Relation between filter paper activity and saccharifying ability of a Trichoderma cellulase system

Summary Tests made to study the relation between filter paper activity and actual saccharifying ability of Trichoderma cellulases show that 30 IU/g of cellulose were sufficient to achieve over 80% hydrolysis of a 25 g/L cellulose suspension in 24 h. With the same enzyme/substrate ratio, but double the concentration of substrate, about 60% hydrolysis was achieved. End- product inhibition is one factor which seriously limits the degree of hydrolysis and therefore the concentration of sugars achievable by enzymatic hydrolysis at high levels of substrate concentration or enzyme/substrate ratio.     

Standardization of the filter paper activity assay for solid substrate fermentation

The conditions of the filter paper activity (FPA) assay were standardized for solid substrate fermentation (SSF). The FPA is a relative measure of the overall cellulose hydrolysing capacity of microbial cellulase preparations, thus reliable and comparable data may be obtained only under standardized conditions. The standardization developed for submerged fermentation (SF) cannot be translated directly to SSF. In SSF, the FPA is strongly dependent on the extraction volume and on the dilution of the enzyme in the assay. The optimal extraction volume was substrate dependent in SSF of corn fiber, spent brewing grains and wheat straw for cellulase production by Trichoderma reesei Rut C30. Other cellulolytic enzyme assays (endoglucanase, beta-glucosidase and xylanase) were much less sensitive to the extraction volume.     

A new solvent system for the separation of neutral glycosphingolipids

A solvent system and a column for high performance liquid chromatography for the separation of glycosphingolipids without derivatization is described. A column pakced with porous silica gel (latrobeads) and eluted with a mixture of isopropanol-hexane-water with increasing water content and decreasing hexane content was used. Glycosphingolipids with mono- to dodeca- or tetrakaidecasaccharides were separated within 60 min and the separation pattern was highly reproducible. The method was applied for preparative separation of highly complex glycolipids with blood group activity.     

Development and testing of a filter system for isolation of Giardia lamblia cysts from water

An inexpensive, practical, and reliable method for isolation of Giardia lamblia cysts from potable and environmental water has been developed from commercially available components. This system was successfully used to isolate cysts from well water associated with a family outbreak of giardiasis.     

Sulfhydrylcellulose: a new medium for chromatography of mercurated polynucleotides

We have synthesized a new medium, sulfhydrylcellulose, for affinity chromatography of mercurated polynucleotides. It is the product of reaction between aminoethylcellulose and N-acetylhomocysteine thiolactone. Sulfhydrylcellulose carries up to 90 mumol of SH groups/g and is inexpensive, easy to prepare, and stable. Because it binds mercurated RNA specifically and reversibly and exhibits no size discrimination, sulfhydrylcellulose should have wide applications.     

Electrophoresis of proteins on filter paper

A simplified procedure for filter paper electrophoresis is described in which disturbing factors such as evaporation, heating, buffer concentration gradients, and pH changes in the electrode vessels were reduced to a minimum. Artificial mixtures of highly purified proteins could be separated and the components isolated. The application of the method to a variety of studies on serum proteins is demonstrated. Protein concentration in paper segments was determined by two different methods of protein estimation. Curves were obtained showing the same five major peaks for normal serum as found by the classical methods of free electrophoresis. Comparisons were made of the areas of the various components under the curves obtained with the different methods. Two dimensional electrophoresis was applied to serum and serum components. It proved of value in demonstrating the heterogeneity of fractions such as the gamma-globulin of serum. The polysaccharide dextran was used as an index of the extent of electro-osmotic flow during the course of the various experiments. The ratio of the distance of electroosmotic flow and the distance of protein migration was shown to be constant for a given type of paper. For serum albumin on Munktell 20 paper this ratio was 0.35. A formula for mobilities applicable to liquid in a highly porous supporting medium is presented. Mobility values for human serum albumin at various pH levels on paper showed approximate agreement with those obtained in free solution giving a similar isoelectric point.     

Development and testing of a filter system for isolation of Giardia lamblia cysts from water.

An inexpensive, practical, and reliable method for isolation of Giardia lamblia cysts from potable and environmental water has been developed from commercially available components. This system was successfully used to isolate cysts from well water associated with a family outbreak of giardiasis.     

Bioelectrochemical system accelerates microbial growth and degradation of filter paper

Bioelectrochemical reactors (BERs) with a cathodic working potential of −0.6 or −0.8 V more efficiently degraded cellulosic material, i.e., filter paper (57.4–74.1% in 3 days and 95.9–96.3% in 7 days) than did control reactors without giving exogenous potential (15.4% in 3 days and 64.2% in 7 days). At the same time, resultant conversions to methane and carbon dioxide in cathodic working chamber of BERs by application of electrochemical reduction in 3 days of operation were larger than control reactors. However, cumulative methane production in cathodic BERs was similar to those in control reactors after 7 days of operation. Microscopic observation and 16S rRNA gene analysis showed that microbial growth in the entire consortium was higher after 2 days of operation of cathodic BERs as compared with the control reactors. In addition, the number of methanogenic 16S rRNA gene copies in cathodic BERs was higher than in control reactors. Moreover, archaeal community structures constructed in cathodic BERs consisted of hydrogenotrophic methanogen-related organisms and differed from those in control reactors after 2 days of operation. Specifically, the amount of Methanothermobacter species in cathodic BERs was higher within archaeal communities than in those control reactors after 2 days of operation. Electrochemical reduction may be effective for accelerating microbial growth in the start-up period and thereby increasing microbial treatment of cellulosic waste and methane production.