耐碱性芽孢杆菌碱性淀粉酶的研究：Ⅱ.菌株的诱变与产酶条件

耐碱性巨大芽孢杆菌（Ｂａｃｉｌｌｕｓ ｍｅｇａｔｅｒｉｕｍ）９－Ａ２经紫外线、甲基磺酸乙酯和硫酸二乙酯等多次诱变处理,获得一株产碱性α－淀粉酶能力较高的变异株Ｌ－４９。Ｌ－４９菌株比出发株的产酶能力提高近２．５倍,当以酵母膏为氮源,可溶性淀粉为碳源,初始ｐＨ９￣１０,种龄１８ｈ,接种量５％（Ｖ／Ｖ）,３０℃培养４８ｈ,酶活力可达７３０μ／ｍｌ。     

PARP1 rs1136410 C/C genotype associated with an increased risk of esophageal cancer in smokers

Molecular Biology Reports - DNA repair system plays a crucial role in maintaining genomic integrity and stability and in protecting against cancer. Poly(ADP-ribose) polymerase 1 (PARP1) functions...     

Understanding the “SMART” features of hematopoietic stem cells and beyond

Science China Life Sciences - Since the huge success of bone marrow transplantation technology in clinical practice, hematopoietic stem cells (HSCs) have become the gold standard for defining the...     

DFT investigation on the decarboxylation mechanism of <Emphasis Type="Italic">ortho</Emphasis> hydroxy benzoic acids with acid catalysis

A density functional theory (DFT) study was performed to explore the mechanisms of the acid-catalyzed decarboxylation reaction of salicylic acids using the B3LYP method with 6-31++G(d,p) basis set in both gas phase and aqueous environment. The α-protonated cation of carboxylate acid was formed during the decarboxylation process in acidic conditions, and the presence of hydrogen ions promotes decarboxylation greatly by significantly decreasing the overall reaction energy barriers to 20.98 kcal mol^?1 in gas phase and 20.93 kcal mol^?1 in water, respectively. The hydrogen in the α-carbon came directly from the acid rather than from the carboxyl group in neutral state. Compared with the reaction in gas phase, water in aqueous state causes the reaction to occur more easily. Substituents of methyl group, chlorine and fluorine at the ortho-position to the carboxyl of salicylic acid could further lower the decarboxylation energy barriers and facilitate the reaction.     

产鼠李糖脂生物表面活性剂大肠杆菌的构建与优化

目前鼠李糖脂生物表面活性剂主要由条件致病的铜绿假单胞菌生产获得,从而影响工业应用。为了开发一种相对安全的鼠李糖脂生产菌,将带有不同强度组成型合成启动子的鼠李糖基转移酶基因(Rhamnosyltransferase gene,rhl AB)以单、中、高3种拷贝数分别在大肠杆菌ATCC 8739中异源表达,实现了不同产量的鼠李糖脂异源合成。对rhl AB基因和rha BDAC基因簇(TDP-L-鼠李糖合成的基因簇)进一步利用合成启动子进行组合调控,筛选获得了最优生产鼠李糖脂工程菌——大肠杆菌TIB-RAB226。对大肠杆菌TIB-RAB226进行发酵温度优化,鼠李糖脂产量达到124.3 mg/L,是优化前的1.17倍。通过分批补料发酵,12h时鼠李糖脂产量达到209.2 mg/L。对发酵产物进行高效液相色谱-质谱联用技术分析,共检出相对含量变化的5类质核比不同的鼠李糖脂同系物。本研究可为异源合成产鼠李糖脂提供重要参考。     

Morphology,cell growth,and polysaccharide production of <Emphasis Type="Italic">Nostoc flagelliforme</Emphasis> in liquid suspension culture at different agitation rates

Noscoc flagelliforme is a terrestrial macroscopic cyanobacterium with high economic value. Free-living cells that were separated from a natural colony of N. flagelliforme were cultivated in a 20-L photobioreactor for 16 days at five agitation rates with impeller tip speeds at 0.3, 0., 0.8, 1.0, and 1.5 m·s⁻¹. With different impeller tip speeds there were significant differences in the cell growth and polysaccharide production, and different types of cell colonies appeared because of different shear forces caused by agitation. At harvest time, cell concentrations with tip speeds of 0.8 and 1.0 m·s⁻¹ were clearly higher than those with the other three tip speeds, but dry cell weights with the tip speeds of 0.3, 0.5, 0.8, and 1.0 m·s⁻¹ were almost the same. The highest RPS (polysaccharide that released into liquid medium) production was obtained with the tip speeds of 0.8 and 1.0 m·s⁻¹, while the highest EPS (polysaccharide that formed capsule or slime layer) production was obtained with the tip speed of 0.5 m·s⁻¹. The tip speed of 1.5 m·s⁻¹ was harmful for both cell growth and polysaccharide production, indicating that an appropriate shear force was needed in the liquid suspension culture of N. flagelliforme.     

Enhanced bacterial cellulose production by Gluconacetobacter xylinus via expression of Vitreoscilla hemoglobin and oxygen tension regulation

Applied Microbiology and Biotechnology - Oxygen plays a key role during bacterial cellulose (BC) biosynthesis by Gluconacetobacter xylinus. In this study, the Vitreoscilla hemoglobin (VHb)-encoding...     

发状念珠藻胞外多糖的纯化与性质分析

采用DEAE阴离子交换层析和Sephadex G100凝胶层析对液体悬浮培养发状念珠藻胞外多糖进行纯化, 得到两个组分NFPS1和NFPS2。对组分NFPS2进行理化性质分析, 并与野生发状念珠藻多糖NFPS0的性质进行对比。结果表明二者具有相似的单糖组成, 均为葡萄糖、木糖、半乳糖、甘露糖; 表观分子量分别为2.79×105、2.26×105; 均不含核酸、蛋白质等物质, 是非硫酸化多糖; 有较高的热稳定性, 其降解温度在245oC左右。但在微观结构上, 两者存在一定差别。     

Growth characteristics of the cyanobacterium Nostoc flagelliforme in photoautotrophic, mixotrophic and heterotrophic cultivation

Nostoc flagelliforme is a terrestrial cyanobacterium with high economic value. Dissociated cells separated from a natural colony of N. flagelliforme were cultivated for 7 days under either phototrophic, mixotrophic or heterotrophic culture conditions. The highest biomass, 1.67 g L⁻¹ cell concentration, was obtained under mixotrophic culture, representing 4.98 and 2.28 times the biomass obtained in phototrophic and heterotrophic cultures, respectively. The biomass in mixotrophic culture was not the sum as that in photoautotrophic and heterotrophic cultures. During the first 4 days of culture, the cell concentration in mixotrophic culture was lower than the sum of those in photoautotrophic and heterotrophic cultures. However, from the 5th day, the cell concentration in mixotrophic culture surpassed the sum of those obtained from the other two trophic modes. Although the inhibitor of photosynthetic electron transport DCMU [3-(3,4-dichlorophenyl)-1,1-dimethylurea] efficiently inhibited autotrophic growth of N. flagelliforme cells, under mixotrophic culture they could grow by using glucose. The addition of glucose changed the response of N.flagelliforme cells to light. The maximal photosynthetic rate, dark respiration rate and light compensation point in mixotrophic culture were higher than those in photoautotrophic cultures. These results suggest that photoautotrophic (photosynthesis) and heterotrophic (oxidative metabolism of glucose) growth interact in mixotrophic growth of N. flagelliforme cells.     

利用代谢工程改善大肠杆菌的3-脱氢莽草酸生产

3-脱氢莽草酸是芳香族氨基酸合成代谢途径中的一种重要中间产物。除可作为一种高效的抗氧化剂,还可用于合成己二酸、香草醛等一些重要的化工产品,具有重要的应用价值。相关研究证明具有去酪氨酸反馈抑制的3-脱氧-D-阿拉伯庚酮糖-7-磷酸合成酶基因aroFFBR以及转酮醇酶基因tktA可以有效影响3-脱氢莽草酸的过量合成。通过增加aroFFBR和tktA串联过量表达的拷贝数,可使工程菌株在摇瓶发酵条件下3-脱氢莽草酸产量提高2.93倍。通过同源重组无痕基因敲除技术依次敲除出发菌大肠杆菌Escherichia coli AB2834的乳酸、乙酸、乙醇等副产物合成途径中的重要基因ldhA、ackA-pta和adhE,可使工程菌株的3-脱氢莽草酸产量进一步提高,达到了1.83 g/L,是初始出发菌株大肠杆菌E.coli AB2834产量的6.7倍。利用5 L发酵罐进行分批补料发酵,62 h后工程菌株3-脱氢莽草酸产量达到了25.48 g/L。本研究可为构建有应用前景的3-脱氢莽草酸生产菌株提供重要参考。