Measurement of filter paper activities of cellulase with microplatebased assay article pdf available in saudi journal of biological sciences 441 july 2015 with 756 reads how we measure reads. Analytical scientists have developed a number of alternative methods. This procedure is only appropriate for the determination of fpu activity in a cellulase preparation as defined by the iupac procedure as outlined above. Microbial cellulases production, applications and challenges rajeev k sukumaran, reeta rani singhania and ashok pandey biotechnology division, regional research laboratory, industrial estate p o, trivandrum 695 019 microbial cellulases find applications in various industries and constitute a major group of the industrial enzymes. Measuring cellulase activities 87 9 9 methods for measuring cellulase activities by thomas m. Measurement of cellulase activities laboratory analytical procedure lap. Enzymatic hydrolysis of cellulose requires mixtures of hydrolytic enzymes including endoglucanases, exoglucanases cellobiohydrolases, and. The synergistic action of endoglucanases, exoglucanases, and.
After 160 h of cultivation, carboxymethyl cellulase reached 9. For other cellulase enzymes, the ph and the assay temperature may be different. The assay conditions must be defined when reporting results. A rational and governing equation for crude cellulase assay was derived, and reliable results for quantitatively estimating the saccharifying activities of crude cellulases during the progress of hydrolysis of several cellulosic substrates were obtained. A fast, sensitive and easy colorimetric assay for chitinase. Methods for measuring cellulase activities sciencedirect.
Europe pmc is a service of the europe pmc funders group, in partnership with the european bioinformatics institute. The life and lifework of mary mandels first lady of cellulase research the upsurge in research and funding in the biomasstobiofuels arena has attracted an enormous following of new researchers to the field of cellulases and related plant cell walldegrading enzymes. For a practical measurement of saccharifying cellulase measurement of endo b glucanase or of any other single component is unsatisfactory. Pdf measurement of filter paper activities of cellulase. Sep 01, 2009 for a practical measurement of saccharifying cellulase measurement of endo. Cellulase activity in solid state fermentation of palm kernel cake with trichoderma sp. Hydrolysis of insoluble cellulase by a complete cellulase from trichoderma viride 5% cellulase incubated at ph 4. Experiments have shown that carboxymethyl cellulose is the best source of carbon, and ammonium sulfate is the best source of nitrogen. Measurement of filter paper activities of cellulase with. Mar 10, 2014 most of the current colorimetric methods for detection of chitinase or cellulase activities on the insoluble natural polymers chitin and cellulose depend on a chemical redox reaction. Measurement of cellulase activities 259 in the face of these difficulties, and in view of the applied nature of most cellulase work, it is understand able that investigators in different laboratories have each developed a series of empirical assay proce. Cellulase activity in solid state fermentation of palm kernel. The reaction involves the reducing ends of the hydrolytic products. Measurement of cellulase activities laboratory analytical procedure 1.
Microbial cellulases production, applications and challenges. A cellulolytic fungus, trichoderma pseudokoningii s38, was isolated previously by our laboratory and used for production of cellulase ma 1990. The effect of ph on the production of cellulases in. A new approach to measurement of saccharifying capacities of crude cellulase, biores. It includes content provided to the pmc international archive by participating publishers. Engineering the oleaginous red yeast rhodotorula glutinis. The rate of synthesis of cm and fpcellulases decreased after 90 h of cultivation but. A practical, quantitative approach has been designed, which makes it possible to accurately estimate the saccharifying activities of crude cellulase preparations for insoluble cellulosics. Thus, an accurate and reproducible assay for the measurement of cellulase hydrolysis rate is still required wu et al. Europe pmc is an elixir core data resource learn more. Citeseerx which permits unrestricted use, distribution, and.
Other organisms such as pestalotiopsis westerdijkii may yield as much as 400 cmc unitsml of culture filtrate. In the face of these difficulties, and in view of the applied nature of most cellulase work, it is understand able that investigators in different laboratories have each developed a series of empirical assay proce dures. The optimal ph for the production of extracellular cellulolytic enzymes in the wild strain of aspergillus terreus was shown to be ph 5. After each cellulase was diluted gradiently, the whatman no.
Measurement of saccharification by cellulases rosanne mullings department of biochemistry and unit of molecular genetics, university of bristol, university walk, bristol bs8 1 td, uk summary. The cellulase activity was screened using fastcellulase screening 4% sigmacell cellulose type 20 as the carbon source and the data showed that almost all transformants had higher cellulase. The method is based on measurement of reducing moieties released by the enzymes from arabinoxylan. She worked for forty years at the head of the us armys national bioconversion studies.
Hemicellulose is generally considered a negative factor for enzymatic cellulose hydrolysis, but it was proved to be a positive one for the enzymatic action of delignified pulp in this study. Cellulase production from species of fungi and bacteria from. As the native substrate, cellulose, is a waterinsoluble polymer, traditional reducing sugar assays using this substrate can not be employed for the measurement of cellulase activity. Apr 11, 2016 microbial cellulases have been receiving worldwide attention, as they have enormous potential to process the most abundant cellulosic biomass on this planet and transform it into sustainable biofuels and other value added products. Measurement of saccharification by cellulases sciencedirect. One of the most used methods is ghoses cellulase and endoglucanase assay, developed by the international union of pure and applied chemistry in 1987. Measurement of saccharifying cellulase biotechnology for. A simple, robust and highly reproducible method for the determination of xylanase.
Measurement of saccharifying cellulase pdf paperity. Cellulose, the major constituent of all plant materials and the most abundant organic molecule on the earth, is a linear biopolymer of glucose molecules, connected by. For a practical measurement of saccharifying cellulase measurement of endo. Production of extracellular cellulase by lunulospora curvula. The life and lifework of mary mandels first lady of. Rahman1, nurul jannah hasnan1, moftah massaud ben name1, hideyuki nagao2, mohd omar ab kadir3 1school of distance education, universiti sains malaysia usm, 11800 minden, penang, malaysia. Measurement of filter paper cellulase units per ml. A new approach to measurement of saccharifying capacities of.
Carrying out this method demands high volumes of reagents and generation of high. A new approach to measurement of saccharifying capacities of crude cellulase bin wua, yue zhaoa, pei ji gaoa a practical, quantitative approach has been designed, which makes it possible to accurately estimate the saccharifying activities of crude cellulase preparations for insoluble cellulosics. In conclusion, the measurement of cellulase is complex and there is no absolute unit as can be measured for a single enzyme acting on a soluble substrate. Douglas e eveleigh, mary mandels, ra ymond andreotti and charles roche.
Measurement of cellulase activities 259 in the face of these difficulties, and in view of the applied nature of most cellulase work, it is understandable that investigators in different laboratories have each developed a series of empirical assay procedures. The unit value will depend on the substrate chosen, its concentration, and the extent of conversion. New studies on cellulolytic enzymes aiming to improve biofuels production lead to a concern over the assaying methods commonly applied to measure their activity. Mahalingeshwara bhat the physical heterogeneity of the substrate and the complexity of the cellulase enzyme system present those wishing to measure cellulase activ ities with formidable problems. Mary elizabeth hickox mandels 12 september 1917 17 february 2008 was an early advocate for converting waste biomass to fuels and chemicals.
Researchers have mainly focused on improving methods 61 for measurement of cellulase activity which have already been widely used. Measurement of saccharifying cellulase europe pmc article. Fpas of six commercial cellulases were measured with the iupac method in this work. For this reason the filter paper assay was introduced. The procedure has been designed to measure cellulase activity in terms of filterpaper units fpu milliliterperof. Studies on the production of extracellular cellulase in two aquatic hyphomyceteslunulospora curvula andflagellospora penicillioides have shown that several factors such as carbon source, nitrogen source, ph and temperature affect the production of the enzyme. An approach is presented for obtaining relative filter paper activities for enzyme preparations having activities below that required for application of the traditional international union of pure and applied chemistry filter paper assay.