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Engineering of

Ruizhao Wang,Xiaoli Gu,Mingdong Yao,Caihui Pan,Hong Liu,Wenhai Xiao,Ying Wang,Yingjin Yuan

《化学科学与工程前沿（英文）》 2017年第11卷第1期页码 89-99 doi: 10.1007/s11705-017-1628-0

摘要： The conversion of -carotene to astaxanthin is a complex pathway network, in which two steps of hydroxylation and two steps of ketolation are catalyzed by -carotene hydroxylase (CrtZ) and -carotene ketolase (CrtW) respectively. Here, astaxanthin biosynthesis pathway was constructed in by introducing heterologous CrtZ and CrtW into an existing high -carotene producing strain. Both genes and were codon optimized and expressed under the control of constitutive promoters. Through combinatorial expression of CrtZ and CrtW from diverse species, nine strains in dark red were visually chosen from thirty combinations. In all the selected strains, strain SyBE_Sc118060 with CrtW from DC263 and CrtZ from sp. strain PC-1 achieved the highest astaxanthin yield of 3.1 mg/g DCW. Protein phylogenetic analysis shows that the shorter evolutionary distance of CrtW is, the higher astaxanthin titer is. Further, when the promoter of in strain SyBE_Sc118060 was replaced from FBA1p to TEF1p, the astaxanthin yield was increased by 30.4% (from 3.4 to 4.5 mg/g DCW). In the meanwhile, 33.5-fold increase on transcription level and 39.1-fold enhancement on the transcriptional ratio of to were observed at early exponential phase in medium with 4% (w/v) glucose. Otherwise, although the ratio of to were increased at mid-, late-exponential phases in medium with 2% (w/v) glucose, the transcription level of both and were actually decreased during the whole time course, consequently leading to no significant improvement on astaxanthin production. Finally, through high cell density fed-batch fermentation using a carbon source restriction strategy, the production of astaxanthin in a 5-L bioreactor reached to 81.0 mg/L, which was the highest astaxanthin titer reported in yeast. This study provides a reference to greatly enhance desired compounds accumulation by employing the key enzyme(s) in microbes.

关键词： synthetic biology astaxanthin β-carotene hydroxylase β-carotene ketolase Saccharomyces cerevisiae

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Improving prodeoxyviolacein production via multiplex SCRaMbLE iterative cycles

Juan Wang, Bin Jia, Zexiong Xie, Yunxiang Li, Yingjin Yuan

《化学科学与工程前沿（英文）》 2018年第12卷第4期页码 806-814 doi: 10.1007/s11705-018-1739-2

摘要：

The synthetic chromosome rearrangement and modification by loxP-mediated evolution (SCRaMbLE) system has been used to improve prodeoxyviolacein (PDV) production in haploid yeast containing chromosome synV. To rapidly and continuously generate genome diversification with the desired phenotype, the multiplex SCRaMbLE iterative cycle strategy has been developed for the screening of high PDV production strains. Whole-genome sequencing analysis reveals large duplications, deletions, and even the whole genome duplications. The deletion of YER151C is proved to be responsible for the increase. This study demonstrates that artificial DNA rearrangement can be used to accelerate microbial evolution and the production of biobased chemicals.

关键词： synthetic biology genome rearrangement prodeoxyviolacein SCRaMbLE Saccharomyces cerevisiae

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Exposure-response of Cr(III)-organic complexes to Saccharomyces cerevisiae

Nivedita CHATTERJEE, Zejiao LUO,

《环境科学与工程前沿（英文）》 2010年第4卷第2期页码 196-202 doi: 10.1007/s11783-010-0008-5

摘要： Hexavalent chromium [Cr(VI)] bioreduction produces soluble Cr(III)-organic complexes. The Cr(III)-organic complexes are relatively stable once they are formed, and no data about their toxicity were reported. Therefore, this study aims to investigate the bioavailability and toxicity of the soluble Cr(III)-organic complexes. L-1 wild type yeast strain was chosen as the model organism and Cr(III)-citrate was selected as the representative compound of the Cr(III)-organic complexes. The short-term chronic aquatic toxicity tests of the Cr(III)-citrate was explored by measuring growth inhibition, direct viable cell count, dry biomass, biosorption, and the amount of CO production. Cr(III)-citrate exerted a toxicity of 51mg/L with an , which was calculated from the percent growth inhibition. These toxicity data would be helpful to define the toxic potential of the organo-chromium-III compounds in the environment.

关键词： Cr(III)-organic complexes Saccharomyces cerevisiae toxicity EC50 bioavailability

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Biosorption of Cd

Fengyu ZAN, Shouliang HUO, Beidou XI, Xiulan ZHAO

《环境科学与工程前沿（英文）》 2012年第6卷第1期页码 51-58 doi: 10.1007/s11783-011-0206-9

摘要： The biosorption of Cd and Cu onto the immobilized ( ) was investigated in this study. Adsorption kinetics, isotherms and the effect of pH were studied. The results indicated that the biosorption of Cd and Cu on the immobilized was fast at initial stage and then became slow. The maximum biosorption of heavy metal ions on immobilized were observed at pH 4 for Cd and Cu . by the pseudo-second-order model described the sorption kinetic data well according to the high correlation coefficient ( ) obtained. The biosorption isotherm was fitted well by the Langmuir model, indicating possible mono-layer biosorption of Cd and Cu on the immobilized . Moreover, the immobilized after the sorption of Cd and Cu could be regenerated and reused.

关键词： Saccharomyces cerevisiae (S. cerevisiae) biosorption heavy metals immobilization desorption

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Assembly of biosynthetic pathways in

Lidan Ye,Xiaomei Lv,Hongwei Yu

《化学科学与工程前沿（英文）》 2017年第11卷第1期页码 126-132 doi: 10.1007/s11705-016-1597-8

摘要： A robust and versatile tool for multigene pathway assembly is a key to the biosynthesis of high-value chemicals. Here we report the rapid construction of biosynthetic pathways in using a marker recyclable integrative toolbox (pUMRI) developed in our research group, which has features of ready-to-use, convenient marker recycling, arbitrary element replacement, shuttle plasmid, auxotrophic marker independence, GAL regulation, and decentralized assembly. Functional isoprenoid biosynthesis pathways containing 4–11 genes with lengths ranging from ~10 to ~22 kb were assembled using this toolbox within 1–5 rounds of reiterative recombination. In combination with GAL-regulated metabolic engineering, high production of isoprenoids (e.g., 16.3 mg?g dcw carotenoids) was achieved. These results demonstrate the wide range of application and the efficiency of the pUMRI toolbox in multigene pathway construction of .

关键词： pathway assembly toolbox reiterative recombination S. cerevisiae biosynthesis

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利用异源酶组合构建酿酒酵母中咖啡酸的生物合成 Article

刘岚清, 刘宏, 张伟, 姚明东, 李炳志, 刘夺, 元英进

《工程（英文）》 2019年第5卷第2期页码 287-295 doi: 10.1016/j.eng.2018.11.029

摘要：

微生物中构建植物源天然产物的生物合成面临着许多挑战，尤其是当需要表达与激活植物源细胞色素P450酶的时候。通过从几种细菌中筛选HpaB和HpaC两种酶，本文在酿酒酵母中构建了具有活性的4-羟基苯乙酸3-羟化酶（4 HPA3H），它可以发挥植物源细胞色素P450酶的相似作用，用于生产咖啡酸。在与一个共同的酪氨酸氨裂解酶（TAL）协同作用下，不同的异源HpaB酶和HpaC酶组合在将底物L-酪氨酸转化为目标产物咖啡酸上展现出不同能力。以铜绿假单胞菌的HpaB酶和肠沙门氏菌的HpaC酶的异源酶组合可生产最高咖啡酸产量，摇瓶培养下可达到（289.4 ± 4.6）mg·L⁻¹。酵母底盘细胞与异源酶的相容性得到了有效的改善，咖啡酸的产量比初始值提高了40倍。铜绿假单胞菌HpaB酶中黄素腺嘌呤二核苷酸（FAD）结合域周围的6个关键氨基酸残基与其他细菌来源的HpaB酶有明显区别，可能在影响酶活性方面起关键作用。综上，我们建立了一种有效的方法来构建高效的酵母系统用于合成非天然羟基苯丙烷类化合物。

关键词：酿酒酵母咖啡酸异源酶细胞色素P450酶合成生物学

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Comparative lipidomic analysis of

Bin QIAO, Hong-Chi TIAN, Ying-Jin YUAN

《化学科学与工程前沿（英文）》 2012年第6卷第4期页码 461-469 doi: 10.1007/s11705-012-1223-3

摘要： Variations in the composition and level of phospholipids (PLs) in yeast cells during industrial ethanol fermentation processes were analyzed. A comparative lipidomic method was used to investigate the changes in total cellular PLs during continuous and fed-batch/batch processes. The phospholipid metabolism in yeast changed during both processes, mainly due to the presence of long-chain poly unsaturated fatty acids (PUFA) that contained phosphatidyglycerol (PG), phosphatidylethanolamine (PE) and phosphatidylserine (PS). The complexity of the media affected the growth of the yeast and the membrane composition. Yeast incorporated lots of exogenous saturated and PUFAs from the feedstock during the fermentations. During the continuous fermentation, there was an increase in PLs with shorter chains as the fermentation progressed and early in process there were more long-chains. During the fed-batch/batch process, the PG species increased as the fermentation progressed. This is probably due to an inositol deficiency in the earlier part of the fermentation.

关键词： lipidomics systems biology phospholipids stirred-tank saccharomyces cerevisiae biorefinery engineering

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Development of a new method for RMR and Q classification method to optimize support system in tunneling

Asghar RAHMATI,Lohrasb FARAMARZI,Manouchehr SANEI

《结构与土木工程前沿（英文）》 2014年第8卷第4期页码 448-455 doi: 10.1007/s11709-014-0262-x

摘要： Rock mass classification system is very suitable for various engineering design and stability analysis. classification method is confirmed by Japan Highway Public Corporation that this method can figure out either strength or deformability of rock mass, further appropriating the amount of rock bolts, thickness of shotcrete, and size of pitch of steel ribs just after the blasting procedure. Based on these advantages of method, in this study, according to data of five deep and long tunnels in Iran, two equations for estimating the value of method from and classification systems were developed. These equations as a new method were able to optimize the support system for and classification systems. From classification and its application in these case studies, it is pointed out that the method for the design of support systems in underground working is more reliable than the and classification systems.

关键词： JH classification Q and RMR classification new method

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抗白粉病基因 Pm40i> 在我国“后 Pm21i> 时代”小麦育种中的重要作用 Perspective

唐圣雯,胡玉亭,钟胜福,罗培高

《工程（英文）》 2018年第4卷第4期页码 500-506 doi: 10.1016/j.eng.2018.06.004

摘要：

由小麦白粉菌[Blumeria graminis f. sp. tritici（Bgt）] 引起的小麦白粉病是一种重要的小麦叶部病害，对小麦的产量产生很大的影响。20 世纪80 年代，通过簇毛麦（Heuchera villosa）的6VS 染色体与小麦6AL 染色体易位将抗白粉病基因Pm21 转移到普通小麦中。最近，在一些地方发现了对Pm21 有毒的Bgt，虽然这些菌株的病理学特性还有待研究，但这一现象提醒小麦育种者应注意应用Pm21 的风险。来源于普通小麦与中间偃麦草（Thinopyrum intermedium）杂种后代的抗白粉病基因Pm40，被定位在小麦7BS 染色体上，对Bgt 具有广谱和持久的抗性。通过细胞学研究，并未在Pm40 的载体品种中发现大片段外缘染色体。过去几年的研究发现，Pm40 的载体品种具有优良的农艺性状。因此，我们相信在未来的育种工程中，Pm40 将会在Pm21 的抗性被克服之后起巨大的作用。另外，Pm21 和Pm40 都来源于外缘物种，这暗示着外源基因的抗性可能比小麦本身的基因更为持久和有效。

关键词：小麦白粉病 Pm21i> Pm40i> 外源基因内源抗性

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NiFeRuO_xi> nanosheets on Ni foam as an electrocatalyst for efficient

《化学科学与工程前沿（英文）》 2023年第17卷第11期页码 1698-1706 doi: 10.1007/s11705-023-2334-8

摘要： The electrocatalyst NiFeRuO_x/NF, comprised of NiFeRuO_x nanosheets grown on Ni foam, was synthesized using a hydrothermal process followed by thermal annealing. NiFeRuO_x/NF displays high electrocatalytic activity and stability for overall alkaline seawater splitting: 98 mV@ 10 mA∙cm⁻² in hydrogen evolution reaction, 318 mV@ 50 mA∙cm⁻² in oxygen evolution reaction, and a cell voltage of 1.53 V@ 10 mA∙cm⁻², as well as 20 h of durability. A solar-driven system containing such a bifunctional NiFeRuO_x/NF has an almost 100% Faradaic efficiency. The NiFeRuO_x coating around Ni foam is an anti-corrosion layer and also a critical factor for enhancement of bifunctional performances.

关键词： NiFeRuO_x nanosheets Ni foam electrocatalysis overall seawater splitting solar-driven system

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2014—2016年食品动物源RE-cmeABCi>阳性弯曲菌流行与耐药性现状 Article

刘德俊, 刘蔚雯, 李星, 姚红, 沈张奇, 汪洋, 沈建忠

《工程（英文）》 2020年第6卷第1期页码 34-39 doi: 10.1016/j.eng.2019.10.013

摘要：

弯曲菌是引起人类胃肠炎的一类重要食源性致病菌。RE-CmeABC是近年来弯曲菌中新发现的一种增强型耐药外排泵，可介导对兽医和人医临床治疗弯曲菌感染的重要药物，如氟喹诺酮类、酰胺醇类、大环内酯类及四环素类等药物的高水平耐药，而目前尚未有对该外排泵的大范围流行病学调查。因此，本研究调查了2014—2016年连续三年间我国山东、上海和广东等三个重点养殖区食品动物源RE-cmeABC阳性弯曲菌的流行与耐药性现状。结果显示，在1088株弯曲菌（931株结肠弯曲菌与157株空肠弯曲菌）中共检测出122株（11.2%）RE-cmeABC阳性菌株，包括111株（70.7%）空肠弯曲菌与11株（1.2%）结肠弯曲菌，空肠弯曲菌中RE-cmeABC的阳性率显著高于结肠弯曲菌。111株RE-cmeABC阳性空肠弯曲菌与46株RE-cmeABC阴性空肠弯曲菌相比在氟苯尼考、克林霉素和红霉素的耐药率上具有显著差异（P < 0.05），而在环丙沙星、四环素和庆大霉素的耐药率上无显著差异。然而，携带RE-cmeABC的菌株可以使环丙沙星与四环素对弯曲菌的MIC值分布趋于更高水平范围。脉冲场凝胶电泳分型（PFGE）结果显示RE-cmeABC阳性菌株在上海和广东地区以水平传播为主，在山东地区以克隆传播为主；三株分离于上海和广东的阳性菌为同一克隆型，4株分离于上海和山东的阳性菌为同一克隆型。本研究表明RE-cmeABC已在我国食品动物源弯曲菌中广泛流行，对公共卫生安全具有潜在威胁。

关键词： RE- cmeABC i> 弯曲菌环丙沙星氟苯尼考

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A highly efficient methodology for the preparation of

Yongxin Zhang, Shucheng Wang, Yaodong Huang

《化学科学与工程前沿（英文）》 2021年第15卷第3期页码 679-686 doi: 10.1007/s11705-020-1979-9

摘要： A convenient and highly efficient method is described for the synthesis of -methoxycarbazole derivatives, including those with sterically demanding, benzannulated, or strongly electron-donating or -withdrawing substituents. Various -methoxycarbazole derivatives were directly prepared in good-to-moderate yields by the Pd (dba) CHCl /9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene-catalyzed reactions of the corresponding dibromobiphenyl compounds and methoxya-mine. Based on this methodology, the first total synthesis of 3,3′-[oxybis(methylene)]bis(9-methoxy-9 -carbazole), an antimicrobial dimeric carbazole alkaloid previously isolated from the stem bark of , was achieved in 18% yield over seven steps from 1,2-dibromobenzene.

关键词： N-methoxyl carbazole dimeric N-methoxyl carbazole alkaloid total synthesis double N-arylation of methoxyamine

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通过 B i>位Ta置换对 AB i> O₄型EuNbO₄的热学和力学性质进行优化 Article

陈琳, 冯晶

《工程（英文）》 2020年第6卷第2期页码 177-184 doi: 10.1016/j.eng.2019.12.006

摘要：

铁弹性的ABO₄型RETaO₄陶瓷和RENbO₄陶瓷（RE表示稀土）是具有潜在应用价值的热障涂层（TBC）材料，研究人员正在对其进行大量研究。结果显示， RETaO₄陶瓷的力学性质优于RENbO₄陶瓷的力学性质。在本研究中，我们通过B位钽（Ta）置换对利用固相法（SSR）制备的EuNbO₄陶瓷的热学和力学性质进行优化。我们使用X射线衍射（XRD）法和拉曼光谱技术对所制备的晶体结构进行确认，并利用扫描电子显微镜（SEM）对结构表面的微观形貌进行观察。结果表明，通过B位Ta置换可以有效提高EuNbO₄陶瓷的杨氏模量和热膨胀系数（TEC），且其最大值分别为169 GPa和1.12×10⁻⁵ K⁻¹（1200 ℃）； EuNbO₄陶瓷的热导率被降低至1.52 W·K^–1·m^–1（700 ℃），且材料的抗热辐射能力得到了改善。我们建立了声子热扩散系数与温度之间的关系，用以通过消除热辐射效应来确定声子本征热导率。研究结果表明，通过B位Ta置换能够有效优化EuNbO₄的热学和力学性质，从而使得此种材料在将来可成为一种高温结构陶瓷材料。

关键词：热障涂层，稀土铌酸盐，置换，热导率，热膨胀系数，杨氏模量

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Traces and echoes of De Architecturai> by Marcus Vitruvius Pollio in the work of Xu Guangqi in 17th

Michela CIGOLA,Yibing FANG

《机械工程前沿（英文）》 2016年第11卷第1期页码 3-11 doi: 10.1007/s11465-015-0359-5

摘要：

This study aims to investigate the role played by Xu Guangqi (1562–1633), minister of the Ming Dynasty, in the development of European scientific and technical knowledge in China between the 16th and 17th centuries by analyzing a book of Western technology that he wrote, namely, Taixi Shuifa (On Western Hydraulics). Several Western books related to machine knowledge are searched to trace the source of the illustrations in Taixi Shuifa. We found that Archimedes’ screw and Ctesibius’ machine, which are included in Vitruvius’ De Architectura volumes, also appear in the work of Xu Guangqi.

关键词： Vitruvius’ De Architectura Xu Guangqi’s Taixi Shuifa Jesuit missionaries Italian Renaissance drawing representation

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Accounting for the uncertainties in the estimation of average shear wave velocity using – correlations

《结构与土木工程前沿（英文）》 2021年第15卷第5期页码 1199-1208 doi: 10.1007/s11709-021-0749-1

摘要： Site-specific seismic hazard analysis is crucial for designing earthquake resistance structures, particularly in seismically active regions. Shear wave velocity ( V _S) is a key parameter in such analysis, although the economy and other factors restrict its direct field measurement in many cases. Various V _S–SPT– N correlations are routinely incorporated in seismic hazard analysis to estimate the value of V _S. However, many uncertainties question the reliability of these estimated V _S values. This paper comes up with a statistical approach to take care of such uncertainties involved in V _S calculations. The measured SPT– N values from all the critical boreholes were converted into statistical parameters and passed through various correlations to estimate V _S at different depths. The effect of different soil layers in the boreholes on the Vs estimation was also taken into account. Further, the average shear wave velocity of the top 30 m soil cover ( V _S30) is estimated after accounting for various epistemic and aleatoric uncertainties. The scattering nature of the V _S values estimated using different V _S– N correlations was reduced significantly with the application of the methodology. Study results further clearly demonstrated the potential of the approach to eliminate various uncertainties involved in the estimation of V _S30 using general and soil-specific correlations.

关键词： uncertainties V _S– N correlations V _S30 SPT data statistical methodology