Studies on the ɛ-Lysine Acylase

?-Lysine acylase of Achromobacter pestifer EA was purified by fractionations with ammonium sulfate and acetone, and by vertical zone electrophoresis. As a result, this bacterial ?-lysine acylase was obtained as an electrophoretically homogeneous protein, specific activity of which is the highest among ?-lysine acylases ever reported.     

Studies on the ɛ-Lysine Acylase

Since Achromobacter pestifer EA isolated from soils shows markedly high ?-Iysine acylase activity compared with those of the other microorganisms ever tested, cultural conditions for the production of this enzyme were investigated.

As a result, it was confirmed that simple medium containing 1% peptone, 5% glucose and some inorganic salts is most suitable for the enzyme production and that much more ?-Iysine acylase is produced by shaken culture or submerged culture in jar fermentor than by stationary culture. α-Amino acylase activity in this organism was also studied.     

Studies on the Flavors in Saké

A very small amount of vanillin was found in Saké, but the mechanism of its formation during Saké brewing has not yet been elucidated. Therefore, shaking culture of a Saké yeast (Kyokai No. 7 strain) was carried out in the Hayduck’s solution containing ferulic acid which was considered to be a precursor of vanillin. By the analysis of the fermentation products, formation of p-hydroxybenzoic acid and vanillic acid was elucidated. On the other hand, in the similar experiment using vanillin in place of ferulic acid, p-hydroxybenzoic acid, p-hydroxybenzaldehyde and vanillic acid were identified.

On these results, it was suggested that vanillin might be formed as an intermediate of the degradation reaction of ferulic acid, and also, the demethoxylation of vanillin might be occurred in the fermentation of yeast.     

Studies on the Fluorescence of Saké

By spectrofluorophotometric investigation on various kinds of Saké it was found that they have at least two kinds of fluorescent colors, the one is blue, the other yellowish green. The former is always more dominant than the latter, but is unstable although the intensities of both color decrease remarkably by treatment of active charcoal. Ferulic acid and harman as the blue fluorescent components are isolated, the former from Saké in young, the latter from Saké kept for a long time under direct sun light.     

Studies on the oxidation–reduction systems of the erythrocyte

1. Starvation for 3 days produces a decrease in methaemoglobin-reductase and glutathione-reductase activities, but it does not alter the glucose 6-phosphate-dehydrogenase activity of the rat erythrocyte. 2. The feeding of a protein-free diet for 11 days causes greater changes in the first two enzymes and also a diminution of the third. Under this experimental condition slight decreases in protein and haemoglobin contents were noted. 3. The experimental animals did not show methaemoglobinaemia, probably because the activity of methaemoglobin diaphorase is preserved. 4. The GSH content was not affected but the stability of the tripeptide in the presence of an oxidizing agent was diminished.     

Studies on the thermostability of the α-amylase of Bacillus caldovelox

Summary Molecular mechanisms of thermoinactivation of the thermostable -amylase of Bacillus caldovelox were examined. Monomolecular conformational processes were found to be the major causes of thermoinactivation at both pH 4.5 and 8.0. The enzyme possessed considerable additional thermostability at pH 8.0, with half-lives of 0.75 and 7.0 min at 90° C and pH 4.5 and 8.0, respectively. The amino acid composition was examined with respect to the underlying thermostability exhibited by this enzyme. The inherent thermostability exhibited may be due to the high proline content (4.47 mol%), but more likely due to the high content of residues forming hydrophobic bonds (60.89 mol%) allied to a low content of residues responsible for ionic interactions (28.34 mol%). Offprint requests to: C. T. Kelly     

Studies on the cyanophytes of Cuba 4–6

The article comprises three subsequently published studies of the taxonomy of Cuban cyanophyte/cyanobacterial flora: (4)Lyngbyopsis willei: This oscillatorialean genus and species, described byGardner (1927) from mountain creeks in Puerto Rico, was found more than 50 years later in similar localities in Cuba. The morphological variability of the Cuban populations is described and similarity with the genusSchizothrix (sect.Inactis) discussed. —(5)Cylindrospermum-species: The morphological variability of twenty-oneCylindrospermum-populations collected in Cuba was studied, documented by graphical methods and compared with the published data. Four new taxa were recognized (C. minutissimum v.rinoi, C. zonatum, C. bourrellyi, andC. desikacharyi). The other populations belong to the variation ranges ofC. breve, C. minutissimum, C. michailovskoense, andC. muscicola v.kashmiriense.— (6)Gomphosphaerioideae-species: Fifty eight populations (9 species) of the subfamilyGomphosphaerioideae (Microcystaceae, Chroococcales) from freshwater biotopes of Cuba were evaluated: Four planktic species with a probable cosmopolitan distribution were found (Coelosphaerium kuetzinginianum, C. minutissimum, Snowella lacustris andCoelomoron pusillus), and from the genusCoelomoron Buell two new species,C. microcystoides andC. vestitus were described. The tropical planktic speciesWoronichinia fremyi forms occasionally water blooms in larger reservoirs. Two tropical species from the genusGomphosphaeria Kütz, were recognized,G. multiplex (Nyg.) c. n. andG. semen-vitis sp. n.     

Studies on the Chemical Components of the Tacca chanteraeri André

A steroidal saponin has been isolated from the roots of Taccachanteraeri Andre', a folk medicine in Yunnan, which identified as diosgenin 3β-O-α-L-rhamnopyranosyl-(1→2)- O-α-L-rhamnopyranosyl-( 1 → 3 )-O-β-D-glucopyranoside ( 1 ) with stigmasterol (2) and daucosterin (3). Our studies support I. H. Burkill and A. Takhtajan's idea according to which Taccaceae would be the closest to Dioscoreaceae and be included in Liliales.     

Studies on the histochemical „Leucine aminopeptidase“ reaction

Summary Since it was recently observed that purified cathepsin B will hydrolyse the chromogenic substrate LNA the obvious possibility of demonstrating the activity of this enzyme in tissue sections was explored. Chemical and histochemical experiments suggested that all aminopeptidase activity could be excluded by choosing a sufficiently acid pH for incubation, whereby the selective demonstration of cathepsin B activity was possible. This conclusion was further supported by different attempts to influence the histochemical reaction by preincubation with suitable activating and/or inhibiting reagents. This more specific procedure was used on sections from liver, kidney and granulomas rich in active macrophages. The suggested cathepsin B activity was demonstrated in autophagic vacuoles, and in other lysosome-like enlarged vacuoles or granules, appearing as a consequence of cellular injury as well as in actively phagocytosing macrophages.The following Abbreviations have been used AM Pase(s) aminopeptidase(s) in general - NA naphthylamides in general. — Substrates - LA Leucine amide - GPA Gly-Phe-amide - LNA L-leucyl--naphthylamide - Arg-NA L-arginyl--naphthylamide - BANA Benzoyl-dl-arginine--naphthylamide     

Studies on the Cell Wall of Piricularia oryzae†

The chemical constituents of the cell wall of Piricularia oryzae, the pathogenic fungus of rice blast disease, were studied with the aids of chemical analysis, X-ray diffraction, infra-red absorption and enzymatic degradation. The sugar constituents were identified by chromatography as glucose (62%), mannose (4%), galactose (0.5%), and hexosamine (13%). The acidic amino acid rich protein was comprised 4.6% in the cell wall. The cell wall consists of at least three different polysaccharide complexes: a) α-Heteropolysaccharide protein complex containing mannose, glucose and galactose, b) β-1,3-Glucan containing β-1, 6-linked branch, c) Chitin like substance.     

Studies on the histochemical “leucine aminopeptidase” reaction

Summary Fractionation experiments on fresh beef spleen homogenates have been performed with the aim to separate the LNA-splitting group of enzymes from the regular aminopeptidases and cathepsin C. The results indicate that the LNA-splitting enzymes are distinct from the LA-splitting aminopeptidases, and further that the group of LNA-splitting enzymes is heterogeneous with respect to the effect of added cysteine + EDTA.With 2 Figures in the Text     

Studies on the histochemical “leucine aminopeptidase” reaction

Summary A correlative chemical and histochemical study on the leucyl--naphthyl-amide-splitting activity is presented.In the first part comparison is made between homogenates and smears of strain L cells and ELD ascites tumor cells. It was concluded that only the available enzyme activity could be visualized by histochemical means. The growing tumor cells appeared to have more activity available than the strain L cells. The intensity of the histochemical LNAse reaction bore no correlation to the total enzymatic activity extractable. Most of the available activity seemed to reside in lysosome-like structures. Cell damage by repeated freezing and thawing increased the amount of activity available to substrate interaction.In the second part a comparison is made between chemical and histochemical results in four types of progressively growing transplanted or induced mouse tumors. Chemical data showed the occurrence of a mixed pattern of LNA-splitting enzymes, among which the previously described metal-dependent group constituted the main component in the tumor cells. In all cases and types of tumors a positive histochemical LNAse reaction was noticed both in growing tumor cells as well as in different parts of the stromal compartment. The intracellular reaction in tumor cells seemed confined mainly to lysosomal-like bodies. A positive stromal reaction seemed associated with the destructive (cytolytic and collagenolytic) activity at the tumor periphery. The stromal LNAse reaction was probably due to enzymes partly different from the intracellular ones. Additional enzymatic terms, including a chymotrypsin-like and/or a carboxypeptidase activity, may originate from various host cells. These contributions may be greatly increased by accumulation of inflammatory host cells as previously noted by Hess and Mottet.     

Endemism in the Flora of China—Studies on the Endemic Genera

China, under highly varied ecological conditions resulted from wide latitudinal and altitudinal ranges and from the adequate precipitation, has developed a very rich flora of great diversity. As far as flowering plants are concerned, there are 2980 genera, 214 of which, belonging to 64 families, are endemic. Among these endemic genera, there are 9 genera of taxads and conifers, 19 genera of monocots and others of dicots. Of the approximately 129 herbaceous endemic genera in the Chinese flora as a whole, about 22 (17%) are annual and 107 (83%) are biennial or perennial. In the present paper the ecological distribution, the nature of endemic genera and the centers of endemism are discussed. 1. Three types of endemic genera are distinguished, neoendemics, palaeoendemics and active epibiotics, The endemic genera in the flora of China are, for the most part, considered to be very old ones, and most of them are of temperate nature. 2. the degree of endemism in our 22 floristic regions is shown in Figure 1. The areas richest in endemic genera in the Chinese flora as a whole are the 13, 16 and 17 regions. The poorest are the 2, 4, 9 and 10 regions, and no one in the 1 and 3 regions These results on floristic richness are of general applicability. As shown in table 1, the difference in the degree of endemism among the seven Chinese floristic subkingdoms are most pronounced. 101 endemic genera are known to occur in one subkingdom, 72 to occur in two subkingdoms, and 3 to occur in four subkingdoms, only one genus widely distributed in five subkingdoms. However, there is no genus occurring in seven subkingdoms. The difference in the degree of endemism in each subkingdom reveals that the distribution of endemic genera is not well-distributed in the Chinese flora as a whole. Analysis of the vertical distribution of the 200 endemic genera of the Chinese flora bears out that there is no evident increase in endemism as a whole with altitude. 3. Three centers of endemism are found (Fig. 2). These are as follows: a). Eastern Sichuan-western Hubei center. b). Southeastern Yunnan-western Guangxi center. c). Western Sichuan-northwestern Yunnan center. The degree of endemism andcharacters of endemic genera in each center are discussed.     

Studies on sodium-potassium-activated adenosinetriphosphatase—XV the rectal gland of the elasmobranchs

• 1.1. The enzyme NaK activated adenosinetriphosphatase (NaK ATPase) was found in high activity in the rectal gland of nine elasmobranch species.
• 2.2. Species with a radial arrangement of tubules in the gland had higher activities per kg body weight than species with lobular division of the glandular parenchyma.
• 3.3. The properties of the NaK ATPase system suggest that it has a primary, rate-limiting role in the NaCl secretion by this gland.

     

Studies on the Origin of the Traditional Chinese Drug “Jinxiong”

A comparison of morphology, karyotype and the chemical compounds among “Jinxiong”, Ligasticum chuanxiong Hort. and L. chuanxiong cv. Fuxiong shows that “Jin-xiong”, like L. chuanxiong cv. Fuxiong, is a triploid and its karyotype is similar to that of L. chuanxiong with the formula K(2n)=33=24m+6sm+3st(SAT). Thin-layer chromatography of essential oils extracted from stem tubers confirms that the three taxa are extremely alike in chemical compounds. Using the features of leaves and the forms of stem tubers, they can be easily distinguished. A new triploid cultivar of L. chuanxiong Hort. i.e.L. chuan-xiong cv. Jinxiong is described for “Jinxiong” in the present paper.