Measurement of saccharifying cellulase

Abstract

This article sets forth a simple cellulase assay procedure. Cellulose is variable in nature, insoluble and resistant to enzymatic attack. As a result there have been a bevy of bewildering cellulase assays published that yielded irrational results. Certain protocols focused on the rapidity of the assay while ignoring that only the most readily susceptible cellulose regions were being hydrolyzed. Other assays simplified the system by using modified soluble substrates and yielded results that bore no relationship to the real world hydrolysis of insoluble cellulose. In this study Mandels, Andreotti and Roche utilized a common substrate, Whatman filter paper. Hydrolysis of a 50 mg sample of the paper was followed to roughly 4% degradation, which circumvented the problems of attack of only the most susceptible zones. This common hydrolysis target range also resulted in some balance with regard to the interaction of the several cellulase components. The method was subsequently widely adopted.

Newly isolated Penicillium oxalicum A592-4B secretes enzymes that degrade milled rice straw with high efficiency

“…Avicel, CMC, and xylan are known as the substrates for cellobiohydrases (hereinafter, CBH), endo-β-1,4-glucanase (hereinafter, EG), 18) and endo-xylanase, 19) respectively. Since Avicel, xylan, and CMC exhibit different degradability, their reaction times were set as 2, 1, and 0.5 h, respectively.…”

Characteristics of cellulase in cellulose-degrading bacterium strain Clostridium straminisolvens (CSK1)

“…It was used as the substrates to determine the activity of CMCase. Basically, the CMC was degraded by the cellulase to produce free reducing sugars, which quantity was then determined by the DNS method to evaluate the cellulase activity (Aoyama et al, 2015;Eveleigh et al, 2009;Wang et al, 2014). In operation, the substrate was suspended in 1/15 mol/L disodium hydrogen phosphate-potassiumdihydrogen phosphate buffer (pH 6.24) at 1% (w/v).…”

Characterization of a Cellulose and Xylose Utilization Bacillus cereus Yc-A1

“…FPase activity were measured according to the method of Eveleigh with minor modification [11]. Briefly, 0.5 ml enzyme solution and 1.5 ml sodium citrate buffer (0.05 mol/L, pH4.8) was filled in 15 cm tube, then sterilized Whatman No.…”

“…CMC solid medium contain 2% agar in CMC-Na liquid medium. Filter paper hydrolyzing medium contain 1×6 cm filter paper strips, 0.02% CaCL 2 .2H 2 O, 0.2% (NH 4 ) 2 SO 4 , 0.1% KH 2 PO 4 , 0.05% MgSO 4 .7H 2 O, 0.15%Na 2 SO 4 , 0.05% peptone, 0.05% yeast extract [11]. YPD medium contain 2% glucose, 2% peptone, 1% yeast extract, 0.25% agar.…”