Nickel Inhibition of the Growth of a Sulfur-oxidizing Bacterium Isolated from Corroded Concrete

A sulfur-oxidizing bacterium strain NB1-3 isolated from corroded concrete was a Gram negative, non-spore-forming, and rod-shaped bacterium (0.5–1.0x 1.5–2.0μm) with a polar flagellum. Strain NB1-3 had its optimum temperature and pH for growth at 30°C and 3.0–4.0, respectively. Strain NB1-3 had enzyme activities that oxidized elemental sulfur, thiosulfate, tetrathionate, and sulfide and the activity to incorporate ¹⁴CO₂ into the cells. The mean G+C content of the DNA was 52.9 mol%. These results indicate that strain NB1-3 is Thiobacillus thiooxidans. Since nickel has been known to protect concrete from corrosion, the effect of Ni on the growth of strain NB1-3 was studied. The cell growth on tiosulfate-, elemental sulfur-, or tetrathionate-medium was completely inhibited by 0.1% metal nickel or 5mM NiSO₄. Both cellular activities of elemental sulfur oxidation and CO₂ incorporation were strongly inhibited by 5mM NiSO₄. The amounts of Ni in cells with or without nickel treatment were 1.7 and 160.0 nmol/mg protein, respectively. These results indicate that nickel binds to strain NB1-3 cells and inhibits enzymes involved in sulfur oxidation of this bacterium, and as a result, inhibits cell growth.     

Mutational Analysis of the Multicopy hao Gene Coding for Hydroxylamine Oxidoreductase in Nitrosomonas sp. Strain ENI-11

The ammonia-oxidizing bacterium Nitrosomonas sp. strain ENI-11 contains three copies of the hao gene (hao ₁, hao ₂, and hao ₃) coding for hydroxylamine oxidoreductase (HAO). Three single mutants (hao ₁::kan, hao ₂::kan, or hao ₃::kan) had 68 to 75% of the wild-type growth rate and 58 to 89% of the wild-type HAO activity when grown under the same conditions. A double mutant (hao ₁::kan and hao ₃::amp) also had 68% of the wild-type growth and 37% of the wild-type HAO activity.     

Characteristics of the Kabicidin Resistant Mutant of Fusarium sp. Having a High Productivity of Alkaline Protease

In order to elucidate the biochemical mechanism of the alkaline protease accumulation from n-paraffins by a kabicidin-resistant mutant of Fusarium sp., the cell constituents and the extracellular products of the mutant strain were compared with those of the parent strain. No prominent differences in the cell constituents were observed between the parent and the mutant. From the analysis of the extracellular products, however the mutant was found to have a high productivity of some hydrolytic enzymes, such as amylase and ribonuclease, and ergosterol which is a structural constituent of fungal cell membrane. The relationship of secretion of ergosterol, resistance to kabicidin and accumulation of alkaline protease is discussed.     

Biochemical characteristics of cellulose and a green alga degradation by Gilvimarinus japonicas 12-2T,and its application potential for seaweed saccharification

ABSTRACT

Cellulose is one of the major constituents of seaweeds, but reports of mechanisms in microbial seaweed degradation in marine environment are limited, in contrast to the multitude of reports for lignocellulose degradation in terrestrial environment. We studied the biochemical characteristics for marine cellulolytic bacterium Gilvimarinus japonicas 12-2^T in seaweed degradation. The bacterial strain was found to degrade green and red algae, but not brown algae. It was shown that the bacterial strain employs various polysaccharide hydrolases (endocellulase, agarase, carrageenanase, xylanase, and laminarinase) to degrade seaweed polysaccharides. Electrophoretic analysis and peptide sequencing showed that the major protein bands on the electrophoresis gel were homologous to known glucanases and glycoside hydrolases. A seaweed hydrolysate harvested from the bacterial culture was found useful as a substrate for yeasts to produce ethanol. These findings will provide insights into possible seaweed decomposition mechanisms of Gilvimarinus, and its biotechnological potential for ethanol production from inedible seaweeds.     

Identification of a Compound in Chamaecyparis taiwanensis Inhibiting the Ice-nucleating Activity of Pseudomonas fluorescens KUIN-1

Inactivation of the ice-nucleating activity of Pseudomonas fluorescens KUIN-1 by compounds in the leaves from coniferous trees were investigated, and the inactivated material was identified. Intact cells of the strain KUIN-1 and the acetone or methanol extracts of leaves of various coniferous trees were allowed to react for 30 min at 18°C. Antinucleation compounds were obtained from Chamaecyparis taiwanensis. When the acetone extract from the leaves of coniferous trees was added to the cell suspension (about 10⁶ cells/ml) in 50 mM potassium phosphate buffer (pH 7.0), the ice nucleating temperature, T₅₀, was significantly decreased (T₅₀<-5°C). This inhibitor was isolated by using TLC, then identified as hinokitiol based on UV-VIS, IR, and mass spectral data. When intact cells of the strain KUIN-1 were incubated with hinokitiol, limonene, and α-pinene of the principal constituent of the leaves of coniferous trees in 50 mM potassium phosphate buffer (pH 7.0), the ice-nucleating activity decreased, but not in α-terpinene. Furthermore, the ice-nucleating activities from other ice-nucleating bacteria also decreased in the presence of hinokitiol. This inhibition was proportional to the concentration of hinokitinol. The pH and thermal stabilities of the ice-nucleating activity of the cells were changed by the addition of hinokitiol (10 mM).     

Chemical Synthesis and Characterization of α-N-Octanoyl and Other α-N-Acyl Colistin Nonapeptide Derivatives

Eight α-N-acyl colistin nonapeptide derivatives including three aliphatic, four aromatic and one alicyclic derivatives were synthesized by the reaction of colistin nonapeptide with corresponding acid chlorides. This acylation reaction was carried out under the condition kept restrictedly at pH 5,0 in order to introduce an acyl group only to α-amino group but not to γ-amino group existing in a colistin nonapeptide molecule. Synthetic method and several physico-chemical natures of these acyl colistin nonapeptide derivatives are given in this paper.

All of the acylated derivatives thus synthesized exhibited characteristic antimicrobial activities. Antimicrobial spectra were substantially based upon a gram-negative type and not so much altered by chemical structures of acyl groups which were considerably differentiated from each other such as cyclic or chain form. Thus, more possible response of carbon size than its structure to the antimicrobial effectiveness was inferred. In spite of almost no toxicity and feeble antimicrobial activity of colistin nonapeptide itself, these acylated colistin nonapeptide derivatives showed a toxicity against mice at a dose of 16.9~70 mg/kg in LD₅₀, which, however, was inferior to the toxicity of colistin sulfate, possibly correspondent to their much weaker antimicrobial activities, as a whole. Hence, it seems likely that acyl part of these acylated colistin nonapeptide derivatives including that of colistin is seriously responsible for the biological activities.     

Lactobacillus helveticus SBT2171 alleviates allergic symptoms in a murine model for pollen allergy

ABSTRACT

Lactic acid bacteria are known to have various health-promoting effects and are highly expected to find applications in anti-allergic food materials. In this study, we focused on Lactobacillus helveticus SBT2171 (LH2171), which reportedly modifies some unique immune responses and ameliorated symptoms of patients allergic to mites and house dust in the previous studies. We examined the effect of LH2171 on cytokine production by antigen-stimulated murine naïve splenocytes in vitro and demonstrated that it inhibited IL-4 and IL-13 production while enhancing IFN-γ and IL-10 production. Then, we examined the anti-allergic effect of LH2171 in vivo using a murine model of pollen allergy and found that LH2171 reduced the sneezing frequency when orally administered to mice. We successfully confirmed the immune modulatory activity of LH2171 and its anti-allergic activity against inhaled antigens. These evidences would contribute to identifying the anti-allergic mechanism of LH2171.

Abbreviations: ALDH: aldehyde dehydrogenase; EGCG: epigallocatechin gallate; LAB: lactic acid bacteria; LH2171: Lactobacillus helveticus SBT2171; NALT: nasal-associated lymphoid tissue; OVA: ovalbumin     

The reproductive phenology of nine species of Rhodophyta in the subtidal region of the Isle of Man

Monthly samples of about 40 separate plants of each species were collected from 1 to 3 m below lowest astronomical tide on Port Erin breakwater, Isle of Man, Irish Sea. In three species growing on rock, Plocamium cartilagineum, Cryptopleura ramosa and Callophyllis laciniata, about 90% of the plants were fertile in late summer but less than 10% in spring although some fertile plants were always present. Delesseria sanguinea and Odonthalia dentata, also epilithic, had a winter sporing season, Odonthalia extending into late spring, and all plants were sterile in summer. Three species growing epiphytically, Palmaria palmata, Membranoptera alata and Phycodrys rubens, reproduced maximally in the first half of the year at the time when the stipes of the host species, Laminaria hyperborea, grow fastest. Only Palmaria had a sterile season, late summer. The encrusting Cruoria pellita showed little seasonality. The first three species, which reproduce mainly when the sea temperature is above average, are in the northern part of their geographical range. The remaining species (apart from Cruoria) reproduce mainly at low temperatures and are in the southern half of their ranges. Male plants appear to be in a minority in all species, presumably because they were manifest for a shorter period than carposporic plants. They appeared first after sterile periods and were absent as sporing declined. Plocamium and to a lesser extent Cryptopleura show an extremely high preponderance of tetrasporophytes in the population. This is attributed to perennation and some factor disallowing the survival of most of the tetraspores.     

联苯菊酯降解菌株BF-3的分离筛选、鉴定和降解特性

【背景】联苯菊酯是人工合成的类似天然除虫菊素的一种仿生杀虫剂,近年来被广泛应用于农业病虫害的防治。联苯菊酯化学性质较稳定,在环境中的残留期长,是我国出口果蔬、茶叶中残留较严重的农药之一。微生物降解具有降解速度快、无二次污染等优点,被认为是有效去除农药残留的绿色生产技术。【方法】通过富集驯化,从农药厂排污口的污泥中筛选分离能够降解联苯菊酯的菌株,经形态学观察、生理生化特性测定及16SrRNA序列分析对其进行鉴定,通过单因素试验对菌株的降解特性进行研究。【结果】分离到1株联苯菊酯的降解菌株BF-3。菌株BF-3为革兰氏阳性菌,被鉴定为蜡状芽孢杆菌;该菌株在联苯菊酯质量浓度为100mg·L^-1的无机盐液体培养基（MSM）中呈现s型生长,7d后对联苯菊酯的降解率达到87．9％。单因素试验表明,菌株最适降解条件为pH7．0、30oC,初始菌株D600am=1．0。【结论与意义】蜡状芽孢杆菌BF．3能够有效降解联苯菊酯,在环境中联苯菊酯残留的生物修复方面具有应用潜力。