Photochemical reaction of 7,8-dihydrorhodopsin at low temperatures

The photoreaction of 9-cis-7,8-dihydrorhodopsin was examined at liquid nitrogen temperatures (-180 degrees C) in order to elucidate the photochemical events in visual pigments. This rhodopsin analog was prepared by incubating 9-cis-7,8-dihydroretinal with bovine opsin in the dark. 9-cis-7,8-Dihydrorhodopsin (lambda max = 427 nm) was cooled to -180 degrees C, and then irradiated at -180 degrees C with a 390 nm light, resulting in formation of its bathochromic product (lambda max = 465 nm). This result indicates that the presence of four double-bonds adjacent to the Schiff base nitrogen is sufficient to allow formation of a bathochromic product. Thus, the mechanism of formation of bathorhodopsin (in bovine rhodopsin system) may be considered as some change of the interaction between the conjugated double-bond system from C-9 to the Schiff base nitrogen and its surrounding charges in opsin, caused by rotation of 11-12 double-bond.     

D-泛解酸内酯水解酶产生菌的筛选及产酶条件研究

筛选到一株产D-泛解酸内酯水解酶的菌株,经鉴定为串珠镶孢霉菌（Fusarium monili-forme)SW-902。产酶条件研究表明,用甘油作碳源,蛋白胨作氮源,初始pH8.0,温度26℃,摇瓶培养3d,产酶量最高,在60L发酵罐中通风发酵45h,产菌丝体生物量7.18g干菌体/L,D-泛解酸内酯水解酶酶活力达到0.92IU/g干菌体。     

Desmosome assembly in MDCK cells: transport of precursors to the cell surface occurs by two phases of vesicular traffic and involves major changes in centrosome and Golgi location during a Ca(2+) shift

Desmosome formation in MDCK cells was investigated using a Ca(2+) shift. Following preliminary treatment with cycloheximide at 37 degrees C, continued surface transport and subsequent endocytosis were minimized by incubating cells at 19 degrees C to trap nascent glycoproteins within the Golgi body. Release into high Ca(2+) medium (HCM) at 37 degrees C resulted in junction formation as well as relocation of the Golgi body and centrosomes to a subapical location. Desmosome formation occurred in two stages over 2 h, the first occurring within 30 min of the shift to HCM, in 60-nm vesicles containing chiefly Dsc2 and lower concentrations of Dsg and E-cadherin distributed to the entire cell surface. Much of this material was subsequently endocytosed. The second stage involved transport of Dsg, E-cadherin, plakoglobin, and beta-catenin, in more complex vesicles some 200 nm in size, directed to possible nucleation sites on the developing basolateral surface. Plaque proteins such as desmoplakin I/II were added subsequently. Stage-two vesicles, but possibly not those of stage one, were accessible to endocytic markers via retrograde transport from multivesicular bodies prelabeled at 19 degrees C.     

影响被孢霉产生含γ-亚麻酸油脂的几种因素

对培养条件,如氮源种类、C／N比率、pH、种子种龄与接种量对被孢霉(Mortierellasp.)M14菌株的细胞生长和油脂形成的影响作了研究。结果表明,二级发酵优于一级发酵。M14菌株二级发酵时,种子种龄以49h为适,接种量宜大(30％)。无机氮源有利于不饱和脂肪酸的产生,有机氮源有利于细胞的增殖。低C／N比率有利于菌丝体产量的提高,提高C／N比率则能促进菌体细胞内的油脂合成。pH对菌体生长、油脂产生以及γ-亚麻酸含量都有显著的影响。     

The effects of temperature, incubation atmosphere, and medium composition on arthrospore formation in the fungus Trichophyton mentagrophytes.

Several growth conditions were found to allow abundant arthrospore formation in T. mentagrophytes. These included growth at 32--37 degrees C on Sabouraud's medium (1% neopeptone, 4% glucose) and growth at temperatures below 32 degrees C solely on neopeptone or other complex peptide sources without the addition of glucose, a supplementary carbon source. Sabouraud's medium did not allow arthropsore formation at 30 degrees C under normal atmospheric conditions. However, if oxygen tension were reduced by partial replacement of air with either N2 or CO2 arthrosporulation did occur on Sabouraud's medium at 30 degrees C. The rate of germ tube elongation was lower under those conditions which supported arthrospore formation, suggesting a correlation between decreased rate of hyphal extension and arthrospore formation. Stimulation of arthrospore formation by sublethal concentrations of several antifungal agents tends to support this hypothesis.     

Regulation of flagellar motility by temperature-dependent phosphorylation of a 43 kDa axonemal protein in fowl spermatozoa.

Phosphorylation of fowl sperm proteins was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) after incubating the demembranated spermatozoa with [gamma-32P]ATP at 30 degrees C or 40 degrees C. A marked difference of phosphorylation between 30 degrees C and 40 degrees C was observed in a 43 kDa protein. This protein was slightly phosphorylated at 40 degrees C, but strongly phosphorylated at 30 degrees C in a cAMP-independent manner. The motility of demembranated spermatozoa was negligible at 40 degrees C, but vigorous movement was observed at 30 degrees C. These results showed that phosphorylation of a 43 kDa protein is likely to be a regulatory step in the maintenance of fowl sperm motility.     

Viscoelastic and light scattering studies on thermally induced sol to gel phase transition in fish myosin solutions

Viscoelastic (VE) and dynamic light scattering (DLS) analyses of fish (white croaker) myosin solutions were performed at myosin concentrations of 30 mg/mL for VE and 0.1 mg/mL for DLS at 0.6M KCl and pH 7.0 to clarify thermally induced gelation. The hydrodynamic radius R(h) considerably decreased around 30-35 degrees C. The shear modulus G was constant below 25 degrees C and increased by incubating the sample at 30 degrees C. G further increased as the temperature of the incubated sample decreased. The curves of G vs T for different time courses showed a sharp peak around 35 degrees C and a moderate peak around 60 degrees C in the heating process, while a stepwise increase in G was observed around 30 degrees C in the cooling process when the temperature was elevated to not more than 60 degrees C. No distinct stepwise change was observed once the temperature of the sample exceeded 60 degrees C. The absolute value of G strongly depended on the maximum elevated temperature and the incubation time at that temperature. The corresponding behavior of the viscosity eta was observed for each time course. Based on these results, the mechanism of thermally induced gelation of myosin solutions is discussed in view of S-S bridge formation in the head and tail portions and unwinding/rewinding of coiled-coil alpha-helices in the tail portion.     

假丝酵母尿酸酶形成条件

选出了一株产尿酸酶的产朊候丝酵母(Candida utilis)AS2．117。此菌株尿酸酶形成条件的研究表明：尿酸、黄嘌呤和鸟嘌呤对酶形成起诱导作用;玉米浆对菌株生长和酶形成起十分重要的作用;蔗糖、葡萄塘、D-甘露糖和果糖是酶形成的适合碳源;生物素对酶产生有促进作用;在含有玉米浆培养基中加入无机氮源对产酶无作用,添加有机氮略增加产酶量。尿酸酶形成最适培养基组成为(％)：蔗糖;,玉米浆3,尿酸0．1,蛋白胨0．1,生物素0．05,KCI0．1,NaCl 0.1。最适pH为6．2。在250ml三角瓶中装30ml培养基为最适。在200r／min的旋转摇床上25℃振荡培养21h,在此条件下最终酶活力可达0．6u／ml。     

Circumstantial evidence for phytoalexin involvement in the resistance of peanuts to Aspergillus flavus

Three stilbene phytoalexins, elicited by slicing and incubating imbibed peanut kernels under aerobic conditions, inhibited spore germination and hyphal extension of Aspergillus flavus with ED50 values in the range 4.9-12.8 micrograms ml-1. Phytoalexin yield was dependent on cultivar, conditions and duration of incubation after slicing, and crop history. The yield of phytoalexin from ten cultivars studied, after slicing and incubating at 25 degrees C for 24 h, ranged from 28 to 935 micrograms per g fresh weight and was negatively correlated with dry kernel colonization by A. flavus [r = -0.868 when plotted as 1n (phytoalexin concn) against 1n (percentage peanut colonization)]. When the incubation period was extended to 96 h there was no such correlation. Reduced phytoalexin yields were obtained when sliced kernels of one cultivar studied were incubated in water or at 37 degrees C, and no phytoalexin was obtained when the slices were incubated under nitrogen gas or frozen before aerobic incubation. Drought stress during pod development in four cultivars studied reduced phytoalexin yields of sliced kernels incubated at 25 degrees C for 24 h by 17-65% compared with non-stressed controls.     

Field and laboratory studies of the effect of urea on ascospore production of Venturia inaequalis (Cke.) Wint

Applications of urea after harvest but before leaf-fall restricted perithecial production by Venturia inaequalis. Immersion of detached leaves in urea appeared to be the most effective method of preventing perithecial formation, although spraying attached leaves was equally effective when leaf abscission occurred within a week of treatment. A high nitrogen content within the leaf was one of the major factors contributing to suppression. Urea-treated leaves decomposed rapidly, thus destroying the overwintering substrate for the fungus. When apple plants (clone M. 111) were sprayed in autumn with 5 % urea, followed by a second (pre-bud-burst) application at 2 %, ascospore production in the spring was suppressed. The second treatment appeared to prevent the release of ascospores from mature perithecia.     

Avirulent isolates of Corynebacterium fascians that are unable to utilize agmatine and proline

Growth of a highly virulent strain of the phytopathogen Corynebacterium fascians on rich media at 37 degrees C resulted in a loss of virulence in a majority of the population within 10 generations. Strains retained virulence during cultivation at 30 degrees C on a minimal medium with ammonia as a nitrogen source. Populations of avirulent strains on the surfaces of pea seedlings decreased, whereas the number of cells of the virulent strain increased 1,000-fold during a 3-week period. All avirulent mutants isolated by growth on rich media at 37 degrees C were unable to grow on media containing agmatine or proline as sole sources of nitrogen. The ability of the mutants to grow on pea seedlings and cause fasciation disease appeared to be related to their ability to utilize nitrogen sources available on plant surfaces.     

Avirulent isolates of Corynebacterium fascians that are unable to utilize agmatine and proline.

Growth of a highly virulent strain of the phytopathogen Corynebacterium fascians on rich media at 37 degrees C resulted in a loss of virulence in a majority of the population within 10 generations. Strains retained virulence during cultivation at 30 degrees C on a minimal medium with ammonia as a nitrogen source. Populations of avirulent strains on the surfaces of pea seedlings decreased, whereas the number of cells of the virulent strain increased 1,000-fold during a 3-week period. All avirulent mutants isolated by growth on rich media at 37 degrees C were unable to grow on media containing agmatine or proline as sole sources of nitrogen. The ability of the mutants to grow on pea seedlings and cause fasciation disease appeared to be related to their ability to utilize nitrogen sources available on plant surfaces.     

Production and properties of hemicellulases by a Thermomyces lanuginosus strain

Thermophilic fungi producing extremely high beta-xylanase and their associated hemicellulases have attracted considerable attention because of potential industrial applications. Thermomyces lanuginosus strain SSBP isolated from soil, produced beta-xylanase activity of 59 600 nkat ml-1 when cultivated on a medium containing corn cobs as substrate and yeast extract as nitrogen source. Lower beta-xylanase activities were produced after growth on other xylan substrates, sugars and soluble starch. Other hemicellulases were produced extracellularly at significantly lower levels than the beta-xylanase activity produced on corn cobs. No cellulase activity was observed. The optimal conditions for beta-xylanase production were 50 degrees C and pH 6.5, whereas 70 degrees C and between pH 5. 5 and 9.5 were optimal for beta-xylanase activity. The temperature optima for other hemicellulases were less than the xylanase with the exception of beta-mannosidase. The pH optima of the other hemicellulases were between 5.0 and 6.5. Xylanase was stable up to 70 degrees C and between pH 5.5 and 9.0 for 30 min whereas the other hemicellulase were less stable. These results suggest that the most suitable conditions for hydrolysis of hemicellulose by these enzymes would be at 50 degrees C and pH 6.0.     

Spore differentiation in a clinical strain of Trichophyton mentagrophytes

Spore differentiation and, in particular, arthrosporogenesis in a clinical strain of T. mentagrophytes was investigated using a variety of methods and by altering environmental conditions. Results are discussed with reference to the in vivo situation. Arthrospores were obtained in the presence of increased CO2 tension but not increased N2 tension. High humidity was necessary for arthrospore formation but maturity (i.e. crops of single spores) was associated with conditions of reduced humidity. Desiccation reduced arthrospore viability. Glucose and peptone based media were suitable for arthrospore formation. Arthrospores were produced at 30 degrees C and 37 degrees C, but 30 degrees C is preferred since chlamydospores were prevalent at 37 degrees C. Conditions for production of arthrospore, microconidial and mycelial suspensions are presented.     

Reproduction of rock pigeon exposed to extreme ambient temperatures

1. The breeding biology of rock pigeon (Columba livia) exposed to ambient temperatures (Ta) between 50 and 60 degrees C was investigated. 2. Four families accomplished three complete life cycles after long term daily exposure to extreme Ta, with about 100% success. 3. The steady state temperatures in the nest were 60, 58, 53 and 44.6 degrees C in the air, substrate surface, underwing, and in the egg's microenvironment, respectively. 4. At thermal conditions between 30 and 60 degrees C, egg temperature (Tegg) was regulated between 36.8 +/- 0.8 (S.D.) and 41.7 +/- 0.4 (S.D.). Tegg increases by 0.163 degrees C/1 degree C rise in Ta. 5. Mean Tb of the nonincubating parent exposed to 30-60 degrees C is 41.6 +/- 0.6 degrees C (S.D.). Under the same conditions the incubating parent regulated a significantly (P less than 0.01) lower Tb (38.8 degrees C) at 45 degrees C Ta and about 1 degree C lower Tb at 30 and 60 degrees C Ta, respectively. 6. By comparing the differences between fast (5 min) cooling of hot egg (44.8 degrees C) to slow heating (60-90 min), we could demonstrate the high sensitivity of the incubating parent to the danger of embryo overheating. 7. The significance of the adaptive behavioral and physiological mechanisms in breeding under extreme thermal conditions are discussed.     

On the activation of bovine chymotrypsinogen A. Preparation of alanine-neochymotrypsinogen and its activation to alpha-chymotrypsin

Alanine-neochymotrypsinogen was prepared by incubating 20 parts bovine pancreas chymotrypsinogen A with one part alpha-chymotrypsin in a solution containing 1 M (NH4)2SO4, 0.1 M sodium acetate, 0.05 M Tris buffer (pH 8.0) and 0.5 mg/ml soybean trypsin inhibitor. Optimal yields of NH2-terminal alanine were obtained after 60 h incubation at 4 degrees C. Ala-neochymotrypsinogen was isolated from the reaction mixture by affinity chromatography and ion-exchange chromatography on carboxymethyl-cellulose. As expected, the purified preparation was enzymatically inactive and, compared to chymotrypsinogen, had one additional NH2-terminal group identified as alanine. Ala-neochymotrypsinogen was activated by incubating with trypsin at a zymogen : trypsin ratio of 30 : 1 in 0.1 M phosphate buffer, pH 7.6 at 4 degrees C for 1 h. The fully active, stable species was identified as alpha-chymotrypsin.     

Effect of media supplements and culture conditions on inulinase production by an actinomycete strain

Streptomyces sp. GNDU 1 produced high levels of extra-cellular inulinase (0.552 IU/ml) after 24 h at pH 7.5, temperature 46 degrees C in the presence of 1% inulin. The optimum temperature and pH for enzyme activity were 60 degrees C and 5.5 respectively. Yeast extract as a nitrogen source was found to be most suitable one for inulinase production whereas ammonium ion was inhibitory to the enzymatic production. All these conditions make Streptomyces sp. GNDU 1, a potential candidate for industrial enzymatic production of fructose from inulin.     

无需覆土的蘑菇属食用菌——中国美味蘑菇

采集于新疆的“中国美味蘑菇”是一种个体巨大的野生蘑菇,迄今未有人工栽培报道。本文已成功对其进行驯化栽培并完成了它的生物学特性研究,可为今后商业化栽培提供科学依据。结果表明,中国美味蘑菇菌丝生长的最佳碳源是葡萄糖,最佳氮源是大豆蛋白胨;菌丝生长最适温度是25℃,最适pH值为6。该种蘑菇可利用稻草、麦杆、芦苇等基质进行栽培;它还有一重要的特点就是出菇可不需覆土,而覆土为一般栽培的蘑菇属种类生产中的必须环节。初步驯化表明,该种蘑菇以芦苇为基质比以稻草为基质的生物学效率高15.01%。     

Enhanced production of Penicillium expansum PED-03 lipase through control of culture conditions and application of the crude enzyme in kinetic resolution of racemic Allethrolone

Alkaline lipase production was performed in submerged fermentation by Penicillium expansum PED-03. It was found that the suitable carbon source and nitrogen source for lipase production were 0.5% starch and 4.0% soybean meal, respectively. The maximal lipase activity (850 U/mL) of production was achieved at initial pH 5.5-6.0, 26 degrees C, 72 h. Tween-80 was an effective enhancer for lipase production. Agitation speed of the fermentor played an important role, and the suitable agitation speed for lipase production was 500 r/min. The lipase was stable within the range of pH 7.0-10.0 and 20-40 degrees C, and the optimum conditions for the enzymatic reaction were 35 degrees C and pH 9.5. The enzymatic resolution of racemic allethrolone (4-hydroxy-3-methyl-2-(2-propenyl)-2- cyclopenten-1-one) was carried out by the lipase from P. expansum PED-03, and the conversion reached 48% with excellent enantioselectivity (E > 100), which showed a good application potential in the production of optically pure allethrolone.     

The Influence of n-Propanol on the Growth and Conidiation of Pestalotia rhododendri

Pestalotia rhododendri was exposed to vapours from 1 ml propanol solution in water and linear growth, formation of aerial hyphae and production of conidia were determined. A special Petri dish technique was used and maximum stimulation of conidial formation was induced by the vapours from a propanol concentration of 3–4 % (v/v) at 25°C. When propanol was added directly to the medium, a concentration of 1.2 × 10^?2M was optimal for growth and sporulation at 30°C. Sporulation stimulated by propanol was observed at temperatures from 20–32°C, with an optimum at 30°C. Certain observations indicated that an exposure to propanol for 24 hours was enough to induce a stimulated spore production. The stimulation was noticed on different media at 25°C, and was more pronounced at 30°C. One exception was observed. Propanol did not promote sporulation when the fungus was grown on maltagar at 30°C. Propanol 3 ° (v/v) in combination with the standard medium containing (NH₄)₂-tartrate as sole nitrogen source, inhibited the linear growth at 15–20°C, was inactive at 22.5° and 25°C, and stimulated growth at 27.5–31°C. The stimulatory effect was maximal at 30°C. Other media were tested at 25° and 30°C. At both temperatures stimulations of linear growth caused by propanol were observed with a medium containing KNO₃ as sole nitrogen source, and inhibitions with maltagar and another medium containing l -asparaginc as sole nitrogen source. The linear growth could be either inhibited or stimulated while the sporulation was stimulated.