十溴联苯醚降解菌群的降解特性与组成分析

[目的]针对水体沉积物中日益严重的多溴联苯醚污染问题,以电子垃圾污染河床沉积物为种源富集驯化获得的菌群Cf3,研究该菌群对十溴联苯醚的降解特性以及其菌群结构组成.[方法]通过GC-MS分析十溴联苯醚降解后低溴代产物组成,并测定其降解率;通过DGGE技术分析了该BDE-209降解菌群的结构组成.[结果]菌群Cf3具有较强降解BDE-209的能力,经过120 d的培养,初始量为2.6 μmol的BDE-209降解率达到80.03％,OD600从0.01增长到0.21,pH由初始的6.93增加到反应结束时的8.50.菌群Cf3经过单菌落分离,共获得10株可培养细菌,通过16S rRNA基因序列比对发现,其中6株与柠檬酸杆菌属(Citrobacter spp.)具有较高同源性,其余4株与产碱杆菌属(Alcaligenes spp.)较相似.进一步采用DGGE分析菌群Cf3的结构组成时发现,除了分离得到的2个菌属外,该菌群中还含有拟杆菌属(Wolinella spp.)、氨基酸球菌属(Acidaminococcus spp.),以及随着降解时间延长而消失的脱硫弧菌属(Desulfovibrio spp.)和醋杆菌属(Acetobacterium spp.).[结论]获得了具有较强多溴联苯醚降解能力的菌群,并分析了其降解特性和群落组成,为进一步开展溴代阻燃剂等持久性有机污染物的生物修复提供宝贵的菌种资源和科学数据.     

利用Pichia pastoris生产S-腺苷甲硫氨酸的发酵工艺

在摇瓶中考察了重组Pichia pastoris发酵的诱导剂量,L-甲硫氨酸,以及pH对腺苷甲硫氨酸产量的影响.放大到3.7 L发酵罐和30 L发酵罐后,研究了重组细胞的发酵过程变化,对S-腺苷甲硫氨酸初步纯化.摇瓶中优化后的发酵条件是:每天添加1%甲醇诱导,L-甲硫氨酸为50mmol/L,培养基pH 5.0.培养144 h后SAM产量达到2.32 g/L.3.7 L发酵罐中发酵251 h后细胞浓度为120 g/L,SAM总量为15.18 g.放大到30 L发酵罐中,发酵225.5 h后细胞浓度约为120 g/L,SAM总量为145.05 g.纯化后SAM的纯度为93.5%,回收率为84.5%.     

他克莫司发酵培养基的响应面优化设计

用两水平因子设计考察培养基中的所有七个成分对他克莫司产量的影响大小,选出其中影响较大的葡萄糖、酵母粉、花生粉和NaCl进行最佳浓度范围考察,然后用中心组合方法及响应面分析确定最佳配比,他克莫司的产量提高了55%左右.     

蛋白酶生产和应用的进展

蛋白酶是一类极重要的酶,广泛应用于各个方面.本文概述了蛋白酶的分类与专一性以及国内外有关蛋白酶的生产和应用的进展.     

大豆膨胀素基因GmEXPB5和GmEXPB7生物信息学及表达分析

膨胀素(expansin,EXP)通过调控细胞壁的松弛在植物应对环境胁迫过程中起着重要作用。为研究EXP基因在大豆应对非生物胁迫过程中的作用,该文对大豆中的两个EXP基因(GmEXPB5和GmEXPB7)及其蛋白序列进行生物信息学分析,通过实时荧光定量PCR(qRT-PCR)检测基因表达量。结果表明:(1)GmEXPB5和GmEXPB7分别位于大豆第10号和第12号染色体上,编码的蛋白序列长度分别为272和267个氨基酸。GmEXPB5蛋白分子量为29.07 kD,理论等电点为7.51; GmEXPB7蛋白分子量为29.09 kD,理论等电点为8.66。GmEXPB5和GmEXPB7均为稳定的亲水蛋白且定位于细胞壁中。GmEXPB5和GmEXPB7蛋白均含有一段信号肽序列和一个保守的DPBB_1结构域。(2)GmEXPB5蛋白与鹰嘴豆CaEXPB15蛋白亲缘关系最近,GmEXPB7蛋白与密花豆、赤豆和豇豆的EXPB3蛋白有着较近的亲缘关系。(3)GmEXPB5和GmEXPB7在大豆根、茎和叶中均有表达且它们在根和叶中的表达量均显著高于茎中的表达量。(4)GmEXPB5和GmEXPB7在大豆幼苗中可以响应盐、干旱和低温胁迫。(5)GmEXPB5启动子区域含有2种与逆境相关的顺式作用元件(ABRE和ARE); GmEXPB7启动子区域含有5种与逆境相关的顺式作用元件(ABRE、ARE、CGTCA-motif、TC-rich repeats和MBS)。综上所述,GmEXPB5和GmEXPB7能够参与大豆对非生物胁迫的应答。     

Staining Sulfated Mucopolysaccharides in Sections by Means of Acriflavine

Acriflavine gave insoluble salts with sulfated esters. Frozen or paraffin sections (fixed in 10% formol or Carnoy's solution) were stained in M/20 acriflavine solution and excess dye was rinsed in 95% alcohol. Then nuclei were stained with Meyer's haemalum. Thereafter the sections were washed in water, dehydrated in alcohol, cleared in xylene and mounted in balsam. Sulfated esters in the tissue sections were colored yellow or orange-yellow, generally more densely in frozen than in paraffin sections.     

Experimental Approach Reveals the Role of alx1 in the Evolution of the Echinoderm Larval Skeleton

Over the course of evolution, the acquisition of novel structures has ultimately led to wide variation in morphology among extant multicellular organisms. Thus, the origins of genetic systems for new morphological structures are a subject of great interest in evolutionary biology. The larval skeleton is a novel structure acquired in some echinoderm lineages via the activation of the adult skeletogenic machinery. Previously, VEGF signaling was suggested to have played an important role in the acquisition of the larval skeleton. In the present study, we compared expression patterns of Alx genes among echinoderm classes to further explore the factors involved in the acquisition of a larval skeleton. We found that the alx1 gene, originally described as crucial for sea urchin skeletogenesis, may have also played an essential role in the evolution of the larval skeleton. Unlike those echinoderms that have a larval skeleton, we found that alx1 of starfish was barely expressed in early larvae that have no skeleton. When alx1 overexpression was induced via injection of alx1 mRNA into starfish eggs, the expression patterns of certain genes, including those possibly involved in skeletogenesis, were altered. This suggested that a portion of the skeletogenic program was induced solely by alx1. However, we observed no obvious external phenotype or skeleton. We concluded that alx1 was necessary but not sufficient for the acquisition of the larval skeleton, which, in fact, requires several genetic events. Based on these results, we discuss how the larval expression of alx1 contributed to the acquisition of the larval skeleton in the putative ancestral lineage of echinoderms.     

Extensive size homoplasy at a microsatellite locus in the Japanese bumblebee, Bombus diversus

An extensive size homoplasy was found at microsatellite locus B11 of the bumblebee, Bombus diversus, in northern to central Honshu, Japan. A total of 16 alleles of different nucleotide sequences in five length morphs was obtained at B11 for this species. Of these alleles, five were 141 base pairs (bp) in length, five were 137 bp and four were 133 bp. Allele diversity in each length morph was high compared with previous studies. It is noteworthy that this extensive size homoplasy was found in a relatively small geographic area, in contrast to results from previous studies. Reconstruction of a median‐joining network revealed the complicated evolutionary process of the locus, involving insertion/deletion and point mutations. Preliminary estimation of the mutation rate of the B11 locus in B. diversus gives a value comparable to those estimated from experimental Drosophila populations. Effects of the extensive size homoplasy in population genetic studies is discussed.