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1.
Nature ; 606(7913): 414-419, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35650436

RESUMO

All known triterpenes are generated by triterpene synthases (TrTSs) from squalene or oxidosqualene1. This approach is fundamentally different from the biosynthesis of short-chain (C10-C25) terpenes that are formed from polyisoprenyl diphosphates2-4. In this study, two fungal chimeric class I TrTSs, Talaromyces verruculosus talaropentaene synthase (TvTS) and Macrophomina phaseolina macrophomene synthase (MpMS), were characterized. Both enzymes use dimethylallyl diphosphate and isopentenyl diphosphate or hexaprenyl diphosphate as substrates, representing the first examples, to our knowledge, of non-squalene-dependent triterpene biosynthesis. The cyclization mechanisms of TvTS and MpMS and the absolute configurations of their products were investigated in isotopic labelling experiments. Structural analyses of the terpene cyclase domain of TvTS and full-length MpMS provide detailed insights into their catalytic mechanisms. An AlphaFold2-based screening platform was developed to mine a third TrTS, Colletotrichum gloeosporioides colleterpenol synthase (CgCS). Our findings identify a new enzymatic mechanism for the biosynthesis of triterpenes and enhance understanding of terpene biosynthesis in nature.


Assuntos
Ascomicetos , Talaromyces , Triterpenos , Ascomicetos/enzimologia , Colletotrichum/enzimologia , Ciclização , Difosfatos/metabolismo , Esqualeno/química , Especificidade por Substrato , Talaromyces/enzimologia , Triterpenos/química , Triterpenos/metabolismo
2.
New Phytol ; 241(2): 779-792, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37933426

RESUMO

(+)-Nootkatone is a natural sesquiterpene ketone widely used in food, cosmetics, pharmaceuticals, and agriculture. It is also regarded as one of the most valuable terpenes used commercially. However, plants contain trace amounts of (+)-nootkatone, and extraction from plants is insufficient to meet market demand. Alpinia oxyphylla is a well-known medicinal plant in China, and (+)-nootkatone is one of the main components within the fruits. By transcriptome mining and functional screening using a precursor-providing yeast chassis, the complete (+)-nootkatone biosynthetic pathway in Alpinia oxyphylla was identified. A (+)-valencene synthase (AoVS) was identified as a novel monocot-derived valencene synthase; three (+)-valencene oxidases AoCYP6 (CYP71BB2), AoCYP9 (CYP71CX8), and AoCYP18 (CYP701A170) were identified by constructing a valencene-providing yeast strain. With further characterisation of a cytochrome P450 reductase (AoCPR1) and three dehydrogenases (AoSDR1/2/3), we successfully reconstructed the (+)-nootkatone biosynthetic pathway in Saccharomyces cerevisiae, representing a basis for its biotechnological production. Identifying the biosynthetic pathway of (+)-nootkatone in A. oxyphylla unravelled the molecular mechanism underlying its formation in planta and also supported the bioengineering production of (+)-nootkatone. The highly efficient yeast chassis screening method could be used to elucidate the complete biosynthetic pathway of other valuable plant natural products in future.


Assuntos
Alpinia , Plantas Medicinais , Sesquiterpenos , Alpinia/metabolismo , Saccharomyces cerevisiae/metabolismo , Sesquiterpenos/metabolismo , Plantas Medicinais/metabolismo
3.
Proc Natl Acad Sci U S A ; 118(29)2021 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-34257153

RESUMO

Chimeric terpene synthases, which consist of C-terminal prenyltransferase (PT) and N-terminal class I terpene synthase (TS) domains (termed PTTSs here), is unique to fungi and produces structurally diverse di- and sesterterpenes. Prior to this study, 20 PTTSs had been functionally characterized. Our understanding of the origin and functional evolution of PTTS genes is limited. Our systematic search of sequenced fungal genomes among diverse taxa revealed that PTTS genes were restricted to Dikarya. Phylogenetic findings indicated different potential models of the origin and evolution of PTTS genes. One was that PTTS genes originated in the common Dikarya ancestor and then underwent frequent gene loss among various subsequent lineages. To understand their functional evolution, we selected 74 PTTS genes for biochemical characterization in an efficient precursor-providing yeast system employing chassis-based, robot-assisted, high-throughput automatic assembly. We found 34 PTTS genes that encoded active enzymes and collectively produced 24 di- and sesterterpenes. About half of these di- and sesterterpenes were also the products of the 20 known PTTSs, indicating functional conservation, whereas the PTTS products included the previously unknown sesterterpenes, sesterevisene (1), and sesterorbiculene (2), suggesting that a diversity of PTTS products awaits discovery. Separating functional PTTSs into two monophyletic groups implied that an early gene duplication event occurred during the evolution of the PTTS family followed by functional divergence with the characteristics of distinct cyclization mechanisms.


Assuntos
Alquil e Aril Transferases/genética , Proteínas Fúngicas/genética , Proteínas Mutantes Quiméricas/genética , Alquil e Aril Transferases/metabolismo , Diterpenos/química , Diterpenos/metabolismo , Evolução Molecular , Proteínas Fúngicas/metabolismo , Fungos/classificação , Fungos/enzimologia , Fungos/genética , Genoma Fúngico/genética , Estrutura Molecular , Proteínas Mutantes Quiméricas/metabolismo , Mutação , Filogenia , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Sesterterpenos/química , Sesterterpenos/metabolismo
4.
Metab Eng ; 69: 122-133, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34781019

RESUMO

Plant-derived natural active products have attracted increasing attention for use in flavors and perfumes. These compounds also have applications in insect pest control because of their environment-friendly properties. Holy basil (Ocimum sanctum), a famous herb used in Ayurveda in India, is a natural source of medical healing agents and insecticidal repellents. Despite the available genomic sequences and genome-wide bioinformatic analysis of terpene synthase genes, the functionality of the sesquiterpene genes involved in the unique fragrance and insecticidal activities of Holy basil are largely unknown. In this study, we systematically screened the sesquiterpenoid biosynthesis genes in this plant using a precursor-providing yeast system. The enzymes that synthesize ß-caryophyllene and its close isomer α-humulene were successfully identified. The enzymatic product of OsaTPS07 was characterized by in vivo mining, in vitro reaction, and NMR detection. This product was revealed as (-)-eremophilene. We created a mutant yeast strain that can achieve a high-yield titer by adjusting the gene copy number and FPP precursor enhancement. An optimized two-stage fed-batch fermentation method achieved high biosynthetic capacity, with a titer of 34.6 g/L cyclic sesquiterpene bioproduction in a 15-L bioreactor. Further insect-repelling assays demonstrated that (-)-eremophilene repelled the insect pest, fall leafworm, suggesting the potential of (-)-eremophilene as an alternative to synthetic chemicals for agricultural pest control. This study highlights the potential of our microbial platform for the bulk mining of plant-derived ingredients and provides an impressive cornerstone for their industrial utilization.


Assuntos
Ocimum sanctum , Sesquiterpenos , Reatores Biológicos , Fermentação , Saccharomyces cerevisiae/genética
5.
Angew Chem Int Ed Engl ; 61(32): e202207132, 2022 08 08.
Artigo em Inglês | MEDLINE | ID: mdl-35653160

RESUMO

Microbial cell factories reinvigorate current industries by producing complex fine chemicals at low costs. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) is the main reducing power to drive the biosynthetic pathways in microorganisms. However, insufficient intrinsic NADPH limits the productivity of microorganisms. Here, we report that supplying microorganisms with long-lived electrons from persistent phosphor mesoporous Al2 O3 (meso-Al2 O3 ) can elevate the NADPH level to facilitate efficient fine chemical production. The defects in meso-Al2 O3 were demonstrated to be highly efficient in prolonging electrons' lifetime. The long-lived electrons in meso-Al2 O3 can pass the material-microorganism interface and power the biosynthetic pathways of E. coli to produce jet fuel farnesene. This work represents a reliable strategy to design photo-biosynthesis systems to improve the productivity of microorganisms with solar energy.


Assuntos
Biocombustíveis , Energia Solar , Escherichia coli , NADP , Luz Solar
6.
J Am Chem Soc ; 142(6): 2760-2765, 2020 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-31999448

RESUMO

Herein, we report a short semisynthesis of the potent transient receptor potential canonical (TRPC) channel agonist englerin A (EA) and the related guaianes oxyphyllol and orientalol E. The guaia-6,10(14)-diene starting material was systematically engineered in Escherichia coli and Saccharomyces cerevisiae using the CRISPR/Cas9 system and was produced with high titers. The potentially scalable approach combines the advantages of synthetic biology and chemical synthesis providing an efficient and economical method for producing EA and analogues.


Assuntos
Engenharia Metabólica , Plantas/química , Sesquiterpenos de Guaiano/química , Sistemas CRISPR-Cas , Escherichia coli/genética , Saccharomyces cerevisiae/genética , Sesquiterpenos de Guaiano/síntese química
7.
Chembiochem ; 21(4): 486-491, 2020 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-31476106

RESUMO

The absolute configuration of fusaterpenol (GJ1012E) has been revised by an enantioselective deuteration strategy. A bifunctional enzyme with a terpene synthase and a prenyltransferase domain from Aspergillus brasiliensis was characterised as variediene synthase, and the absolute configuration of its product was elucidated. The uniform absolute configurations of these and structurally related di- and sesterterpenes together with a common stereochemical course for the geminal methyl groups of GGPP unravel a similar conformational fold of the substrate in the active sites of the terpene synthases. For variediene, a thermal reaction observed during GC/MS analysis was studied in detail for which a surprising mechanism was uncovered.


Assuntos
Aspergillus/enzimologia , Bactérias/química , Fungos/química , Sesterterpenos/química , Alquil e Aril Transferases/química , Proteínas Fúngicas/química , Estrutura Molecular
8.
Chembiochem ; 20(5): 677-682, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30484946

RESUMO

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.


Assuntos
Fusarium/metabolismo , Sesquiterpenos/química , Carbono-Carbono Liases/química , Clonagem Molecular , Família 26 do Citocromo P450/química , Escherichia coli/genética
9.
Beilstein J Org Chem ; 15: 2052-2058, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31501673

RESUMO

Sesquiterpene synthases in Trichoderma viride have been seldom studied, despite the efficiency of filamentous fungi for terpenoid production. Using the farnesyl diphosphate-overexpressing Saccharomyces cerevisiae platform to produce diverse terpenoids, we herein identified an unknown sesquiterpene synthase from T. viride by genome mining and determined the structure of its corresponding products. One new 5/6 bicyclic sesquiterpene and its esterified derivative were characterised by GC-MS and 1D and 2D NMR spectroscopy. To the best of our knowledge, this is the first well-identified sesquiterpene synthase from T. viride to date.

10.
Angew Chem Int Ed Engl ; 57(48): 15887-15890, 2018 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-30277637

RESUMO

Based on a terpenoid overproduction platform in yeast for genome mining, a chimeric diterpene synthase from the endophytic fungus Colletotrichum gloeosporioides ES026 was characterized as the (5R,12R,14S)-dolasta-1(15),8-diene synthase. The absolute configuration was independently verified through the use of enantioselectively deuterated terpene precursors, which unequivocally established the predicted C1-III-IV cyclization mode for this first characterized clade II-D enzyme. Extensive isotopic labeling experiments and isolation of the intermediate (1R)-δ-araneosene supported the proposed cyclization mechanism.


Assuntos
Alquil e Aril Transferases/metabolismo , Colletotrichum/enzimologia , Alquil e Aril Transferases/química , Ciclização , Estrutura Molecular , Estereoisomerismo
11.
Metab Eng ; 42: 1-8, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28438645

RESUMO

Terpenoids represent the largest family of natural products. Their structural diversity is largely due to variable skeletons generated by terpene synthases. However, terpene skeletons found in nature are much more than those generated from known terpene synthases. Most promiscuous terpene synthases (i.e. those that can generate more than one product) have not been comprehensively characterised. Here, we first demonstrated that the promiscuous terpene synthases can produce more variable terpenoids in vivo by converting precursor polyisoprenoid diphosphates of different lengths (C10, C15, C20, C25). To release the synthetic potential of these enzymes, we integrated the engineered MVA pathway, combinatorial biosynthesis, and point mutagenesis to depict the comprehensive product profiles. In total, eight new terpenoids were characterised by NMR and three new skeletons were revealed. This work highlights the key role of metabolic engineering for natural product discovery.


Assuntos
Alquil e Aril Transferases/química , Proteínas de Bactérias/química , Mutação Puntual , Fosfatos de Poli-Isoprenil/química , Alquil e Aril Transferases/genética , Proteínas de Bactérias/genética
12.
Mol Carcinog ; 52(12): 983-96, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22821714

RESUMO

Actinomycin D (Act D), a well known of clinical antitumor drug, has been used for the treatment of some highly malignant tumors, however, the clinical application was limited by its extreme cytotoxicity. In the present study, we reported that methylated actinomycin D (mAct D), a novel actinomycin D analog isolated from Streptomyces sp. KLBMP 2541 in our previous study, could not only exert stronger inhibitory effects on several human cancer cells than Act D in dose- and time-dependent manner at ng concentrations, especially on HepG2 cells, but also lower cytotoxicity in normal cells (HL-7702). Base on these results, HepG2 cells were treated for further study to illustrate the potential mechanism of mAct D. The results of nuclei morphology examination, DNA fragmentation detection, sub-G1 analysis, annexin V-FITC/PI staining and activation of caspase-3 indicated mAct D significantly induced HepG2 cells apoptosis. Semiquantitative RT-PCR and Western blot analysis revealed that mAct D induced apoptosis in HepG2 cells through mitochondria-dependent pathway by increasing levels of caspase-9, Bax, Bak while decreasing levels of Bcl-2, Bid, and Fas-dependent pathway by increasing levels of Fas, FasL, FADD, and caspase-8. Subsequently, pretreatment with specific inhibitor of caspase-8 Z-LEHD-FMK and caspase-9 Z-LEHD-FMK significantly attenuated caspase-3 activity, the cleavage of caspase-3 and PARP, meanwhile increased the cell viability. In addition, p53 and mitochondrial transcription factor A (mtTFA) were also upregulated. Taken together, ng concentrations mAct D induces the apoptosis of HepG2 through Fas- and mitochondria-mediated pathway and presents a potential novel alternative agent for the treatment of human hepatic carcinoma.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Dactinomicina/análogos & derivados , Mitocôndrias/efeitos dos fármacos , Receptor fas/metabolismo , Antineoplásicos/administração & dosagem , Inibidores de Caspase/farmacologia , Caspases/metabolismo , Linhagem Celular Tumoral/efeitos dos fármacos , Fragmentação do DNA/efeitos dos fármacos , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Dactinomicina/farmacologia , Relação Dose-Resposta a Droga , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Genes p53 , Células Hep G2/efeitos dos fármacos , Humanos , Mitocôndrias/metabolismo , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Oligopeptídeos/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
13.
Int J Syst Evol Microbiol ; 63(Pt 8): 2770-2775, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23291896

RESUMO

A novel actinomycete, designated KLBMP 1284(T), was isolated from the surface-sterilized stems of a coastal halophyte Tamarix chinensis Lour. collected from the city of Nantong, Jiangsu Province, east China. The strain was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. Analysis of the 16S rRNA gene sequence of strain KLBMP 1284(T) revealed that the strain formed a distinct clade within the phylogenetic tree based on 16S rRNA gene sequences and the highest sequence similarity (99.43 %) was to Streptomyces sulphureus NRRL B-1627(T). 16S rRNA gene sequence similarity to other species of the genus Streptomyces was lower than 97 %. Based on DNA-DNA hybridization values and comparison of morphological and phenotypic data, KLBMP 1284(T) could be distinguished from the closest phylogenetically related species, Streptomyces sulphureus NRRL B-1627(T). Thus, based on these data, it is evident that strain KLBMP 1284(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces halophytocola sp. nov. is proposed. The type strain is KLBMP 1284(T) (= KCTC 19890(T) = NBRC 108770(T)).


Assuntos
Filogenia , Plantas Tolerantes a Sal/microbiologia , Streptomyces/classificação , Tamaricaceae/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Endófitos/classificação , Endófitos/genética , Endófitos/isolamento & purificação , Ácidos Graxos/análise , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Fosfolipídeos/análise , Caules de Planta/microbiologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Streptomyces/genética , Streptomyces/isolamento & purificação
14.
Int J Syst Evol Microbiol ; 63(Pt 6): 2197-2202, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23148095

RESUMO

A novel actinomycete, designated strain KLBMP 1279(T), was isolated from surface-sterilized roots of a coastal halophyte, Salicornia europaea Linn., collected from Jiangsu Province, in the east of China. The taxonomic status of this organism was established using a polyphasic approach. 16S rRNA gene sequence analysis indicated that strain KLBMP 1279(T) was closely related to Modestobacter marinus 42H12-1(T) (99.5% 16S rRNA gene sequence similarity), Modestobacter versicolor CP153-2(T) (98.4%) and Modestobacter multiseptatus AA-826(T) (97.5%). Chemotaxonomic characteristics were consistent with its assignment to the genus Modestobacter in that the isolate had meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall, MK-9(H4) as major menaquinone and a polar lipid profile containing diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides, two unknown aminophospholipids and an unknown phospholipid. The predominant fatty acids were iso-C16:0, iso-C15:0 and C17:1ω8c. The DNA G+C content was 71.7 mol%. However, DNA-DNA hybridization assays as well as physiological and biochemical analyses differentiated strain KLBMP 1279(T) from its closest phylogenetic relatives. On the basis of phenotypic, chemotaxonomic and phylogenetic evidence, the isolate KLBMP 1279(T) represents a novel species of the genus Modestobacter, for which the name Modestobacter roseus sp. nov. is proposed; the type strain is KLBMP 1279(T) (=KCTC 19887(T)=NBRC 108673(T)=DSM 45764(T)). An emended description of the genus Modestobacter is also proposed.


Assuntos
Actinomycetales/classificação , Chenopodiaceae/microbiologia , Filogenia , Actinomycetales/genética , Actinomycetales/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/análise , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Raízes de Plantas/microbiologia , RNA Ribossômico 16S/genética , Plantas Tolerantes a Sal/microbiologia , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/análise
15.
Antonie Van Leeuwenhoek ; 103(2): 433-9, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23053697

RESUMO

A novel actinomycete, designated strain KLBMP 1262(T), was isolated from a coastal plant Dendranthema indicum (Linn.) Des Moul collected from the coastal region of Nantong, Jiangsu Province, in east China and was studied in detail for its taxonomic position. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain KLBMP 1262(T) is a member of the genus Amycolatopsis. The 16S rRNA gene sequence similarity indicated that strain KLBMP 1262(T) is most closely related to Amycolatopsis sulphurea DSM 46092(T) (97.96 %), Amycolatopsis ultiminotia RP-AC36(T) (97.50 %) and Amycolatopsis jejuensis N7-3(T) (97.44 %); similarity to other type strains of the genus Amycolatopsis was less than 97.0 %. The organism was determined to have chemical and morphological features consistent with its classification in the genus Amycolatopsis such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan and arabinose and galactose as the diagnostic sugars. The predominant menaquinone was determined to be MK-9 (H(4)). The polar lipids detected were phosphatidylmethylethanolamine, phosphatidylethanolamine, an unknown aminophospholipid, two unknown glycolipids and several unknown lipids. The major fatty acids were found to be C(16:0), iso-C(16:0) and iso-C(15:0). DNA-DNA relatedness data, together with phenotypic differences, clearly distinguished the isolate from its closest relatives. On the basis of these phenotypic and genotypic data, the isolate is considered to represent a novel species, for which the name A. jiangsuensis sp. nov. is proposed. The type strain is KLBMP 1262(T) (=KCTC 19885 (T) = NBRC 108679(T)).


Assuntos
Actinomycetales/classificação , Actinomycetales/isolamento & purificação , Chrysanthemum/microbiologia , Endófitos/classificação , Endófitos/isolamento & purificação , Técnicas de Tipagem Bacteriana , Carboidratos/análise , Parede Celular/química , China , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácido Diaminopimélico/análise , Ácidos Graxos/análise , Microscopia , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Fosfolipídeos/análise , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análise
16.
Antonie Van Leeuwenhoek ; 103(6): 1369-76, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23559043

RESUMO

A halotolerant actinomycete strain, designated strain KLBMP 1305(T), was isolated from a salt marsh plant Dendranthema indicum (Linn.) Des Moul collected from the coastal region of Nantong, Jiangsu Province, in east China and was studied in detail for its taxonomic position. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain KLBMP 1305(T) is a member of the genus Saccharopolyspora. The 16S rRNA gene sequence similarity indicated that strain KLBMP 1305(T) was most closely related to 'Saccharopolyspora pathumthaniensis' S582(T) (99.31 %), 'Saccharopolyspora endophytica' YIM 61095(T) (99.17 %) and Saccharopolyspora tripterygii YIM 65359(T) (99.15 %); similarity to other type strains of the genus Saccharopolyspora was <97.2 %. The organism had chemical and morphological features consistent with its classification in the genus Saccharopolyspora such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan and arabinose and galactose as the diagnostic sugars. The predominant menaquinone was MK-9(H4). The polar lipids detected were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unknown glycolipid and an unknown lipid. The major fatty acids were iso-C16:0, iso-C15:0, anteiso-C15:0, anteiso-C17:0 and sum in feature 8 (18:1ω7c/18:1ω6c). The G+C content of the genomic DNA of the type strain was 68.7 mol%. DNA-DNA relatedness data, together with phenotypic differences, clearly distinguished the isolate from its closest relatives. On the basis of these phenotypic and genotypic data, the isolate represents a novel species, for which the name Saccharopolyspora dendranthemae sp. nov. is proposed. The type strain is KLBMP 1305(T) (=KCTC 19889(T) = NBRC 108675(T)).


Assuntos
Chrysanthemum/microbiologia , Endófitos/isolamento & purificação , Endófitos/fisiologia , Saccharopolyspora/isolamento & purificação , Saccharopolyspora/fisiologia , Tolerância ao Sal , Áreas Alagadas , Técnicas de Tipagem Bacteriana , Composição de Bases , Sequência de Bases , China , DNA Bacteriano/genética , Ácido Diaminopimélico/metabolismo , Endófitos/classificação , Endófitos/genética , Metabolismo Energético , Ácidos Graxos , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Saccharopolyspora/classificação , Saccharopolyspora/genética , Análise de Sequência de DNA , Cloreto de Sódio , Vitamina K 2/análise
17.
Adv Mater ; : e2305828, 2023 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-37677048

RESUMO

At the intersection of synthetic biology and materials science, engineered living materials (ELMs) exhibit unprecedented potential. Possessing unique "living" attributes, ELMs represent a significant paradigm shift in material design, showcasing self-organization, self-repair, adaptability, and evolvability, surpassing conventional synthetic materials. This review focuses on reviewing the applications of ELMs derived from bacteria, fungi, and plants in environmental remediation, eco-friendly architecture, and sustainable energy. The review provides a comprehensive overview of the latest research progress and emerging design strategies for ELMs in various application fields from the perspectives of synthetic biology and materials science. In addition, the review provides valuable references for the design of novel ELMs, extending the potential applications of future ELMs. The investigation into the synergistic application possibilities amongst different species of ELMs offers beneficial reference information for researchers and practitioners in this field. Finally, future trends and development challenges of synthetic biology for ELMs in the coming years are discussed in detail.

18.
Nat Commun ; 14(1): 6800, 2023 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-37884498

RESUMO

Microbial communication can drive coordinated functions through sensing, analyzing and processing signal information, playing critical roles in biomanufacturing and life evolution. However, it is still a great challenge to develop effective methods to construct a microbial communication system with coordinated behaviors. Here, we report an electron transfer triggered redox communication network consisting of three building blocks including signal router, optical verifier and bio-actuator for microbial metabolism regulation and coordination. In the redox communication network, the Fe3+/Fe2+ redox signal can be dynamically and reversibly transduced, channeling electrons directly and specifically into bio-actuator cells through iron oxidation pathway. The redox communication network drives gene expression of electron transfer proteins and simultaneously facilitates the critical reducing power regeneration in the bio-actuator, thus enabling regulation of microbial metabolism. In this way, the redox communication system efficiently promotes the biomanufacturing yield and CO2 fixation rate of bio-actuator. Furthermore, the results demonstrate that this redox communication strategy is applicable both in co-culture and microbial consortia. The proposed electron transfer triggered redox communication strategy in this work could provide an approach for reducing power regeneration and metabolic optimization and could offer insights into improving biomanufacturing efficiency.


Assuntos
Ferro , Consórcios Microbianos , Biocatálise , Oxirredução , Transporte de Elétrons
19.
Antonie Van Leeuwenhoek ; 102(2): 353-60, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22552630

RESUMO

A novel Gram-positive, aerobic, rod-shaped and mycelia-producing bacterial strain, designated KLBMP 1050(T), was isolated from the stem of the oil-seed plant Jatropha curcas L. collected from Sichuan Province, south-west China. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the isolate KLBMP 1050(T) belonged to the genus Nocardioides, with the highest sequence similarity to Nocardioides albus KCTC 9186(T) (99.38 %) and Nocardioides luteus KCTC 9575(T) (99.03 %). However, the DNA-DNA relatedness of isolate KLBMP 1050(T) to these two type strains were 37.5 ± 3.5 and 33 ± 2.3 %, respectively. Strain KLBMP 1050(T) grew at the pH range 6-11, temperature range 10-32 °C and with 0-12 % NaCl. The physiological properties of strain KLBMP 1050(T) differ from those of N. albus KCTC 9186(T) and N. luteus KCTC 9575(T). The cell-wall peptidoglycan contained LL: -diaminopimelic acid and MK-8(H(4)) was the major respiratory quinone. The predominant cellular fatty acid of strain KLBMP 1050(T) was iso-C(16:0) (23.3 %). The total DNA G+C content was 70.1 mol%. On the basis of the phenotypic, chemotaxonomic and phylogenetic data, strain KLBMP 1050(T) represents a novel species of the genus Nocardioides, for which the name Nocardioides panzhihuaensis sp. nov. is proposed. The type strain is KLBMP 1050(T) (= KCTC 19888(T) = NBRC 108680(T)).


Assuntos
Actinomycetales/isolamento & purificação , Endófitos/isolamento & purificação , Jatropha/microbiologia , Plantas Medicinais/microbiologia , Actinomycetales/classificação , Actinomycetales/genética , Actinomycetales/metabolismo , Composição de Bases , Endófitos/classificação , Endófitos/genética , Endófitos/metabolismo , Ácidos Graxos/metabolismo , Dados de Sequência Molecular , Filogenia
20.
Antonie Van Leeuwenhoek ; 102(2): 289-96, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22527624

RESUMO

A novel actinomycete, designated strain KLBMP 4601(T), was isolated from the root of the medicinal plant Curcuma phaeocaulis collected from Sichuan Province, south-west China. The strain produced extensively branched substrate and aerial hyphae that carried straight to flexuous spore chains. Chemotaxonomic properties of this strain were consistent with those of members of the genus Streptomyces. The cell wall of strain KLBMP 4601(T) contained LL-diaminopimelic acid as the characteristic diamino acid. The major menaquinone was MK-9(H(4)), with minor amounts of MK-9(H(6)), MK-9(H(8)) and MK-10(H(2)). The major fatty acids were C(16:0), iso-C(16:0), C(18:1)ω9c and C(16:1), iso G. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain KLBMP 4601(T) belongs to the genus Streptomyces and is most closely related to Streptomyces armeniacus JCM 3070(T) (97.9 %), Streptomyces pharmamarensis PM267(T) (97.6 %) and Streptomyces artemisiae YIM 63135(T) (97.5 %). The 16S rRNA gene sequence similarity between strain KLBMP 4601(T) and other members of this genus were lower than 97.5 %. DNA-DNA hybridization studies of strain KLBMP 4601(T) with the three closest species showed relatedness values of 36.3 ± 4.2 %, 27.3 ± 0.6 %, and 30.9 ± 2.5 %, respectively. On the basis of chemotaxonomic, phenotypic and genotypic characteristics, it is evident that strain KLBMP 4601(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces phytohabitans sp. nov. is proposed. The type strain is KLBMP 4601(T) (=KCTC 19892(T) = NBRC 108772(T)).


Assuntos
Curcuma/microbiologia , Endófitos/isolamento & purificação , Plantas Medicinais/microbiologia , Streptomyces/isolamento & purificação , China , Curcuma/fisiologia , Endófitos/classificação , Endófitos/genética , Endófitos/fisiologia , Ácidos Graxos/metabolismo , Dados de Sequência Molecular , Filogenia , Plantas Medicinais/fisiologia , Streptomyces/classificação , Streptomyces/genética , Streptomyces/fisiologia
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