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Electronic signals are electrogenetically relayed to control cell growth and co-culture composition 电子信号通过电学传递来控制细胞生长和共培养成分
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-12-01 DOI: 10.1016/j.mec.2021.e00176
Kristina Stephens , Fauziah Rahma Zakaria , Eric VanArsdale , Gregory F. Payne , William E. Bentley

There is much to be gained by enabling electronic interrogation and control of biological function. While the benefits of bioelectronics that rely on potential-driven ionic flows are well known (electrocardiograms, defibrillators, neural prostheses, etc) there are relatively few advances targeting nonionic molecular networks, including genetic circuits. Redox activities combine connectivity to electronics with the potential for specific genetic control in cells. Here, electrode-generated hydrogen peroxide is used to actuate an electrogenetic “relay” cell population, which interprets the redox cue and synthesizes a bacterial signaling molecule (quorum sensing autoinducer AI-1) that, in turn, signals increased growth rate in a second population. The dramatically increased growth rate of the second population is enabled by expression of a phosphotransferase system protein, HPr, which is important for glucose transport. The potential to electronically modulate cell growth via direct genetic control will enable new opportunities in the treatment of disease and manufacture of biological therapeutics and other molecules.

使电子审讯和控制生物功能成为可能将大有裨益。虽然依赖于电位驱动离子流的生物电子学的好处是众所周知的(心电图、除颤器、神经假体等),但针对非离子分子网络(包括遗传电路)的进展相对较少。氧化还原活动结合了与电子的连接以及细胞中特定遗传控制的潜力。在这里,电极产生的过氧化氢被用来驱动一个电遗传“中继”细胞群,它解释氧化还原信号并合成一种细菌信号分子(群体感应自诱导剂AI-1),反过来,在第二个群体中发出增长速度的信号。第二种群的生长速度急剧增加是由于磷酸转移酶系统蛋白HPr的表达,这对葡萄糖运输很重要。通过直接遗传控制电子调节细胞生长的潜力将为疾病治疗和生物疗法和其他分子的制造提供新的机会。
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引用次数: 8
Erratum regarding previously published articles in volumes 9, 10 and 11 关于第9卷、第10卷和第11卷以前发表的文章的勘误
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-12-01 DOI: 10.1016/j.mec.2021.e00186
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引用次数: 0
Identification of novel inhibitory metabolites and impact verification on growth and protein synthesis in mammalian cells 新型抑制代谢物的鉴定及其对哺乳动物细胞生长和蛋白质合成的影响验证
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-12-01 DOI: 10.1016/j.mec.2021.e00182
Bingyu Kuang , Venkata Gayatri Dhara , Duc Hoang , Jack Jenkins , Pranay Ladiwala , Yanglan Tan , Scott A. Shaffer , Shaun C. Galbraith , Michael J. Betenbaugh , Seongkyu Yoon

Mammalian cells consume large amount of nutrients during growth and production. However, endogenous metabolic inefficiencies often prevent cells to fully utilize nutrients to support growth and protein production. Instead, significant fraction of fed nutrients is diverted into extracellular accumulation of waste by-products and metabolites, further inhibiting proliferation and protein synthesis. In this study, an LC-MS/MS based metabolomics pipeline was used to screen Chinese hamster ovary (CHO) extracellular metabolites. Six out of eight identified inhibitory metabolites, caused by the inefficient cell metabolism, were not previously studied in CHO cells: aconitic acid, 2-hydroxyisocaproic acid, methylsuccinic acid, cytidine monophosphate, trigonelline, and n-acetyl putrescine. When supplemented back into a fed-batch culture, significant reduction in cellular growth was observed in the presence of each metabolite and all the identified metabolites were shown to impact the glycosylation of a model secreted antibody, with seven of these also reducing CHO cellular productivity (titer) and all eight inhibiting the formation of mono-galactosylated biantennary (G1F) and biantennary galactosylated (G2F) N-glycans. These inhibitory metabolites further impact the metabolism of cells, leading to a significant reduction in CHO cellular growth and specific productivity in fed-batch culture (maximum reductions of 27.2% and 40.6% respectively). In-depth pathway analysis revealed that these metabolites are produced when cells utilize major energy sources such as glucose and select amino acids (tryptophan, arginine, isoleucine, and leucine) for growth, maintenance, and protein production. Furthermore, these novel inhibitory metabolites were observed to accumulate in multiple CHO cell lines (CHO–K1 and CHO-GS) as well as HEK293 cell line. This study provides a robust and holistic methodology to incorporate global metabolomic analysis into cell culture studies for elucidation and structural verification of novel metabolites that participate in key metabolic pathways to growth, production, and post-translational modification in biopharmaceutical production.

哺乳动物细胞在生长和生产过程中消耗大量的营养物质。然而,内源性代谢效率低下往往阻止细胞充分利用营养来支持生长和蛋白质生产。相反,大量的营养物质被转移到细胞外积累的废物副产品和代谢物中,进一步抑制了增殖和蛋白质合成。本研究采用基于LC-MS/MS的代谢组学方法筛选中国仓鼠卵巢(CHO)细胞外代谢物。由细胞代谢效率低下引起的8种已确定的抑制性代谢物中有6种以前未在CHO细胞中研究过:乌头酸、2-羟基异己酸、甲基琥珀酸、单磷酸胞苷、葫芦巴碱和n-乙酰腐胺。当补充回补料批培养时,观察到每种代谢物存在时细胞生长显著减少,所有鉴定的代谢物都显示影响模型分泌抗体的糖基化,其中7种也降低CHO细胞生产力(滴度),所有8种都抑制单半乳糖化双天线(G1F)和双天线半乳糖化(G2F) n -聚糖的形成。这些抑制性代谢物进一步影响细胞的代谢,导致CHO细胞生长和比产率显著降低(最大降幅分别为27.2%和40.6%)。深入的途径分析表明,当细胞利用葡萄糖等主要能量来源和选择氨基酸(色氨酸、精氨酸、异亮氨酸和亮氨酸)进行生长、维持和蛋白质生产时,这些代谢物就会产生。此外,这些新的抑制性代谢物被观察到在多种CHO细胞系(CHO - k1和CHO- gs)以及HEK293细胞系中积累。本研究提供了一种强大而全面的方法,将全球代谢组学分析纳入细胞培养研究,以阐明和结构验证参与生物制药生产中生长,生产和翻译后修饰关键代谢途径的新型代谢物。
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引用次数: 7
Production of thermostable phycocyanin in a mesophilic cyanobacterium 嗜温蓝藻菌中耐热藻蓝蛋白的产生
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-12-01 DOI: 10.1016/j.mec.2021.e00175
Anton Puzorjov , Katherine E. Dunn , Alistair J. McCormick

Phycocyanin (PC) is a soluble phycobiliprotein found within the light-harvesting phycobilisome complex of cyanobacteria and red algae, and is considered a high-value product due to its brilliant blue colour and fluorescent properties. However, commercially available PC has a relatively low temperature stability. Thermophilic species produce more thermostable variants of PC, but are challenging and energetically expensive to cultivate. Here, we show that the PC operon from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 (cpcBACD) is functional in the mesophile Synechocystis sp. PCC 6803. Expression of cpcBACD in an ‘Olive’ mutant strain of Synechocystis lacking endogenous PC resulted in high yields of thermostable PC (112 ± 1 mg g−1 DW) comparable to that of endogenous PC in wild-type cells. Heterologous PC also improved the growth of the Olive mutant, which was further supported by evidence of a functional interaction with the endogenous allophycocyanin core of the phycobilisome complex. The thermostability properties of the heterologous PC were comparable to those of PC from T. elongatus, and could be purified from the Olive mutant using a low-cost heat treatment method. Finally, we developed a scalable model to calculate the energetic benefits of producing PC from T. elongatus in Synechocystis cultures. Our model showed that the higher yields and lower cultivation temperatures of Synechocystis resulted in a 3.5-fold increase in energy efficiency compared to T. elongatus, indicating that producing thermostable PC in non-native hosts is a cost-effective strategy for scaling to commercial production.

藻蓝蛋白(PC)是一种可溶性藻胆蛋白,存在于蓝藻和红藻的光收集藻胆体复合体中,由于其明亮的蓝色和荧光特性而被认为是一种高价值的产品。然而,商用PC具有相对较低的温度稳定性。嗜热物种产生更耐热的PC变种,但具有挑战性和能量昂贵的培养。本研究表明,来自嗜热蓝藻热共生球菌(Thermosynechococcus elongatus) BP-1 (cpcBACD)的PC操纵子在嗜热共生菌(Synechocystis sp. PCC 6803)中具有功能。在缺乏内源性PC的聚囊菌' Olive '突变株中,cpcBACD的表达导致耐热PC的高产(112±1 mg g−1 DW),与野生型细胞中的内源性PC相当。外源PC也促进了橄榄突变体的生长,这进一步得到了与藻胆异构体的内源异藻蓝蛋白核心的功能相互作用的证据的支持。外源PC的热稳定性与长叶葡萄的PC相当,可以用低成本的热处理方法从橄榄突变体中纯化得到。最后,我们建立了一个可扩展的模型来计算在聚囊藻培养物中从长形霉中生产PC的能量效益。我们的模型显示,与T. elongatus相比,更高的产量和更低的栽培温度导致能量效率提高3.5倍,这表明在非本地宿主中生产耐热PC是一种成本效益高的规模化商业生产策略。
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引用次数: 12
Erratum regarding missing Declaration of competing interest statements in previously published articles 关于先前发表的文章中缺少竞争利益声明的勘误表
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-12-01 DOI: 10.1016/j.mec.2021.e00189
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引用次数: 0
Diversion of metabolic flux towards 5-deoxy(iso)flavonoid production via enzyme self-assembly in Escherichia coli 大肠杆菌通过酶自组装将代谢通量转向生产5-脱氧(异)类黄酮
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-12-01 DOI: 10.1016/j.mec.2021.e00185
Jianhua Li , Fanglin Xu , Dongni Ji , Chenfei Tian , Yuwei Sun , Ishmael Mutanda , Yuhong Ren , Yong Wang

5-Deoxy(iso)flavonoids are structural representatives of phenylpropanoid-derived compounds and play critical roles in plant ecophysiology. Recently, 5-deoxy(iso)flavonoids gained significant interest due to their potential applications as pharmaceuticals, nutraceuticals, and food additives. Given the difficulties in their isolation from native plant sources, engineered biosynthesis of 5-deoxy(iso)flavonoids in a microbial host is a highly promising alternative approach. However, the production of 5-deoxy(iso)flavonoids is hindered by metabolic flux imbalances that result in a product profile predominated by non-reduced analogues. In this study, GmCHS7 (chalcone synthase from Glycine max) and GuCHR (chalcone reductase from Glycyrrhizza uralensis) were preliminarily utilized to improve the CHR ratio (CHR product to total CHS product). The use of this enzyme combination improved the final CHR ratio from 39.7% to 50.3%. For further optimization, a protein-protein interaction strategy was employed, basing on the spatial adhesion of GmCHS7:PDZ and GuCHR:PDZlig. This strategy further increased the ratio towards the CHR-derived product (54.7%), suggesting partial success of redirecting metabolic flux towards the reduced branch. To further increase the total carbon metabolic flux, 15 protein scaffolds were programmed with stoichiometric arrangement of the three sequential catalysts GmCHS7, GuCHR and MsCHI (chalcone isomerase from Medicago sativa), resulting in a 1.4-fold increase in total flavanone production, from 69.4 mg/L to 97.0 mg/L in shake flasks. The protein self-assembly strategy also improved the production and direction of the lineage-specific compounds 7,4′-dihydroxyflavone and daidzein in Escherichia coli. This study presents a significant advancement of 5-deoxy(iso)flavonoid production and provides the foundation for production of value-added 5-deoxy(iso)flavonoids in microbial hosts.

5-Deoxy(iso)类黄酮是苯丙类化合物的结构代表,在植物生理生态中起着重要作用。近年来,5-脱氧(iso)类黄酮因其在药物、营养食品和食品添加剂方面的潜在应用而引起了人们的极大兴趣。考虑到从原生植物中分离5-脱氧黄酮类化合物的困难,在微生物宿主中工程生物合成5-脱氧黄酮类化合物是一种非常有前途的替代方法。然而,5-脱氧(iso)类黄酮的生产受到代谢通量不平衡的阻碍,导致产品轮廓以非还原类似物为主。本研究初步利用GmCHS7 (Glycine max查尔酮合成酶)和GuCHR (Glycyrrhizza uralensis查尔酮还原酶)提高CHR (CHR产物与总CHS产物的比值)。使用该酶组合后,最终CHR比由39.7%提高到50.3%。为了进一步优化,采用基于GmCHS7:PDZ和GuCHR:PDZlig空间粘附的蛋白-蛋白相互作用策略。这一策略进一步增加了chr衍生产物的比例(54.7%),表明部分成功地将代谢通量重定向到减少的分支。为了进一步提高总碳代谢通量,我们对15个蛋白质支架进行了化学量学排列,将三种顺序的催化剂GmCHS7、GuCHR和MsCHI(紫花苜蓿查尔酮异构酶)进行了编程,使摇瓶中总黄酮的产量增加了1.4倍,从69.4 mg/L增加到97.0 mg/L。蛋白质自组装策略还改善了大肠杆菌中具有谱系特异性的化合物7,4 ' -二羟黄酮和大豆苷元的产生和方向。该研究为5-脱氧(iso)类黄酮的生产提供了重要进展,为微生物宿主生产增值5-脱氧(iso)类黄酮提供了基础。
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引用次数: 1
A lysate proteome engineering strategy for enhancing cell-free metabolite production 提高细胞游离代谢物产生的裂解物蛋白质组工程策略
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-06-01 DOI: 10.1016/j.mec.2021.e00162
David C. Garcia , Jaime Lorenzo N. Dinglasan , Him Shrestha , Paul E. Abraham , Robert L. Hettich , Mitchel J. Doktycz

Cell-free systems present a significant opportunity to harness the metabolic potential of diverse organisms. Removing the cellular context provides the ability to produce biological products without the need to maintain cell viability and enables metabolic engineers to explore novel chemical transformation systems. Crude extracts maintain much of a cell’s capabilities. However, only limited tools are available for engineering the contents of the extracts used for cell-free systems. Thus, our ability to take full advantage of the potential of crude extracts for cell-free metabolic engineering is constrained. Here, we employ Multiplex Automated Genomic Engineering (MAGE) to tag proteins for selective depletion from crude extracts so as to specifically direct chemical production. Specific edits to central metabolism are possible without significantly impacting cell growth. Selective removal of pyruvate degrading enzymes resulted in engineered crude lysates that are capable of up to 40-fold increases in pyruvate production when compared to the non-engineered extract. The described approach melds the tools of systems and synthetic biology to showcase the effectiveness of cell-free metabolic engineering for applications like bioprototyping and bioproduction.

无细胞系统为利用各种生物体的代谢潜力提供了一个重要的机会。去除细胞环境提供了在不需要维持细胞活力的情况下生产生物产品的能力,并使代谢工程师能够探索新的化学转化系统。粗提取物保留了细胞的大部分功能。然而,只有有限的工具可用于工程内容的提取物用于无细胞系统。因此,我们充分利用粗提取物的潜力进行无细胞代谢工程的能力受到限制。在这里,我们使用多重自动基因组工程(MAGE)来标记蛋白质,以便从粗提取物中选择性地去除,从而特异性地指导化学生产。在不显著影响细胞生长的情况下,对中枢代谢进行特异性编辑是可能的。丙酮酸降解酶的选择性去除导致工程粗裂解物的丙酮酸产量比非工程提取物增加了40倍。所描述的方法融合了系统和合成生物学的工具,以展示无细胞代谢工程在生物原型和生物生产等应用中的有效性。
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引用次数: 6
Development of antisense RNA-mediated quantifiable inhibition for metabolic regulation 反义rna介导的可量化代谢调控抑制的发展
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-06-01 DOI: 10.1016/j.mec.2021.e00168
Ruihua Zhang, Yan Zhang, Jian Wang, Yaping Yang, Yajun Yan

Trans-regulating elements such as noncoding RNAs are crucial in modifying cells, and has shown broad application in synthetic biology, metabolic engineering and RNA therapies. Although effective, titration of the regulatory levels of such elements is less explored. Encouraged by the need of fine-tuning cellular functions, we studied key parameters of the antisense RNA design including oligonucleotide length, targeting region and relative dosage to achieve differentiated inhibition. We determined a 30-nucleotide configuration that renders efficient and robust inhibition. We found that by targeting the core RBS region proportionally, quantifiable inhibition levels can be rationally obtained. A mathematic model was established accordingly with refined energy terms and successfully validated by depicting the inhibition levels for genomic targets. Additionally, we applied this fine-tuning approach for 4-hydroxycoumarin biosynthesis by simultaneous and quantifiable knockdown of multiple targets, resulting in a 3.58-fold increase in titer of the engineered strain comparing to that of the non-regulated. We believe the developed tool is broadly compatible and provides an extra layer of control in modifying living systems.

非编码RNA等反式调控元件在细胞修饰中起着至关重要的作用,在合成生物学、代谢工程和RNA治疗中有着广泛的应用。虽然有效,但对这些元素的调节水平的滴定研究较少。由于需要微调细胞功能,我们研究了反义RNA设计的关键参数,包括寡核苷酸长度、靶向区域和相对剂量,以实现差异化抑制。我们确定了一个30个核苷酸的配置,使有效和强大的抑制。我们发现,通过比例靶向核心RBS区域,可以合理地获得可量化的抑制水平。建立了具有精细能量项的数学模型,并成功地通过描述基因组靶点的抑制水平进行了验证。此外,我们将这种微调方法应用于4-羟基香豆素的生物合成,通过同时可量化地敲除多个靶点,使工程菌株的滴度比未调节菌株提高3.58倍。我们相信开发的工具是广泛兼容的,并为修改生命系统提供了额外的控制层。
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引用次数: 4
Co-expressing Eranthis hyemalis lysophosphatidic acid acyltransferase 2 and elongase improves two very long chain polyunsaturated fatty acid production in Brassica carinata 共表达Eranthis hyemalis溶血磷脂酸酰基转移酶2和延长酶提高了两种超长链多不饱和脂肪酸的产量
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-06-01 DOI: 10.1016/j.mec.2021.e00171
Dauenpen Meesapyodsuk , Yi Chen , Shengjian Ye , Robert G. Chapman , Xiao Qiu

Docosadienoic acid (DDA, 22:2–13,16) and docosatrienoic acid (DTA, 22:3–13,16,19) are two very long chain polyunsaturated fatty acids (VLCPUFAs) that are recently shown to possess strong anti-inflammatory and antitumor properties. An ELO type elongase (EhELO1) from wild plant Eranthis hyemalis can synthesize the two fatty acids by sequential elongation of linoleic acid and alpha-linolenic acid, respectively. Seed-specific expression of this gene in oilseed crop Brassica carinata produced a considerable amount of DDA and DTA in transgenic seeds. However, these fatty acids were excluded from the sn-2 position of triacylglycerols (TAGs). To improve the production level and nutrition value of the VLCPUFAs in the transgenic oilseed crop, a cytoplasmic lysophosphatidic acid acyltransferase (EhLPAAT2) for the incorporation of the two fatty acids into the sn-2 position of triacylglycerols was identified from E. hyemalis. RT-PCR analysis showed that it was preferentially expressed in developing seeds where EhELO1 was exclusively expressed in E. hyemalis. Seed specific expression of EhLPAAT2 along with EhELO1 in B. carinata resulted in the effective incorporation of DDA and DTA at the sn-2 position of TAGs, thereby increasing the total amount of DDA and DTA in transgenic seeds. To our knowledge, this is the first plant LPAAT that can incorporate VLCPUFAs into TAGs. Improved production of DDA and DTA in the oilseed crop using EhLPAAT2 and EhELO1 provides a real commercial opportunity for high value agriculture products for nutraceutical uses.

二十二碳二烯酸(DDA, 22:2 - 13,16)和二十二碳三烯酸(DTA, 22:3 - 13,16,19)是两种非常长链的多不饱和脂肪酸(VLCPUFAs),最近被证明具有很强的抗炎和抗肿瘤特性。野生植物叶毛花ELO型延长酶EhELO1可以分别通过亚油酸和α -亚麻酸的顺序延长合成这两种脂肪酸。该基因在油料作物芸苔(Brassica carinata)中的种子特异性表达可在转基因种子中产生大量的DDA和DTA。然而,这些脂肪酸被排除在三酰基甘油(TAGs)的sn-2位置之外。为了提高转基因油籽作物中VLCPUFAs的生产水平和营养价值,从叶毛孢中鉴定出一种细胞质溶血磷脂酸酰基转移酶(EhLPAAT2),该酶能将这两种脂肪酸转移到三酰基甘油的n-2位置。RT-PCR分析结果显示,EhELO1基因在萌发种子中优先表达,而EhELO1基因仅在胚膜菊中表达。EhLPAAT2和EhELO1的种子特异性表达使DDA和DTA在tag的sn-2位置有效结合,从而增加了转基因种子中DDA和DTA的总量。据我们所知,这是第一个可以将VLCPUFAs整合到tag中的植物LPAAT。利用EhLPAAT2和EhELO1在油籽作物中提高DDA和DTA的产量,为高价值的营养保健农产品提供了真正的商业机会。
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引用次数: 5
Isobutene production in Synechocystis sp. PCC 6803 by introducing α-ketoisocaproate dioxygenase from Rattus norvegicus 引入褐家鼠α-酮异己酸双加氧酶对褐家鼠PCC 6803产异丁烯的影响
IF 5.2 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-06-01 DOI: 10.1016/j.mec.2021.e00163
Henna Mustila , Amit Kugler, Karin Stensjö

Cyanobacteria can be utilized as a platform for direct phototrophic conversion of CO2 to produce several types of carbon-neutral biofuels. One promising compound to be produced photobiologically in cyanobacteria is isobutene. As a volatile compound, isobutene will quickly escape the cells without building up to toxic levels in growth medium or get caught in the membranes. Unlike liquid biofuels, gaseous isobutene may be collected from the headspace and thus avoid the costly extraction of a chemical from culture medium or from cells. Here we investigate a putative synthetic pathway for isobutene production suitable for a photoautotrophic host. First, we expressed α-ketoisocaproate dioxygenase from Rattus norvegicus (RnKICD) in Escherichia coli. We discovered isobutene formation with the purified RnKICD with the rate of 104.6 ​± ​9 ​ng (mg protein)-1 min-1 using α-ketoisocaproate as a substrate. We further demonstrate isobutene production in the cyanobacterium Synechocystis sp. PCC 6803 by introducing the RnKICD enzyme. Synechocystis strain heterologously expressing the RnKICD produced 91 ​ng ​l−1 OD750−1 ​h−1. Thus, we demonstrate a novel sustainable platform for cyanobacterial production of an important building block chemical, isobutene. These results indicate that RnKICD can be used to further optimize the synthetic isobutene pathway by protein and metabolic engineering efforts.

蓝藻可以作为直接光养转换二氧化碳的平台,以生产几种碳中性生物燃料。异丁烯是一种在蓝藻中产生光生物学的有前途的化合物。作为一种挥发性化合物,异丁烯会迅速逃离细胞,而不会在生长培养基中形成毒性水平,也不会被细胞膜捕获。与液体生物燃料不同,气态异丁烯可以从顶空收集,从而避免了从培养基或细胞中提取化学物质的昂贵费用。在这里,我们研究了一种适用于光自养寄主的异丁烯合成途径。首先,我们在大肠杆菌中表达褐家鼠α-酮异己酸双加氧酶(RnKICD)。我们发现纯化的RnKICD以α-酮异己酸酯为底物形成异丁烯的速率为104.6±9 ng (mg蛋白)-1 min-1。通过引入RnKICD酶,我们进一步证明了蓝细菌synnechocystis sp. PCC 6803中异丁烯的产生。异源表达RnKICD的胞囊菌产生91 ng l−1 OD750−1 h−1。因此,我们展示了一个新的可持续平台的蓝藻生产的一个重要的构建块化学品,异丁烯。这些结果表明,RnKICD可以通过蛋白质和代谢工程的努力进一步优化合成异丁烯途径。
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引用次数: 11
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Metabolic Engineering Communications
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