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streptomyces lydicus A_(01) affects soil microbial diversity, improving growth and resilience in tomato

Journal of Integrative Plant Biology 2019年第2期61卷 182-196页

作者： Qiong Wu Caige Lu Mi Ni Hongli Wang Weicheng Liu Jie Chen The Key laboratory of Urban(South)Agriculture School of Agriculture and Biology Shanghai Jiao Tong University Laboratory of Biological Microbiology Institute of Plant and Environment Protection Beijing Academy of Agriculture and Forestry Sciences

The actinomycete streptomyces lydicus A_(01) promotes tomato seedling growth; however, the underlying mechanism is unclear. In this study, we investigated whether changes in soil microbial diversity, following streptomyces lydicus A_(01) treatment, were responsible for the increased tomato seedling growth. Eukaryotic 18 S ribosomal DNA(rDNA) sequencing showed that S. lydicus A_(01)-treated and untreated soil shared 193 operational taxonomic units(OTUs), whereas bacterial 16 S rDNA sequencing identified 1,219 shared OTUs between the treated and untreated soil. Of the 42 dominant eukaryotic OTUs, eight were significantly increased and six were significantly decreased after A_(01) treatment. Of the 25 dominant bacterial OTUs, 12 were significantly increased and eight were significantly decreased after A_(01) *** of the eukaryotes and bacteria that increased in abundance exhibited growth promoting characteristics,which were mainly predicted to be associated with mineralization of nitrogen and phosphorus, phosphate solubilization, nutrient accumulation, and secretion of auxin, whereas some were related to plant protection,such as the degradation of toxic and hazardous substances. Soil composition tests showed that S. lydicus A_(01) treatment enhanced the utilization of nitrogen, phosphorus, and potassium in tomato seedlings. Thus, microbial fertilizers based on S. lydicus A_(01) may improve plant growth, without the detriment effects of chemical fertilizers.

关键词： OTU streptomyces lydicus A affects soil microbial diversity improving growth and resilience in tomato

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Identification and Characterization of hmr19 Gene Encoding a Multidrug Resistance Efflux Protein from streptomyces hygroscopicus *** Strain 10-22

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 519-528页

作者： Lei QIN , Heng-An WANG1 , Zhong-Qin WU, Xiao-Feng ZHANG, Mei-Lei JIN,Zi-Xin DENG2, and Guo-Ping ZHAOShanghaiResearch Center of Biotechnology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences,Graduate School of the Chinese Academy of Sciences, Shanghai 200233, China 1School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 201101, China 2School of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai 200030, China ShanghaiResearch Center of Biotechnology Shanghai Institute for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences School of Agriculture and Biology Shanghai Jiaotong University

The hmr19 gene was cloned from streptomyces hygroscopicus subsp. yingchengensis strain 10- 22, a bacterium strain producing agricultural antibiotics. Sequence similarity comparison indicates that hmr19 gene may encode a predicted protein with 14 putative transmembrane α-helical spanners, belonging to the drug:H+ antiporter-2 family of the major facilitator superfamily. The expression of hmr19 in the mycelium of strain 10-22 was detected by Western blotting analysis. Gene replacement technology was employed to construct an hmr19 disruption mutant. The growth inhibition test against different antibiotics indicated that the mutant strain was 5–20 fold more susceptible to tetracycline, vancomycin and mitomycin C than the parental wild type strain. The mutant took up tetracycline much faster and accumulated more antibiotics than the wild type strain 10-22. While with the addition of an energy uncoupler, carbonyl cyanide m- chlorophenylhydrazone, the characteristics of the accumulation of [3H]tetracycline in these two strains were almost the same. It was thus concluded that hmr19 encoded a multidrug resistance efflux protein.

关键词： multidrug resistance (MDR) gene replacement (knockout) streptomyces streptomyces hygroscopicus subsp. yingchengensis strain 10-22

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Location and PCR analysis of catabolic genes in a novel streptomyces sp. DUT_AHX capable of degrading nitrobenzene

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Journal of Environmental Sciences 2008年第7期20卷 865-870页

作者： Al Haixin Zhou Jiti Lv Hong Wang Jing Guo Hanb Liu Guangfei Qu Yuanyuan Dalian Univ Technol Sch Environm & Biol Sci & Technol Dalian 116024 Peoples R China Hebei Univ Sci & Technol Sch Environm Sci & Engn Shijiazhuang 050018 Peoples R China

A novel strain of streptomyces ***_AHX was isolated from sludge contaminated with nitrobenzene and identified on the basis of physiological and biochemical tests and 16S ribosomal DNA (rDNA) sequence *** optimal degradation conditions were as follows:temperature 30℃,pH 7.0-8.0,shaking speed 150-180 r/min,and inocula 10%(V/V).The strain,which possessed a partial reductive pathway with the release of ammonia,was also able to grow on mineral salts basal (MSB) medium plates with 2-aminophenol, pheno...

关键词： streptomyces nitrobenzene degradation plasmid curing PCR

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Multiplex genome editing using a dCas9-cytidine deaminase fusion in streptomyces

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Science China(Life Sciences) 2020年第7期63卷 1053-1062页

作者： Yawei Zhao Jinzhong Tian Guosong Zheng Jun Chen Chuanwen Sun Zhongyi Yang Andrei A.Zimin Weihong Jiang Zixin Deng Zhijun Wang Yinhua Lu College of Life Sciences.Shanghai Normal University Shanghai 200234China State Key Laboratory of Microbial Metabolism School of Life Sciences and Biotechnology Shanghai Jiao Tong UniversityShanghai 200030China Key Laboratory of Synthetic Biology CAS Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghai 200032China School of Life Science Taizhou UniversityTaizhou 318000China Institute of Biochemistry and Physiology of Microorganisms Russian Academy of SciencesPushchino 142290Russia Shanghai Engineering Research Center of Plant Germplasm Resources College of Life SciencesShanghai Normal UniversityShanghai 200234China

CRISPR/Cas-mediated genome editing has greatly facilitated the study of gene function in streptomyces. However, it could not be efficiently employed in streptomycetes with low homologous recombination(HR) ability. Here, a deaminase-assisted base editor d Cas9-CDA-ULstr was developed in streptomyces, which comprises the nuclease-deficient Cas9(dCas9), the cytidine deaminase from Petromyzon marinus(PmCDA1), the uracil DNA glycosylase inhibitor(UGI) and the protein degradation tag(LVA tag). Using d Cas9-CDA-ULstr , we achieved single-, double-and triple-point mutations(cytosine-to-thymine substitutions)at target sites in streptomyces coelicolor with efficiency up to 100%, 60% and 20%, respectively. This base editor was also demonstrated to be highly efficient for base editing in the industrial strain, streptomyces rapamycinicus, which produces the immunosuppressive agent rapamycin. Compared with base editors derived from the cytidine deaminase rAPOBEC1, the PmCDA1-assisted base editor dCas9-CDA-ULstr could edit cytosines preceded by guanosines with high efficiency, which is a great advantage for editing streptomyces genomes(with high GC content). Collectively, the base editor dCas9-CDA-ULstr could be employed for efficient multiplex genome editing in streptomyces. Since the d Cas9-CDA-ULstr -based genome editing is independent of HR-mediated DNA repair, we believe this technology will greatly facilitate functional genome research and metabolic engineering in streptomyces strains with weak HR ability.

关键词： base editors dCas9-CDA-ULstr multiplex editing streptomyces

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Purification and properties of glutamate synthase from streptomyces lincolnensis

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Science China(Life Sciences) 1998年第1期41卷 37-46页

作者：金詟焦瑞身 1. Shanghai Institute of Plant Physiology Chinese Academy of Sciences 200032 Shanghai China

Glutamate synthase (EC 1 4 1 14) was purified to homogeneity from a cell\|free extract of streptomyces lincolnensis by precipitation with streptomycin sulfate and ammonium sulfate, and column chromatography on DEAE\| cellulose, Sepharose 6B, DEAE\|sephadex A\|50, hydroxyapatite and Sephadex G\|150. The enzyme activity is stabilized by addition of α ketoglutarate, PMSF,EDTA, β mercaptoethanol and glycerol. The native enzyme has a molecular weight of 138 000 and is composed of two nonidentical subunits with molecular weights of 81 000 and 57 000. Spectroscopic examination of the enzyme gave absorption maximum at 280 and none at 380 and 440 nm, indicating the absence of iron and flavin. The enzyme shows optimum activity at pH 7.2 and 30℃. Km values for α ketoglutarate, L\|glutamine and NADH were 417, 435, and 52.1 μmol/L, respectively. When NADPH was substituted for NADH as reductant, there was approximately 13% of the control activity. The activity of this glutamate synthase is inhibited by its products (i.e. glutamate and NAD), several metal ions, amino acids and tricarboxylic acid cycle intermediates.

关键词： streptomyces lincolnensis glutamate synthase purification properties.

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Multi-omics Data Reveal the Effect of Sodium Butyrate on Gene Expression and Protein Modification in streptomyces

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Genomics, Proteomics & Bioinformatics 2023年第6期21卷 1149-1162页

作者： Jiazhen Zheng Yue Li Ning Liu Jihui Zhang Shuangjiang Liu Huarong Tan State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of SciencesBeijing 100101China College of Life Sciences University of Chinese Academy of SciencesBeijing 100049China State Key Laboratory of Microbial Biotechnology Shandong UniversityQingdao 266237China

Streptomycetes possess numerous gene clusters and the potential to produce a large amount of natural *** deacetylase(HDAC)inhibitors play an important role in the regulation of histone modifications in fungi,but their roles in prokaryotes remain poorly ***,we investigated the global effects of the HDAC inhibitor,sodium butyrate(SB),on marine-derived streptomyces olivaceus FXJ 8.021,particularly focusing on the activation of secondary metabolite *** antiSMASH analysis revealed 33 secondary metabolite biosynthetic gene clusters(BGCs)in strain FXJ 8.021,among which the silent lobophorin BGC was activated by *** data showed that the expression of genes involved in lobophorin biosynthesis(ge00097–ge00139)and CoA-ester formation(e.g.,ge02824),as well as the glycolysis/gluconeogenesis pathway(e.g.,ge01661),was significantly up-regulated in the presence of *** CoA-ester analysis confirmed that SB triggered the biosynthesis of CoA-ester,thereby increasing the precursor supply for lobophorin *** acetylomic analysis revealed that the acetylation levels on 218 sites of 190 proteins were up-regulated and those on 411 sites of 310 proteins were *** acetylated proteins were particularly enriched in transcriptional and translational machinery components(e.g.,elongation factor GE04399),and their correlations with the proteins involved in lobophorin biosynthesis were established by protein–protein interaction network analysis,suggesting that SB might function via a complex hierarchical.

关键词： Silent gene cluster streptomyces Sodium butyrate Protein modification Acetylome

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Cangumycins A-F, six new angucyclinone analogues with immunosuppressive activity from streptomyces

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Chinese Journal of Natural Medicines 2019年第12期17卷 982-987页

作者： WANG Lei WANG Li ZHOU Zhi WANG Yong-Jiang HUANG Jian-Ping MA Ya-Tuan LIU Yang HUANG Sheng-Xiong State Key Laboratory of Phytochemistry and Plant Resources in West China and CAS Center for Excellence in Molecular Plant SciencesKunming Institute of BotanyChinese Academy of SciencesKunming 650201China College of Chemistry and Pharmacy Northwest Agriculture and Forestry UniversityYangling 712100China Research Center Chengdu Medical CollegeChengdu 610500China Savaid Medical School University of Chinese Academy of SciencesBeijing 100049China

Cangumycins A-F(1–6), six new angucyclinone analogues, together with two known ones(7 and 8), were isolated from the fermentation broth of a soil-derived streptomyces sp. KIB-M10. Structures of these compounds were elucidated via a joint use of spectroscopic analyses and single-crystal X-ray diffractions. Among them, cangumycins E(5) and F(6) share a C-ring cleaved backbone, and cangumycins B(2) and E(5) exhibit potent immunosuppressive activity(IC50 8.1 and 2.7 μmol·L^-1, respectively) against human T cell proliferation at a non-cytotoxic concentration.

关键词： streptomyces Angucyclinones Benz[a]anthracenes Natural products Immunosuppressive activity

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Cyclic and Linear Thiopeptides from Soil-Derived streptomyces *** 203702 with Antiviral and Antibacterial Activities

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Chinese Journal of Chemistry 2021年第12期39卷 3277-3284页

作者： Dewu Zhang Yujia Wang Xinxin Hu Xiaoyu Wang Linzi Li Guowei Gu Bingyuan Zhang Shan Cen Xuefu You Liyan Yu Division for Medicinal Microorganisms Related Strains CAMS Collection Center of Pathogenic Microorganisms Institute of Medicinal BiotechnologyChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China College of Health and Environment Beijing Union UniversityBeijing 100023China School of Pharmacy Yantai UniversityYantaiShangdong 264005China

A new cyclic thiopeptide,berninamycin F(3),three new linear thiopeptides,berninamycins G—I(4-6),and two known berninamycin derivatives,berninamycins C and D(1 and 2)were isolated from streptomyces *** *** structures were elucidated through HRESIMS and one-and two-dimensional NMR data,and the absolute configurations were assigned using Marfey's *** 4-6 are the first examples of linear berninamycin ***,compound 4 is the first example of linear thiopeptide with a 1-(oxazol-2-yl)ethan-1-one group,compound 6 is the first linear thiopeptide derivative with methylated oxoacetate *** 1-6 exhibited excellent anti-Zika virus activities with IC_(50) values in the range of 4.4-10.5μmol/L,which were superior to that of the positive control ribavirin(IC_(50)=49.2μmol/L).Compounds 1 and 3 showed strong anti-influenza A virus activities with the IC_(50) values of 15.6 and 3.2μmol/L,*** 1 exhibited good antibacterial activities against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus ***.

关键词： Thiopeptide streptomyces Natural products Structure elucidation Antiviral agents

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Rational construction of genome-minimized streptomyces host for the expression of secondary metabolite gene clusters

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Synthetic and Systems Biotechnology 2024年第3期9卷 600-608页

作者： Hui Li Sheng Gao Sanyuan Shi Xiaomin Zhao Haoyu Ye Yunzi Luo Department of Gastroenterology State Key Laboratory of BiotherapyWest China HospitalSichuan UniversityChengdu610041China Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering(Ministry of Education) School of Chemical Engineering and TechnologyTianjin UniversityTianjin300072China Georgia Tech Shenzhen Institute Tianjin UniversityTangxing Road 133Nanshan DistrictShenzhen518071China Haihe Laboratory of Sustainable Chemical Transformations Tianjin300192China

streptomyces offer a wealth of naturally occurring compounds with diverse structures,many of which possess significant pharmaceutical ***,new product exploration and increased yield of specific compounds in streptomyces have been technically challenging due to their slow growth rate,complex culture conditions and intricate genetic *** this study,we screened dozens of streptomyces strains inhabiting in a plant rhizosphere for fast-growing candidates,and further employed CRISPR/Cas-based engineering techniques for stepwise refinement of a particular strain,streptomyces sp.A-14 that harbors a 7.47 Mb *** strategic removal of nonessential genomic regions and most gene clusters,we reduced its genome size to 6.13 Mb,while preserving its growth rate to the greatest *** further demonstrated that cleaner metabolic background of this engineered strain was well suited for the expression and characterization of heterologous gene clusters,including the biosynthetic pathways of actinorhodin and polycyclic tetramate ***,this streamlined genome is anticipated to facilitate directing the metabolic flux towards the production of desired compounds and increasing their yields.

关键词： streptomyces Chassis engineering CRISPR Natural products Synthetic biology

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Characterization of a LAL-type regulator NemR in nemadectin biosynthesis and its application for increasing nemadectin production in streptomyces cyaneogriseus

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Science China(Life Sciences) 2019年第3期62卷 394-405页

作者： Chuang Li Hairong He Jiabin Wang Hui Liu Haiyan Wang Yajie Zhu Xiangjing Wang Yanyan Zhang Wensheng Xiang State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences School of Life Science Northeast Agricultural University College of Food and Bioengineering Qiqihar University

Nemadectin, a macrocyclic lactone antibiotic, is produced by streptomyces cyaneogriseus ssp. noncyanogenus. A methoxime derivative of nemadectin, moxdectin, has been widely used to control insect and helminth in animal health. Despite the importance of nemadectin, little attention has been paid to the regulation of nemadectin biosynthesis, which has hindered efforts to improve nemadectin production via genetic manipulation of regulatory genes. Here, we characterize the function of nemR, the cluster-situated regulatory gene encoding a LAL-family transcriptional regulator, in the nemadectin biosynthesis gene cluster of S. cyaneogriseus ssp. noncyanogenus NMWT1. NemR is shown to be essential for nemadectin production and found to directly activate the transcription of nemA1-1/A1-2/A2, nemC and nemA4/A3/E/D operons, but indirectly activate that of nemG and nemF. A highly conserved sequence 5′-TGGGGTGKATAGGGGGTA-3′(K=T/G) is verified to be essential for NemR ***, four novel targets of NemR, including genes encoding an SsgA-like protein(TU94_12730), a methylmalonyl-CoA mutase(TU94_19950), a thioesterase of oligomycin biosynthesis(TU94_22425) and a MFS family transporter(TU94_24835)are identified. Overexpression of nemR significantly increased nemadectin production by 79.9%, in comparison with NMWT1,suggesting that nemR plays an important role in the nemadectin biosynthesis.

关键词： nemadectin streptomyces cyaneogriseus ssp.noncyanogenus nemR LAL overexpression

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