology.

Extraction and Partial Purification of Protease from Bilimbi (Averrhoa bilimbi L.)

2013 ◽  
Vol 10 (2) ◽  
pp. 29
Author(s):  
Normah Ismail ◽  
Nur' Ain Mohamad Kharoe
Keyword(s):  
Molecular Weight ◽  
Protein Content ◽  
Ammonium Sulfate ◽  
Molecular Weight Distribution ◽  
Weight Distribution ◽  
Protease Activity ◽  
Protein Concentration ◽  
Temperature Stability ◽  
Partial Purification ◽  
Ammonium Sulfate Precipitation

Unripe and ripe bilimbi (Averrhoa bilimbi L.) were ground and the extracted juices were partially purified by ammonium sulfate precipitation at the concentrations of 40 and 60% (w/v). The collected proteases were analysed for pH, temperature stability, storage stability, molecular weight distribution, protein concentration and protein content. Protein content of bilimbi fruit was 0.89 g. Protease activity of both the unripe and ripe fruit were optimum at pH 4 and 40°C when the juice were purified at 40 and 60% ammonium sulfate precipitation. A decreased in protease activity was observed during the seven days of storage at 4°C. Molecular weight distribution indicated that the proteases protein bands fall between IO to 220 kDa. Protein bands were observed at 25, 50 and 160 kDa in both the unripe and ripe bilimbi proteases purified with 40% ammonium sulfate, however, the bands were more intense in those from unripe bilimbi. No protein bands were seen in proteases purified with 60% ammonium sulfate. Protein concentration was higher for proteases extracted with 40% ammonium sulfate at both ripening stages. Thus, purification using 40% ammonium sulfate precipitation could be a successful method to partially purify proteases from bilimbi especially from the unripe stage. 

A simple method to construct a low-cost immunosensor based on a dithiol-functionalized polydopamine platform

2021 ◽  
Author(s):  
Bo Liu ◽  
Luanying Yang ◽  
Gang Wang ◽  
Sha He ◽  
Xiaobo Wang ◽  
...  
Keyword(s):  
Detection Limit ◽  
Linear Response ◽  
Low Cost ◽  
Covalent Immobilization ◽  
The Self ◽  
Simple Method ◽  
Low Detection Limit

A simple and low-cost electrochemical CEA immunosensor was investigated via the self-polymerization of dopamine and a dithiol compound spacer for the covalent immobilization of antibodies. The designed CEA immunosensor exhibited a linear response and a low detection limit.

scholarly journals Deconstruction of Lignin: From Enzymes to Microorganisms

Molecules ◽  
2021 ◽  
Vol 26 (8) ◽  
pp. 2299
Author(s):  
Jéssica P. Silva ◽  
Alonso R. P. Ticona ◽  
Pedro R. V. Hamann ◽  
Betania F. Quirino ◽  
Eliane F. Noronha
Keyword(s):  
Microbial Communities ◽  
Metabolic Pathways ◽  
Low Cost ◽  
Direct Analysis ◽  
Industrial Applications ◽  
Natural Environments ◽  
Lignocellulosic Residues ◽  
Complementary Techniques ◽  
Plant Polymer ◽  
Powerful Strategy

Lignocellulosic residues are low-cost abundant feedstocks that can be used for industrial applications. However, their recalcitrance currently makes lignocellulose use limited. In natural environments, microbial communities can completely deconstruct lignocellulose by synergistic action of a set of enzymes and proteins. Microbial degradation of lignin by fungi, important lignin degraders in nature, has been intensively studied. More recently, bacteria have also been described as able to break down lignin, and to have a central role in recycling this plant polymer. Nevertheless, bacterial deconstruction of lignin has not been fully elucidated yet. Direct analysis of environmental samples using metagenomics, metatranscriptomics, and metaproteomics approaches is a powerful strategy to describe/discover enzymes, metabolic pathways, and microorganisms involved in lignin breakdown. Indeed, the use of these complementary techniques leads to a better understanding of the composition, function, and dynamics of microbial communities involved in lignin deconstruction. We focus on omics approaches and their contribution to the discovery of new enzymes and reactions that impact the development of lignin-based bioprocesses.

Preparation of Human Immunoglobulin by Ammonium Sulfate Precipitation

Vox Sanguinis ◽  
1976 ◽  
Vol 31 (6) ◽  
pp. 423-434
Author(s):  
A.F.S.A. Habeeb ◽  
Robert D. Francis
Keyword(s):  
Ammonium Sulfate ◽  
Human Immunoglobulin ◽  
Ammonium Sulfate Precipitation

scholarly journals A simple method for isolating chicken egg yolk immunoglobulin using effective delipidation solution and ammonium sulfate

2015 ◽  
Vol 94 (1) ◽  
pp. 104-110 ◽  
Author(s):  
Chenyao Tong ◽  
Fang Geng ◽  
Zhenjiao He ◽  
Zhaoxia Cai ◽  
Meihu Ma
Keyword(s):  
Ammonium Sulfate ◽  
Egg Yolk ◽  
Simple Method ◽  
Chicken Egg ◽  
Egg Yolk Immunoglobulin ◽  
Chicken Egg Yolk

scholarly journals A20 DIAGNOSTIC ACCURACY OF A LOW COST, WIDELY AVAILABLE TEST STRIP FOR PREDICTING A POSITIVE FECAL CALPROTECTIN TEST

2021 ◽  
Vol 4 (Supplement_1) ◽  
pp. 210-212
Author(s):  
R Trasolini ◽  
S Wong ◽  
B Salh
Keyword(s):  
Sample Size ◽  
Likelihood Ratio ◽  
Gold Standard ◽  
Low Cost ◽  
Positive Likelihood Ratio ◽  
Negative Likelihood Ratio ◽  
Rapid Test ◽  
Test Strip ◽  
Fecal Calprotectin ◽  
Type I

Abstract Background Fecal calprotectin is a non-invasive test of colonic inflammation used for monitoring inflammatory bowel disease activity and for risk stratifying non-specific colonic symptoms. Calprotectin is a leukocyte specific enzyme. A similar test, leukocyte esterase is used to detect leukocytes in urine and is widely available as a low-cost point-of-care test strip. We hypothesize that an unmodified version of the urine test strip would be highly accurate in predicting a positive fecal calprotectin test in a real world sample of patients. Aims To explore a low cost, rapid alternative to the fecal calprotectin test Methods All inpatient and outpatient stool samples tested for calprotectin by the Vancouver General Hospital laboratory from February 2020 to November 2020 were included prospectively. Samples were simultaneously tested for fecal leukocyte esterase using an unmodified Roche Cobas Chemstrip urinalysis test strip by central lab personnel. An identical aliquot was sent to LifeLabs for calprotectin as per standard protocol. All samples were suspended in buffer using established laboratory protocols prior to testing. Fecal leukocyte esterase results were reported as 0–4+ based on visual interpretation, calprotectin results were reported as mcg/g of stool. REB review and approval was obtained prior to data collection. Sensitivity, Specificity and AUROC were calculated using Microsoft Excel and JROCFIT. Results 26 samples were collected. Using a fecal calprotectin greater than 120 mcg/g as a gold standard an AUROC of 0.89 (SE= .06) was calculated. A leukocyte esterase reading of 2+ or greater had the best test characteristics based on ROC curve analysis. Using this cutoff, 21/26 samples were concordant, giving an accuracy of 80.8%, sensitivity of 90.9% and specificity of 73.3%. Positive likelihood ratio was 8.07 and negative likelihood ratio was 0.29. Assuming an AUROC of 0.8, the sample size N=26 is 90% powered (β=0.9) to predict the true AUROC within 0.1 with a type I error rate of .05 (α<.05). Conclusions This study suggests application of a prepared stool sample to a urinalysis test strip gives a result highly predictive of a positive fecal calprotectin test. Further results are being collected prospectively to improve the robustness of these preliminary data. Secondary outcomes including comparison to endoscopy and biopsy results where available are planned if an adequate sample size can be accrued. Future studies justifying independent clinical use of leukocyte esterase would require a common gold standard comparator such as endoscopy. Fecal calprotectin testing is not universally insured and is not available as a rapid test strip. Use of fecal leukocyte esterase may reduce costs and shorten time to results if proven to be independently reliable. Funding Agencies None

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