Screening Cellulolytic Fungi Isolated From Malaysia Cocoa Pod Husk and Its Culture Conditions for Cellulases Production

The aim of this study was to screen few fungal isolates from local cocoa pod husks (CPH) which able to secrete cellulases. The isolates were plated on carboxymethyl cellulose (CMC) agar plates which then incubated for two days at 28ºC. Then, these plates were stained with congo red dye for 0.5-1 h followed by destaining with 1 M NaCl solution for 15-20 minutes to observe its cellulolytic activity. One isolates which exposed the largest cellulolytic zone on CMC agar plate was selected for further study. In this study, culture conditions with respect to pH, incubation time, amount of substrate (CPH) and temperature were screened using Design expert @version 8.0 by employing two-level factorial design. The selected fungus isolate was cultured in shake flask at 37°C with agitation of 200 rpm for 5 days in incubator shaker. During fermentation period, samples were collected every day for fungal-cellulases activity of filter paper activity (FPase) and carboxymethyl cellulase (CMCase) activity. Analysis of variance (ANOVA) of this study showed that the most significant parameters that affects the production of cellulases from the selected fungi isolates were the amount of substrate (CPH) used followed by the interaction of amount of substrate with pH (p< 0.05). It showed that the cellulases activity was high when the pH 9 with more amount of substrate used. However, it was observed that less significant changes of celllulases activity when different amount of substrate was used at same pH of 3. Based on the microscopic observation of isolate, it morphology was closed to Rhizopus sp.. In conclusion, it is suggested to optimize the selected culture parameters obtained in this study in order to maximize the activity of cellulases from the selected isolates.

A Simplified Procedure for Cellulase Filter Paper Assay

A new procedures to minimize labor intensiveness and complexity that has long been recognized in cellulase filter paper activity measurement (FPAase) described by the international union of pure and applied chemistry (IUPAC) was developed. It follows the main idea of IUPAC finding at least two cellulase dilutions have optical densities slightly more and less than a reference optical density of an arbitrary fixed 2 mg absolute glucose amount after a red-ox color reaction and due to cellulase-filter paper hydrolysis. The yielding glucose amount difference as compared to this reference is expressed in terms of absorbance difference percent determined by 3,5-dinitrosalicylic acid (DNS), in case of cellulase is cellubiase rich. If not, an external supplemental portion should be added. The intersection of the line of these two cellulase dilutions with abscissa intersect a vertical at a hypothetical 0% absorbance difference percent corresponding to a critical cellulase dilution exactly release this fixed arbitrary 2 mg glucose amount value. The factor 0.37 of this critical cellulase dilution equals to its filter paper units expressed in FPU per ml. A cellulase Cellic Ctech2 from novozymes has been tested with this methodology giving a satisfactory results with IUPAC procedures.

A Simplified Procedures for Cellulase Filter Paper Assay

A new procedures to minimize labor intensiveness and complexity that has long been recognized in cellulase filter paper activity measurement (FPAase) described by the international union of pure and applied chemistry (IUPAC) was developed. It follows the main idea of IUPAC with only exception finding at least two cellulase dilutions have optical densities slightly more and less than a reference optical density of an arbitrary fixed 2 mg absolute glucose amount after a red-ox color reaction and due to cellulase-filter paper hydrolysis. The yielding glucose amount difference as compared to this reference is expressed in terms of absorbance difference percent determined by 3,5-dinitrosalicylic acid (DNS), in case of cellulase is cellubiase rich. If not, an external supplemental portion should be added. The intersection of the line of these two cellulase dilutions with abscissa intersect a vertical at a hypothetical 0% absorbance difference percent corresponding to a critical cellulase dilution exactly release this fixed arbitrary 2 mg glucose amount value. The factor 0.37 of this critical cellulase dilution equals to its filter paper units expressed in FPU per ml.

A Simplified Procedures for Cellulase Filter Paper Assay

<p>A new procedures to minimize labor intensiveness and complexity that has long been recognized in cellulase filter paper activity measurement (FPAase) described by the international union of pure and applied chemistry (IUPAC)was developed. It follows the main idea of IUPAC with only exception finding at least two cellulase dilutions have optical densities slightly more and less than a reference optical density of an arbitrary fixed 2 mg absolute glucose amount after a red-ox color reaction and due to cellulase-filter paper hydrolysis. The yielding glucose amount difference as compared to this reference is expressed in terms of absorbance difference percent determined by 3,5-dinitrosalicylic acid (DNS), in case of cellulase is cellubiase rich. If not, an external supplemental portion should be added. The intersection of the line of these two cellulase dilutions with abscissa intersect a vertical at a hypothetical 0% absorbance difference percent corresponding to a critical cellulase dilution exactly release this fixed arbitrary 2 mg glucose amount value. The factor 0.37 of this critical cellulase dilution equals to its filter paper units expressed in FPU per ml.</p><br>

A Simplified Procedures for Cellulase Filter Paper Assay

<p>A new procedures to minimize labor intensiveness and complexity that has long been recognized in cellulase filter paper activity measurement (FPAase) described by the international union of pure and applied chemistry (IUPAC)was developed. It follows the main idea of IUPAC with only exception finding at least two cellulase dilutions have optical densities slightly more and less than a reference optical density of an arbitrary fixed 2 mg absolute glucose amount after a red-ox color reaction and due to cellulase-filter paper hydrolysis. The yielding glucose amount difference as compared to this reference is expressed in terms of absorbance difference percent determined by 3,5-dinitrosalicylic acid (DNS), in case of cellulase is cellubiase rich. If not, an external supplemental portion should be added. The intersection of the line of these two cellulase dilutions with abscissa intersect a vertical at a hypothetical 0% absorbance difference percent corresponding to a critical cellulase dilution exactly release this fixed arbitrary 2 mg glucose amount value. The factor 0.37 of this critical cellulase dilution equals to its filter paper units expressed in FPU per ml.</p><br>

Newly Isolated Penicillium ramulosum N1 Is Excellent for Producing Protease-Resistant Acidophilic Xylanase

<i>Penicillium ramulosum</i> N1 was isolated from decaying wood. This strain produces extracellular xylanases and cellulases. The highest activities of xylanases (250 U/ml) and carboxymethyl cellulose (CMCase; 6.5 U/ml) were produced when 1% barley straw was added as a carbon source. The optimum temperature and pH for xylanase activity was 55 and 3.0°C, respectively. The xylanases exhibited strong protease resistance. CMCase revealed maximum activities at pH 3.0 and in the range of 60-70°C. Filter paper activity was optimally active at pH 5.0 and 55°C. The zymograms produced by the SDS-PAGE resolution of the crude enzymes indicated that there are four bands of protein with xylanase activity and three bands of proteins with endoglucanase. The results revealed that <i>P. ramulosum</i> N1 is a promising acidophilic and protease-resistant xylanase-producing microorganism that has great potential to be used in animal feed and food industry applications.

Morphogenesis and Production of Enzymes byPenicillium echinulatumin Response to Different Carbon Sources

The effect of different carbon sources on morphology and cellulase and xylanase production ofPenicillium echinulatumwas evaluated in this work. Among the six carbon sources studied, cellulose and sugar cane bagasse were the most suitable for the production of filter paper activity, endoglucanases, xylanases, andβ-glucosidases. However, sucrose and glucose showedβ-glucosidase activities similar to those obtained with the insoluble sources. The polyacrylamide gels proved the enzymatic activity, since different standards bands were detected in the media mentioned above. Regarding morphology, it was observed that the mycelium in a dispersed form provided the greatest enzymatic activity, possibly due to greater interaction between the substrate and hyphae. These data are important in understanding the physiology of fungi and could contribute to obtaining enzyme with potential application in the technology of second generation ethanol.

Cellulase and Xylanase Production by Penicillium echinulatum in Submerged Media Containing Cellulose Amended with Sorbitol

The present work investigated the use of sorbitol as a soluble carbon source, in association with cellulose, to produce cellulases and xylanases in submerged cultures of Penicillium echinulatum 9A02S1. Because cellulose is an insoluble carbon source, in cellulase production, there are some problems with rheology and oxygen transfer. The submerged fermentations containing media composed of 0, 0.25, 0.5, 0.75, and 1% (w/v) sorbitol and cellulose that were added at different times during the cultivation; 0.2% (w/v) soy bran; 0.1% (w/v) wheat bran; and a solution of salts. The highest filter paper activity (FPA) ( IU·mL−1) was obtained on the seventh day in the medium containing 0.5% (w/v) sorbitol and 0.5% (w/v) cellulose added 24 h after the start of cultivation. However, the CMCases showed an activity peak on the sixth day ( IU·mL−1) in the medium containing 0.75% (w/v) sorbitol and 0.75% (w/v) cellulose added after 12 h of cultivation. The xylanases showed the highest activity in the medium with 0.75% (w/v) sorbitol and 0.25% (w/v) cellulose added 36 h after the start of cultivation. This strategy enables the reduction of the cellulose concentration, which in high concentrations can cause rheological and oxygen transfer problems.