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Xylanase production from a Bacillus subtilis gene cloned into a strain of Escherichia coli was monitored. Although this gene was expressed in E. coli at several temperatures, efficient xylanase secretion did not occur; the observed protein release apparently depended on cell leakage or lysis. Screening for a better microbial protein secretor free of cellulase selected Bacillus cereus No. 518. A bidirectional vector plasmid (pMK3) was employed to carry the cloned gene into this B. cereus strain. Transformation was carried out by electroporation. Total xylanase production by the new pMK3-borne gene in B. cereus was similar to that from E. coli but the xylanase was shown to be normally secreted. The xylanase gene products from the E. coli and B. cereus hosts were shown to function identically. Both xylanases improved the delignification of unbleached softwood and hardwood kraft pulps, thus reducing the Cl2 required to achieve a given degree of bleaching, without altering the physical properties of the fibers. Using a target kappa number (lignin content) of 5, xylanase pretreatment of aspen kraft (chemical) pulp led to a 22% savings of chlorine. Adsorbable organic halogens in the bleachery effluent were also lowered by more than 50%.Key words: Bacillus subtilis, endoxylanase gene, bleaching, kraft pulp properties.

A mixed culture in which cellulose is capable of being converted to methane and carbon dioxide was obtained from an inoculum procured from a sewage-treatment plant and maintained in a synthetic medium containing tissue paper and an inorganic salt and vitamin mixture. The culture was tested for its ability to degrade 12 different paper and cotton products under batch conditions in 3-ℓ anaerobic fermenters. This culture degraded 6–8 mmol/ℓ per week of cellulose, expressed as glucose equivalents, with total gas yields of 0.3 m3/kg of cellulose degraded. The gas produced contained between 56 and 59% of methane. Maximum cellulose degradation occurred at chemical oxygen demand: nitrogen: phosphorus level of 80:5:1 and was adversely affected by high stirring rate. Also the presence of higher proportions of lignin in cellulose products adversely affected the ability of this culture to degrade cellulose.

Multiple electrophoretic species of arylsulfatase were demonstrated in sonicates of Proteus rettgeri by paper electrophoresis. Subsequent treatment of the sonicates with streptomycin sulfate and DEAE-cellulose chromatography resolved enzyme activity into four fractions. Electrophoretic examination of each chromatographic fraction revealed sulfatase heterogeneity comparable to that of the initial extract. These electrophoretic components differed in their general response to a number of arylsulfatase inhibitors. The results are discussed in relation to the finding of multiple species of arylsulfatase in other biological systems.

The reaction of Paramecium caudatum to solutions containing the ash of cigarette paper was consistent with the response of Paramecium aurelia previously reported. After this confirmation, both these species of Paramecium were used as test organisms during a search for the causative agents for the "Paramecium reaction". A qualitative spectrochemical analysis of cigarette paper ash showed that its chief constituent is calcium (97%) with 12 other detected elements present at levels too low to elicit any response from paramecia. Hydroxyl ions in concentrations equivalent to 0.001 M Ca(OH)2, 0.0008 M KOH, or 0.002 M NaOH simulated but did not duplicate the effect of standard cigarette paper ash solution (as of one cigarette paper in 20 ml water). While neutralization caused the three hydroxide solutions to lose their toxicity, the toxicity of cigarette paper ash solution was not appreciably affected by neutralization. Equivalent solutions of facial tissue, filter paper, and a paper towel, burned the same way as cigarette paper, were used as controls. Each of them affected paramecia differently and none simulated the Paramecium reaction. Further, a comparison between ashes from a paperless cigarette, a whole cigarette, and one with two extra roll-your-own wrappers demonstrated a dependence of the Paramecium reaction on cigarette paper ash as well as a direct correlation between the reaction intensity and the amount of cigarette paper ash present.