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 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 Improved Neomycin Sulfate Potency in Streptomyces fradiae Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization

2022 ◽  
Vol 10 (1) ◽  
pp. 94
Author(s):  
Fei Yu ◽  
Min Zhang ◽  
Junfeng Sun ◽  
Fang Wang ◽  
Xiangfei Li ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Room Temperature ◽  
Screening Method ◽  
Fermentation Medium ◽  
Soluble Starch ◽  
High Yield ◽  
Streptomyces Fradiae ◽  
Temperature Plasma ◽  
Artp Mutagenesis ◽  
Neomycin Sulfate

To improve the screening efficiency of high-yield neomycin sulfate (NM) Streptomyces fradiae strains after mutagenesis, a high-throughput screening method using streptomycin resistance prescreening (8 μg/mL) and a 24-deep well plates/microplate reader (trypan blue spectrophotometry) rescreening strategy was developed. Using this approach, we identified a high-producing NM mutant strain, Sf6-2, via six rounds of atmospheric and room temperature plasma (ARTP) mutagenesis and screening. The mutant displayed a NM potency of 7780 ± 110 U/mL and remarkably stable genetic properties over six generations. Furthermore, the key components (soluble starch, peptone, and (NH4)2SO4) affecting NM potency in fermentation medium were selected using Plackett-Burman and optimized by Box-Behnken designs. Finally, the NM potency of Sf6-2 was increased to 10,849 ± 141 U/mL at the optimal concentration of each factor (73.98 g/L, 9.23 g/L, and 5.99 g/L, respectively), and it exhibited about a 40% and 100% enhancement when compared with before optimization conditions and the wild-type strain, respectively. In this study, we provide a new S. fradiae NM production strategy and generate valuable insights for the breeding and screening of other microorganisms.

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.

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