JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY
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2018
·Vol. 94
·期 4
·1220-1229
·
IF 2.587
Weide Xiong,Xianhai Zeng,Shih-Hsin Ho,Xueping Ling,Liang Shen,Chuanyi Yao,Yinghua Lu
BACKGROUND Phospholipase D (PLD) has great potential in the pharmaceutical and food industries for manufacturing bioactive phospholipids. Overproduction of PLD in Escherichia coli is difficult due to its high cytotoxicity which causes plasmid instability, short-term PLD synthesis, cell lysis and PLD leakage. To address this issue, integrated optimization of PLD production in upstream and downstream processing was performed. RESULTS Effective strategies in upstream processing include the use of plasmids containing the pSC101 par region in recA − E. coli , the use of moderately strong and tightly regulated promoter P BAD , optimization of codon usage and amino acid supplementation, maintaining the best cellular state by supplementing nutrition, and saturation induction at low temperature. Using these strategies, a large amount of PLD (1.2 × 10 5 U L −1 , 81.5 mg L −1 ) was obtained in the batch culture and PLD synthesis lasted 9 h. Additionally, cell lysis did not occur –99% of the recombinant PLD remained in the periplasm. Ultimately, the PLD was purified 327-fold by a four-step downstream process comprising extraction with 10 mmol L −1 Ethylenediaminetetraacetic acid and 0.05% (w/v) sodium deoxycholate, ammonium sulfate precipitation, anion exchange and Ni 2+ -affinity chromatography. CONCLUSION This study provides the basis for large-scale production of PLD in E. coli . © 2018 Society of Chemical Industry
研究领域:
生物技术
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微生物学
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制药业
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食品工业
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DOI
高影响因子
MOLECULES
·
2016
·Vol. 21
·期 8
·970
·
IF 2.465
Ying Han,Erli Tian,Dongbo Xu,Min Ma,Zixin Deng,Kui Hong
During our search for interesting bioactive secondary metabolites from mangrove actinomycetes, the strainStreptomycessp. 219807 which produced a high elaiophylin yield of 4486 mg/L was obtained. A new elaiophylin derivative, halichoblelide D (1), along with seven known analogues2–8was isolated and identified from the culture broth. Their chemical structures were determined by detailed analysis of 1D and 2D NMR and HRMS data. The absolute configuration of halichoblelide D (1) was confirmed by comparing the CD spectrum with those of the reported analogues. Compounds1–7exhibited potent cytotoxic activities against HeLa and MCF-7 cells with IC50values ranging from 0.19 to 2.12 μM.Keywords:mangrove actinomycetes;Streptomycessp. 219807;elaiophylin;cytotoxic
研究领域:
微生物学
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药物化学
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肿瘤学
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DOI
PubMed
高影响因子
JOURNAL OF FISH DISEASES
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2026
·
IF 2.3
Qianhao Shao,Yan Zhou,Xinhui Tong,Huan Qi,Shiwen Sun,Xutin Guan,Jidong Wang,Dawei Song
The highly pathogenic Micropterus salmoides rhabdovirus (MSRV) poses a serious threat to largemouth bass aquaculture, leading to considerable economic losses. We assessed the anti-MSRV activity of milbemycin A4 (A4), a macrolide derived from Streptomyces bingchenggensis in both cell-based and animal models. Cytotoxicity assays revealed that A4 was not toxic to EPC cells at concentrations up to 6.4 mg/L. It significantly reduced viral N gene expression by approximately 90.25%. A4 also suppressed MSRV-induced apoptosis, decreasing the apoptotic cell population by about 21.88%, and preserved mitochondrial membrane potential. Investigations of the viral life cycle demonstrated that A4 directly targets virions, impairing their binding and internalization, and additionally inhibits early post-entry replication events. In vivo analyses further revealed that A4 treatment led to significant induction of antiviral-related genes ( IRF3 , IRF7 and IFN-γ ), which was accompanied by enhanced survival of infected fish (54%), along with reduced viral accumulation in the liver and spleen and alleviated histopathological damage. These findings suggest that A4 exhibits strong anti-MSRV activity by targeting early infection stages, protecting mitochondrial integrity and enhancing host innate immunity, highlighting its potential for application in aquaculture disease control.
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PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2024
·
IF 4.2
Jiahao Lai,Bing Liu,Guihong Xiong,Qian Luo,Shuilin Song,Junxi Jiang,Hongyi Wei,Junwen Wang
Actinomycetes have emerged as significant biocontrol resources due to their rich array of bioactive natural products. While much research has historically focused on secondary metabolites isolated from their fermentation broth, there remains a dearth of reports on their volatile organic compounds (VOCs). Here, strain ML27, isolated from soil, was identified as Streptomyces albidoflavus based on morphological features, physiological, biochemical, and molecular characteristics ( 16S rRNA , atpD , recA , and rpoB gene sequences). VOCs from S. albidoflavus strain ML27 were effectively captured using solid-phase microextraction (SPME) and tentatively identified through gas chromatography–mass spectrometry (GC/MS). Among these compounds, 4-ethyl-1,2-dimethoxybenzene exhibited broad-spectrum antifungal activity and demonstrated efficacy in controlling citrus anthracnose, with a control efficacy of 86.67%. Furthermore, the inhibitory mechanism of 4-ethyl-1,2-dimethoxybenzene against Colletotrichum gloeosporioides was revealed. Results indicated that 4-ethyl-1,2-dimethoxybenzene induced swelling, deformity, and breakage in C . gloeosporioides mycelia, and significantly inhibited spore germination. Transcriptome analysis revealed that 4-ethyl-1,2-dimethoxybenzene inhibited the growth and development of C. gloeosporioides primarily by disrupting energy metabolism and the integrity of the cell wall and membrane. Based on these results, it is promising to develop 4-ethyl-1,2-dimethoxybenzene as a novel biopesticide for controlling citrus anthracnose.
研究领域:
微生物学
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植物病理学
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分子生物学
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生物技术
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产品关联
DOI
PubMed
高影响因子
Frontiers in Microbiology
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2019
·
IF 4.259
Wu Hao,Guo Ye,Chen Lei,Chen Guiguang,Liang Zhiqun
This study characterized the biosynthetic pathway of the secondary metabolite 1-deoxynojirimycin (DNJ) from Streptomyces lavendulae. The results revealed that glucose was a preferable precursor for DNJ synthesis, and its carbon skeleton underwent a C2-N-C6 cyclization reaction during synthesis. The biosynthetic pathway was related to the glycolysis pathway, and started from fructose-6-phosphate, and involved amination, dephosphorylation, oxidation, cyclization, dehydration, and reduction reaction steps, yielding DNJ. Then, based on clarified biosynthetic pathway information, precursors, analogs, and metabolism inhibitors were used as novel regulators to enhance the production of DNJ. The results demonstrated that the titer of DNJ could reach 296.56 mg/L, which was 3.3-fold higher than that of a control group (90 mg/L) when sodium citrate (0 h, 5 g/L), sorbose (0 h, 1 g/L), iodoacetic acid (20 h, 50 mg/L), and glucose (26 h, 7 g/L) were added during the fermentation process. This study provides a new understanding of the biosynthetic pathway of DNJ, and also provides an efficient strategy to regulate the production of DNJ based on this biosynthetic pathway, which is a new perspective for the regulation of other secondary metabolites.
研究领域:
微生物学
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代谢工程
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DOI
PubMed
高影响因子
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
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2022
·
IF 3.141
Dong Zhang,Yingling Bao,Zhi Ma,Jiawei Zhou,Hanchi Chen,Yuele Lu,Linjiang Zhu,Xiaolong Chen
Valinomycin is a cyclodepsipeptide antibiotic with a broad spectrum of biological activities, such as antiviral, antitumor, and antifungal activities. However, the low yield of valinomycin often limits its applications in medicine, agriculture, and industry. In our previous report, Streptomyces sp. ZJUT-IFE-354 was identified as a high-yielding strain of valinomycin. In this study, Plackett–Burman design (PBD) and response surface methodology (RSM) were used to optimize components of medium. The optimal medium contained 31 g/L glucose, 22 g/L soybean meal, and 1.6 g/L K2HPO4·3H2O, which could generate 262.47 ± 4.28 mg/L of valinomycin. Then, the culture conditions were optimized by a one-factor-at-a-time (OFAT) approach. The optimal conditions for the strain included a seed age of 24 h, an inoculum size of 8% (v/v), an incubation temperature of 28 °C, an initial pH of 7.2, an elicitor of 0.1% Bacillus cereus feeding at 24 h cultivation, and the feeding of 0.6% L-valine at 36 h cultivation. The final valinomycin production increased to 457.23 ± 9.52 mg/L, which was the highest yield ever reported. It highlights that RSM and OFAT may be efficient methods to enhance valinomycin production by Streptomyces sp. ZJUT-IFE-354.
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高影响因子
Marine Drugs
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2021
·Vol. 19
·期 6
·300
·
IF 5.118
Tong Chen,Gong Cheng,Siming Jiao,Lishi Ren,Chuanfang Zhao,Jinhua Wei,Juntian Han,Meishan Pei,Yuguang Du,Jian-Jun Li
It is known that bioactivities of chitooligosaccharide (COS) are closely related to the degree of polymerization (DP); therefore, it is essential to prepare COS with controllable DP, such as chitobiose showing high antioxidant and antihyperlipidemia activities. In this study, BLAST, sequence alignment and phylogenetic analysis of characterized glycoside hydrolase (GH) 46endo-chitosanases revealed that a chitosanase Sn1-CSN fromStreptomyces niveuswas different from others. Sn1-CSN was overexpressed inE. coli, purified and characterized in detail. It showed the highest activity at pH 6.0 and exhibited superior stability between pH 4.0 and pH 11.0. Sn1-CSN displayed the highest activity at 50 °C and was fairly stable at ≤45 °C. Its apparent kinetic parameters against chitosan (DDA: degree of deacetylation, >94%) were determined, withKmandkcatvalues of 1.8 mg/mL and 88.3 s−1, respectively. Cu2+enhanced the activity of Sn1-CSN by 54.2%, whereas Fe3+inhibited activity by 15.1%. Hydrolysis products of chitosan (DDA > 94%) by Sn1-CSN were mainly composed of chitobiose (87.3%), whereas partially acetylated chitosan with DDA 69% was mainly converted into partially acetylated COS with DP 2-13. Thisendo-chitosanase has great potential to be used for the preparation of chitobiose and partially acetylated COS with different DPs.Keywords:endo-chitosanases;Streptomyces niveus;pH and temperature-rate profile;pH and thermal stability;chitobiose;partially acetylated chitooligosaccharide;structure modelling
研究领域:
生物化学
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微生物学
·
酶学
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PubMed
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GENOMICS PROTEOMICS & BIOINFORMATICS
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2022
·
IF 6.409
Jiazhen Zheng,Yue Li,Ning Liu,Jihui Zhang,Shuangjiang Liu,Huarong Tan
Streptomycetes possess numerous gene clusters and the potential to produce large amounts of natural products. Histone deacetylase (HDAC) inhibitors play an important role in the regulation of histone modifications in fungi, but little is known about the roles of HDAC in prokaryotes. Here, we described the global effects of the HDAC inhibitor, sodium butyrate (SB), on marine-derived Streptomyces olivaceus FXJ 8.021, particularly on the activation of secondary metabolites biosynthesis. antiSMASH analysis revealed 33 secondary metabolite biosynthetic gene clusters (BGCs) in strain FXJ 8.021, among which the silent lobophorin BGC was activated by SB. Transcriptomic data showed that the expression of genes involved in lobophorin biosynthesis ( ge00097–ge00139 ) and CoA-ester formation ( e.g. , ge02824 ), as well as glycolysis/gluconeogenesis pathway ( e.g ., ge01661 ), was mainly up-regulated in the presence of SB. Intracellular CoA-ester analysis confirmed that SB triggered the biosynthesis of CoA-ester, increasing the precursor supply for lobophorin biosynthesis. Further acetylome analysis revealed that the acetylation levels on 218 acetylation sites of 190 proteins were up-regulated and those on 411 sites of 310 proteins were down-regulated. These 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 regulation to activate expression of lobophorin BGC. These findings provide solid evidence that acetylated proteins triggered by SB could affect the expression of genes in the biosynthesis of primary and secondary metabolites in prokaryotes.
研究领域:
微生物学
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分子生物学
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生物化学
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基因组学
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DOI
PubMed
高影响因子
Bioresources and Bioprocessing
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2017
·Vol. 4
·期 1
·1-7
·
IF 0
Jiang Ying,Shang Yue-Peng,Li Hao,Zhang Chao,Pan Jiang,Bai Yun-Peng,Li Chun-Xiu,Xu Jian-He
To improve the fermentation production of transglutaminase (TGase) from Streptomyces mobaraensis for applications in the food industry, the atmospheric and room-temperature plasma (ARTP) mutagenesis was applied to breed S. mobaraensis mutants with increased TGase production.After eight rounds of iterative ARTP mutagenesis, four genetically stable mutants, Sm5-V1, Sm6-V13, Sm2-V10, and Sm7-V12, were identified, which showed increased TGase production by 27, 24, 24, and 19%, respectively. The best mutant Sm5-V1 exhibited a maximum TGase activity of 5.85 U/mL during flask fermentation. Compared to the wild-type strain, the transcription levels of the zymogen TGase genes in the mutants increased significantly as indicated by quantitative real-time PCR, while the gene nucleotide sequences of the mutants did not change at all. It was shown that the overexpression of TGase zymogen gene in the mutants contributes to the increase in TGase production.ARTP is a potentially efficient tool for microbial mutation breeding to bring some significant changes required for the industrial applications.
研究领域:
微生物学
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食品科学
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工业生物技术
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DOI
PubMed
Journal of Water Process Engineering
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2025
·
IF 6.7
Mi Feng,Junjie Zhao,Yanqin Lu,Xuan Luo,Xinghui Feng,Yao Huang,Zijun Chen,Dongxia Fu,Jianhua Chen
Streptomyces sp. FW-32, an actinobacterial strain capable of degrading 3,6-dibromocarbazole (3,6-DBCZ), was isolated from polluted soils. >85 % of 3,6-DBCZ was degraded within 5 d under optimal conditions (glucose concentration of 5 g/L, pH of 7, initial 3,6-DBCZ concentration of 1 mg/L, temperature of 35 °C). The biodegradation of 3,6-DBCZ followed the first-order kinetic model with the degradation rate constant of 0.01713 h −1 . Biodegradation pathways of 3,6-DBCZ by Streptomyces sp. FW-32 were proposed, including debromination, oxidation, and aromatic ring cleavage. The intracellular enzymes of Streptomyces sp. FW-32, such as cytochrome P450, played an important role in 3,6-DBCZ biodegradation. In the earlier stage of 3,6-DBCZ biodegradation (1–2 d), the activities of SOD and CAT were enhanced by 3,6-DBCZ (≤10 mg/L), but 10 mg/L of 3,6-DBCZ inhibited the activities of these two antioxidant enzymes at the later period (4–6 days). Cell viability of strain FW-32 was stimulated by 3,6-DBCZ at 1 mg/L in the first two days of treatment, but it was suppressed by 10 mg/L of 3,6-DBCZ after 3 days of exposure. Strain FW-32 was able to efficiently degrade 3,6-DBCZ and its key intermediate carbazole in real river water. The results of this study illuminated the mechanism of 3,6-DBCZ biodegradation by a newly isolated strain FW-32 and indicated that strain FW-32 might be suitable for the elimination of 3,6-DBCZ from contaminated water. Overall, these findings were significant for developing microbial remediation to control 3,6-DBCZ pollution in water environments.
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TETRAHEDRON
·
2019
·
IF 2.379
Wenwen Yi,Qiao Li,Tengfei Song,Lei Chen,Xing-Cong Li,Zhizhen Zhang,Xiao-Yuan Lian
New indole alkaloids streptoprenylindoles A–C ( 1 – 3 ) and diterpenoids 18-acetyl-cyclooctatin ( 8 ), 5,18-dedihydroxy-cyclooctatin ( 9 ), and 5-dehydroxy-cyclooctatin ( 10 ) were isolated from the culture of marine-derived Streptomyces sp. ZZ820, along with known 3-cyanomethyl-6-[3-methyl-2-butenyl]indole ( 4 ), N -(2-(1 H -indol-3-yl)ethylacetamide ( 5 ), 1-acetyl-β-carboline ( 6 ), indole-3-methylethanoate ( 7 ), cyclooctatin ( 11 ), and chromomycin A 3 ( 12 ). Their structures were elucidated by a combination of extensive spectroscopic analyses, ECD calculation, and the Mosher's method. Streptoprenylindoles A ( 1 ) and B ( 2 ) are enantiomers that were separated through the preparation of their Mosher esters. Three new diterpenoids ( 8 – 10 ) showed antibacterial activities against methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli with MIC values of 24.11–55.12 μM, while chromomycin A 3 ( 12 ) showed potent antibacterial activities against MRSA (MIC: 0.59 μM) and E. coli (MIC 0.04 μM).
研究领域:
微生物学
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天然产物化学
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药理学
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DOI
高影响因子
ACS Synthetic Biology
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2021
·
IF 5.11
Ming Zhao,Mingrui Wang,Shuiling Wang,Liangbin Xiong,Bei Gao,Min Liu,Xinyi Tao,Feng-Qing Wang,Dongzhi Wei
Streptomyces species possess strong secondary metabolism, the switches of which from the primary metabolism are complex and thus a challenge to holistically optimize their productivities. To avoid the complex switches and to reduce the limitations of different metabolic stages on the synthesis of metabolites, we designed a Streptomyces self-sustained system (StSS) that contains two functional modules, the primary metabolism module (PM) and the secondary metabolism module (SM). The PM includes endogenous housekeeping sigma factor σhrdB and σhrdB-dependent promoters, which are used to express target genes in the primary metabolism phase. SM consists of the expression cassette of σhrdB under the control of a secondary metabolism promoter, which maintains continuous activity of the σhrdB-dependent promoters in the secondary metabolism phase. As a proof-of-principle, the StSS was used to boost the production of some non-toxic metabolites, including indigoidine, undecylprodigiosin (UDP), ergothioneine, and avermectin, in Streptomyces. All these metabolites can undergo a continuous production process spanning the primary and secondary metabolism stages instead of being limited to a specific stage. Scale-up of UDP fermentation in a 4 L fermentor indicated that the StSS is a stable and robust system, the titer of which was enhanced to 1.1 g/L, the highest at present. This study demonstrated that the StSS is a simple but powerful strategy to rationally engineer Streptomyces cell factories for the efficient production of non-toxic metabolites via reconstructing the relationships between primary and secondary metabolism.
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