Purification and characterization of extracellular chitinase from a novel strain Aspergillus fumigatus CS-01

Twelve samples derived from different locations in south central area of China are treated by enrichment and spread-plate technique for initial screening. Seven chitinase-producing strains are isolated. The chitinase present in the culture supernatant of strain CS-01 possesses the maximum activity of 0.118 U/mL. Analysis of the morphological feature and the ITS rDNA sequence reveals that strain CS-01 belongs to Aspergillus fumigatus. Production of the chitinase is regulated by a inducible way and the maximum activity appears at 36 h in colloidal chitin culture. Purification of the chitinase is carried out by salting out, gel filtrate chromatography and anion exchange chromatography sequentially. Native-PAGE and SDS-PAGE indicate that the chitinase from A. fumigatus CS-01 is a monomer with the relative molecular mass estimated to be 4.50×10⁴. Its maximum activity appears at pH 5 and 55 °C. The chitinase is stable at pH 4.0–7.5 and below 45 °C. Foundation item: Projects(50621063, 50674101) supported by the National Natural Science Foundation of China     

Identification and fermentation optimization of protopectinase-overproducing strain Aspergillus niger CD-01 for pectin production

In order to solve the citrus peel resource waste problem and minimize the drawbacks of chemical extraction of pectin, a protopectinase-overproducing strain CD-01 for pectin production was isolated from a pit soil dumped with perished orange in Changde City, Hunan Province of China. The strain CD-01 had the same morphology and 28S rRNA gene sequence (FJ184995) as that of Aspergillus niger (ATCC 64028). It was thus identified and named as Aspergillus niger CD-01. The fermentation condition was optimized based on L₉(3⁴) orthogonal experimental design and the variances analyses. The results show that the optimal condition for producing pectin is as follows: time 36 h, temperature 35 °C, pH 5, and urea as the nitrogen source. Under this condition, the pectin yield can reach up to 24.5%. This shows a great potential of Aspergillus niger CD-01 in pectin extraction from citrus. Foundation item: Projects(50621063, 50674101) supported by the National Natural Science Foundation of China     

海参肠碱性磷酸酶的提取及粗酶的特性研究

碱性磷酸酶(Alkaline phosphatase,ALP)是最重要的磷酸酶之一,在生物体内直接参与磷酸基团的转移和代谢过程。本文研究了海参肠碱性磷酸酶粗酶的提取,并分析了其酶学特性。经含有0.2%Triton X-100的Tris-HCl缓冲液(pH 8.9)浸提、正丁醇处理、70%硫酸铵沉淀和超滤等步骤获得海参肠ALP粗酶,纯化倍数达到2.74倍,得率为63.32%。ALP粗酶的最适反应pH为11.0,在pH 10.0～12.0稳定性较好;最适反应温度为45℃,在20～45℃具有很高的稳定性。金属离子Mg2+(1～30mmol/L)、Zn2+(<10mmol/L)对海参肠ALP粗酶具有显著的激活作用;Fe3+、Fe2+、Cu2+和Mn2+(10mmol/L)可明显抑制该酶活力。EDTA-Na2、DTT、Na2WO4及Na2HPO4对海参肠ALP粗酶有明显的抑制作用,抑制程度依次为:EDTA-Na2>DTT>Na2WO4>Na2HPO4。     

Production and characterization of biosurfactant from Bacillus subtilis CCTCC AB93108

The production and properties of the biosurfactant synthesized by Bacillus subtilis CCTCC AB93108 were studied. The maximum concentration of the surfactant is 1.64 g/L when the bacteria grow in a medium supplemented with glucose as carbon sources. The isolated biosurfactant is a complex of protein and polysaccharide without lipids. It reduces the surface tension of distilled water to 45.9 mN/m, and its critical micelle concentration (CMC) is 2.96 g/L. It can stabilize emulsions of several aromatic and aliphatic hydrocarbons, such as benzene, xylene, n-pentane, n-nonane, gasoline and diesel oil. It presents high emulsification activity and stability in a wide range of temperature (4-100℃) and a long period of duration.     

Isolation and characterization of YNTC-1, a novel Alicyclobacillus sendaiensis strain

A heterotrophic acidothermophilic bacterial strain, YNTC-1, was isolated from an acidic hot spring in Tengchong, Yunan, China. YNTC-1 grows at pH value of 1.5-8.0 and temperature of 40-70 ℃, with optimal pH and temperature at 3.0 and 55 ℃, respectively. The cells of the strain are in shape of short rod, with 1.0-1.2 μm in length and 0.7-0.8 μm in diameter, and with distinct spores at both poles of each cell. The predominant fatty acids in cellular membrane of the strain are C18：1 ω7c. 16s rRNA gene analysis reveals that this strain is closely related to Alicyclobacillus sendaiensis, with over 99% sequence similarity. Based on phenotypic and genotypic analyses, YNTC-1 is identified as a member ofA. sendaiensis. Considering some important morphological and biochemical differences between strain YNTC-1 and A. sendaiensis ATCC 27009T, YNTC-1 may be proposed to be a novel subspecies of A. sendaiensis. However, this viewpoint has to be confirmed by further studies. Co-bioleaching of pyrite and chalcopyrite with strain YN22, Sulfobacillus thermosulfidooxidans, shows that strain YNTC-1 has no evident influence on bioleaching rates of these two sulphide minerals.     

Fractionation and characterization of polysaccharides from cyanobacterium Spirulina （Arthrospira） maxima in nitrogen-limited batch culture

By a sequentially selective extraction procedure, polysaccharides (PS) from cyanobacterium Spirulina (Arthrospira) maxima that can be bio-functionally healthy components were fractionated in four parts including the culture medium (PSCM), the external layers of the cell (PSEL), the cell wall (PSCW) and storage granules (PSSG). The four fractionated parts of polysaccharides were characterized by gas-liquid chromatography (GLC) equipped with FID and fused-silica capillary column (15 m×0.53 mm i. d.). The contents of PSCM, PSEL, PSCW and PSSG were about 2.0%, 2.6%, 10.0% and 52.0% to cell dry matter, respectively. Glucose was almost the only monosaccharide in PSSG and PSCW and most predominant in PSEL, while in PSCM, xylose, rhamnose and glucose were the main ones. Two uronic acids represented by glucuronic acid and galacturoic acid, six neutral monosaccharides including fucose, rhamnose, xylose, mannose, galactose and glucose, and two possible unidentified sugars were found in PSEL and PSCM. These data are valuable for the selective productions of high-added value sugars from Spirulina. Foundation item: Scholarship for Ph. D at FPMs, Belgium Biography of the first author: NIE Zhen-yuan, Engineer in biomedicine, born in 1963, majoring in biotechnology and bioengineering.     

Isolation, identification and characterization of cadmium-resistant Pseudomonas aeruginosa strain E1

Strain E₁ with resistance to 18 mmol/L cadmium (Cd), isolated from Cd-contaminated soil was identified by morphological observation, biochemical and physiological characterization and 16S rDNA sequence analysis. The resistance to heavy metals Cd, Cu, Co, Mn, Pb, Zn and 12 antibiotics was examined. The ability of removing Cd from solution was studied. The characterizations show that strain E₁ is affiliated to Pseudomonas aeruginosa (P. aeruginosa). Strain E₁ has high resistance to heavy metals and the order is found to be Cd>Mn>Zn>Cu>Pb>Co in solid media. Strain E₁ also exhibits the resistance to 12 antibiotics. Both living and non-living cells of strain E₁ can remove Cd from solution, and living cell has better biosorption than non-living cell. Foundation item: Project (50621063) supported by the National Natural Science Foundation of China; Project (2004CB619204) supported by the Major State Basic Research and Development Program of China     

Isolation and characterization of Acidithiobacillus caldus from several typical environments in China

Six strains of moderately thermophilic sulfur-oxidizing bacteria were isolated from several different typical environments in China. The identities of the isolates were confirmed by analyses of their 16S rRNA genes, and some key physiological traits. The isolates are Gram negative, rod-shaped bacteria, their optimal temperature and pH value for growth are 45-50 ℃ and 2.5-3.5 respectively. They are autotrophic and used＇elemental sulfur, sodium thiosulfate and potassium tetrathionate as electron donor, while a little glucose stimulated their growth. 16S rDNA sequences analysis reveals that the strains are phylogenetically clustered to Acidithiobacillus caldus.     

多汁乳菇多糖的分离纯化及三种食用菌多糖的抗氧化研究

以多汁乳菇子实体为原料,采用水提醇沉辅助超声破碎提取多糖,经Sevage除蛋白得到多汁乳菇多糖,并通过体外实验比较多汁乳菇多糖(refined polysaccharide from Lactarius volemus Fr.,RPLV)、白玉菇多糖(refined polysaccharide from White Hypsizigus marmoreus,RPHM)、姬菇多糖(refined polysaccharide from Pleurotus cornucopiae,RPPC)对·OH、·O^-₂、DPPH自由基的清除率及还原力来评价三者的抗氧化活性. 结果显示,RPLV的糖含量为66.44%,蛋白质含量为3.57%; RPLV、RPHM、RPPC均可较好地清除·OH、·O^-₂、DPPH自由基、增强还原力,且均随着浓度的增加而增强; 抗氧化活性从强到弱依次是RPPC、RPHM、RPLV. 研究结果表明,3种食用菌多糖尤其是RPPC均有望开发成良好的天然抗氧化剂.