scholarly journals Multi-omics Comparative Analysis of Streptomyces Mutants Obtained by Iterative Atmosphere and Room-Temperature Plasma Mutagenesis

2021 ◽  
Vol 11 ◽  
Author(s):  
Tan Liu ◽  
Zhiyong Huang ◽  
Xi Gui ◽  
Wei Xiang ◽  
Yubo Jin ◽  
...  
Keyword(s):  
Comparative Analysis ◽  
Room Temperature ◽  
Gene Clusters ◽  
Treatment Method ◽  
Relative Molecular Mass ◽  
Type I ◽  
Wild Type ◽  
Temperature Plasma ◽  
Artp Mutagenesis ◽  
Type Strains

Sponges, the most primitive multicellular animals, contain a large number of unique microbial communities. Sponge-associated microorganisms, particularly actinomyces, have the potential to produce diverse active natural products. However, a large number of silent secondary metabolic gene clusters have failed to be revived under laboratory culture conditions. In this study, iterative atmospheric room-temperature plasma. (ARTP) mutagenesis coupled with multi-omics conjoint analysis was adopted to activate the inactive wild Streptomyces strain. The desirable exposure time employed in this study was 75 s to obtain the appropriate lethality rate (94%) and mutation positive rate (40.94%). After three iterations of ARTP mutagenesis, the proportion of mutants exhibiting antibacterial activities significantly increased by 75%. Transcriptome analysis further demonstrated that the differential gene expression levels of encoding type I lasso peptide aborycin had a significant upward trend in active mutants compared with wild-type strains, which was confirmed by LC-MS results with a relative molecular mass of 1082.43 ([M + 2H]2+ at m/z = 2164.86). Moreover, metabolome comparative analysis of the mutant and wild-type strains showed that four spectra or mass peaks presented obvious differences in terms of the total ion count or extracting ion current profiles with each peak corresponding to a specific compound exhibiting moderate antibacterial activity against Gram-positive indicators. Taken together, our data suggest that the ARTP treatment method coupled with multi-omics profiling analysis could be used to estimate the valid active molecules of metabolites from microbial crudes without requiring a time-consuming isolation process.

scholarly journals Combination of Atmospheric and Room Temperature Plasma (ARTP) mutagenesis, Genome shuffling and Dimethyl sulfoxide (DMSO) feeding to improve FK506 production in Streptomyces tsukubaensis

2021 ◽  
Author(s):  
Liting Ye ◽  
Ruifang Ye ◽  
Fengxian Hu ◽  
Guozhu Wang
Keyword(s):  
Type Strain ◽  
Room Temperature ◽  
Genome Shuffling ◽  
Specific Gene ◽  
Wild Type ◽  
Temperature Plasma ◽  
Random Mutation ◽  
Expression Levels ◽  
Streptomyces Tsukubaensis ◽  
Artp Mutagenesis

Abstract FK506 is a clinically important macrocyclic polyketide with immunosuppressive activity produced by Streptomyces tsukubaensis . However, the production capacity of the strain is very low. To improve production, atmospheric and room temperature plasma (ARTP)mutagenesis was adopted to get the initial strains used in genome shuffling (GS). After three rounds of GS, S. tsukubaensis R3-C4 was the most productive strain, resultingin a FK506 concentration of 335 μg/mL, 2.6 times than that of the originalwild-type strain. Moreover, exogenous DMSO 4 % (v/v) addition could induce efflux of FK506 and increased FK506 production by 27.9% to 429 μg/mL . Finally, analyses of the differences in morphology, fermentation characteristics and specific gene expression levels between S. tsukubaensis R3-C4 and the wild-type strain revealed that R3-C4 strain: has hampered spore differentiation , thicker mycelia and more red pigment, which are likely related to the downregulation of bldD and cdgB expression. In addition, the expression levels of fkbO , fkbP , dahp , pccB and prpE all showed up-regulation at diverse degrees compared to the wild-type S. tsukubaensis . Overall, these results show that a combined approach involving classical random mutation and exogenous feeding canbe applied to increase FK506 biosynthesis andmay be applied also tothe improvement of other important secondary metabolites.

scholarly journals Polysaccharides and virulence of Burkholderia pseudomallei

2007 ◽  
Vol 56 (8) ◽  
pp. 1005-1010 ◽  
Author(s):  
M. Sarkar-Tyson ◽  
J. E. Thwaite ◽  
S. V. Harding ◽  
S. J. Smither ◽  
P. C. F. Oyston ◽  
...  
Keyword(s):  
Burkholderia Pseudomallei ◽  
Capsular Polysaccharide ◽  
Gene Clusters ◽  
Type I ◽  
Wild Type ◽  
Time To Death ◽  
Type Iii ◽  
Type Iv ◽  
Mutant Strains ◽  
Delayed Time

Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease of humans and animals. Gene clusters which encode capsular polysaccharide (type I O-PS) and LPS (type II O-PS), both of which play roles in virulence, have previously been identified. Here, the identification of two further putative clusters, type III O-PS and type IV O-PS, is reported. Mice challenged with type III O-PS or type IV O-PS mutants showed increased mean times to death (7.8 and 11.6 days) compared to those challenged with wild-type B. pseudomallei (3 days). To investigate the possible roles of polysaccharides in protection, mice were immunized with killed cells of wild-type B. pseudomallei or killed cells of B. pseudomallei with mutations in the O antigen, capsular polysaccharide, type III O-PS or type IV O-PS gene clusters. Immunization with all polysaccharide mutant strains resulted in delayed time to death compared to the naïve controls, following challenge with wild-type B. pseudomallei strain K96243. However, immunization with killed polysaccharide mutant strains conferred different degrees of protection, demonstrating the immunological importance of the polysaccharide clusters on the surface of B. pseudomallei.

scholarly journals Limited In Vivo Production of Type I or Type III Interferon After Infection of Macaques with Vaccine or Wild-Type Strains of Measles Virus

2015 ◽  
Vol 35 (4) ◽  
pp. 292-301 ◽  
Author(s):  
Rupak Shivakoti ◽  
Debra Hauer ◽  
Robert J. Adams ◽  
Wen-Hsuan W. Lin ◽  
William Paul Duprex ◽  
...  
Keyword(s):  
Measles Virus ◽  
Type I ◽  
Wild Type ◽  
Type Iii ◽  
Type Iii Interferon ◽  
Type Strains

scholarly journals Metabolic Analysis of Schizochytrium Mutants With High DHA Content Achieved With ARTP Mutagenesis Combined With Iodoacetic Acid and Dehydroepiandrosterone Screening

2021 ◽  
Vol 9 ◽  
Author(s):  
Lei Zeng ◽  
Yanqi Bi ◽  
Pengfei Guo ◽  
Yali Bi ◽  
Tiantian Wang ◽  
...  
Keyword(s):  
Room Temperature ◽  
Strain Improvement ◽  
Tca Cycle ◽  
Iodoacetic Acid ◽  
Metabolomic Analysis ◽  
Improvement Strategy ◽  
Temperature Plasma ◽  
Metabolic Analysis ◽  
Artp Mutagenesis ◽  
Dha Production

High DHA production cost caused by low DHA titer and productivity of the current Schizochytrium strains is a bottleneck for its application in competition with traditional fish-oil based approach. In this study, atmospheric and room-temperature plasma with iodoacetic acid and dehydroepiandrosterone screening led to three mutants, 6–8, 6–16 and 6–23 all with increased growth and DHA accumulations. A LC/MS metabolomic analysis revealed the increased metabolism in PPP and EMP as well as the decreased TCA cycle might be relevant to the increased growth and DHA biosynthesis in the mutants. Finally, the mutant 6–23, which achieved the highest growth and DHA accumulation among all mutants, was evaluated in a 5 L fermentor. The results showed that the DHA concentration and productivity in mutant 6–23 were 41.4 g/L and 430.7 mg/L/h in fermentation for 96 h, respectively, which is the highest reported so far in literature. The study provides a novel strain improvement strategy for DHA-producing Schizochytrium.

scholarly journals Effects of Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis on Physicochemical Characteristics and Immune Activity In Vitro of Hericium erinaceus Polysaccharides

Molecules ◽  
2019 ◽  
Vol 24 (2) ◽  
pp. 262 ◽  
Author(s):  
Lingli Zhu ◽  
Di Wu ◽  
Henan Zhang ◽  
Qiaozhen Li ◽  
Zhong Zhang ◽  
...  
Keyword(s):  
Room Temperature ◽  
Practical Method ◽  
Physicochemical Characteristics ◽  
Original Strain ◽  
Temperature Plasma ◽  
Hericium Erinaceus ◽  
Polysaccharide Content ◽  
Raw264.7 Macrophages ◽  
Artp Mutagenesis

The polysaccharide is the main active substance contained in Hericium erinaceus and is commonly used in the treatment of neurasthenia, tumors, and digestive diseases. Six intracellular polysaccharide components were obtained from H. erinaceus fruiting bodies cultivated by ARTP (atmospheric and room temperature plasma) mutagenic strain (321) and the original strain (0605), respectively. This study was designed to investigate the physicochemical characteristics of these polysaccharide components and their potential immunomodulatory activities on RAW264.7 macrophages. The results showed that the yield of fruiting body cultivated by mutated strain increased by 22% and the polysaccharide content improved by 16% compared with the original one owing to ARTP mutagenesis. The molecular weight distribution and the monosaccharide compositions of polysaccharide components from H. erinaceus induced by ARTP mutagenesis were significantly different from that of the original one. The NO, IL-6, IL-10, IL-1β, and TNF-α production activities of macrophages were enhanced by stimulation of 20% ethanol precipitated polysaccharides from H. erinaceus induced by ARTP mutagenesis. These results indicated that ARTP is an efficient and practical method for high polysaccharide content breeding of the H. erinaceus strain and this provided a reference for obtaining high quality resources and healthy product development from H. erinaceus.

scholarly journals Putative Role of the Aldo-Keto Reductase from Trypanosoma cruzi in Benznidazole Metabolism

2016 ◽  
Vol 60 (5) ◽  
pp. 2664-2670 ◽  
Author(s):  
Patricia Andrea Garavaglia ◽  
Marc Laverrière ◽  
Joaquín J. B. Cannata ◽  
Gabriela Andrea García
Keyword(s):  
Trypanosoma Cruzi ◽  
Inhibitory Concentration ◽  
Reductase Activity ◽  
Differential Susceptibility ◽  
Type I ◽  
Wild Type ◽  
Content Type ◽  
Type Strains ◽  
Dependent Type

ABSTRACTBenznidazole (Bz), the drug used for treatment of Chagas' disease (caused by the protozoanTrypanosoma cruzi), is activated by a parasitic NADH-dependent type I nitroreductase (NTR I). However, several studies have shown that other enzymes are involved. The aim of this study was to evaluate whether the aldo-keto reductase fromT. cruzi(TcAKR), a NADPH-dependent oxido-reductase previously described by our group, uses Bz as the substrate. We demonstrated that both recombinant and nativeTcAKR enzymes reduce Bz by using NADPH, but not NADH, as a cofactor.TcAKR-overexpressing epimastigotes showed higher NADPH-dependent Bz reductase activity and a 50% inhibitory concentration (IC50) value for Bz 1.8-fold higher than that of the controls, suggesting thatTcAKR is involved in Bz detoxification instead of activation. To understand the role ofTcAKR in Bz metabolism, we studiedTcAKR expression and NADPH/NADH-dependent Bz reductase activities in twoT. cruzistrains with differential susceptibility to Bz: CL Brener and Nicaragua. Taking into account the results obtained withTcAKR-overexpressing epimastigotes, we expected the more resistant strain, Nicaragua, to have higherTcAKR levels than CL Brener. However, the results were the opposite. CL Brener showed 2-fold higherTcAKR expression and 5.7-fold higher NADPH-Bz reduction than the Nicaragua strain. In addition, NADH-dependent Bz reductase activity, characteristic of NTR I, was also higher in CL Brener than in Nicaragua. We conclude that althoughTcAKR uses Bz as the substrate,TcAKR activity is not a determinant of Bz resistance in wild-type strains and may be overcome by other enzymes involved in Bz activation, such as NADPH- and NADH-dependent reductases.

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