scholarly journals Anthocyanin extract from Lycium ruthenicum enhanced production of biomass and polysaccharides during submerged fermentation of Agaricus bitorquis (Quél.) Sacc. Chaidam

2021 ◽  
Author(s):  
Shan Wu ◽  
Hong-Yun Lu ◽  
Qi-He Chen ◽  
Hui-Chun Xie ◽  
Ying-Chun Jiao
Keyword(s):  
Crude Extract ◽  
Submerged Fermentation ◽  
Biologically Active ◽  
Differentially Expressed ◽  
Tibet Plateau ◽  
Shake Flask Culture ◽  
Mycelial Biomass ◽  
Enhanced Production ◽  
Lycium Ruthenicum ◽  
Agaricus Bitorquis

AbstractAgaricus bitorquis (Quél.) Sacc. Chaidam (ABSC) is a wild edible fungus uniquely found in the Tibet Plateau. ABSC is rich in polysaccharides that are considered biologically active. This study aimed to determine the feasibility of enhancing exopolysaccharide (EPS) production by ABSC in shake flask culture by supplementing the fermentation medium with anthocyanin extract. Different concentrations of Lycium ruthenicum Murr. (LRM) anthocyanin crude extract were tested on ABSC fermentation. The activity of phosphoglucose isomerase (PGI), phosphoglucose mutase (PGM), and phosphomannose isomerase (PMI), enzymes presumably involved in EPS synthesis by ABSC, was determined. ABSC transcriptomic profile in response to the presence of anthocyanins during fermentation was also investigated. LRM anthocyanin crude extract (0.06 mg/mL) was most effective in increasing EPS content and mycelial biomass (by 208.10% and 105.30%, respectively, P < 0.01). The activity of PGI, PGM, and PMI was increased in a medium where LRM anthocyanin extract and its main components (proanthocyanidins and petunia anthocyanin) were added. RNA-Seq analysis showed that 349 genes of ABSC were differentially expressed during fermentation in the medium containing anthocyanin extract of LRM; 93 genes were up-regulated and 256 genes down-regulated. From gene ontology enrichment analysis, differentially expressed genes were mostly assigned to carbohydrate metabolism and signal transduction categories. Collectively, LRM anthocyanins extract positively affected EPS production and mycelial biomass during ABSC fermentation. Our study provides a novel strategy for improving EPS production and mycelial growth during ABSC liquid submerged fermentation.

scholarly journals Analytical Methods for Quantification and Identification of Intact Glucosinolates in Arabidopsis Roots Using LC-QqQ(LIT)-MS/MS

Metabolites ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 47
Author(s):  
Kourosh Hooshmand ◽  
Inge S. Fomsgaard
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Crude Extract ◽  
Matrix Effects ◽  
Critical Role ◽  
Reversed Phase ◽  
Biologically Active ◽  
Natural Soil ◽  
Chromatography Method ◽  
Arabidopsis Root

Glucosinolates are biologically active secondary metabolites in Brassicaceae plants that play a critical role in positive and negative interactions between plants and root-associated microbial communities. The aim of this study was to develop a reversed-phase liquid chromatography method to quantify and identify intact glucosinolates in the root of Arabidopsis thaliana (Arabidopsis) grown in non-sterile natural soil by using liquid chromatography-hybrid triple quadruple-linear ion trap (LC-QqQ(LIT)) mass spectrometry. The Synergi Fusion C18-based column was found to be effective for sufficient retention and separation of nine intact glucosinolates without the need for time-consuming desulfation or ion-pairing steps. Method validation results showed satisfactory inter-day and intra-day precision for all glucosinolates except for 4-hydroxyglucobrassicin. Good inter-day and intra-day accuracy and recovery results were observed for glucoiberin, gluconapin, glucobrassicin, 4-methoxyglucobrassicin and neoglucobrassicin. However, for 4-hydroxyglucobrassicin, glucoraphanin and glucoerucin corrections to quantification results might be necessary since the recovery and accuracy results were not optimal. Matrix effects were shown to have a negligible effect on the ionization of all target analytes. The established liquid chromatography–tandem mass spectrometry (LC-MS/MS) method was applied to quantify target intact glucosinolates in Arabidopsis root crude extract of four different wild-type accessions where differences in terms of concentration and composition of intact glucosinolates were observed. Employment of sensitive and selective precursor ion survey scan of m/z 97 in combination with the information-dependent acquisition (IDA) of the enhanced product ion (EPI) dependent scan (Prec97-IDA-EPI) using LC-QqQ(LIT) provided high confidence in structural characterization of diverse intact glucosinolate profiles in complex Arabidopsis root crude extract.

scholarly journals Identifying Candidate Genes for Hypoxia Adaptation of Tibet Chicken Embryos by Selection Signature Analyses and RNA Sequencing

Genes ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 823
Author(s):  
Xiayi Liu ◽  
Xiaochen Wang ◽  
Jing Liu ◽  
Xiangyu Wang ◽  
Haigang Bao
Keyword(s):  
Candidate Genes ◽  
Rna Sequencing ◽  
Differentially Expressed Genes ◽  
Gallus Gallus ◽  
Differentially Expressed ◽  
Tibet Plateau ◽  
Embryonic Liver ◽  
Selection Signature ◽  
Chicken Embryos ◽  
Hypoxia Adaptation

The Tibet chicken (Gallus gallus) lives on the Qinghai–Tibet Plateau and adapts to the hypoxic environment very well. The objectives of this study was to obtain candidate genes associated with hypoxia adaptation in the Tibet chicken embryos. In the present study, we used the fixation index (Fst) and cross population extended haplotype homozygosity (XPEHH) statistical methods to detect signatures of positive selection of the Tibet chicken, and analyzed the RNA sequencing data from the embryonic liver and heart with HISAT, StringTie and Ballgown for differentially expressed genes between the Tibet chicken and White leghorn (Gallus gallus, a kind of lowland chicken) embryos hatched under hypoxia condition. Genes which were screened out by both selection signature analysis and RNA sequencing analysis could be regarded as candidate genes for hypoxia adaptation of chicken embryos. We screened out 1772 genes by XPEHH and 601 genes by Fst, and obtained 384 and 353 differentially expressed genes in embryonic liver and heart, respectively. Among these genes, 89 genes were considered as candidate genes for hypoxia adaptation in chicken embryos. ARNT, AHR, GSTK1 and FGFR1 could be considered the most important candidate genes. Our findings provide references to elucidate the molecular mechanism of hypoxia adaptation in Tibet chicken embryos.

scholarly journals Different Expressions of Pericardial Fluid MicroRNAs in Patients With Arrhythmogenic Right Ventricular Cardiomyopathy and Ischemic Heart Disease Undergoing Ventricular Tachycardia Ablation

2021 ◽  
Vol 8 ◽  
Author(s):  
Aleksandr A. Khudiakov ◽  
Daniil D. Panshin ◽  
Yulia V. Fomicheva ◽  
Anastasia A. Knyazeva ◽  
Ksenia A. Simonova ◽  
...  
Keyword(s):  
Heart Disease ◽  
Ventricular Tachycardia ◽  
Right Ventricular ◽  
Arrhythmogenic Right Ventricular Cardiomyopathy ◽  
Pericardial Fluid ◽  
Current Data ◽  
Biologically Active ◽  
Differentially Expressed ◽  
Ventricular Cardiomyopathy ◽  
Ventricular Tachycardia Ablation

Introduction: Pericardial fluid is enriched with biologically active molecules of cardiovascular origin including microRNAs. Investigation of the disease-specific extracellular microRNAs could shed light on the molecular processes underlying disease development. Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited heart disease characterized by life-threatening arrhythmias and progressive heart failure development. The current data about the association between microRNAs and ARVC development are limited.Methods and Results: We performed small RNA sequence analysis of microRNAs of pericardial fluid samples obtained during transcutaneous epicardial access for ventricular tachycardia (VT) ablation of six patients with definite ARVC and three post-infarction VT patients. Disease-associated microRNAs of pericardial fluid were identified. Five microRNAs (hsa-miR-1-3p, hsa-miR-21-5p, hsa-miR-122-5p, hsa-miR-206, and hsa-miR-3679-5p) were found to be differentially expressed between patients with ARVC and patients with post-infarction VT. Enrichment analysis of differentially expressed microRNAs revealed their close linkage to cardiac diseases.Conclusion: Our data extend the knowledge of pericardial fluid microRNA composition and highlight five pericardial fluid microRNAs potentially linked to ARVC pathogenesis. Further studies are required to confirm the use of pericardial fluid RNA sequencing in differential diagnosis of ARVC.

scholarly journals Enhanced production of recombinant serratiopeptidase in Escherichia coli and its characterization as a potential biosimilar to native biotherapeutic counterpart

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Vishal Srivastava ◽  
Shivam Mishra ◽  
Tapan K. Chaudhuri
Keyword(s):  
Protein Expression ◽  
Process Parameters ◽  
Recombinant Expression ◽  
Cellular Protein ◽  
Biologically Active ◽  
Fixed Dose ◽  
Antibiofilm Activity ◽  
Protein Accumulation ◽  
E Coli ◽  
Enhanced Production

Abstract Background Serratia marcescens, a Gram-negative nosocomial pathogen secretes a 50 kDa multi-domain zinc metalloprotease called serratiopeptidase. Broad substrate specificity of serratiopeptidase makes it suitable for detergent and food processing industries The protein shows potent anti-inflammatory, anti-edemic, analgesic, antibiofilm activity and sold as an individual or fixed-dose enteric-coated tablets combined with other drugs. Although controversial, serratiopeptidase as drug is used in the treatment of chronic sinusitis, carpal tunnel syndrome, sprains, torn ligaments, and postoperative inflammation. Since the native producer of serratiopeptidase is a pathogenic microorganism, the current production methods need to be replaced by alternative approaches. Heterologous expression of serratiopeptidase in E. coli was tried before but not found suitable due to the limited yield, and other expression related issues due to its inherent proteolytic activity such as cytotoxicity, cell death, no expression, minimal expression, or inactive protein accumulation. Results Recombinant expression of mature form serratiopeptidase in E. coli seems toxic and resulted in the failure of transformation and other expression related issues. Although E. coli C43(DE3) cells, express protein correctly, the yield was compromised severely. Optimization of protein expression process parameters such as nutrient composition, induction point, inducer concentration, post-induction duration, etc., caused significant enhancement in serratiopeptidase production (57.9 ± 0.73% of total cellular protein). Expressed protein formed insoluble, enzymatically inactive inclusion bodies, and gave 40–45 mg/l homogenous (> 98% purity) biologically active and conformationally similar serratiopeptidase to the commercial counterpart upon refolding and purification. Conclusion Expression of mature serratiopeptidase in E. coli C43(DE3) cells eliminated the protein expression associated with toxicity issues. Further optimization of process parameters significantly enhanced the overexpression of protein resulting in the higher yield of pure and functionally active recombinant serratiopeptidase. The biological activity and conformational features of recombinant serratiopeptidase were very similar to the commercially available counterpart suggesting it-a potential biosimilar of therapeutic and industrial relevance.

scholarly journals Enhanced production of lipstatin from mutant of Streptomyces toxytricini and fed-batch strategies under submerged fermentation

3 Biotech ◽  
2020 ◽  
Vol 10 (4) ◽  
Author(s):  
Khushboo ◽  
Pinki ◽  
Punit Kumar ◽  
Archana Tripathi ◽  
Umesh Luthra ◽  
...  
Keyword(s):  
Submerged Fermentation ◽  
Fed Batch ◽  
Enhanced Production ◽  
Streptomyces Toxytricini

scholarly journals EVALUATION OF DIFFERENT CULTURE MEDIA FOR ENHANCED PRODUCTION OF PSEUDOMONAS AERUGINOSA (MTCC NO 2453) BIOMASS AND ITS PROTEINS

2016 ◽  
Vol 8 (11) ◽  
pp. 120 ◽  
Author(s):  
P. Neeraja ◽  
T. Parthasarathy ◽  
M. Sudhakar
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Protein Production ◽  
Culture Media ◽  
Batch Cultivation ◽  
Luria Bertani ◽  
Shake Flask Culture ◽  
Cell Biomass ◽  
Enhanced Production ◽  
Anti Cancer ◽  
The Media

Objective: Microorganisms, especially bacteria and its proteins have proven to be potential anti-cancer agents as they selectively attack the tumor cells or tumor micro-environments. The extract of Pseudomonas aeruginosa found to contain proteins that have shown promising anticancer activity. In this work, it was attempted to increase the biomass and trigger the total protein fraction of Pseudomonas aeruginosa (MTCC 2453).Methods: The organism was cultivated in three different such as Luria-Bertani (LB) broth, minimal medium9 (M9), super broth medium (SB) and asparagine-proline (AP) broth. Asparagine proline broth was selected as it has shown high cell growth rate. The media was further optimized by the addition of NaHCO3 and copper sulphate to trigger the protein production. Optimized Aspergine proline broth has achieved highest cell biomass. After the shake flask culture, the overnight grown culture in optimized AP medium was further grown in a 5 L bioreactor by fed-batch cultivation to achieve higher cell densities.Results: The highest protein production was achieved at 40 ° C. Highest biomass and protein content was observed at pH 8 while lowest biomass was produced at pH 2. A gradual increase in biomass content observed from 12 h towards to 48 h.Conclusion: High biomass and proteins content and of Pseudomonas aeruginosa (MTCC 2453) can be produced in optimized asparagine-proline broth. Further the extract is purified to produce novel anti-cancer proteins.

scholarly journals Ultrasound enhanced production of mycelia and exopolysaccharide by Agaricus bitorquis (Quél.) Sacc. Chaidam

2020 ◽  
Vol 64 ◽  
pp. 105040 ◽  
Author(s):  
Hongyun Lu ◽  
Hanghang Lou ◽  
Tianyu Wei ◽  
Zhengjie Liu ◽  
Yingchun Jiao ◽  
...  
Keyword(s):  
Enhanced Production ◽  
Agaricus Bitorquis

scholarly journals Microarray screening of suppression subtractive hybridization-PCR cDNA libraries identifies novel RNAs regulated by dehydration in the rat supraoptic nucleus

2006 ◽  
Vol 24 (2) ◽  
pp. 163-172 ◽  
Author(s):  
Mohamed T. Ghorbel ◽  
Greig Sharman ◽  
Charles Hindmarch ◽  
Kevin G. Becker ◽  
Tanya Barrett ◽  
...  
Keyword(s):  
General Circulation ◽  
Supraoptic Nucleus ◽  
Subtractive Hybridization ◽  
Biologically Active ◽  
Differentially Expressed ◽  
Cdna Libraries ◽  
Suppressive Subtractive Hybridization ◽  
Cdna Clones ◽  
Microarray Screening ◽  
Mechanistic Basis

The magnocellular neurons (MCNs) of the supraoptic nucleus (SON) and paraventricular nucleus (PVN) of the hypothalamus are the principal site of biosynthesis of prepropeptide precursor of the antidiuretic hormone vasopressin (VP). This precursor is processed during anterograde axonal transportation to terminals in the posterior pituitary gland, where biologically active VP is stored until release into the general circulation in response to physiological activation of the SON by osmotic cues. By binding to V2-type receptors located in the kidney, VP decreases the amount of water lost in urine. Osmotic activation of the SON is accompanied by a dramatic morphological and functional remodeling. We have sought to understand the mechanistic basis of this plasticity in terms of the differential expression of genes. To identify such genes, we adopted an unbiased global approach based on suppressive subtractive hybridization-polymerase chain reaction (SSH-PCR) Using this method, we generated libraries of clones putatively differentially expressed in control vs. dehydrated SON. To rapidly screen these libraries, 1,152 clones were subjected to microarray analysis, resulting in the identification of 459 differentially expressed transcripts. cDNA clones corresponding to 56 of these RNAs were sequenced, revealing many of them to be novel expressed sequence tags (ESTs). Four transcripts were shown by in situ hybridization (ISH) to be significantly up- or downregulated in the SON after dehydration. These genes may represent novel effectors or mediators of SON physiological remodeling.

Sign in / Sign up

Export Citation Format

Share Document