Isolation and partial characterization of basic proteinases from stem bromelain

Crude bromelain extracts from pineapple stems (Ananas comosus) were fractionated by two-step FPLC-cation-exchange chromatography. At least eight basic proteolytically active components were detected. The two main components F4 and F5 together with the most active proteinase fraction F9 were characterized by SDS-PAGE, mass spectroscopy, multizonal cathodal electrophoresis, partial amino acid sequence, and monosaccharide composition analysis. F9 amounts to about 2% of the total protein and has a 15 times higher specific activity against the substratel-pyroglutamyl-l-phenylanalyl-l-leucine-p-nitroanilide (PFLNA) than the main component F4. The molecular masses of F4, F5, and F9 were determined to 24,397, 24,472, and 23,427, respectively, by mass spectroscopy. Partial N-terminal amino acid sequence analysis (20 amino acids) revealed that F9 differs from the determined sequence of F4 and F5 by an exchange at position 10 (tyrosineserine) and position 20 (asparagine glycine). F4 and F5 contained fucose, N-acetylglucosamine, xylose, and mannose in ratio of 1.02.01.02.0, but only 50% of the proteins seem to be glycosylated, whereas F9 was found to be unglycosylated. Polyclonal antibodies (IgG) against F9 detected F4 and F5 with tenfold reduced reactivity. ThepH optimum of F4 and F5 was betweenpH4.0 and 4.5 and for F9 close to neutralpH. The kinetic parameters for PFLNA hydrolysis were similar for F4 (K _m 2.30 mM,k _cat 0.87 sec^–1 and F5 (K _m 2.42 mM,k _cat 0.68 sec^–1), and differed greatly from F9 (K _m 0.40 mM,k _cat 3.94 sec^–1).Dedicated to H. Tschesche, Bielefeld, Germany, on behalf of his 60th anniversary.     

Regulation of the xylan-degrading apparatus of Cellvibrio japonicus by a novel two-component system

The microbial degradation of lignocellulose biomass is not only an important biological process but is of increasing industrial significance in the bioenergy sector. The mechanism by which the plant cell wall, an insoluble composite structure, activates the extensive repertoire of microbial hydrolytic enzymes required to catalyze its degradation is poorly understood. Here we have used a transposon mutagenesis strategy to identify a genetic locus, consisting of two genes that modulate the expression of xylan side chain-degrading enzymes in the saprophytic bacterium Cellvibrio japonicus. Significantly, the locus encodes a two-component signaling system, designated AbfS (sensor histidine kinase) and AbfR (response regulator). The AbfR/S two-component system is required to activate the expression of the suite of enzymes that remove the numerous side chains from xylan, but not the xylanases that hydrolyze the beta1,4-linked xylose polymeric backbone of this polysaccharide. Studies on the recombinant sensor domain of AbfS (AbfS(SD)) showed that it bound to decorated xylans and arabinoxylo-oligosaccharides, but not to undecorated xylo-oligosaccharides or other plant structural polysaccharides/oligosaccharides. The crystal structure of AbfS(SD) was determined to a resolution of 2.6A(.) The overall fold of AbfS(SD) is that of a classical Per Arndt Sim domain with a central antiparallel four-stranded beta-sheet flanked by alpha-helices. Our data expand the number of molecules known to bind to the sensor domain of two-component histidine kinases to include complex carbohydrates. The biological rationale for a regulatory system that induces enzymes that remove the side chains of xylan, but not the hydrolases that cleave the backbone of the polysaccharide, is discussed.     

A new dermatophyte,Thallomicrosporon kuehnii Benedek, 1963

Summary A new species of a dermatophyte,Thallomicrosporon kuehnii Benedek, 1963, is described. In contradistinction to all other known dermatophytes, it reproduces exclusively by thallospores: 1) large, fusiform, macroconidia-like structures, 2) arthrospores and, 3) chlamydospores. Microconidia are present, rodlike or pyriform. The elicitation of a perfect form of fructification in hair-on-soil cultures was unsuccessful.

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Zusammenfassung Eine neue Art der Dermatophyten,Thallomicrosporon kuehnii Benedek, 1963, wird beschrieben. Sie unterscheidet sich von allen anderen Hauptilzen durch den Umstand, dass sie sich ausschliesslich durch Thallosporen reproduziert: 1) Formen, ähnlich fusiformen Makrokonidien; 2) Arthrosporen und 3) Chlamydosporen. Mikrokonidien werden gebildet. Das Hervorbringen der perfekten Phase der Fruktifikation in Kulturen von Haar an Humus ist nicht gelungen.

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Résumé Une espèce nouvelle des dermatophytes,Thallomicrosporon kuehnii Benedek, 1963, est présentée. Elle est distincte de tous les autres dermatophytes par le fait de sa reproduction exclusive par des thallospores: 1) des structures imitant des macroconidies fusiformes; 2) des arthrospores et 3) des chlamydospores. Des microconidies sont présentes. L'élicitation de la phase perfecte de fructification en culture des poils en sol n'a pas réussi.

     

Selective,chemical epilation in the treatment of tinea capitis as contrasted with the diffuse X-ray epilation

Summary After almost a decade of practical experience with the selective, chemical epilation as a most advanced method in the treatment of tinea capitis, a progress report is presented. The author's additional material covered 153 more cases of tinea microsporina, while DoctorGimby's material encompassed 849 cases during an epidemic of tinea microsporina at Sault St. Marie, Ontario, Canada.The formula of the Compound for selective, chemical epilation is given in the body of the paper. Despite the fact that the in vitro tests with its single ingredients (Copper sulfate, Chloral hydrate) revealed some fungistatic properties, the practical, clinical application of this preparation has nothing to do with its possible antifungal effect.The efficiency of the Compound is based solely upon its selective, chemical, epilatory power on fungus infected hairs only. The Compound completely penetrates the hair follicle in its whole depth and saturates the aerial and root portions of the hair from within. The infected hairs — and only the infected ones — are separated from the papila pili so that they can be simply brushed out from the scalp. The fungi are not killed by this method, because they cannot be killed in human tissues by any method.The Compound is neither an irritant, nor a sensitizer; it has no systemic effect. The total duration of the treatment with the Compound, if correctly and conscientiously carried out, is from two to four weeks.Simple instructions for the guardians are presented for the purpose to properly carry out the home-treatment.

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Zusammenfassung Nach der praktischen Erfahrung von beinahe einem Jahrzehnt mit der selektiven, chemischen Epilation, als die meist entwickelte Methode in der Behandlung der mykotischen Infektion der behaarten Kopfhaut, wird dieser Fortschrittsbericht erstattet. Des Autors Krankengut umfasste 153 zusätzliche Fälle von Tinea microsporina, während auf 849 Fälle sich DoktorGimby's Krankengut während einer Epidemie dieser Krankheit in Sault St. Marie, Ontario, Canada erstreckte.Die Formel des Mittels für die selektive, chemische Epilation ist in der Arbeit gegeben. Wie wohl die in vitro Tests mit den einzelnen Bestandteilen des Mittels (Kupfersulfat, Chloralhydrat) etwas fungistatische Wirkung gezeigt hatten, hat die praktische, klinische Anwendung dieses Mittels mit der möglichen, antifungalen Wirkung nichts zu tun.Die Wirksamkeit des Mittels beruht allein auf seiner selektiven, chemischen Eingenschaft, lediglich die Pilz-infizierten Haare von der Kopfhaut zu entfernen. Das selektive Mittel dringt in den Haarfollikelapparat in seine ganze Tiefe ein und durchtränkt den Haarschaft und die Haarwurzel in ihrer ganzen Dicke. Die infizierten Haare, — und nur die infizierten, — werden von der Papilla pili getrennt, so dass sie von der Kopfhaut einfach herausgebürstet werden können. Die Pilze werden dabei nicht getötet, weil man sie mit keiner Methode in dem menschlichen Gewebe töten kann.Weder reizt das selektive Mittel das Gewebe, noch macht es empfindlich. Es hat keine Systemwirkung. Die Dauer der Behandlung mit dem selektiven Mittel, wenn es genau und gewissenhaft angewendet wird, ist zwei bis vier Wochen.Eine einfache Anweisung für die Eltern für die genaue Ausführung der Behandlung ist gegeben.