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萜类化合物具有广泛的生理活性与重要的经济价值,利用酿酒酵母进行萜类合成具有低价、高效等优势。然而部分植物源合成萜类的关键酶在酿酒酵母中难表达、产量低,难以工业应用,因此有效的调控策略显得至关重要。本文从萜类化合物在酿酒酵母中的合成途径入手,介绍了关键酶、代谢途径、CRISPR基因编辑系统和人工合成染色体技术4个方面的调控策略在酿酒酵母合成萜类化合物中的应用。阐述了关键酶的筛选、改造,理性与非理性设计,MVA途径、乙酰辅酶A合成途径与亚细胞结构的代谢途径改造的优势。指出了多重调控策略组合调控的方式是实现酿酒酵母高效合成萜类化合物的有效方法。此外,CRISPR基因编辑系统与人工合成染色体技术的快速发展将为酿酒酵母细胞工厂的深入开发与利用提供有力工具。 相似文献
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酿酒酵母作为细胞工厂被用来生产多种萜类化合物。乙酰辅酶A为合成萜类化合物的基本前体,细胞质乙酰辅酶A供应不足会导致目标产物产量较低,调控乙酰辅酶A合成是构建目标萜类化合物高产合成途径的重要手段。本文介绍了酿酒酵母乙酰辅酶A作为重要中心碳代谢分子,主要在细胞核组蛋白乙酰化、细胞质丙酮酸脱氢酶支路、线粒体三羧酸循环和过氧化物酶体乙醛酸循环中参与的代谢过程。总结了通过强化酿酒酵母内源丙酮酸脱氢酶支路,引入低三磷酸腺苷(ATP)消耗的异源乙酰辅酶A合成途径,增加辅酶A合成和利用线粒体乙酰辅酶A含量高且对其不渗透的特性进行区域化合成以提高乙酰辅酶A含量的代谢工程策略,旨在为酿酒酵母萜类化合物的高效合成提供借鉴。 相似文献
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萜类化合物具有可观的经济价值,但是目前的生产过程复杂、产量低。酿酒酵母甲羟戊酸途径为萜类化合物的合成提供直接前体,因此酿酒酵母细胞具有合成异源萜类化合物的天然优势。对酿酒酵母甲羟戊酸途径的清晰认识是对其进行有效利用的基础,本文从代谢途径、关键酶的特点和全局调控机制3个方面对该途径进行了介绍。从代谢途径的构建和优化、模块与底盘细胞的适配、模块构建及组装方式的角度概述了酿酒酵母细胞异源合成单萜、倍半烯萜、二萜、三萜类化合物的研究进展。指出实现酿酒酵母高效合成萜类化合物所需要解决的基础问题是对酿酒酵母甲羟戊酸途径进行更为全面了解和对萜类化合物的天然代谢途径进行明确解析;另外,合成生物学的进一步发展也将为此提供应用基础。 相似文献
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萜类化合物在自然界存在广泛,许多已被用于香料工业,萜类香料也是芳香产业关注的热点之一。生物合成可以利用廉价和可再生资源合成萜类化合物,提供一种独立于天然来源的可持续获取所需产品的替代方法,与植物提取和化学合成相比是一种更有前途的香料生产途径。随着香精香料市场的不断增长,合成生物学指导下的萜类化合物生产在推动未来市场前景方面显示出巨大的潜力。综述了合成生物学在萜类香料生产的应用、萜类化合物的微生物细胞工厂构建及工程菌株技术平台等,同时展望了生物合成萜类香料的应用前景。 相似文献
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三萜类化合物在天然产物中广泛存在,含量丰富,是许多药用植物的主要活性成分之一。早在上世纪30年代就有关于三萜的研究,该类物质以其显著的活性与广泛的药理作用受到研究者的青睐。近年来关于三萜及其衍生物的合成是受人关注的研究热点。三萜的结构修饰多以四环三萜及五环三萜为起始物,针对四环三萜的修饰位点主要在A环C-2、C-3位及D环上的侧链,多引入羟基、酯基或含氮基团等。针对五环三萜的修饰主要在A环C-2、C-3位及C-28位,主要有酯化、酰胺化及引入含氮结构等修饰,还有部分非常见的修饰位点同样对活性有重要影响。综述了三萜及其衍生物的结构修饰与活性变化研究进展,为三萜的合成修饰与进一步开发利用提供参考。 相似文献
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三萜皂苷类化合物通常由一个或多个糖基连接在疏水的皂苷元上构成,其作为传统中草药的活性成分有着广泛应用。三萜皂苷类化合物的传统获取方法是从植物中提取,由于植物中三萜皂苷类化合物含量较低,植物生长受到土地资源、气候、环境等因素影响较大,严重制约了其大规模推广和应用。利用合成生物学技术,设计构建微生物细胞工厂合成三萜皂苷类化合物被认为是一种变革性的生产方法,成为新的研究热点。在三萜皂苷类化合物的合成过程中,糖基转移酶起着重要作用。阐述了利用糖基转移酶合成三萜皂苷类化合物的研究进展,为三萜皂苷类化合物的进一步应用提供参考。 相似文献
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三萜皂苷类化合物通常由一个或多个糖基连接在疏水的皂苷元上构成,其作为传统中草药的活性成分有着广泛应用。三萜皂苷类化合物的传统获取方法是从植物中提取,由于植物中三萜皂苷类化合物含量较低,植物生长受到土地资源、气候、环境等因素影响较大,严重制约了其大规模推广和应用。利用合成生物学技术,设计构建微生物细胞工厂合成三萜皂苷类化合物被认为是一种变革性的生产方法,成为新的研究热点。在三萜皂苷类化合物的合成过程中,糖基转移酶起着重要作用。阐述了利用糖基转移酶合成三萜皂苷类化合物的研究进展,为三萜皂苷类化合物的进一步应用提供参考。 相似文献
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天然产物泛指来源于植物、动物、微生物、海洋生物和矿物的化合物。天然产物的来源有多种途径,从天然产物中寻找有活性的化合物作为先导化合物创制新药是世界药学工作者公认的有效途径之一。天然来源药物可分为原始天然化合物、经半合成或全合成得到的化合物。 相似文献
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聚酮化合物具有广泛的药用活性和极高的经济价值,但如何高效、经济、绿色、环保地合成聚酮化合物是目前急需解决的问题。随着合成生物学的发展及分子生物学技术的进步,不断有新的技术和策略被用于聚酮化合物的生物制造。本文介绍了聚酮化合物生物制造中的关键酶、前体物质及代谢途径等,分析了通过CRISPR技术及翻译后修饰代谢工程优化代谢调控网络;通过替换及优化启动子等手段改造与优化代谢途径;通过构建简单、高效的异源表达系统等策略提高聚酮化合物的生物制造效率等。在此基础上对红霉素、阿维菌素、多杀菌素的合成生物学研究的最新进展进行了总结,进而对当前聚酮化合物生物制造面临的产量及效率低下等问题和可能的解决途径,如平衡初级代谢与次级代谢,构建新型、优势底盘细胞及代谢网络的重新设计与改造等进行了展望。 相似文献
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Yujie Yuan Dr. Martin Litzenburger Shu Cheng Dr. Guangkai Bian Dr. Ben Hu Pan Yan Dr. Yousheng Cai Prof. Zixin Deng Prof. Dr. Rita Bernhardt Prof. Dr. Tiangang Liu 《Chembiochem : a European journal of chemical biology》2019,20(5):677-682
Sesquiterpenes represent a class of important terpenoids with high structural diversity and a wide range of applications. The cyclized core skeletons are generated by sesquiterpene cyclases, and the structural diversity is further increased by a series of modification steps. Cytochromes P450 (P450s) are a class of monooxygenases and one of the main contributors to the structural diversity of natural products. Some of these P450s show a broad substrate range and might be promising candidates for the implementation of cascade reactions. In this study, a combinatorial biosynthesis approach was utilized by the combination of a promiscuous myxobacterial P450 (CYP260B1) with two sesquiterpene cyclases (FgJ01056, FgJ09920) of filamentous fungi. Two oxygenated products, culmorin and culmorone, and a new compound, koraidiol, were successfully generated and characterized. This approach suggests the potential use of noncognate P450s to produce novel oxygenated terpenoids, or to generate a novel biosynthetic route for known terpenoids by a combinatorial biosynthesis strategy. 相似文献
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Jianrong Wang Yangyuan Li Danni Liu 《International journal of molecular sciences》2014,15(12):22188-22202
Poria cocos (P. cocos) has long been used as traditional Chinese medicine and triterpenoids are the most important pharmacologically active constituents of this fungus. Farnesyl pyrophosphate synthase (FPS) is a key enzyme of triterpenoids biosynthesis. The gene encoding FPS was cloned from P. cocos by degenerate PCR, inverse PCR and cassette PCR. The open reading frame of the gene is 1086 bp in length, corresponding to a predicted polypeptide of 361 amino acid residues with a molecular weight of 41.2 kDa. Comparison of the P. cocos FPS deduced amino acid sequence with other species showed the highest identity with Ganoderma lucidum (74%). The predicted P. cocos FPS shares at least four conserved regions involved in the enzymatic activity with the FPSs of varied species. The recombinant protein was expressed in Pichia pastoris and purified. Gas chromatography analysis showed that the recombinant FPS could catalyze the formation of farnesyl diphosphate (FPP) from geranyl diphosphate (GPP) and isopentenyl diphosphate (IPP). Furthermore, the expression profile of the FPS gene and content of total triterpenoids under different stages of development and methyl jasmonate treatments were determined. The results indicated that there is a positive correlation between the activity of FPS and the amount of total triterpenoids produced in P. cocos. 相似文献
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利用工程化微生物生产天然药物如紫杉醇在近年来受到研究者广泛关注。本工作研究如何设计和构建人工酵母以生产紫杉醇生物合成途径中第一个由细胞色素P450酶催化的羟化产物--5α羟化紫杉二烯醇(taxadien-5α-ol)。利用组合设计原理,选用3种不同红豆杉来源的紫杉二烯5α羟化酶和2种不同植物源的细胞色素P450还原酶,分别对其进行N端穿膜区域的预测和截短,并对还原酶使用2种不同强度启动子进行调控,所得羟化酶模块和还原酶模块之间分别表达共产生72种组合方式。然后将其分别整合入紫杉二烯生产菌株基因组中,最终在72个组合中筛选到48个可以生产5α羟化紫杉二烯醇的菌株,最高产量67.3 μg·L-1,在酿酒酵母中实现紫杉二烯C5位羟化产物的从头合成。研究结果表明,利用组合设计策略研究模块间相互作用及其与底盘间适配性将为在微生物细胞中实现细胞色素P450酶系介导的催化反应过程提供重要参考。 相似文献
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RM Domingues EL Oliveira CS Freire RM Couto PC Simões CP Neto AJ Silvestre CM Silva 《International journal of molecular sciences》2012,13(6):7648-7662
Eucalyptus bark contains significant amounts of triterpenoids with demonstrated bioactivity, namely triterpenic acids and their acetyl derivatives (ursolic, betulinic, oleanolic, betulonic, 3-acetylursolic, and 3-acetyloleanolic acids). In this work, the supercritical fluid extraction (SFE) of Eucalyptus globulus deciduous bark was carried out with pure and modified carbon dioxide to recover this fraction, and the results were compared with those obtained by Soxhlet extraction with dichloromethane. The effects of pressure (100-200 bar), co-solvent (ethanol) content (0, 5 and 8% wt), and multistep operation were studied in order to evaluate the applicability of SFE for their selective and efficient production. The individual extraction curves of the main families of compounds were measured, and the extracts analyzed by GC-MS. Results pointed out the influence of pressure and the important role played by the co-solvent. Ethanol can be used with advantage, since its effect is more important than increasing pressure by several tens of bar. At 160 bar and 40 °C, the introduction of 8% (wt) of ethanol greatly improves the yield of triterpenoids more than threefold. 相似文献
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Chunhua Song Yalin Guan Dongrui Zhang Xun Tang Ying Chang 《International journal of molecular sciences》2022,23(10)
Fragrant woodfern (Dryopteris fragrans) is a medicinal plant rich in terpenoids. Ultraviolet-B (UV–B) light could increase concentration of terpenoids. The aim of this study was to analyze how UV–B regulates the terpenoid synthesis of the molecular regulatory mechanism in fragrant woodfern. In this study, compared with the control group, the content of the terpenes was significantly higher in fragrant woodfern leaves under UV–B treatment for 4 days (d). In order to identify how UV–B regulates the terpenoid metabolic mechanism in fragrant woodfern, we examined the mRNAs and small RNAs in fragrant woodfern leaves under UV–B treatment. mRNA and miRNA–seq identified 4533 DEGs and 17 DEMs in the control group compared with fragrant woodfern leaves under UV–B treatment for 4 d. mRNA–miRNA analysis identified miRNA target gene pairs consisting of 8 DEMs and 115 miRNAs. The target genes were subjected to GO and KEGG analyses. The results showed that the target genes were mainly enriched in diterpene biosynthesis, terpenoid backbone biosynthesis, plant hormone signal transduction, MEP pathway and MVA pathway, in which miR156 and miR160 regulate these pathways by targeting DfSPL and DfARF, respectively. The mRNA and miRNA datasets identified a subset of candidate genes. It provides the theoretical basis that UV–B regulates the terpenoid synthesis of the molecular regulatory mechanism in fragrant woodfern. 相似文献
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Brock NL Tudzynski B Dickschat JS 《Chembiochem : a European journal of chemical biology》2011,12(17):2667-2676
The fungus Fusarium fujikuroi IMI58289 emits a complex pattern of volatile terpenoids including two major compounds, the sesquiterpene alcohol α-acorenol and the diterpene ent-kaurene. ent-Kaurene is the precursor for the phytohormone gibberellic acid (GA(3)) and is produced from geranylgeranyl diphosphate (GGPP) via ent-copalyl diphosphate by the bifunctional ent-copalyl diphosphate/ent-kaurene synthase (CPS/KS). Several structurally related diterpenes were identified as side products of the CPS/KS. Deletion of the cps/ks gene or the whole GA(3) biosynthetic gene cluster resulted in completely abolished diterpene production. Mutants with deletions of the cytochrome P450 monooxygenase gene P450-4, which is responsible for the three oxidation steps from ent-kaurene to ent-kaurenoic acid en route to GA(3), accumulate diterpene hydrocarbons. Feeding with [6,6,6-(2) H(3)] mevalonolactone gave insights into the stereochemistry of the GGPP cyclisation, which operates with a chair-chair-"antipodal" fold. A rational biosynthetic scheme for all identified sesquiterpenes demonstrated their formation from farnesyl diphosphate (FPP) via three alternative initial cyclisations. Genome sequencing revealed the presence of five putative sesquiterpene synthase genes in the F. fujikuroi genome. The structures of several trace compounds from other classes have been identified as new natural products; these were delineated from their mass spectra and unambiguously assigned by comparison to synthetic references. 相似文献
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Xiaobin Shang Kai Oliver Bker Shahed Taheri Thelonius Hawellek Wolfgang Lehmann Arndt F. Schilling 《International journal of molecular sciences》2021,22(18)
Osteoarthritis (OA) is a chronic disease affecting the whole joint, which still lacks a disease-modifying treatment. This suggests an incomplete understanding of underlying molecular mechanisms. The Wnt/β-catenin pathway is involved in different pathophysiological processes of OA. Interestingly, both excessive stimulation and suppression of this pathway can contribute to the pathogenesis of OA. microRNAs have been shown to regulate different cellular processes in different diseases, including the metabolic activity of chondrocytes and osteocytes. To bridge these findings, here we attempt to give a conclusive overview of microRNA regulation of the Wnt/β-catenin pathway in bone and cartilage, which may provide insights to advance the development of miRNA-based therapeutics for OA treatment. 相似文献