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合成生物学 1

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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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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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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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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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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