scholarly journals Comparative transcriptomic analysis of two Saccharopolyspora spinosa strains reveals the relationships between primary metabolism and spinosad production

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yunpeng Zhang ◽  
Xiaomeng Liu ◽  
Tie Yin ◽  
Qi Li ◽  
Qiulong Zou ◽  
...  
Keyword(s):  
Metabolic Pathways ◽  
Genetic Manipulation ◽  
Fermentation Medium ◽  
Transcriptomic Analysis ◽  
High Yield ◽  
Primary Metabolism ◽  
Yield Strain ◽  
Saccharopolyspora Spinosa ◽  
Large Numbers ◽  
Close Relationship

AbstractSaccharopolyspora spinosa is a well-known actinomycete for producing the secondary metabolites, spinosad, which is a potent insecticides possessing both efficiency and safety. In the previous researches, great efforts, including physical mutagenesis, fermentation optimization, genetic manipulation and other methods, have been employed to increase the yield of spinosad to hundreds of folds from the low-yield strain. However, the metabolic network in S. spinosa still remained un-revealed. In this study, two S. spinosa strains with different spinosad production capability were fermented and sampled at three fermentation periods. Then the total RNA of these samples was isolated and sequenced to construct the transcriptome libraries. Through transcriptomic analysis, large numbers of differentially expressed genes were identified and classified according to their different functions. According to the results, spnI and spnP were suggested as the bottleneck during spinosad biosynthesis. Primary metabolic pathways such as carbon metabolic pathways exhibited close relationship with spinosad formation, as pyruvate and phosphoenolpyruvic acid were suggested to accumulate in spinosad high-yield strain during fermentation. The addition of soybean oil in the fermentation medium activated the lipid metabolism pathway, enhancing spinosad production. Glutamic acid and aspartic acid were suggested to be the most important amino acids and might participate in spinosad biosynthesis.

Improvement of Spinosad Production upon Utilization of Oils and Manipulation of β-Oxidation in a High-Producing Saccharopolyspora spinosa Strain

2018 ◽  
Vol 28 (2) ◽  
pp. 53-64
Author(s):  
Ying Huang ◽  
Xiaolin Zhang ◽  
Chen Zhao ◽  
Xuhui  Zhuang ◽  
Lin Zhu ◽  
...  
Keyword(s):  
Streptomyces Coelicolor ◽  
Building Blocks ◽  
Genetically Engineered ◽  
High Yield ◽  
Control Group ◽  
Yield Strain ◽  
Saccharopolyspora Spinosa ◽  
Camellia Oil ◽  
Expression Of Genes ◽  
And Oxidative Stress

Spinosad, a member of polyketide-derived macrolides produced in the actinomycete <i>Saccharopolyspora spinosa</i>, has been developed as a broad-spectrum and effective insecticide. The β-oxidation pathway could be an important source of building blocks for the biosynthesis of spinosad, thus the effect of vegetable oils on the production of spinosad in a high-yield strain was investigated. The spinosad production increased significantly with the addition of strawberry seed oil (511.64 mg/L) and camellia oil (520.07 mg/L) compared to the control group without oil (285.76 mg/L) and soybean oil group (398.11 mg/L). It also revealed that the addition of oils would affect the expression of genes involved in fatty acid metabolism, precursor supply, and oxidative stress. The genetically engineered strain, in which <i>fadD1</i> and <i>fadE</i> genes of <i>Streptomyces coelicolor</i> were inserted, produced spinosad up to 784.72 mg/L in the medium containing camellia oil, while a higher spinosad production level (843.40 mg/L) was detected with the addition of 0.01 mM of thiourea.

scholarly journals Acyltransferase AniI as tailoring enzyme with broad substrate tolerance for high-level production of anisomycin

2021 ◽  
Author(s):  
Qing Wang ◽  
Lingxin Kong ◽  
Xiaoqing Zheng ◽  
Jufang Shen ◽  
Junbo Wang ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Cellular Localization ◽  
Genetic Manipulation ◽  
Structural Diversity ◽  
Biosynthetic Gene Cluster ◽  
Hydroxyl Group ◽  
High Yield ◽  
Pyrrolidine Ring ◽  
Yield Strain ◽  
Acyl Donors

Anisomycin (1), a pyrrolidine antibiotic, exhibits diverse biological and pharmacologic activities. The biosynthetic gene cluster of 1 has been identified previously and the multistep assembly of the core benzylpyrrolidine scaffold was characterized. However, enzymatic modifications, such as acylation involved in 1 biosynthesis are unknown. In this study, the genetic manipulation of aniI proved that it encoded indispensable acetyltransferase for 1 biosynthesis. Bioinformatics analysis suggested AniI as a member of LbH-MAT-GAT sugar O-acetyltransferase, but the biochemical assay identified that its target site was the hydroxyl group of the pyrrolidine ring. AniI was found to be tolerant of acyl donors with different chain length for the biosynthesis of 1 and derivatives 12 and 13 with butyryl and isovaleryl groups, respectively. Meanwhile, it showed comparable activity towards biosynthetic intermediates and synthesized analogues, suggesting promiscuity to the pyrrolidine ring structure of 1. These data may inspire new viable synthetic routes for the construction of more complex pyrrolidine ring scaffolds in 1. Finally, the overexpression of aniI under the control of strong promoters contributed to the higher productivities of 1 and its analogues. These findings reported here not only improved the understanding of anisomycin biosynthesis but also expand the substrate scope of O-acetyltransferase working on the pyrrolidine ring and pave the way for future metabolic engineering construction of high-yield strain. IMPORTANCE Acylation is an important tailoring reaction during natural products biosynthesis. Acylation could increase the structural diversity, affect the chemical stability, volatility, biological activity and even the cellular localization of specialized compounds. Many acetyltransferases have been reported in natural product biosynthesis. The typical example of LbH-MAT-GAT sugar O-acetyltransferase subfamily was reported to catalyze the CoA-dependent acetylation of the 6-hydroxyl group of sugars. However, no protein of this family has been characterized to acetylate non-sugar secondary metabolic product. Here, AniI was found to catalyze the acylation of the hydroxyl group of the pyrrolidine ring and be tolerant of diverse acyl donors and acceptors, which made the biosynthesis more efficient and exclusive for 1 and its derivatives biosynthesis. Moreover, the overexpression of aniI serves as a successful example of genetic manipulation of a modification gene for the high production of final products and might set the stage for future metabolic engineering.

scholarly journals Transcriptomic resources for prairie grass (Bromus catharticus): expressed transcripts, tissue-specific genes, and identification and validation of EST-SSR markers

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Ming Sun ◽  
Zhixiao Dong ◽  
Jian Yang ◽  
Wendan Wu ◽  
Chenglin Zhang ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Ssr Markers ◽  
Large Scale ◽  
Genetic Research ◽  
High Yield ◽  
Yield Strain ◽  
Tissue Samples ◽  
Prairie Grass ◽  
The Future ◽  
Genomic Resource

Abstract Background Prairie grass (Bromus catharticus) is a typical cool-season forage crop with high biomass production and fast growth rate during winter and spring. However, its genetic research and breeding has remained stagnant due to limited available genomic resources. The aim of this study was to generate large-scale genomic data using high-throughput transcriptome sequencing, and perform a preliminary validation of EST-SSR markers of B. catharticus. Results Eleven tissue samples including seeds, leaves, and stems were collected from a new high-yield strain of prairie grass BCS1103. A total of 257,773 unigenes were obtained, of which 193,082 (74.90%) were annotated. Comparison analysis between tissues identified 1803, 3030, and 1570 genes specifically and highly expressed in seed, leaf, and stem, respectively. A total of 37,288 EST-SSRs were identified from unigene sequences, and more than 80,000 primer pairs were designed. We synthesized 420 primer pairs and selected 52 ones with high polymorphisms to estimate genetic diversity and population structure in 24 B. catharticus accessions worldwide. Despite low diversity indicated by an average genetic distance of 0.364, the accessions from South America and Asia and wild accessions showed higher genetic diversity. Moreover, South American accessions showed a pure ancestry, while Asian accessions demonstrated mixed internal relationships, which indicated a different probability of gene flow. Phylogenetic analysis clustered the studied accessions into four clades, being consistent with phenotypic clustering results. Finally, Mantel analysis suggested the total phenotypic variation was mostly contributed by genetic component. Stem diameter, plant height, leaf width, and biomass yield were significantly correlated with genetic data (r > 0.6, P < 0.001), and might be used in the future selection and breeding. Conclusion A genomic resource was generated that could benefit genetic and taxonomic studies, as well as molecular breeding for B. catharticus and its relatives in the future.

scholarly journals To Stretch the Boundary of Secondary Metabolite Production in Plant Cell-Based Bioprocessing: Anthocyanin as a Case Study

2004 ◽  
Vol 2004 (5) ◽  
pp. 264-271 ◽  
Author(s):  
Wei Zhang ◽  
Chris Franco ◽  
Chris Curtin ◽  
Simon Conn
Keyword(s):  
Secondary Metabolites ◽  
Plant Cell ◽  
Metabolic Pathways ◽  
Anthocyanin Biosynthesis ◽  
Rational Approach ◽  
Great Promise ◽  
Primary Metabolism ◽  
Global Correlation ◽  
Empirical Level

Plant cells and tissue cultures hold great promise for controlled production of a myriad of useful secondary metabolites on demand. The current yield and productivity cannot fulfill the commercial goal of a plant cell-based bioprocess for the production of most secondary metabolites. In order to stretch the boundary, recent advances, new directions and opportunities in plant cell-based bioprocessing, have been critically examined for the 10 years from 1992 to 2002. A review of the literature indicated that most of the R&D work was devoted predominantly to studies at an empirical level. A rational approach to molecular plant cell bioprocessing based on the fundamental understanding of metabolic pathways and their regulations is urgently required to stimulate further advances; however, the strategies and technical framework are still being developed. It is the aim of this review to take a step forward in framing workable strategies and technologies for molecular plant cell-based bioprocessing. Using anthocyanin biosynthesis as a case study, an integrated postgenomic approach has been proposed. This combines the functional analysis of metabolic pathways for biosynthesis of a particular metabolite from profiling of gene expression and protein expression to metabolic profiling. A global correlation not only can thus be established at the three molecular levels, but also places emphasis on the interactions between primary metabolism and secondary metabolism; between competing and/or complimentary pathways; and between biosynthetic and post-biosynthetic events.

Optimization of the Fermentation Conditions for 1-Deoxynojirimycin Production by Streptomyces lawendulae Applying the Response Surface Methodology

2011 ◽  
Vol 7 (3) ◽  
Author(s):  
Zhao-Jun Wei ◽  
Le-Chun Zhou ◽  
Hua Chen ◽  
Gui-Hai Chen
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Starch Content ◽  
Ultra Violet ◽  
Soluble Starch ◽  
High Yield ◽  
Yield Strain ◽  
Fermentation Time ◽  
Ethyl Sulfate ◽  
Initial Ph

Moranoline (1-Deoxynojirimycin, DNJ) is a piperidine alkaloid, and shows high inhibit activities to glucoamylase and ?-glucosidase. One DNJ high-yield strain of Streptomyces lawendulae was obtained after isolated form soil and mutated with the ultra violet (UV) and ethyl sulfate (DES), which named as TB-412, and can produce DNJ with 35.925 mg/L. Response surface methodology (RSM) was applied to optimize the parameters of DNJ yield from S. lawendulae TB-412. The effects of independent variables of fermentation, including time, temperature, initial pH and the soluble starch content were investigated. The statistical analysis showed that the fermentation time, pH and the soluble starch content, and the quadratics of time, temperature, pH and the soluble starch content, as well as the interactions between fermentation time and pH, and time and the soluble starch content, showed significant effects on DNJ yield. The optimal process parameters for DNJ production within the experimental range of the variables researched was at 11d, 27 °C, pH 7.5, and 8% soluble starch content. At this condition, the DNJ yield was predicted to be 42.875 mg/L.

scholarly journals Uncovering and Engineering a Mini-Regulatory Network of the TetR-Family Regulator SACE_0303 for Yield Improvement of Erythromycin in Saccharopolyspora erythraea

2021 ◽  
Vol 9 ◽  
Author(s):  
Ying Liu ◽  
Sabir Khan ◽  
Panpan Wu ◽  
Bowen Li ◽  
Lanlan Liu ◽  
...  
Keyword(s):  
Regulatory Network ◽  
Target Genes ◽  
Antibacterial Activities ◽  
Saccharopolyspora Erythraea ◽  
High Yield ◽  
Yield Strain ◽  
Yield Improvement ◽  
Erythromycin Production

Erythromycins produced by Saccharopolyspora erythraea have broad-spectrum antibacterial activities. Recently, several TetR-family transcriptional regulators (TFRs) were identified to control erythromycin production by multiplex control modes; however, their regulatory network remains poorly understood. In this study, we report a novel TFR, SACE_0303, positively correlated with erythromycin production in Sac. erythraea. It directly represses its adjacent gene SACE_0304 encoding a MarR-family regulator and indirectly stimulates the erythromycin biosynthetic gene eryAI and resistance gene ermE. SACE_0304 negatively regulates erythromycin biosynthesis by directly inhibiting SACE_0303 as well as eryAI and indirectly repressing ermE. Then, the SACE_0303 binding site within the SACE_0303-SACE_0304 intergenic region was defined. Through genome scanning combined with in vivo and in vitro experiments, three additional SACE_0303 target genes (SACE_2467 encoding cation-transporting ATPase, SACE_3156 encoding a large transcriptional regulator, SACE_5222 encoding α-ketoglutarate permease) were identified and proved to negatively affect erythromycin production. Finally, by coupling CRISPRi-based repression of those three targets with SACE_0304 deletion and SACE_0303 overexpression, we performed stepwise engineering of the SACE_0303-mediated mini-regulatory network in a high-yield strain, resulting in enhanced erythromycin production by 67%. In conclusion, the present study uncovered the regulatory network of a novel TFR for control of erythromycin production and provides a multiplex tactic to facilitate the engineering of industrial actinomycetes for yield improvement of antibiotics.

High-Throughput System for Screening of Cephalosporin C High-Yield Strain by 48-Deep-Well Microtiter Plates

2013 ◽  
Vol 169 (5) ◽  
pp. 1683-1695 ◽  
Author(s):  
Jun Tan ◽  
Ju Chu ◽  
Yuyou Hao ◽  
Yuanxin Guo ◽  
Yingping Zhuang ◽  
...  
Keyword(s):  
High Throughput ◽  
High Yield ◽  
Cephalosporin C ◽  
Yield Strain ◽  
Deep Well ◽  
Microtiter Plates

Growth and reproductive biology of Joffrea speirsii gen. et sp. nov., a Cercidiphyllum-like plant from the Late Paleocene of Alberta, Canada

1985 ◽  
Vol 63 (2) ◽  
pp. 340-364 ◽  
Author(s):  
Peter R. Crane ◽  
Ruth A. Stockey
Keyword(s):  
Upper Cretaceous ◽  
Constant Field ◽  
Short Shoot ◽  
Early Tertiary ◽  
Late Paleocene ◽  
Large Numbers ◽  
Close Relationship ◽  
Field Association ◽  
Short Shoots ◽  
Floodplain Habitats

Six kinds of angiosperm compression fossils are described from the Paskapoo Formation (Late Paleocene) at Joffre Bridge near Red Deer, Alta. Pistillate inflorescences with attached carpels, folliculate infructescences, seeds, seedlings, leaves, and shoots arc all assigned to Joffrea speirsii gen. et sp. nov. Crane and Stockey. Evidence for treating the different organs under a single binomial includes attachment of inflorescences and leaf petioles to long and short shoot systems, morphological intermediates between carpels and follicles, follicles preserved expelling seeds, seeds preserved during germination, stages in seedling development, similarity between seedling and adult foliage, and constant field association. Joffrea is similar in many respects to the widespread Upper Cretaceous and early Tertiary fossil Nyssidium arcticum (Heer) Iljinskaja. Comparison of Joffrea to the extant genera Cercidiphyllum Siebold et Zuccarini, Trochodendron Siebold et Zuccarini, and Tetracentron Oliver indicates a close relationship to Cercidiphyllum; and Joffrea is interpreted as an extinct representative of the family Cercidiphyllaceae. The fossil material clarifies morphological interpretations of the pistillate reproductive structures in extant Cercidiphyllum. The probable staminate inflorescences of Joffrea, are also similar to extant Cercidiphyllum. They are budlike and composed of 10–20 bracts, some of which have stamens in their axils. Pollen has not been recovered from the anthers. Pistillate inflorescences developed from axillary buds on the short shoots and were probably wind-pollinated. The production of numerous small winged seeds, epigeal germination, and the preservation of large numbers of in situ seedlings suggest that Joffrea speirsii and similar extinct Upper Cretaceous and early Tertiary species may have been "weedy" plants capable of rapidly colonizing open floodplain habitats.

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