Chemical Studies on Amino—Carbonyl Reaction

When N-n-butyl-D-xylosylamine was heated with acetic acid in methanol at 55~70°G, it decomposed to N-n-butyIpyrrole-2-aldehyde,** through 3-deoxy-d-pentosulose as an intermediate. d-Xylose and methylamine in neutralized aqueous solution at 65~100°C also formed N-methylpyrrole-2-aldehyde. N-n-Butyl-l-rhamnosylamine, in a mixture of methanol and acetic acid, formed the corresponding pyrrolealdehyde, l-n-butyl-5-methylpyrrole-2- aldehyde, at the almost same rate as did N-xyloside. On the contrary, N-n-butyl-d-glucosylamine, under the same condition, did not form any detectable amount of the corresponding pyrrolealdehyde, but formed complicated products. A formation mechanism of the pyrrolealdehydes from 3-deoxyosulose and amine was proposed.     

Studies of Antarctic yeast for β-glucosidase production

Moss and lichen samples from the region of the Bulgarian base on Livingston Island, Antarctica were examined for the presence of yeasts. Six pure cultures were obtained. They were screened for -glucosidase production and two of them were selected. These were identified as Cryptococcus albidus AL₂ and C. albidus AL₃, according to their morphology, reproductive behaviour, and growth at different temperatures, salt concentrations, nutritional characteristics and various biochemical tests. These strains were examined for biosynthesis of -glucosidase on different carbon sources under aerobic conditions. High exocellular and endocellular activities were obtained when they were grown on cellobiose, methyl--D-glucopyranoside and salicin. The time course of growth and -glucosidase production of the yeast was examined by cultivation in a medium with cellobiose under aerobic conditions at temperatures 18 and 24 °C for 96 h. Cryptococcus albidus AL₂ and C. albidus AL₃ synthesized exocellular enzyme, respectively 58.33 and 55.83 U/ml and endocellular enzyme 137.75 and 205.34 U/ml at 24 °C for 72 h of the cultivation.     

Theoretical Studies on α-Helix—DNA Interactions

Abstract

The ¹H NMR relaxation effects produced by paramagnetic Cr(III) complexes on nucleoside 5′-mono- and -triphosphates in D₂O solution at Ph′=3 were measured. The paramagnetic probes were [Cr(III)(H₂O) ₆]³⁺, [Cr(III)(H₂O)₃ (HATP)], [Cr(III)(H₂O)₃(HCTP)] and [Cr(III) (H₂O)₃(UTP)^?, while the matrix nucleotides (0.1 M) were H₂AMP, HIMP^?, and H₂ATP^2-. For the aromatic base protons, the ratios of the transverse to longitudinal paramagnetic relaxation rates (R_2p/R_1p) for the [Cr(III)(H₂O)₆]³⁺/H₂ATP^2-, [Cr(III)(H₂O)₃(HATP)]/H₂ATP^2-, [Cr(III)(H₂O)₃(HCTP)]/H₂ATP² and [Cr(III)(H₂O)₃(UTP)]^?/H₂ATP² systems were below 2.33 so the dipolar term predominates. For a given nucleotide, R_1p for the purine H(8) signal was larger than for the H(2) signal with the [Cr(III)(H₂O)₆]³⁺ probe, while R_1p for the H(2) signal was larger with all the other Cr(III) probes. Molecular mechanics computations on the [Cr(III)(H₂O)₄(HPP)(α,β)], [Cr(III)(NH₃)₄(HPP)(α,β)], [Co(III)(NH₃)₃(H₂PPP)(α,βγ)] and [Co(III)(NH₃)₄(HPP)(α,β)] complexes gave calculated energy-minimized geometries in good agreement with those reported in crystal structures. The molecular mechanics force constants found were then used to calculate the geometry of the inner sphere [Cr(III)(H₂O)₆]³⁺ and [Cr(III)(H₂O)₃(HATP)(α,βγ)] complexes as well as the structures of the outer sphere [Cr(III) (H₂O)₆]³⁺-(H₂AMP) and [Cr(III)(H₂O)₆]-(HIMP)^? species. The gas-phase structure of the [Cr(III)(H₂O)₃(HATP)(α,βγ)] complex shows the existence of a hydrogen bond interaction between a water ligand and the adenine N(7) (O…N = 2.82 Å). The structure is also stabilized by intramolecular hydrogen bonds involving the -O(2′)H group and the adenine N(3) (O…N = 2.80 Å) as well as phosphate oxygen atoms and a water molecule (O…O = 2.47 Å). The metal center has an almost regular octahedral coordination geometry.

The structures of the two outer-sphere species reveal that the phosphate group interacts strongly with the hexa-aquochromium probe. In both complexes, the nucleotides have a similar “anti” conformation around the N(9)-C(l′) glycosidic bond. However, a very important difference characterizes the two structures. For the (HIMP)^? complex, strong hydrogen bond interactions exist between one and two water ligands and the inosine N(7) and O(6) atoms, respectively (O…O = 2.63 Å O…N = 2.72, 2.70 Å). For the H₂AMP complex, the [Cr(III) (H₂O)c]³⁺ cation does not interact with N(7) since it is far from the purine system. Hydrogen bonds occur between water ligands and phosphate oxygens. The Cr-H(8) and Cr-H(2) distances revealed by the energy-minimized geometries for the two outer sphere species were used to calculate the R_1p values for the H(8) and H(2) signals for comparison with the observed R_1p values: 0.92(c), 1.04(ob) (H(8)) and 0.06(c), 0.35(ob) (H(2)) for H₂AMP; and 3.76(c), 4.53(ob) (H(8)) and 0.16(c), 0.77(ob) s^?1 (H(2)) for HIMP^?. These results suggest that the dynamic relaxation effects can be only partially understood with molecular mechanics computations, although the success of the geometry calculations suggests that future efforts in the development of computational methods are justified.     

Studies on anther cultures of tomato —Lycopersicon esculentum Mill

The anthers of three genotypes ofLycopersicon esculentum, viz. cv. HS-101, cv. HS-102 and an F₁ hybrid (Montfavet 63-4xHS-101) in different stages of development were cultured in various defined nutritive media. Only anthers containing microspores in the early uninucleate stage were found to respond with the culture medium in the formation of androgenic callus. The DGII medium with 2 mg l⁻¹ NAA and 1 mg 1⁻¹ kinetin was found to be best for callus induction but MS medium supplemented with 2 mg l⁻¹ 2,4-D and 0.1 mg 1⁻¹ BAP favoured proliferation and growth of the callus. The androgenic microspores followed the ‘B’ type pathway of androgenesis in the formation of callus. Induction of tracheids in the callus could be achieved by supplementing the basal medium with NAA and kinetin or 2,4-D and BAP. Initiation of vessel elements and cambium were favoured by addition of NAA and kinetin and that of the phloem in the presence of 2,4-D and BAP in the basal medium, suggesting that the hormonal requirements for production of different elements of the vascular system in androgenic callus are different. Although roots could be induced from the callus, shoot differentiation could not be achieved under cultural conditions.     

Studies on solution NMR structure of brazzein——Secondary structure and molecular scaffold

Brazzein is a sweet-tasting protein isolated from the fruit of West African plant Pentadiplandra brazzeana Baillon. It is the smallest and the most water-soluble sweet protein discovered so far and is highly thermostable. The proton NMR study of brazzein at 600 MHz (pH 3.5, 300 K) is presented. The complete sequence specific assignments of the individual backbone and sideehain proton resonances were achieved using through-bond and through-space eonneetivities obtained from standard two-dimensional NMR techniques. The secondary structure of brazzein contains one α-helix (residues 21—29), one short 3_(10)-helix (residues 14—17), two strands of antiparallel β-sheet (residues 34—39, 44—50) and probably a third strand (residues 5—7) near the N-terminus. A comparative analysis found that brazzein shares a so-called 'eysteine-stabilized alpha-beta' (CSαβ) motif with scorpion neurotoxins, insect defensins and plant γ-thionins. The significance of this multi-function motif, the possible active sites an     

Studies on solution NMR structure of brazzein——Secondary structure and molecular scaffold

Brazzein is a sweet-tasting protein isolated from the fruit of West African plantPentadiplandra brazzeana Baillon. It is the smallest and the most water-soluble sweet protein discovered so far and is highly thermostable. The proton NMR study of brazzein at 600 MHz (pH 3.5, 300 K) is presented. The complete sequence specific assignments of the individual backbone and sidechain proton resonances were achieved using through-bond and through-space connectivities obtained from standard two-dimensional NMR techniques. The secondary structure of brazzein contains one alpha-helix (residues 21-29), one short 3(10)-helix (residues 14-17), two strands of antiparallel beta-sheet (residues 34-39, 44-50) and probably a third strand (residues 5-7) near the N-terminus. A comparative analysis found that brazzein shares a so-called 'cysteine-stabilized alpha-beta' (CSalphabeta) motif with scorpion neurotoxins, insect defensins and plant gamma - thionins. The significance of this multi-function motif, the possible active sites and the structural basis of themostability were discussed.     

Studies on Cow’s Urine

An oil obtained from cow’s urine was examined by means of gas chromatography. Ethylbenzene, phenol, m-cresol, p-cresol, and p-ethylphenol were identified as the major components of the oil, while there were at least four components still remaining unknown.

A hypothesis concerning the degradation of equol,¹⁾ 7-hydroxy-3-(4’-hydroxy) chroman, to p-cresol and p-ethylphenol in the urine was proposed.     

Studies on the ɛ-Lysine Acylase

In order to study the enzymatic properties of the ?-lysine acylase in Achromobacter pestifer EA, experiments were carried out with the pure enzyme preparation. As a result of the investigation, it was found that this enzyme readily hydrolyzes ?-N-acyllysine and shows no α-amino acylase activity, and that the enzyme is not specifically activated by metal ions unlike the other acylases.     

Studies on α-Glucosidase in Rice

Two kinds of αglucosidase which were homogeneous in disc electrophoretic and ultra-centrifugal analysis were isolated from rice seeds by means of ammonium sulfate fractionation and CM-cellulose, Sephadex G–100 and DEAE-cellulose column chromatography and designated as α-glucosidase I and α-glucosidase II.

Both α-glucosidases hydrolyzed maltose and soluble starch to glucose and showed same optimal pH (4.0) on the both substrates. In addition, both enzymes acted on various α-linked gluco-oligosaccharides and soluble starch but little or not on α-linked hetero-glucosides and α-l,6-glucan (dextran).

Activity of the enzymes on maltose and soluble starch was inhibited by Tris and erythritol. α-Glucosidase II was more sensitive to the inhibitors than α-glucosidase I.

Km value for maltose was 1.1 mM for α-glucosidase I and 2.0 mM for α-glucosidase II.     

Studies on the Chinese Woodsiaceae (3)—Phylogeny and Speciation

The history of the study on the woodsiodes is briefly surveyed in the paper. The family Woodsiaceae is recognized by the present author. The relationships among the species and the probable evolution of the family are discussed based on author's cytological and comparative morphological studies, and are indicated by Wagner's method, with numerical values as the indices. Woodsiaceae may have originated from the common ancestor of modern Dicranopteris and Sticherus of Gleicheniaceae, and evolved from it into two main branches, i.e., Woodsia and Protowoodsia. The origin of species through polyploid series are discussed, and W. andersonii, W. subcordata, W. alpina and Cheilanthopsis indosiosa considered as fertile allopolyploids; the probable way of speciation is also suggested for these species.     

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 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.