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Improvement of Thermostability of Penicillium expansum Lipase by Site-directed Mutagenesis 利用定点诱变提高膨胀青霉脂肪酶的热稳定性
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00043
Cai Shaoli, Zou Youtu, Hu Jianzhong, Lin Lin
In order to improve the thermostability of Penicillium expansum lipase(PEL),the lipase gene was mutated by sitedirected mutagenesis.A recombinant plasmid pAO815-ep8-K115R which contains double mutant genes was constructed by overlap extension PCR using the cDNA of a random-mutant lipase ep8(a single site mutant) as the template and two special primers were used to generate another mutation site K115R.The recombinant vector was transformed into Pichia pastoris GS115 by electroporation and the recombinant mutant GS-pAO815-ep8-K115R can secret the double-mutant lipase PEL-ep8K115R-GS into the medium when it was induced by methanol.Thermostability analysis revealed that the residual activity of the double-mutant lipase PEL-ep8-K115R-GS after incubated at 40 ℃ for 30 min was 54% and 27% higher than that of the wild type lipase PEL-GS and the random-mutant lipase PEL-ep8-GS respectively.Tm of the double-mutant lipase PEL-ep8-K115RGS was 42.2 ℃,3.5 ℃ higher than that of the wild type lipase PEL-GS,and 2.0 ℃ higher than that of the random-mutant lipase PEL-ep8-GS.
为了提高膨胀青霉菌脂肪酶(PEL)的热稳定性,采用定点诱变的方法对脂肪酶基因进行了突变。以随机突变型脂肪酶ep8(单位点突变)cDNA为模板,采用重叠延伸PCR技术构建了含有双突变基因的重组质粒pAO815-ep8-K115R,并利用两种特殊引物构建了另一个突变位点K115R。通过电孔法将重组载体转化为毕赤酵母GS115,重组突变体GS-pAO815-ep8-K115R在甲醇诱导下可将双突变体脂肪酶PEL-ep8K115R-GS分泌到培养基中。热稳定性分析表明,双突变型脂肪酶PEL-ep8-K115R-GS在40℃孵育30 min后的剩余活性分别比野生型和随机突变型脂肪酶PEL-ep8-GS高54%和27%。双突变型脂肪酶PEL-ep8-K115RGS的温度为42.2℃,比野生型脂肪酶PEL-GS高3.5℃,比随机突变型脂肪酶PEL-ep8-GS高2.0℃。
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引用次数: 0
Molecular Cloning and Heterologous Expression of Putative ( S )-norcoclaurine Synthases from Arabidopsis thaliana : Molecular Cloning and Heterologous Expression of Putative ( S )-norcoclaurine Synthases from Arabidopsis thaliana 拟南芥推定(S)- norcoclurine合成酶的分子克隆与异源表达:拟南芥推定(S)- norcoclurine合成酶的分子克隆与异源表达
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00061
Xiang Zhang, Lei Ma, Yongqiang Tian, Guolin Zhang, Yinggang Luo
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引用次数: 1
Genetic Diversity Among Barley Varieties (Lines) Bred in China: Genetic Diversity Among Barley Varieties (Lines) Bred in China 中国大麦品种(系)遗传多样性:中国大麦品种(系)遗传多样性
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00079
Yan Liu, X. Qiang, Jing Huang, X. Zhang, Ai-Tee Tan, Chun-Feng Zhao, J. Chen
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引用次数: 0
Optimization of Butanol Production from Canna edulis Ker by Clostridium acetobutylicum Using Response Surface Methodology: Optimization of Butanol Production from Canna edulis Ker by Clostridium acetobutylicum Using Response Surface Methodology 响应面法优化乙酰丁酸梭菌从美人蕉中提取丁醇的工艺研究
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00054
F. Long, Yanling Jin, Yun Zhao, Xiaofeng Gao, Yuhao Li, Qian Chen, Hai Zhao
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引用次数: 1
Bacterial Community Structure in Lakes on the Northeastern Qinghai-Tibetan Plateau 青藏高原东北部湖泊细菌群落结构
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00750
Li Ming, G. Jia, Shi Zheng-guo, Y. Libin, Yan Hong
The northeast of Qinghai-Tibetan Plateau is located in the convergence zone of the monsoon region of eastern China, northwest arid area and Qinghai-Tibetan Plateau region. During April to May 2012, an investigation was carried out on the bacterial community structure in ten lakes on northeastern Qinghai-Tibetan Plateau. With 16S rRNA sequencing and PCR-DGGE technology, twelve bacteria genus level libraries were constructed with a total of 117 operational taxonomic unit including 27 Betaproteobacteria, 8 Gamaproteobacteria, 21 Alphaproteobacteria, 7 Deltaproteobacteria,16 Acidobacteria, 2Deinococci, 5 Sphingobacteria, 6 Flavobacteria, 5 Nitrospira, 11 Actinobacteria, 6 Gemmatimonadetes and 3 unclassified bacteria. Most sequences in these studied lakes were closer to aquatic bacteria than any other bacterial clusters. The number of operational taxonomic unit in northeastern Qinghai-Tibetan Plateau was 32.1 ± 11.2, Shannon-Wiener diversity index 0.97± 0.29, Pielou's evenness index 0.19 ± 0.04, and Sorenson similarity index between lakes 0.31 ± 0.11, all comparatively low.Notably, the environment factors played an important role in bacteria community, with temperature positively related to OTUs(R = 0.763, P 0.05), to Shannon-Wiener diversity index(R = 0.708, P 0.05), and to Pielou's evenness index(R = 0.641, P 0.05); the lake area was also positively related to OTUs(R= 0.754, P 0.05) and Shannon-Wiener diversity index(R = 0.668, P 0.05). Water chemistry also played an important part in bacteria community. The result is helpful for study of climatic factors of community structure and sustainable development in this region.
青藏高原东北部处于中国东部季风区、西北干旱区和青藏高原地区的交汇地带。2012年4 ~ 5月,对青藏高原东北部10个湖泊的细菌群落结构进行了调查。通过16S rRNA测序和PCR-DGGE技术,构建了12个细菌属水平文库,共有117个操作分类单元,包括27个Betaproteobacteria、8个Gamaproteobacteria、21个Alphaproteobacteria、7个Deltaproteobacteria、16个Acidobacteria、2个deinococci、5个Sphingobacteria、6个Flavobacteria、5个Nitrospira、11个放线菌ia、6个gemmatimonadees和3个未分类细菌。在这些被研究的湖泊中,大多数序列比任何其他细菌簇更接近水生细菌。青藏高原东北部运行分类单元数为32.1±11.2,Shannon-Wiener多样性指数为0.97±0.29,Pielou均匀度指数为0.19±0.04,Sorenson相似性指数为0.31±0.11,均较低。环境因子对细菌群落的影响显著,温度与OTUs(R = 0.763, P 0.05)、Shannon-Wiener多样性指数(R = 0.708, P 0.05)、Pielou均匀度指数(R = 0.641, P 0.05)呈正相关;湖泊面积与OTUs(R= 0.754, P 0.05)和Shannon-Wiener多样性指数(R= 0.668, P 0.05)呈正相关。水化学在细菌群落中也起着重要作用。研究结果对该区群落结构和可持续发展的气候因子研究具有一定的指导意义。
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引用次数: 0
Asymmetric anti -Prelog Reduction of 3,5-bis(trifluoromethyl)-acetophenone by Microbacterium oxydans C3: Asymmetric anti -Prelog Reduction of 3,5-bis(trifluoromethyl)-acetophenone by Microbacterium oxydans C3 氧化微杆菌C3对3,5-二(三氟甲基)-苯乙酮的不对称抗预log还原
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00037
P. Gai, Chuan-Gen Tang, Jingyuan Liu, Yan Liu, Chao Zhang, Z. Wu
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引用次数: 6
Chironomid Larva Community and Indication to Eutrophication Progress in Lake Nansi, Shandong, China: Chironomid Larva Community and Indication to Eutrophication Progress in Lake Nansi, Shandong, China 山东南四湖摇尾蛾幼虫群落及其富营养化进程指示:山东南四湖摇尾蛾幼虫群落及其富营养化进程指示
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00141
F. Shu, L. Kong, Saidi Wang, Fangfang Zi
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引用次数: 2
Cloning and Diversity Analysis of the Conserved Fragments of Pheromone Receptor Genes in Genus Agrocybe: Cloning and Diversity Analysis of the Conserved Fragments of Pheromone Receptor Genes in Genus Agrocybe 农草属信息素受体基因保守片段的克隆与多样性分析&农草属信息素受体基因保守片段的克隆与多样性分析
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00794
Yingying He, Weimin Chen, Yongchang Zhao, Dequn Zhou
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引用次数: 0
Simultaneous Production of Methane and Acetate from Carbon Dioxide with Bioelectrochemical Systems 利用生物电化学系统从二氧化碳中同时生产甲烷和醋酸盐
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00833
J. Yong, S. Min, Z. Yao, Tao Yong, Li Daping
Reduction of CO2 to organics with bioelectrical systems has become a research area of environmental microbiology.This study constructed a bioelectrochemical system(BES) capable of reducing CO2 to CH4 and CH3COOH via abiotical H2 gas production and/or direct extracellular electron transfer. CH4 and CH3COOH were produced at rates of 17.3 mL h-1L-1 and 13.9 mg h-1L-1, respectively(at potential of-1 100 mV). The current capture efficiency reached 94.0% in batch potentiostatic experiments. Microbial characterization revealed that the microbial populations consisted of Methanobacterium palustre,Methanocorpusculum parvum, and acetogen Acetobacterium sp. These results suggested that mixed culture in BESs can convert CO2 into various organic compounds by accepting electrons in different ways.
利用生物电系统将CO2还原为有机物已成为环境微生物学的一个研究领域。本研究构建了一个生物电化学系统(BES),该系统能够通过非生物产氢和/或直接胞外电子转移将CO2还原为CH4和CH3COOH。CH4和CH3COOH的产率分别为17.3 mL h-1L-1和13.9 mg h-1L-1(电位-1 100 mV)。在批量恒电位实验中,电流捕获效率达到94.0%。微生物鉴定结果表明,微生物群主要由palustre Methanobacterium,Methanocorpusculum parvum和Acetobacterium sp.组成。这些结果表明,BESs中混合培养可以通过不同的方式接受电子,将CO2转化为多种有机化合物。
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引用次数: 4
Effect of Two Allelochemicals of Casuarina equisetifolia L. on Chlorophyll Fluorescence Parameters in Hydroponically Cultured Seedling: Effect of Two Allelochemicals of Casuarina equisetifolia L. on Chlorophyll Fluorescence Parameters in Hydroponically Cultured Seedling 木麻黄两种化感物质对水培苗叶绿素荧光参数的影响:木麻黄两种化感物质对水培苗叶绿素荧光参数的影响
Q4 Agricultural and Biological Sciences Pub Date : 2013-12-13 DOI: 10.3724/SP.J.1145.2013.00781
Jian Li, Yi Liu, Cheng-zhen Wu, T. Hong, Yongming Lin, Han Lin, Can Chen, W. Hong
In order to investigate the resistance of Casuarina equisetifolia L. to autotoxicity and to provide scientific basis for continuous planting of C. equisetifolia, clones of ‘Huian No.1’ C. equisetifolia were water cultivated to study the effects of chlorophyll fluorescence in branchlet under allelochemical stress (of seven concentrations: 0, 12.5, 25, 50, 100, 200 and 400 mg L) induced by quercetin-3-α-araboside (allelochemical A) and quercetin-3-β-glucoside (allelochemical B). The membership function of fuzzy mathematics was used to analyze the tolerance abilities of C. equisetifolia seedling under different allelochemicals stress. The results showed slight variations in F0, Fm, Fv/F0 and Fv/Fm of the branchlet under slight allelochemicals stress (12.5, 25, 50 mg L). The Fv/Fm was observed to maintain 78% of normal photosynthesis reaction. The F0, qN, Drate, LPAR of the branchlet significantly (P < 0.05) increased with the concentration of allelochemicals(100, 200, 400 mg L), while the values of Fm, Fv/Fm, Fv/F0, Prate, ΦPSII, qP decreased significantly (P < 0.05). Allelochemicals stress damaged the function of PSII reaction center and decreased the photochemical electron transfer rate, which led to the structural damage of photosynthetic organs and functional degradation in photosynthetic ability. The range of chlorophyll fluorescence parameters in branchlet of C. equisetifolia varied with different allelochemicals,the seedlings had better stress tolerance against allelochemical A than allelochemical B. Fig 3, Tab 1, Ref 28
为了研究木麻黄对自身毒性的抗性,为木麻黄的连作提供科学依据,对“惠安1号”木麻黄无性系进行了水培,研究了7种浓度的化感化学胁迫对木麻黄小枝叶绿素荧光的影响:0、12.5、25、50、100、200和400 mg L槲皮素-3-α-糖苷(化感物质A)和槲皮素-3-β-葡萄糖苷(化感物质B)诱导下的耐药能力。采用模糊隶属函数分析了不同化感物质胁迫下的马叶松幼苗的耐药能力。结果表明,在12.5、25、50 mg L的化感物质胁迫下,小枝的F0、Fm、Fv/F0和Fv/Fm变化不大,Fv/Fm维持正常光合反应的78%。化感物质浓度分别为100、200、400 mg L时,小枝F0、qN、Drate、LPAR显著升高(P < 0.05), Fm、Fv/Fm、Fv/F0、Prate、ΦPSII、qP显著降低(P < 0.05)。化感物质胁迫破坏了PSII反应中心的功能,降低了光化学电子传递速率,导致光合器官的结构损伤和光合能力的功能退化。不同化感物质对马叶松小枝叶绿素荧光参数的影响范围不同,幼苗对化感物质A的耐受性优于化感物质b。图3,表1,参考文献28
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引用次数: 2
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