利用广谱性和特异性组合诱变技术选育辅酶Q10高产菌

为提高辅酶Q10产量，采用了对细胞进行全面的非特异性诱变和针对特定途径的特异性诱变相结合的育种策略，以放射型根瘤菌(Rhizobium radiobacter)WSH2601为出发菌株，选择UV射线和亚硝基胍作为诱变剂，在筛选获得放线菌素D抗性突变株的基础上，通过进一步的诱变处理，分别获得了放线菌素D和L-乙基硫氨酸双抗性突变株、放线菌素D和维生素K双抗性突变株，以及抗放线菌素D和X-gal利用能力提高的突变株．与出发菌株相比，突变株辅酶Q10的产量提高幅度达25％-37％，其中一株放线菌素D和L-乙基硫氨酸双抗性突变株WSH-E25，胞内辅酶Q10含量和辅酶Q10总产量分别达到2．86mg g DCW^-1和40．0mg L^-1，均比出发菌株提高了37％，且遗传稳定性良好，图6表2参8     

氢化可的松高转化菌株的选育及其发酵条件

利用酮康唑抗性突变选育氢化可的松高转化菌株，即将经过紫外线诱变处理的新月弯孢霉原生质体倾注于含有酮康唑最小抑制浓度(10μmoL／L)的培养基平板上，获得了136株酮康唑抗性突变株，氢化可的松转化率高于出发菌株的有14株．其中抗性突变株KA-91，其氢化可的松转化率为出发菌株的1．42倍，达到68．05％．研究了菌株的发酵性能，优化了培养基组成和发酵工艺条件，氢化可的松转化率达到73．6％．图7表3参5     

皮革废物铬革屑的微生物学处理研究Ⅰ:降解铬革屑的微生物筛选及诱变

从不同来源的微生物中筛选出能粗效降解革屑的皱绒青霉 Ａ Ｓ３ ．１０６６ 菌株,可溶性蛋白得率达５３ ．５ ％ ;该株经紫外诱变获得高产蛋白酶诱变株 Ａ Ｓ３ ．１０６６ Ｅ,其蛋白酶活性较原始菌株高３ 倍     

钝顶螺旋藻突变株FBL的生理特性

利用倍频Ｎｄ：ＹＡＧ激光（λ＝５３２ｎｍ,ｐ＝５００ｍＷ,Ｓ＝１６０ｍＷ／ｃｍ＾２）对钝顶螺旋藻进行诱变处理,辐照１０ｍｉｎ,经分离获得藻丝开矿变成直形的钝顶螺旋突变株ＦＢＬ。对该变株ＦＢＬ的生长特性、藻多糖、氨基酸、β－胡萝卜素、叶绿素ａ等生化组成的研究。结果表明：与出发株相比,突变株ＦＢＬ生长速度加快;β－胡萝卜素含量提高２２．３％;藻多糖提高４２．０％;氨基酸增幅５．２％,氨基酸的组成比例发     

利用原生质体诱变技术筛选脱落酸高产菌株

以葡萄孢属TB-3菌株为出发菌株制备其原生质体．在纤维素酶浓度为20g／L,蜗牛酶浓度为3g／L的酶解系统中,25℃酶解3h,其原生质体制备数可达67×105mL-1,在KYM上的再生率为46．1％,在KPDA上的再生率为33．6％,经过3轮原生质体紫外线诱变后回复再生,及对大量再生突变株进行发酵筛选,获得高产稳定株TB－3H8,其发酵液中ABA的质量浓度可达1．4g／L     

卷曲霉素产生菌的改良

以卷曲霉素（ＣＰＭ）产生菌卷曲链霉菌Ｓ．ｃａｐｒｅｏｌｕｓ４００６（效价为６６４０ｕ／ｍＬ）为出发菌,经ＵＶ诱变／五氯酚浓缩,获得一株效价为７９５０ｕ／ｍＬ的营养缺陷型菌株４０６－３５,再经ＥＭＳ诱变,筛选到一株抗５０００ｕ／ｍＬＣＰＭ、效价为８５１３ｕ／ｍＬ的突变株５－４１,经原生质体ＮＴＧ诱变处理,获得一株效价为９０５０ｕ／ｍＬ的高产菌Ｎ１３菌株Ｎ１３连续传代１０代,卷曲霉素效价保持稳定通过单因子试验和多因子正交试验,确定了菌株Ｎ１３的最佳发酵条件菌株Ｎ１３在最佳发酵条件下卷曲霉素效价为９４８３ｕ／ｍＬ,比出发菌Ｓ．ｃａｐｒｅｏｌｕｓ４００６在原有发酵条件的效价提高４２８％     

泰乐菌素降解菌的分离鉴定及其系统发育分析

发酵法生产泰乐菌素过程中产生的大量废弃药渣,因残留泰乐菌素的存在,会对环境带来不利影响.采用微生物法降解药渣中残留的泰乐菌素.结果表明:从堆放泰乐菌素药渣附近土壤中分离筛选到1株高效降解泰乐菌素的菌株,经16S rDNA鉴定为无丙二酸柠檬酸杆菌(Citrobacter amalonaticus).该菌为好氧型革兰氏阴性杆菌,菌落直径0.5~1.5 mm,可利用多种糖类、醇类和氨基酸作为碳源或氮源,具有较强的耐盐、耐酸、耐碱能力,对温度的适应范围较宽.用该菌株在30℃下处理含50 mg/L泰乐菌素培养基72 h,培养基中未检到泰乐菌素的存在.说明利用微生物法可有效降解药渣中残留的泰乐菌素.     

化学诱变结合结构类似物法选育高苯丙氨酸解氨酶菌种

ＥＭＳ诱变结合结构类似物法提高含ＰＡＬ出发菌株红酵母Ｒｈｏｄｏｔｏｒｕｌａ ｓｐ．ＣＩＢＡＳＡ１４０１１００ 生物转化能力阳性率为３５．５ ％ ,部分突变株ＰＡＬ转化能力提高１ 倍以上,其中Ｅ１０５,Ｅ２４０ 具有快速转化能力,反应３ ｈＬＰｈｅ累积浓度达到１ ｇＬ左右．φ１０％ 甘油和３０ ｇＬＧｌｕ 对ＰＡＬ转化具有明显的稳定作用．用含φ１０％ 甘油和３０ ｇＬＧｌｕ 的反应液进行生物转化制备,Ｅ１０５ 和Ｅ２４０ 在２４ ｈ 期间ＬＰｈｅ 质量浓度大于２０ ｇＬ,转化率达８０ ％ ．     

微波诱变选育耐酸高效厌氧产氢菌

为获得厌氧产氢菌的高效突变株,以产氢菌H-8为原始菌株进行微波诱变处理,对微波诱变参数进行了优化,考察了突变株的遗传稳定性、产氢特性及耐酸性.经过微波(低火)物理诱变得到5株高产氢突变株HW7、HW33、HW181、HW184和HW195,多次传代实验表明, HW195是稳定的高产突变株.突变株HW195具有较好的耐酸性,在pH值为2.8时仍能生长.通过间歇发酵实验,其最大产氢量和最大产氢速率分别达到2 460 mL/L培养基和340 mL L-1 h-1,比原始菌分别提高了50.7%和41.7%.图7表2参13     

克拉维酸产生菌的改良

以棒状链霉菌Streptomyces clavu ligerus CCRC11518(ATCC27064)为出发菌,采用经典物理和化学诱变剂处理的微生物诱变技术和现代理性化筛选方法,通过筛选抗终产物结构类似物舒巴坦钠突变株、底物甘油耐受型突变株、营养缺陷型突变株,最后获得一株克拉维酸高产突变株III50,其克拉维酸摇瓶产量为834.8μg/mL,是出发菌株产量(282.4μg/mL)的2.96倍.该突变株在琼脂斜面培养基上连续转接传代8代,克拉维酸的产量保持稳定.表12参8     

Diet of<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> in False Bay,South Africa

The diet of Octopus vulgaris was analysed using instantaneous daytime observations, midden counts, and stomach contents and a total of 39 prey species were identified. From stomach contents, the most important prey species were Plagusia chabrus (64.6% IRI, index of relative importance) and Haliotis midae (21.6% IRI). Crustaceans were the most frequently found prey group in octopus stomachs (63.6% frequency of occurrence), followed by molluscs (37.6%), teleosts (11.2%), and polychaetes (10.8%). Prey size and diversity increased with increasing octopus size. From middens, the mean shell lengths of H. midae consumed by small, medium, and large O. vulgaris were 53.3, 72.6, and 86.0 mm, respectively. Compared with stomach contents, midden counts were 3 times higher for shelled molluscs, but 5 times lower for crustaceans and soft-bodied organisms. Similarly, instantaneous daytime observations were 3 times higher for shelled molluscs, but 5 times lower for crustaceans and 2 times lower for soft-bodied organisms.Communicated by G.F. Humphrey, Sydney     

Statistical interpolation model for the description of ground pollution due to the TCDD produced in the 1976 chemical accident at seveso in the heavily contaminated zone “A”;

The influence of NTA on chromium genetic activity was studied in the D7 strain of Saccharomyces cerevisiae. At low dose (subgenotoxic) of sodium chromate (CrVI) (5mM), NTA increased the point mutation while at higher dose (25 mM) of chromate in the presence of NTA a decrease of point reverse mutation was observed. Probably NTA affected either the uptake of Cr(VI) favouring the intracellular reduction to Cr(III), or the recombinational repair of DNA breaks induced by chromate oxiding activity.

An increase of point (reverse) mutation was obtained in the experiments with NTA and chromium chloride, suggesting the hypothesis that NTA might interact with Cr(III) forming NTA‐Cr(III) complexes able to permeate cellular membranes and bind to DNA. In addition NTA genetic inactivity was confirmed.     

Pleistocene speciation of sister taxa in a North Pacific clade of brooding sea stars (<Emphasis Type="Italic">Leptasterias</Emphasis>)

Although numerous coastal marine species show intra-specific lineage splitting and population divergence times that date to the period of glacial cycles during the Pleistocene epoch, reported instances of recent speciation in the coastal marine environment are relatively rare. Marine organisms with brood-protection and other reproductive modes with limited dispersal potential have been suggested to experience more frequent speciation and extinction events than related species with higher dispersal rates, but few studies have actually estimated divergence times of sister species in these organisms. Here, two mitochondrial gene regions (cytochrome oxidase subunit I, putative control region and upstream tRNAs) and a nuclear gene region (Elongation factor 1α subunit intron 4) provide evidence of recent (0.5–1.2 Mya) cladogenetic events in four pairs of putative sister taxa in a predominantly North Pacific brooding subgenus of sea stars (Leptasterias subgenus Hexasterias). Calibration is obtained from a trans-arctic migration in a related clade of sea stars (Leptasterias subgenera Hexasterias and Nesasterias) that is timed to the opening of the Bering Strait at 3.5 ± 0.25 Mya, and uncertainty in the calibration point is accommodated with a normally-distributed Bayesian prior probability. Similar estimates of population splitting times for two of the pairs of putative sister taxa were obtained by a multilocus coalescent analysis. Estimates of mitochondrial mutation rates (0.01/My) were approximately 50% of the values calibrated for sister species pairs in tropical sea stars and sea urchins. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Enhancement of UV and chromate mutagenesis by nickel ions in the Chinese hamster HGPRT Assay †

1. The HGPRT (Hypoxanthine‐Guanine‐Phospho‐Ribosyl‐Transferase) assay with Chinese Hamster V79 cells was used to measure the mutagenic effects of UV irradiation, potassium dichromate and nickel chloride. The agents were tested separately and in the combinations of UV plus nickel and dichromate plus nickel.

2. UV, Cr(VI) and Ni(II) were confirmed to be mutagenic in the V79 cell assay. The combination of UV(5J/m²) and Ni(II) (0.5 mM) caused a mutation rate 11.2 times above that corresponding to the sum of the individual mutation rates of these agents. The combined action of Cr(VI) (0.1 mM) and Ni(II) (0.5 mM) produced a mutation rate 2.8 fold above that corresponding to the sum of the individual rates of the separate agents.

3. The enhancing effect of nickel chloride on the mutagenicity of UV or Cr(VI) is interpreted by an interference of Ni(II) with the repair of DNA lesions.     

耐热石油烃降解混合菌的筛选及其群落结构分析

对克拉玛依采集的部分石油污染土壤进行了筛选,得到了5组石油烃高效降解混合菌,其中混合菌KL9-1在45℃的条件下,通过7 d的降解,稀油的降解率达到43.27%,稠油的降解率达到20.09%。混合菌KL9-1经过多次分离纯化后,获得3株具有石油烃降解能力的优势单菌,3株单菌对稀油的降解率都在30%以上。结合分离单菌株的形态、生理生化特征和16S rDNA基因序列的分析结果,初步鉴定KL9-1-1为Pseudomonas putida,KL9-1-2和KL9-1-3为Pseudomonas sp.。     

Environmental human silver exposure

Environmental exposure to silver (Ag) was assessed in occupationally non-exposed adult human population by analyzing Ag in the hair (H?·?Ag) and whole blood (WB?·?Ag). H?·?Ag was analyzed in 311 (123 men, M; 188 women, W); while WB?·?Ag was determined in 235 of these individuals (90 M, 145 W). Women had more H?·?Ag than men (M 0.05 vs. W 0.076), whereas WB?·?Ag concentrations in men and women were not significantly different. A natural distribution of the median derivatives was utilized to generate the dataset to fit the logistic sigmoid curve to assess the current human body burden of environmental Ag population exposure for M and W separately. The H?·?Ag (µg?g^?1) below 0.0105 for M and 0.0145 for W, reflects low level of environmental Ag exposure. The adaptive physiological saturation phase followed where H?·?Ag rose rapidly, first for M and then for W in parallel with biological assay. Both parallel saturation curves converged and plateaued at 0.215 for M and 0.965 for W (µg?g^?1). The current level of human environmental Ag exposure is low, but cases of high Ag exposure occurred sporadically. In conjunction with the medical histories overt clinical neural toxicity may be expected for H?·?Ag at 4?µg?g^?1and higher. There were no significant correlation between the H?·?Ag and WB?·?Ag.     

Induction of micronuclei in tadpoles of Odontophrynus americanus (Amphibia: Leptodactylidae) by the pyrethroid insecticide cypermethrin

Cypermethrin (CY) is an active cyano pyrethroid effective against a wide range of pests encountered in agriculture and forestry. Although CY is not mutagenic in in vitro assays for gene mutation, in vivo assays showed conflicting results. In vivo genotoxicity of the synthetic pyrethroid CY in erythrocytes of Odontophrynus americanus tadpoles was examined. The frequency of micronuclei (MN) was recorded in blood smears obtained from tadpoles exposed in vivo to four different nominal concentrations 5, 10, 20 or 40?µg?L^?1 of the compound and fixed at two sampling times 48 and 96?h. As a positive control larvae were exposed to 40?mg?L^?1 of cyclophosphamide (CP). Tadpoles exposed to all CY treatments showed a significant increase in single small MN compared to the negative control group after 48?h and at 5 and 10?µg?L^?1 of CY at 96?h. Results obtained here demonstrated the genotoxic effects of the commercial formulation CY in the anuran larvae analyzed. Thus, data suggest that measurements of MN and other erythrocytes morphological aberrations performed in circulating blood samples of O. americanus tadpoles is a method for detecting cytogenetic damage in other native species.     

不同形态磷对黑藻生长和生理特性的影响

研究了Hoagkand培养液中分别以磷酸二氢钾、焦磷酸钠、六偏磷酸钠、甘油磷酸钠、ATP-Na为磷源时黑藻的生长及生理活性变化.获得了不同形态磷培养黑藻的干重、根系生长以及叶绿素含量、光合、呼吸、磷含量等指标.研究结果表明,磷酸二氢钾对黑藻的生长及生理活性影响最显著,是黑藻吸收的最佳磷形态;有机磷不利于根系生长,叶绿素a/b较高;六偏磷酸钠和焦磷酸钠促进根系早发,光合生产力低于磷酸二氢钾,叶绿素a/b高于磷酸二氢钾试验组.ATP-Na试验组黑藻呼吸率最高,甘油磷酸钠试验组黑藻光合生产力最高.各实验组黑藻磷含量均增加,黑藻对各形态磷均能吸收利用.图5表4参24     

The effects of a no. 2 fuel oil and two crude oils on the growth and photosynthesis of microalgae

Seawater, when equilibrated with a sample of No. 2 fuel oil, becomes toxic in varying degrees to growth of representative types of microalgae, two blue-greens, a diatom, two greens, and a dinoflagellate. For a sensitive organism such as Thalassiosira pseudonana, Strain 3H, 5 ml of seawater equilibrated with fuel oil (containing 15 mg/l of organics) in 20 ml of growth medium is lethal, or roughly in the range of 40 to 400 ppb if the toxic material(s) constitute 1 to 10% water also immediately stops photosynthesis in organism 3H. For other microalgae tested e.g. 580 (a green alga) and PR-6 (a blue-green alga), similar effects on growth and photosynthesis were found, but required higher concentrations of the oil-equilibrated seawater. Water solubles from Kuwait or Southern Louisiana crude oils (when the straight crude oil was equilibrated 1:8 with seawater) were not toxic; however, specific fractions obtained by distillation did show some water-soluble toxicity. Growth experimetns in open or closed growth systems revealed that most organisms were inhibited by varying amounts of these two crude oils when in direct contact with them. Organism 580 would not grow above 5 μl of Southern Louisiana/25 ml of medium, or 10 μl of Kuwait/25 ml of medium (oil in direct contact with algae). With both the seawater equilibrated with fuel oil and the crude oils, the toxic activity is mainly localized in medium and higher boiling fractions derived from distillation cuts from these materials.     

管网生物膜菌株胞外聚合物的提取方法比较

以饮用水管网生物膜样品中一株强成膜能力的菌株Pleomorphomonas oryzae作为研究对象,考察了8种方法(高速离心法、超声法、加热法、EDTA法、H2SO4法、NaOH法、SDS法、甲醛法)对菌株胞外聚合物(EPS)的提取效果,并结合三维荧光光谱(EEM)和傅立叶红外光谱(FTIR)对提取的EPS进行成分分析.结果表明,EDTA法和H2SO4法既能提高EPS的提取效率,提取量分别为64.77 mg.g-1SS和74.43 mg.g-1SS,是离心方法提取量的1.62倍和1.86倍,又不会在提取过程中对菌株细胞造成破坏,是较为理想的EPS提取方法.EEM分析进一步证实,NaOH法对菌株细胞破坏严重,造成EPS成分变化较大.FTIR分析则说明,化学提取方法相较于物理提取方法会引入杂质对组分测定造成干扰.