scholarly journals Biological Stoichiometric Analysis during Substrate Utilization and Secondary Metabolite Production by Non-Saccharomyces Yeasts Using Grape Pomace Extract as Fermentation Medium

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 89
Author(s):  
Maxwell Mewa-Ngongang ◽  
Heinrich W. du Plessis ◽  
Seteno K. O. Ntwampe ◽  
Enoch A. Akinpelu ◽  
Ucrecia F. Hutchinson ◽  
...  
Keyword(s):  
Scale Up ◽  
Cost Effective ◽  
Fermentation Medium ◽  
Grape Pomace ◽  
Yeast Cells ◽  
Biochemical Engineering ◽  
Metabolites Production ◽  
Pichia Kluyveri ◽  
High Density Cell ◽  
Saccharomyces Yeasts

The emerging interest in the search for alternatives to synthetic preservatives has led to various successful research studies exploring the use of yeasts as potential biological control agents and producers of biopreservatives. The findings that yeasts could be used as producers of biopreservatives lacked some engineering considerations regarding cost-effective process design for scale-up, although partial process optimization using renewable agro-waste has been achieved. This study investigated the biological stoichiometry and bioenergetic parameters during yeast growth and secondary metabolites production i.e., biopreservatives from non-Saccharomyces yeasts using grape pomace extract (GPE), a type of agro-waste, as a fermentation medium. This was achieved by reconfirming the optimum production conditions previously found for Candida pyralidae Y1117, Pichia kluyveri Y1125, and Pichia kluyveri Y1164 in GPE broth as a fermentation medium, conditions under which a high amount of yeast cells were obtained. High-density cell cultures were produced, from which the yeast cell pellets were harvested, dried, and combusted for the determination of elemental analysis, heat of combustion, biological stoichiometry, and bioenergetic parameters. This work generated biological stoichiometric models and bioenergetics information that could assist in the design of yeast biochemical conversion system when GPE is used as fermentation medium, thereby, addressing the biochemical engineering aspects that were lacking in a previous biopreservative production study using Candida pyralidae Y1117, Pichia kluyveri Y1125, and Pichia kluyveri Y1164.

scholarly journals Grape Pomace Extracts as Fermentation Medium for the Production of Potential Biopreservation Compounds

Foods ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 51 ◽  
Author(s):  
Maxwell Mewa-Ngongang ◽  
Heinrich du Plessis ◽  
Seteno Ntwampe ◽  
Boredi Chidi ◽  
Ucrecia Hutchinson ◽  
...  
Keyword(s):  
Food Industry ◽  
Isoamyl Alcohol ◽  
Fermentation Medium ◽  
Isoamyl Acetate ◽  
Grape Pomace ◽  
Production Medium ◽  
Zone Of Inhibition ◽  
Food Losses ◽  
Chemical Preservatives ◽  
Pichia Kluyveri

Microbial spoilage causes food losses in the food industry and as such, the use of synthetic chemical preservatives is still required. The current study proposes the use of agro-waste, i.e., grape pomace extracts (GPE), as production medium for biopreservation compounds. Production kinetics, subsequent to optimization using response surface methodology (RSM) for biopreservation compounds production was studied for three yeasts using GPE broth as a fermentation medium. The results showed that the highest volumetric zone of inhibition (VZI) was 1.24 L contaminated solidified media (CSM) per mL biopreservation compounds used (BCU) when Candida pyralidae Y1117 was inoculated in a pH 3-diluted GPE broth (150 g L−1) incubated at 25 °C for 24 h. Similar conditions were applied for Pichia kluyveri Y1125 and P. kluyveri Y1164, albeit under slightly elongated fermentation periods (up to 28 h), prior to the attainment of a maximum VZI of only 0.72 and 0.76 L CSM mL−1 ACU, respectively. The potential biopreservation compounds produced were identified to be isoamyl acetate, isoamyl alcohol, 2-phenyl ethylacetate and 2-phenyl ethanol.

scholarly journals Allogeneic human neural stem cells for improved therapeutic delivery to peritoneal ovarian cancer

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Rachael Mooney ◽  
Wafa Abidi ◽  
Jennifer Batalla-Covello ◽  
Hoi Wa Ngai ◽  
Caitlyn Hyde ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cell Tracking ◽  
Scale Up ◽  
Cancer Therapeutics ◽  
Cost Effective ◽  
Tumor Stroma ◽  
Clinical Safety ◽  
Peritoneal Cancer ◽  
Human Neural Stem Cells ◽  
Link Type

Abstract Background Immortalized, clonal HB1.F3.CD21 human neural stem/progenitor cells (NSCs), loaded with therapeutic cargo prior to intraperitoneal (IP) injection, have been shown to improve the delivery and efficacy of therapeutic agents in pre-clinical models of stage III ovarian cancer. In previous studies, the distribution and efficacy of the NSC-delivered cargo has been examined; however, the fate of the NSCs has not yet been explored. Methods To monitor NSC tropism, we used an unconventional method of quantifying endocytosed gold nanorods to overcome the weaknesses of existing cell-tracking technologies. Results Here, we report efficient tumor tropism of HB1.F3.CD21 NSCs, showing that they primarily distribute to the tumor stroma surrounding individual tumor foci within 3 h after injection, reaching up to 95% of IP metastases without localizing to healthy tissue. Furthermore, we demonstrate that these NSCs are non-tumorigenic and non-immunogenic within the peritoneal setting. Conclusions Their efficient tropism, combined with their promising clinical safety features and potential for cost-effective scale-up, positions this NSC line as a practical, off-the-shelf platform to improve the delivery of a myriad of peritoneal cancer therapeutics.

scholarly journals Pilot-Scale Production of Chito-Oligosaccharides Using an Innovative Recombinant Chitosanase Preparation Approach

Polymers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 290
Author(s):  
Chih-Yu Cheng ◽  
Chia-Huang Tsai ◽  
Pei-Jyun Liou ◽  
Chi-Hang Wang
Keyword(s):  
Cell Density ◽  
Scale Up ◽  
Cost Effective ◽  
Pilot Scale ◽  
Ionic Concentration ◽  
Dihydrogen Phosphate ◽  
Scale Production ◽  
Sodium Dihydrogen Phosphate ◽  
Selective Precipitation ◽  
Pilot Scale Production

For pilot-scale production of chito-oligosaccharides, it must be cost-effective to prepare designable recombinant chitosanase. Herein, an efficient method for preparing recombinant Bacillus chitosanase from Escherichia coli by elimination of undesirable substances as a precipitate is proposed. After an optimized culture with IPTG (Isopropyl β-d-1-thiogalactopyranoside) induction, the harvested cells were resuspended, disrupted by sonication, divided by selective precipitation, and stored using the same solution conditions. Several factors involved in these procedures, including ion types, ionic concentration, pH, and bacterial cell density, were examined. The optimal conditions were inferred to be pH = 4.5, 300 mM sodium dihydrogen phosphate, and cell density below 1011 cells/mL. Finally, recombinant chitosanase was purified to >70% homogeneity with an activity recovery and enzyme yield of 90% and 106 mg/L, respectively. When 10 L of 5% chitosan was hydrolyzed with 2500 units of chitosanase at ambient temperature for 72 h, hydrolyzed products having molar masses of 833 ± 222 g/mol with multiple degrees of polymerization (chito-dimer to tetramer) were obtained. This work provided an economical and eco-friendly preparation of recombinant chitosanase to scale up the hydrolysis of chitosan towards tailored oligosaccharides in the near future.

scholarly journals Scale-Up Preparation of Crocins I and II from Gardeniajasminoides by a Two-Step Chromatographic Approach and Their Inhibitory Activity Against ATP Citrate Lyase

Molecules ◽  
2021 ◽  
Vol 26 (11) ◽  
pp. 3137
Author(s):  
Shuguang Guan ◽  
Qiaoli Pu ◽  
Yinan Liu ◽  
Honghong Wu ◽  
Wenbo Yu ◽  
...  
Keyword(s):  
Inhibitory Activity ◽  
High Speed ◽  
Scale Up ◽  
Cost Effective ◽  
Atp Citrate Lyase ◽  
Gardenia Jasminoides ◽  
Citrate Lyase ◽  
Human Disorders ◽  
Ic50 Values ◽  
Counter Current

Crocins are highly valuable natural compounds for treating human disorders, and they are also high-end spices and colorants in the food industry. Due to the limitation of obtaining this type of highly polar compound, the commercial prices of crocins I and II are expensive. In this study, macroporous resin column chromatography combined with high-speed counter-current chromatography (HSCCC) was used to purify crocins I and II from natural sources. With only two chromatographic steps, both compounds were simultaneously isolated from the dry fruit of Gardenia jasminoides, which is a cheap herbal medicine distributed in a number of countries. In an effort to shorten the isolation time and reduce solvent usage, forward and reverse rotations were successively utilized in the HSCCC isolation procedure. Crocins I and II were simultaneously obtained from a herbal resource with high recoveries of 0.5% and 0.1%, respectively, and high purities of 98.7% and 99.1%, respectively, by HPLC analysis. The optimized preparation method was proven to be highly efficient, convenient, and cost-effective. Crocins I and II exhibited inhibitory activity against ATP citrate lyase, and their IC50 values were determined to be 36.3 ± 6.24 and 29.7 ± 7.41 μM, respectively.

scholarly journals Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alisa Alekseenko ◽  
Donal Barrett ◽  
Yerma Pareja-Sanchez ◽  
Rebecca J. Howard ◽  
Emilia Strandback ◽  
...  
Keyword(s):  
Large Scale ◽  
Direct Detection ◽  
Dna Polymerases ◽  
Scale Up ◽  
Lamp Assay ◽  
Cost Effective ◽  
Reaction Conditions ◽  
Clinical Patient ◽  
Reverse Transcriptases ◽  
Large Scale Testing

AbstractRT-LAMP detection of SARS-CoV-2 has been shown to be a valuable approach to scale up COVID-19 diagnostics and thus contribute to limiting the spread of the disease. Here we present the optimization of highly cost-effective in-house produced enzymes, and we benchmark their performance against commercial alternatives. We explore the compatibility between multiple DNA polymerases with high strand-displacement activity and thermostable reverse transcriptases required for RT-LAMP. We optimize reaction conditions and demonstrate their applicability using both synthetic RNA and clinical patient samples. Finally, we validate the optimized RT-LAMP assay for the detection of SARS-CoV-2 in unextracted heat-inactivated nasopharyngeal samples from 184 patients. We anticipate that optimized and affordable reagents for RT-LAMP will facilitate the expansion of SARS-CoV-2 testing globally, especially in sites and settings where the need for large scale testing cannot be met by commercial alternatives.

scholarly journals Enhanced production of taxadiene in Saccharomyces cerevisiae

2020 ◽  
Author(s):  
Behnaz Nowrouzi ◽  
Rachel Li ◽  
Laura E. Walls ◽  
Leopold d’Espaux ◽  
Koray Malci ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Shake Flask ◽  
Scale Up ◽  
Cost Effective ◽  
Cancer Drug ◽  
List Type ◽  
Taxadiene Synthase ◽  
Enhanced Production ◽  
Anti Cancer ◽  
Positive Effect

AbstractCost-effective production of the highly effective anti-cancer drug, paclitaxel (Taxol®), remains limited despite growing global demands. Low yields of the critical taxadiene precursor remains a key bottleneck in microbial production. In this study, the key challenge of poor taxadiene synthase (TASY) solubility in S. cerevisiae was revealed, and the strains were strategically engineered to relieve this bottleneck. Multi-copy chromosomal integration of TASY harbouring a selection of fusion solubility tags improved taxadiene titres 22-fold, up to 57 ± 3 mg/L at 30 °C at shake flask scale. The scalability of the process was highlighted through achieving similar titres during scale up to 25 mL and 250 mL in shake flask and bioreactor cultivations, respectively. Maximum taxadiene titres of 129 ± 15 mg/L and 119 mg/L were achieved through shake flask and bioreactor cultivation, respectively, of the optimal strain at a reduced temperature of 20 °C. The results highlight the positive effect of coupling molecular biology tools with bioprocess variable optimisation on synthetic pathway development.HighlightsMaximum taxadiene titre of 129 ± 15 mg/L in Saccharomyces cerevisiae at 20 °CIntegrating fusion protein tagged-taxadiene synthase improved taxadiene titre.Consistent taxadiene titres were achieved at the micro-and mini-bioreactor scales.

scholarly journals Optimization of Metal Ions in Sugarcane Bagasse Fermenting Medium for the Production of Streptokinase by Streptococcus equisimilis

2021 ◽  
Vol 9 (2) ◽  
pp. 24-30
Keyword(s):  
Metal Ions ◽  
Sugarcane Bagasse ◽  
Specific Activity ◽  
Gel Filtration ◽  
Value Added ◽  
Cost Effective ◽  
Fermentation Medium ◽  
Exchange Chromatography ◽  
Sds Page ◽  
The Cost

Streptokinase is a fibrinolytic enzyme and a product of β-hemolytic Streptococci strains. This enzyme is used as a medication to break down clots in some cases of heart disease. Streptococcus equisimilis, a species of group C Streptococci, is widely used for the production of streptokinase by fermentation technology. In this study, the sugarcane bagasse fermentation medium was optimized for metal ions (KH2PO4, MgSO4.7H2O, CaCO3 and NaHCO3) at various levels to attain the maximal production of streptokinase. Sugarcane bagasse was used due to its profuse availability and as an ideal substrate for microbial processes for the manufacturing of value-added products. The results showed that maximal streptokinase production was found at 0.04% KH2PO4, 0.04% MgSO4.7H2O, 0.15% NaHCO3 and 0.04% CaCO3. Finally, the optimized medium resulted in 84.75 U/mg specific activity and 74.5% recovery. The purification process was carried out simultaneously using ammonium sulfate precipitation, ion-exchange chromatography, and gel filtration. Finally, a purified sample of streptokinase was run on SDS-PAGE and resolute 47 kDa molecular weight. The use of β-hemolytic Streptococci to obtain streptokinase is not free from health risks and is related to anaphylaxis. This study provides a way forward for the cost-effective ways to obtain streptokinase for the treatment of thrombosis.

scholarly journals NaIO4/Br- as a mild system for the oxidation of 1-methyl-anthra-9,10-quinones

2015 ◽  
Author(s):  
Marcin Cybulski ◽  
Adam Formela ◽  
Katarzyna Sidoryk ◽  
Olga Michalak ◽  
Anna Rosa ◽  
...  
Keyword(s):  
High Pressure ◽  
Carboxylic Acids ◽  
Scale Up ◽  
Lithium Bromide ◽  
Cost Effective ◽  
Building Blocks ◽  
Multidrug Resistant ◽  
Resistant Cell ◽  
Direct Oxidation ◽  
Reaction Conditions

One of the anthraquinone classes comprises compounds with a carbonyl group. These natural or synthetic anthraquinones find their application as building blocks in the synthesis of the compounds with a biological activity. Recently, 4-substituted anthra-9,10-quinone-1-carboxylic acids (2) have been used as key intermediates in the synthesis of patented compounds (3) with anticancer activity against multidrug resistant cell lines. Although 2,7-dihydro-3H-dibenz[de,h]cinnolin-3,7-diones (3) were successfully synthetized in a small laboratory scale, several problems were observed during the preparation of their acid intermediates (2) in a multi-gram scale. The known methods for the preparation of 2 are based on the oxidation of the methyl group in anthra-9,10-quinones (1). The most common are: the oxidation with the diluted nitric acid under high pressure in a sealed tube at the temperature of 195-220 oC, the oxidation in nitrobenzene by passing chlorine gas through the reaction mixture at the temperature of 160-170 oC or in a presence of the fuming sulphuric acid. The mentioned methods require aggressive reagents and specific reaction conditions including high pressure and temperature. Thus, there was a need to find a new efficient, cost-effective and reproducible synthetic method of preparation of 2. While searching literature it was found that the direct oxidation of alkylarenes mediated by the sodium periodate/lithium bromide combination produces benzyl acetates throughout benzyl bromides in the acetic acid, or benzylic acids in the diluted inorganic acid. Based on these results we examined a variety of reaction conditions with or without the bromine source and the oxidizing anion. As a result, a novel procedure for the preparation of highly pure 4-substituted anthra-9,10-quinone-1-carboxylic acids (HPLC > 99.5%) using oxidizing anion/ brominating reagent system was developed. It enabled 2 isolation by the simple filtration of the reaction mixture and was applied in the scale-up of 2,7-dihydro-3H-dibenz[de,h]cinnolin-3,7-dione derivatives.

scholarly journals Modelling to inform the COVID-19 response in Bangladesh

2021 ◽  
Author(s):  
Elaine A Ferguson ◽  
Eric Brum ◽  
Anir Chowdhury ◽  
Shayan Chowdhury ◽  
Mikolaj Kundegorski ◽  
...  
Keyword(s):  
Scale Up ◽  
Cost Effective ◽  
Filtration Efficiency ◽  
Potential Outcomes ◽  
Case Detection ◽  
Effective Response ◽  
Seir Model ◽  
Death Data ◽  
Pharmaceutical Interventions

AbstractBackgroundNon-pharmaceutical interventions (NPIs) used to limit SARS-CoV-2 transmission vary in their feasibility, appropriateness and effectiveness in different contexts. In Bangladesh a national lockdown implemented after the first detected case in early March 2020 rapidly exacerbated poverty and was considered untenable long-term, whilst surging cases in 2021 warrant renewed NPIs. We examine potential outcomes and costs of NPIs considered appropriate and feasible to deploy in Dhaka over the course of the pandemic including challenges of compliance and scale up.MethodsWe developed an SEIR model for application to Dhaka District, parameterised from literature values and calibrated to death data from Bangladesh. We discussed scenarios and parameterizations with policymakers using an interactive app, to guide modelling of lockdown and post-lockdown measures considered feasible to deliver; symptoms-based household quarantining and compulsory mask-wearing. We examined how testing capacity affects case detection and compared deaths, hospitalisations relative to capacity, working days lost from illness and NPI compliance, and cost-effectiveness.ResultsLockdowns alone were predicted to delay the first epidemic peak but were unable to prevent overwhelming of the health service and were extremely costly. Predicted impacts of post-lockdown interventions depended on their reach within communities and levels of compliance: symptoms-based household quarantining alone was unable to prevent hospitalisations exceeding capacity whilst mask-wearing could prevent overwhelming health services and be cost-effective given masks of high filtration efficiency. The modelled combination of these measures was most effective at preventing excess hospitalizations for both medium and high filtration efficiency masks. Even at maximum testing capacity, confirmed cases far underestimate total cases, with saturation limiting reliability for assessing trends. Recalibration to surging cases in 2021 suggests limited immunity from previous infections and the need to re-sensitize communities to increase mask wearing.ConclusionsMasks and symptoms-based household quarantining act synergistically to prevent transmission, and are cost-effective in mitigating impacts. Our interactive app was valuable in supporting decision-making in Bangladesh, where mask-wearing was mandated early, and community teams have been deployed to support household quarantining across Dhaka. This combination of measures likely contributed to averting the worst impacts of a public health disaster as predicted under an unmitigated epidemic, but delivering an effective response at scale has been challenging. Moreover, lack of protection to the B.1.351 variant means messaging to improve mask-wearing is urgently needed in response to surging cases.

Biological Synthesis of Silver Nanoparticles and their Biomedical Activity: A Review

2021 ◽  
Vol 09 ◽  
Author(s):  
Sarvat Zafar ◽  
Aiman Zafar ◽  
Fakhra Jabeen ◽  
Miad Ali Siddiq
Keyword(s):  
Silver Nanoparticles ◽  
Large Scale ◽  
Scale Up ◽  
Cost Effective ◽  
Biological Properties ◽  
Single Step ◽  
Biological Synthesis ◽  
Nanosized Particles ◽  
Environment Friendly ◽  
Physical And Chemical

: Nanotechnology studies the various phenomena of physio-chemical procedures and biological properties for the generation of nanosized particles, and their rising challenges in the various sectors, like medicine, engineering, agriculture, electronic, and environmental studies. The nanosized particles exhibit good anti-microbial, anti-inflammatory, cytotoxic, drug delivery, anti-parasitic, anti-coagulant and catalytic properties because of their unique dimensions with large surface area, chemical stability and higher binding density for the accumulation of various bio-constituents on their surfaces. Biological approaches for the synthesis of silver nanoparticles (AgNPs) have been reviewed because it is an easy and single-step protocol and a viable substitute for the synthetic chemical-based procedures. Physical and chemical approaches for the production of AgNPs are also mentioned herein. Biological synthesis has drawn attention because it is cost-effective, faster, non-pathogenic, environment-friendly, easy to scale-up for large-scale synthesis, and having no demand for usage of high pressure, energy, temperature, or noxious chemical ingredients, and safe for human therapeutic use. Therefore, the collaboration of nanomaterials with bio-green approaches could extend the utilization of biological and cytological properties compatible with AgNPs. In this perspective, there is an immediate need to develop ecofriendly and biocompatible techniques, which strengthen efficacy against microbes and minimize toxicity for human cells. The present study introduces the biological synthesis of silver nanoparticles, and their potential biomedical applications have also been reviewed.

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