Growth Medium Research Articles

Utilization of palm oil and palm oil effluent as a carbon source for the production of polyhydroxybutyrate co‐polyhydroxyhexanoate P(HB‐co‐HHx) a biopolymer in bacillus sp

AbstractBACKGROUNDBiodegradable polymers are gaining a reputation as demand is increasing globally. Amongst these Polyhydroxyalkanoates (PHA) are attracting market attention as a consequence of their properties similar to those of conventional plastics. PHA is a natural polyester that is stored as an intracellular carbon (C) energy reserve of bacteria. This biopolymer can be extracted and used in place of petrochemical plastic. Medium chain‐length (mcl) PHA, specifically poly hydroxybutyrate‐co‐hydroxyhexanoate P (HB‐co‐HHx) copolymer produced by Bacillus, has gained practical attention owing to its superior physicochemical properties compared to the commonly synthesized PHB homopolymer. Co‐polymer production is dependent on C sources provided in the growth medium. In the present study we have employed palm oil and palm oil effluent as C source.RESULTSIn the current study, it was observed that a significant amount of PHA and biomass was produced when the effluent was prepared at a concentration of 100% instead of water for preparing the growth medium. The production of biomass and PHA showed a range of 4.8–8and 1.5–4 g L−1, respectively. This means that the PHA yield obtained using the effluent was 50–80% of the control, whereas the control yielded only 40–50%.CONCLUSIONPalm industry waste can be used as a substrate in bioprocessing industries. Proton nuclear magnetic resonance analysis of PHA obtained herein indicates that it consists of the mcl polymer P (HB‐co‐HHx). © 2024 The Author(s). Journal of Chemical Technology and Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry (SCI).

Screening and evaluation of antibacterial active strains of Actinomycetes isolated from Northern Indian soil for biofilm inhibition against selected ESKAPE pathogens

AbstractThe growing problem of antimicrobial resistance (AMR) globally has warranted the search for new and alternate anti-infective strategies. This study aimed to isolate and screen soil Actinomycetes from three different locations in Northern India for their broad-spectrum antibacterial and antibiofilm activity against certain drug-resistant bacteria from ESKAPE group. A total of 110 isolates of Actinomycetes were screened for antibacterial activity using overlay and agar-well diffusion assay against Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Ninety-five isolates showed antibacterial activity against one or more test bacteria. These isolates were tentatively identified as members of genus Streptomyces. Five isolates (IMA13, IMA25, IMA43, IMA46, and IMA47) demonstrated broad-spectrum activity against test pathogens and were subjected to production of secondary metabolites in liquid medium. Ethyl acetate fraction of culture filtrate showed MIC in the range of 64–512 µg/ml against test bacteria. The most active isolate, IMA-46, was identified by 16srRNA gene sequence analysis as Streptomyces rameus. Further, ethyl acetate fraction from S. rameus-IMA46 was used to inhibit biofilm formation at sub-MIC values. A significant level (50% to 92%) of biofilm inhibition was recorded against test pathogens. Light microscopy and SEM analysis revealed the biofilm structural changes, reduction in cell aggregation and EPS production. LC/MS analysis of the active extract of S. rameus IMA46 showed the presence of many biologically active compounds, including tetracycline and doxycycline. The present study highlights the potential of soil Actinomycetes in production of broad-spectrum anti-infective compounds in combating growing problem of AMR.

Effects of temporal IFNγ exposure on macrophage phenotype and secretory profile: exploring GMP-Compliant production of a novel subtype of regulatory macrophages (MregIFNγ0) for potential cell therapeutic applications

BackgroundMacrophages are involved in tissue homeostasis, angiogenesis and immunomodulation. Proangiogenic and anti-inflammatory macrophages (regulatory macrophages, Mreg) can be differentiated in-vitro from CD14+ monocytes by using a defined cell culture medium and a stimulus of IFNγ.Aim of the studyTo scrutinize the potential impact of temporal IFNγ exposure on macrophage differentiation as such exposure may lead to the emergence of a distinct and novel macrophage subtype.MethodsDifferentiation of human CD14+ monocytes to Mreg was performed using a GMP compliant protocol and administration of IFNγ on day 6. Monocytes from the same donor were in parallel differentiated to MregIFNγ0 using the identical protocol but with administration of IFNγ on day 0. Cell characterization was performed using brightfield microscopy, automated and metabolic cell analysis, transmission electron microscopy, flow cytometry, qPCR and secretome profiling.ResultsMreg and MregIFNγ0 showed no differences in cell size and volume. However, phenotypically MregIFNγ0 exhibited fewer intracellular vesicles/vacuoles but larger pseudopodia-like extensions. MregIFNγ0 revealed reduced expression of IDO and PD-L1 (P < 0.01 for both). They were positive for CD80, CD14, CD16 and CD38 (P < 0.0001vs. Mreg for all), while the majority of MregIFNγ0 did not express CD206, CD56, and CD103 on their cell surface (P < 0.01 vs. Mreg for all). In terms of their secretomes, MregIFNγ0 differed significantly from Mreg. MregIFNγ0 media exhibited reduced levels of ENA-78, Osteopontin and Serpin E1, while the amounts of MIG (CXCL9) and IP10 were increased.ConclusionExposing CD14+ monocytes to an alternatively timed IFNγ stimulation results in a novel macrophage subtype which possess additional M1-like features (MregIFNγ0). MregIFNγ0 may therefore have the potential to serve as cellular therapeutics for clinical applications beyond those covered by M2-like Mreg, including immunomodulation and tumor treatment.

Use of Viscous medium to study anthelmintic drug action in Caenorhabditis elegans

Caenorhabditis elegans is an appealing tool for experimental evolution and for working with antiparasitic drugs, from understanding the molecular mechanisms of drug action and resistance to uncover new drug targets. We present a new methodology for studying the impact of antiparasitic drugs in C. elegans. Viscous medium was initially designed for C. elegans maintenance during long-term evolution experiments. Viscous medium provides a less structured environment than the standard nematode growth media agar, yet the bacteria food source remains suspended. Further, the Viscous medium offers the worm population enough support to move freely, mate, and reproduce at a rate comparable to standard agar cultures. Here, the Viscous medium was adapted for use in antiparasitic research. We observed a similar sensitivity of C. elegans to anthelmintic drugs as in standard liquid media and statistical difference to the standard agar media through a larval development assay. Using Viscous medium in C. elegans studies will considerably improve antiparasitic resistance research, and this medium could be used in studies aimed at understanding long-term multigenerational drug activity.

Development of an Azobenzene-Based Cell Culture Photoresponsive Platform for In Situ Modulation of Surface Topography in Wet Environments.

Azopolymers are light-responsive materials that hold promise to transform in vitro cell culture systems. Through precise light illumination, they facilitate substrate pattern formation and erasure, allowing for the dynamic control and creation of active interfaces between cells and materials. However, these materials exhibit a tendency to locally detach from the supporting glass in the presence of aqueous solutions, such as cell culture media, due to the formation of blisters, which are liquid-filled cavities generated at the azopolymer film-glass interface. These blisters impede precise structurization of the surface of the azomaterial, limiting their usage for surface photoactivation in the presence of cells. In this study, we present a cost-effective and easily implementable method to improve the azopolymer-glass interface stability through silane functionalization of the glass substrate. This method proved to be efficient in preventing blister formation, thereby enabling the dynamic modulation of the azopolymer surface in situ for live-cell experiments. Furthermore, we proved that the light-illumination conditions used to induce azopolymer surface variations do not induce phototoxic effects. Consequently, this approach facilitates the development of a photoswitchable azopolymer cell culture platform for studying the impact of multiple in situ inscription and erasure cycles on cell functions while maintaining a physiological wet microenvironment.

Detergent-resistant α-amylase derived from Anoxybacillus karvacharensis K1 and its production based on whey

In the field of biotechnology, the utilization of agro-industrial waste for generating high-value products, such as microbial biomass and enzymes, holds significant importance. This study aimed to produce recombinant α-amylase from Anoxybacillus karvacharensis strain K1, utilizing whey as an useful growth medium. The purified hexahistidine-tagged α-amylase exhibited remarkable homogeneity, boasting a specific activity of 1069.2 U mg−1. The enzyme displayed its peak activity at 55 °C and pH 6.5, retaining approximately 70% of its activity even after 3 h of incubation at 55 °C. Its molecular weight, as determined via SDS-PAGE, was approximately 69 kDa. The α-amylase demonstrated high activity against wheat starch (1648.8 ± 16.8 U mg−1) while exhibiting comparatively lower activity towards cyclodextrins and amylose (≤ 200.2 ± 16.2 U mg−1). It exhibited exceptional tolerance to salt, withstanding concentrations of up to 2.5 M. Interestingly, metal ions and detergents such as sodium dodecyl sulfate (SDS), Triton 100, Triton 40, and Tween 80, 5,5ʹ-dithio-bis-[2-nitrobenzoic acid (DNTB), β-mercaptoethanol (ME), and dithiothreitol (DTT) had no significant inhibitory effect on the enzyme’s activity, and the presence of CaCl2 (2 mM) even led to a slight activation of the recombinant enzyme (1.4 times). The Michaelis constant (Km) and maximum reaction rate (Vmax), were determined using soluble starch as a substrate, yielding values of 1.2 ± 0.19 mg mL−1 and 1580.3 ± 183.7 μmol mg−1 protein min−1, respectively. Notably, the most favorable conditions for biomass and recombinant α-amylase production were achieved through the treatment of acid whey with β-glucosidase for 24 h.

TOTAL BAKTERI DAN KOLIFORM PADA MEDIA PERTUMBUHAN MAGGOT Black Soldier Fly KOMBINASI FESES SAPI PERAH, ENDAPAN SUSU DAN SAMPAH ORGANIK DAPUR

Livestock industry waste and kitchen waste contain high levels of organic matter and are dangerous if not managed properly. To overcome this problem, bioconversion can be carried out using BSF maggot. This research aims to determine whether the growth media after being degraded by BSF maggots still contains bacteria and coliform that meet the requirements for solid organic fertilizer Kepmentan No. 261 Tahun 2019. The research was carried out experimentally with 4 treatments and 5 repetitions. Treatment consists of P0 = 100% organic kitchen waste, P1 = 50% dairy cow feces and 50% organic kitchen waste, P2 = 50% milk sludge and 50% organic kitchen waste, P3 = 33,33% dairy cow feces, milk sludge 33,33% and kitchen organic waste 33,33%. The data was analyzed using a paired t-test. The results for total bacteria and coliform before and after degradation of BSF maggot experienced a significant decrease (P&lt;0,05) and fulfilled the requirements for Escherichia coli in the coliform group with a total of &lt;102 MPN/g in the BSF maggot growth media.

Recycling spent animal cell culture media using the thermally resistant microalga Chlorella sorokiniana

AbstractCell culture media is a significant contributor to the high cost of bioprocesses. This study explored the potential of algae to re-condition spent cell culture medium, which may reduce the costs of pharmaceutical and lab-grown-meat manufacturing. Chlorella sorokiniana, a thermally resistant microalgal species, exhibited heterotrophic and mixotrophic growth in mammalian cell culture growth media (GM). Spent animal cell media, generated by culturing quail myoblast cells (QM7s) for 4 or 8d (4D-SGM and 8D-SGM, respectively), was harvested for algal culture. Increased algal growth was observed in 4D-SGM, when compared to fresh media after 3d (optical density of 1.39 ± 0.22, 0.47 ± 0.17, p ≤ 0.05). Within 72 h, ammonia and glucose were eliminated from 4D-SGM with algal treatment. After treating 4D-SGM with algae, the treated media was reintroduced into QM7 cell cultures. No cytotoxic effects were observed on QM7 cells grown in algal-treated growth media and QM7 cells exhibited better metabolic activity in algal-treated spent medium than in untreated spent medium (80.85 ± 12.02% and 44.57 ± 10.82% activity of fresh media group, respectively, p ≤ 0.05). These results suggest that C. sorokiniana can be grown in spent media at 37 °C, sequester ammonia, and potentially extend the lifespan of media, thereby enabling more affordable bioprocesses.

Run-and-tumble kinematics of Enterobacter Sp. SM3.

The recent discovery of the peritrichous, swarm-competent bacterium Enterobacter sp. SM3 has offered a new opportunity to investigate the connection between bacterial swimming and swarming. Here, we report the run-and-tumble behavior of SM3 as planktonic swimming cells and as swarming cells diluted in liquid medium, drawing comparison between the two states. Swimming cells of SM3 run for an average of 0.77s with a speed of approximately 30µm/s before tumbling. Tumbles last for a duration of 0.12s on average and cause changes in direction averaging 69^{∘}. Swimming cells exposed to the common chemoattractant serine in bulk solution suppress the frequency of tumbles in the steady state, lengthening the average run duration and decreasing the average tumble angle. When exposed to aspartate, cells do not demonstrate a notable change in run-and-tumble parameters in the steady state. For swarming cells of SM3, the frequency of tumbles is reduced, with the average run duration being 50% longer on average than that of swimming cells in the same liquid medium. Additionally, the average tumble angle of swarming cells is smaller by 35%. These findings reveal that the newly identified species, SM3, performs run-and-tumble motility similar to other species of peritrichous bacteria such as E. coli, both in the swimming and swarming states. We present a simple mechanical model, which provides a physical understanding of the run-and-tumble behavior of peritrichous bacteria.

Enhancement of growth media for extreme iron limitation in Escherichia coli.

Iron is an essential nutrient for microbial growth and bacteria have evolved numerous routes to solubilize and scavenge this biometal, which is often present at very low concentrations in host tissue. We recently used a MOPS-based medium to induce iron limitation in Escherichia coli K-12 during the characterization of novel siderophore-conjugated antibiotics. In this study we confirm that growth media derived from commercially available M9 salts are unsuitable for studies of iron-limited growth, probably through the contamination of the sodium phosphate buffer components with over 100 µM iron. In contrast, MOPS-based media that are treated with metal-binding Chelex resin allow the free iron concentration to be reduced to growth-limiting levels. Despite these measures a small amount of E. coli growth is still observed in these iron-depleted media. By growing E. coli in conditions that theoretically increase the demand for iron-dependent enzymes, namely by replacing the glucose carbon source for acetate and by switching to a microaerobic atmosphere, we can reduce background growth even further. Finally, we demonstrate that by adding an exogeneous siderophore to the growth media which is poorly used by E. coli, we can completely prevent growth, perhaps mimicking the situation in host tissue. In conclusion, this short study provides practical experimental insight into low iron media and how to augment the growth conditions of E. coli for extreme iron-limited growth.

Changes in stresses sensitivity of ohmic heating-induced sublethally injured Staphylococcus aureus during repair: Potential mechanisms at the cellular and molecular levels

Changes in stresses sensitivity of ohmic heating-induced sublethally injured Staphylococcus aureus during repair: Potential mechanisms at the cellular and molecular levels

PAHs Biodegradation by Locally Isolated Phanerochaete chrysosporium and Penicillium citrinum from Liquid and Spiked Soil

In the present study, biodegradation of polycyclic aromatic hydrocarbons (PAHs) was examined using two fungal strains, namely P. chrysosporium and P. citrinum, isolated from locally contaminated soil. These two fungal strains were compared based on degradation properties under standardized conditions (pH 7.0, temperature 30oC, carbon source yeast extract) using PAH sole and a mixture of five different PAHs. In liquid media, PAH degradation was higher as compared to spiked soil by P. chrysosporium, followed by P. citrinum. In liquid culture, maximum degradation was 96.13% phenanathrene, 86.34% fluoranthene, 72.75% pyrene, 52.25% chrysene, and 40.16% benzo(a)pyrene by P. chrysosporium. PAH degradation in spiked soil was 78.5% phenanthrene, 65.91% fluoranthene, 61.73% pyrene, 48.2% chrysene, and 26.82% benzo(a)pyrene within 28 days by P. chrysosporium. Both local fungal isolates showed potential for degradation of PAHs alone and in PAH mixtures.

Exploring the Interplay between Nutrients, Bacteriophages, and Bacterial Lipases in Host- and Bacteria-mediated Pathogenesis.

Pathogenic bacteria and host cells counteract or neutralize each other's effect in two fundamental ways: Direct invasion and secretion of various substances. Among these, lipases secreted by pathogenic bacteria and host cell lysozyme are key actors. Secreted lipases from pathogenic bacterial are suggested as a key player in the pathogen-host interaction. Among the gut microbial energy sources, glucose and fats have been referred to as one of the best inducers and substrates for bacterial lipases. Enrichment of bacterial growth medium with extra glucose or oil has been shown to induce lipase production in pathogenic bacteria. More recently, research has focused on the role of human gut phage alterations in the onset of dysbiosis because the bacteria-phage interactions can be dramatically affected by the nutrient milieu of the gut. However, the reciprocal role of bacterial lipases and phages in this context has not been well studied and there is no data available about how high glucose or fat availability might modulate the cellular milieu of the pathogenic bacteria-phageeukaryotic host cell interface. The purpose of this study was to evaluate the immunologic outcome of pathogenic bacteria-phage interaction under normal, high glucose, and high butter oil conditions to understand how nutrient availability affects lipase activity in pathogenic bacteria and, ultimately, the eukaryotic host cell responses to pathogenic bacteria-phage interaction. 10 groups of co-cultured T84 and HepG2 cells were treated with Pseudomonas aeruginosa strain PAO1 (P.a PAO1) in the presence and absence of its KPP22 phage and incubated in three different growth media (DMEM, DMEM + glucose and DMEM + butter oil). Structural and physiological (barrier function and cell viability), inflammatory (IL-6 and IL-8), metabolic (glucose and triglycerides), and enzymatic (lipases and lysozyme) parameters were determined. Excess glucose or butter oil enhanced additively extracellular lipase activity of P.a PAO1. Excess glucose or butter oil treatments also magnified P. a PAO1- induced secretion of inflammatory signal molecules (IL-1β, IL-6) from co-cultured cells, concomitant with the enhancement of intracellular triglycerides in co-cultured HepG2 cells, these effects being abolished by phage KPP22. The results of the present study imply that KPP22 phage influences the interplay between food substances, gut bacterial lipases, and the gut cellular milieu. This can be applied in two-way interaction: by affecting the microbial uptake of excess free simple sugars and fats from the gut milieu leading to decreased bacterial lipases and by modulating the immune system of the intestinal -liver axis cells. Further studies are needed to see if the biological consequences of these effects also occur in vivo.

Evaluation of hydrolyzed cheese whey medium for enhanced bacterial cellulose production by Komagataeibacter rhaeticus MSCL 1463.

Industrial production of bacterial cellulose (BC) remains challenging due to significant production costs, including the choice of appropriate growth media. This research focuses on optimization of cheese whey (CW) based media for enhanced production of BC. Two modifications were made for CW medium for BC production with Komagataeibacter rhaeticus MSCL 1463. BC production in a medium of enzymatically hydrolyzed CW (final concentration of monosaccharides: glucose 0.13g L-1, galactose 1.24g L-1) was significantly enhanced, achieving a yield of 4.95±0.25g L-1, which markedly surpasses the yields obtained with the standard Hestrin-Schramm (HS) medium containing 20g L-1 glucose and acid-hydrolyzed CW (final concentration of monosaccharides: glucose 1.15g L-1, galactose 2.01g L-1), which yielded 3.29±0.12g L-1 and 1.01±0.14g L-1, respectively. We explored the synergistic effects of combining CW with various agricultural by-products (corn steep liquor (CSL), apple juice, and sugar beet molasses). Notably, the supplementation with 15% corn steep liquor significantly enhanced BC productivity, achieving 6.97±0.17g L-1. A comprehensive analysis of the BC's physical and mechanical properties indicated significant alterations in fiber diameter (62-167nm), crystallinity index (71.1-85.9%), and specific strength (35-82MPa × cm3 g-1), as well as changes in the density (1.1-1.4g cm-3). Hydrolyzed CW medium supplemented by CSL could be used for effective production of BC.

Cultivation of yeasts on liquid digestate to remove organic pollutants and nutrients and for potential application as co-culture with microalgae

In this study, the growth of yeast and yeast-like fungi in the liquid digestate from vegetable wastes was investigated in order to remove nutrients and organic pollutants, and for their application as co-culture members with green microalgae. The studied yeast strains were characterized for their assimilative and enzymatic profiles as well as temperature requirements. In the first experimental stage, the growth dynamics of each strain were determined, allowing to select the best yeasts for further studies. In the subsequent stage, the ability of selectants to remove organic pollutants was assessed. Different cultivation media containing respectively 1:3, 1:1, 3:1 vol ratio of liquid digestate and the basal minimal medium were used. Among all tested yeast strains, Rhodotorula mucilaginosa DSM 70825 showed the most promising results, demonstrating the highest potential for removing organic substrates and nutrients. Depending on the medium, this strain achieved 50–80% sCOD, 45–60% tVFAs, 21–45% TN, 33–52% PO43− reduction rates. Similar results were obtained for the strain Candida sp. OR687571. The high nutrient and organics removal efficiency by these yeasts could likely be linked to their ability to assimilate xylose (being the main source of carbon in the liquid digestate). In culture media containing liquid digestate, both yeast strains achieved good viability and proliferation potential. In the liquid digestate medium, R. mucilaginosa and Candida sp. showed vitality at the level of 51.5% and 45.0%, respectively. These strains seem to be a good starting material for developing effective digestate treatment strategies involving monocultures and/or consortia with other yeasts or green microalgae.

Genetic characteristics of cerebrospinal fluid-derived circulating tumor DNA in brain metastases of lung adenocarcinoma.

e20503 Background: Brain metastases (BM), encompassing parenchymal metastases (PM) and leptomeningeal metastases (LM), are a common and fatal complication of advanced lung adenocarcinoma (LUAD). However, the genomic disparities between PM and LM, as determined by cerebrospinal fluid (CSF)-derived circulating tumor DNA (ctDNA), and its clinical utility as a liquid biopsy medium remain unclear. Methods: We conducted a retrospective study involving 33 patients diagnosed with BM from LUAD, comprising 16 cases with PM and 17 cases with LM, at Shandong Cancer Hospital between March 2018 and June 2021. CSF and matched peripheral blood lymphocyte samples were collected from all patients, and plasma samples were collected from 25 of them. Genomic profiles of CSF and plasma in patients with PM and LM were compared. CSF-ctDNA, plasma-ctDNA, and DNA from matched peripheral blood lymphocytes underwent next-generation sequencing (NGS) based on a 769 cancer-related gene panel. Results: A total of 363 mutations were detected in CSF-ctDNA, with 90.9% (30/33) of all LUAD patients with PM or LM showing mutations. EGFR mutations were the most common, occurring in PM samples from 56.3% (9/16) of patients and in LM samples from 76.5% (13/17) of patients, followed by other frequently mutated driver genes, including TP53 in 19 (57.6%) patients, MLH1 in 9 (27.3%) patients, and KMT2C in 8 (24.2%). Notably, EGFR 19del was more prevalent as a driver mutation in patients with LM compared to those with PM in our study, particularly the p.E746_A750del mutated subtype. Furthermore, the frequency of mutations in the mismatch repair (MMR) pathway was significantly higher in patients with LM than in those with PM. Additionally, CSF exhibited a more comprehensive profile of driver genomic mutations than matched plasma in patients with any form of BM, accompanied by a higher variation burden and detection of druggable mutations. Conclusions: CSF-ctDNA sequencing revealed distinct genomic patterns in patients with BM, particularly in LM, and could potentially enhance the management of BM patients compared to plasma.

Comparative growth response of three rubber (Hevea brasiliensis Muell. Arg) clones in somatic embryo propagation

Abstract Plants have the ability to grow with different responses depending on the clones and varieties of plants. In rubber plants, each clone has specific characteristics or traits that distinguish it from other clones, whether in terms of growth, latex production, or resistance to pests and diseases. The objective of this experiment was to assess the response of three rubber (Hevea brasiliensis Muell. Arg) clones to the same media conditions for the growth and development of somatic embryos (SE). Integuments from three rubber clones, namely PB 260, PB 330, and PB 340, were used as explants to induce embryonic callus on MH media. After four weeks post-induction, the callus was sub-cultured to expressed SE then SE was transferred to the RITA® bio-reactor for plantlets development in liquid MD media. Parameters for each steps, such as the percentage of contamination, percentage of callus response, color of callus, percentage of SE induction, number of stage of SE, and number of shoots were recorded. The results revealed that the highest percentage of viable callus was observed in the PB 260 clone at 23.20%. On SE expression, the highest being in clone PB 340 at 2.04%. On SE development the highest shoot growth was observed in the PB 260 clone, with a total of 5 shoots.

Antibody glycan quality predicted from CHO cell culture media markers and machine learning

N-glycosylation can have a profound effect on the quality of mAb therapeutics. In biomanufacturing, one of the ways to influence N-glycosylation patterns is by altering the media used to grow mAb cell expression systems. Here, we explore the potential of machine learning (ML) to forecast the abundances of N-glycan types based on variables related to the growth media. The ML models exploit a dataset consisting of detailed glycomic characterisation of Anti-HER fed-batch bioreactor cell cultures measured daily under 12 different culture conditions, such as changes in levels of dissolved oxygen, pH, temperature, and the use of two different commercially available media. By performing spent media quantitation and subsequent calculation of pseudo cell consumption rates (termed media markers) as inputs to the ML model, we were able to demonstrate a small subset of media markers (18 selected out of 167 mass spectrometry peaks) in a Chinese Hamster Ovary (CHO) cell cultures are important to model N-glycan relative abundances (Regression - correlations between 0.80–0.92; Classification - AUC between 75.0–97.2). The performances suggest the ML models can infer N-glycan critical quality attributes from extracellular media as a proxy. Given its accuracy, we envisage its potential applications in biomaufactucuring, especially in areas of process development, downstream and upstream bioprocessing.

Endophytic Bacteria Improves the Growth of Tomato Plants: An Initial Trial Towards Sustainable Agriculture

Abstract Microbe-based biofertilizer remains high demand toward sustainable agriculture. Previous study isolated three potential endophytic bacteria from the root of klutuk banana plants namely K2, K8, and K117. The potencies were indicated by IAA production ability. This experiment examined the influence of root inoculation using these endophytic bacteria on the growth of tomato plants. Isolate preparation was initiated by shaking the three isolates in the Nutrient Broth medium for six hours. The seeds of tomato plants were germinated by soaking in sterile water for an hour after surface sterilization. Then, the soaked seeds were incubated for 3 days on the moist paper towel in the petri dish to achieve a high humidity. The germinated tomato seeds were inoculated by soaking the root tip in the bacterial isolates for 10 minutes. Subsequently, the inoculated tomato plants were grown on sterile soils in the greenhouse condition. The height of the plants, leaf count, root length, and quantity of roots were assessed 28 days after inoculation (dai). The t-test analysis indicated two isolates, K2 and K117, significantly improved the height of the plants, leaf count and root lengths compared to the control. In addition, all isolates significantly increased the number of roots of tomato plants. This finding revealed the plant growth improvement caused by root inoculation by these potential endophytic bacteria. This preliminary study gives additional information about a novel biofertilizer for sustainable agriculture.

An Exceptional Valorization of CuO Nanoparticles in Ionic Liquids as an Efficient Medium for the Electrophilic Substitution of Indole Towards the Formation of Bis(indolyl)methanes

Aim: Ionic liquids are promising green solvents with simple but unique structure-related physical properties such as negligible vapour pressure, exceptional thermal conductivity, remarkable thermal stability and their suitability and inertness towards a broad range of catalytic applications. CuO NPs have been addressed as a cost-effective and a reagent of a choice that necessitates only mild reaction conditions to offer a high yield of the desired products with exceptional selectivity in a short duration of time. Therefore, in the present work, attempts have been made to explore the catalytic potentials of CuO NPs in an ionic liquid medium to synthesize biologically important bis(indolyl)methanes. Background: Catalytic explorations of metal oxide nanoparticles in ionic liquids offers a cooperative effect that has a significant impact on the kinetics as well as on the outcome of the reaction. Therefore, such catalytic systems in the present times have seized the scientific community's interest from the perspectives of sustainable development in synthetic organic chemistry. The combination of metal oxide nanoparticles with highly tunable ionic liquids is not only used to synthesize simple organic molecules but also explored in the synthesis of complex organic molecules of high commercial and biological relevance. Objectives: The current work offers a rapid and robust protocol for synthesizing bis(indolyl)methanes via electrophilic substitution reaction between indole and various aldehydes in the presence of a CuO nanoparticles-ionic liquid system. The discussion focuses on the high tolerance of different functionalities by the catalytic system leading to the synthesis of bis(indolyl)methanes. Methods: CuO NPs have been synthesized via the co-precipitation method using ionic liquid. The applicability of metal oxide nanoparticles-IL matrix was further investigated in synthesizing bis(indolyl)methanes. Results: The FT-IR absorption below 600 cm-1 and the XRD pattern showing all the peaks in the diffraction diagram revealed the formation of CuO NPs. FESEM images show the flake-shaped morphology of CuO NPs and are found to be separated from the agglomerated clusters Conclusion: Ionic liquid-CuO NPs matrix reveals good to exceptional catalytic properties, and their advancements as a catalytic system at room temperature open new avenues for synthetic organic chemists.