Inhibitory activity of an extract from a marine bacterium Halomonas sp. HSB07 against the red-tide microalga Gymnodinium sp. （Pyrrophyta）

In recent years, red tides occurred frequently in coastal areas worldwide. Various methods based on the use of clay, copper sulfate, and bacteria have been successful in controlling red tides to some extent. As a new defensive agent, marine microorganisms are important sources of compounds with potent inhibitory bioactivities against red-tide microalgae, such as Gymnodinium sp. （Pyrrophyta）. In this study, we isolated a marine bacterium, HSB07, from seawater collected from Hongsha Bay, Sanya, South China Sea. Based on its 16S rRNA gene sequence and biochemical characteristics, the isolated strain HSB07 was identified as a member of the genus Halomonas. A crude ethyl acetate extract of strain HSB07 showed moderate inhibition activity against Gymnodinium sp. in a bioactive prescreening experiment. The extract was further separated into fractions A, B, and C by silica gel column chromatography. Fractions B and C showed strong inhibition activities against Gymnodinium. This is the first report of inhibitory activity of secondary metabolites of a Halomonas bacterium against a red-tide-causing microalga.     

极端微生物的研究进展

极端微生物是指能够生活在被认为是生命禁区,如极端的酸碱度(pH>8.5,pH<5.0)、温度(>45℃,<15℃)、压力(>50 MPa)、盐度(NaCl浓度>1.0 mol/L)、高浓度金属离子的环境中的生命类群[1].     

超嗜热古菌耐热酸性α-淀粉酶的发酵条件和酶学性质研究

对一株分离自深海热液口的超嗜热古菌(Thermococcus sp.HJ21)菌株进行了α-淀粉酶发酵条件的优化和酶学性质的研究.该菌株发酵9h后到达产酶高峰,产酶温度范围为60-90℃,其最适产酶温度为80℃.产酶pH范围为5.0-9.0,最适产酶pH为7.5.产酶NaCl浓度范围为0.5-4.0%,2.5%为最适宜NaCI浓度.糖原、淀粉、麦芽糖、酵母膏和蛋白胨促进产酶.该菌株产生的α-淀粉酶的分子量为51.4 kDa,酶的最适作用温度为95℃,在100℃仍有60%的酶活力.酶在90℃的半衰期为5 h,在100℃ 2 h仍有40%的残余酶活,酶的热稳定性不依赖Ca2+.酶的最适作用pH为5.0,pH 4.5仍有80%的酶活力,pH在5.5-7.0较稳定(80℃ 4 h).金属离子1 mmol/L的Mg2+、Co2+、Sr2+、Ba2+、K+、Na+对酶有激活作用,Cu2+、Pb2+、Hg2+、zn2+、Al3+对该酶均有抑制作用.     

皮状丝孢酵母利用大米草水解液发酵生产微生物油脂

研究了产油酵母菌皮状丝孢酵母（Trichosporon cutaneum）利用经过简单脱毒处理的大米草水解液，直接发酵生产微生物油脂。首先分别用1％、2％、4％和6％稀硫酸，在122℃水解大米草，大米草／硫酸（固／液比）为10％。结果表明，在6％的酸浓度下水解40rain后总还原糖含量最高。而1％稀硫酸在140℃下水解大米草，2．8h后，水解液中葡萄糖产量达到最高14．2g／L。然后将该条件下大米草水解液冷冻干燥浓缩为不同的倍数，作为皮状丝孢酵母菌的发酵培养基，水解液浓缩倍数为4．0时，产油量最高达6．0g／L。为高效利用大米草，将大米草开发为一种能源植物，提供了新的途径。     

海洋链霉菌分离株M095遗传转化体系的建立

研究了海洋链霉菌分离株M095的基因转移系统。利用属间接合转移将具有oriT的大肠杆菌-链霉菌穿梭质粒pIJ8600转入EscherichiacoliET12567(pUZ8002)中,获得供体菌。将供体菌与预萌发的菌株M095的孢子进行接合转移,将质粒pIJ8600转入菌株M095中,其转化率为1.99×10-4个接合转化子/受体。Southern杂交证明质粒pIJ8600已经整合到菌株M095的染色体上。同时,将来自Spirulinamaxima(Cyanophyta)的别藻蓝蛋白基因(apc)克隆在质粒pIJ8600的XbaI和BglII位点,产生质粒pAPIJ。用接合转移法将质粒pAPIJ转入菌株M095。通过SDS-PAGE分析,得到2个大小为22ku和17ku的蛋白,分别相对应于别藻蓝蛋白的α和β亚基。这些结果进一步证明菌株M095的遗传转化体系已经成功建立起来,这将为其他海洋放线菌遗传转化工作的研究奠定基础。     

Cloning,expression and characterization of a novel esterase from a South China Sea sediment metagenome

Lipolytic enzymes, including esterases and lipases, represent a group of hydrolases that catalyze the cleavage and formation of ester bonds. A novel esterase gene, scsEst01, was cloned from a South China Sea sediment metagenome. The scsEst01 gene consisted of 921 bp encoding 307 amino acid residues. The predicted amino acid sequence shared less than 90% identity with other lipolytic enzymes in the NCBI nonredundant protein database. Scs Est01 was successfully co-expressed in E scherichia coli BL21(DE3) with chaperones(dnaK-dna J-grp E) to prevent the formation of inclusion bodies. The recombinant protein was purified on an immobilized metal ion affinity column containing chelating Sepharose charged with Ni2 +. The enzyme was characterized using p-nitrophenol butyrate as a substrate. Scs Est01 had the highest lipolytic activity at 35℃ and p H 8.0, indicative of a meso-thermophilic alkaline esterase. Scs Est01 was thermostable at 20℃. The lipolytic activity of scs Est01 was strongly increased by Fe2 +, Mn 2+ and 1% Tween 80 or Tween 20.     

响应面法优化重组大肠杆菌生物合成别藻蓝蛋白(holo-apcA)的发酵条件

采用响应面法对重组大肠杆菌生物合成别藻蓝蛋白(holo-apc A)的发酵条件进行优化,提高了蛋白的表达量。以对重组别藻蓝蛋白α亚基在大肠杆菌中表达量影响较大的几种因素用于中心组合设计;中心组合设计试验结果表明诱导温度、培养基初始p H和诱导时机对诱导结果影响显著;经Design Expert 8.0软件优化后的最佳诱导条件为:诱导温度28℃,培养基初始p H 8.5,IPTG终浓度0.1 mmol/L,以及诱导时机3 h。最后验证试验得到的蛋白表达量在已有文献报道结果的基础上提高了70%~580%。中心组合设计-响应面法能够在有限的实验次数下,对影响生物过程的因子进行优化及对其交互作用进行评价,蛋白表达量达20.4 mg/L;IPTG用量由原来的1 mmol/L减少至现在的0.1 mmol/L,降低了发酵成本。     

深海芽孢杆菌E401B03 次生代谢产物分离和结构鉴定

利用硅胶柱、凝胶柱及高效液相(HPLC)等色谱方法,对采自中国南海的深海芽孢杆菌 E401B03发酵液的乙酸乙酯相浸膏进行了分离,通过1H-NMR、13C-NMR、EIMS 等方法鉴定了7个化合物: N-苯乙基乙酰胺、3-(羟基乙酰基)-吲哚、3-(2-羟基乙基)-吲哚、环(缬-缬)二肽、尿嘧啶、胸腺嘧啶、对羟基苯甲醛     

Genetic transformation of Platymonas (Tetraselmis) subcordiformis (Prasinophyceae, Chlorophyta) using particle bombardment and glass-bead agitation

Platymonas (Tetraselmis) subcordiformis is a unicellular marine green alga. It was found to be very sensitive to the herbicide Basta through a sensitivity test indicating it could be employed as a selective agent. The bar gene is a practicable and selectable marker gene. The vector containing the expression cassette of the bar gene was transferred to P. subcordiformis by both particle bombardment and glass-bead agitation and transformants were then selected using Basta. Finally, Southern blotting analysis indicated that the bar gene had been successfully integrated into the nuclear genome of P. subcordiformis using both of the transgenic techniques, with the transformation efficiency of the glass-bead method being slightly higher than that of particle bombardment. This is the first report on stable transformation of P. subcordiformis, and will improve fundamental research and enlarge application of this alga.     

一种精细化的海洋浮标数据质量控制方法

针对海洋浮标数据获取特点及常见数据错情,本文提出了一种精细化的海洋浮标温盐数据质量控制方法。通过范围检验、尖峰检验、莱因达准则等六个步骤实现浮标数据质量控制,其创新性在于有效集成了多种质量检验法,并且研究了一种融合传统尖峰检验与莱因达准则的质量检验新方法;并以中国近海观测研究网络黄、东海站浮标温盐观测数据为例,验证了本方法的有效性和适用性。该方法可广泛用于海洋浮标温盐观测数据的质量控制,识别出长时间序列数据的异常值。经过本方法质控后的浮标观测数据对于海洋科学研究、海洋气象预报、海洋灾害预警以及渔业发展等具有重要意义。