The temporary immersion systems (TISs) have been widely used in plant biotechnology. TISs have different advantages from the point of micropropagation and production of secondary metabolites over other continuous liquid-phase bioreactors. The current work presents the structure, operation mode, configuration type, and micropropagation or secondary metabolite production in TISs. This review deals with the advantages and disadvantages of TISs and the factors affecting their performance. Future research could focus on new designs based on CFD simulation, facilitating sterilization, and combining TISs with other bioreactors (e.g., mist bioreactors) to make a hybrid bioreactor.

Caspases are a family of evolutionary conserved cysteine proteases that play key roles in programmed cell death and inflammation. Among the methods for the detection of caspase activity, biosensors based on luciferases have advantages in genetical encoding and convenience in assay. In this study, we constructed a new set of caspase biosensors based on NanoLuc luciferase. This kind of sensors, named NanoLock, work in dark-to-bright model, with the help of a NanoLuc quencher peptide (HiBiT-R/D) mutated from HiBiT. Optimized NanoLock responded to proteases with high signal to noise ratio (S/N), 1233-fold activation by tobacco etch virus protease in HEK293 cells and > 500-fold induction to caspase 3 in vitro. We constructed NanoLocks for the detection of caspase 1, 3, 6, 7, 8, 9, and 10, and assays in HEK293 cells demonstrated that these sensors performed better than commercial kits in the aspect of S/N and convenience. We further established a cell line stably expressing NanoLock-casp 6 and provided a proof-of-concept for the usage of this cell line in the high throughput screening of caspase 6 modulator.

Sustainable Development Goals (SDGs) have been part of much worldwide cooperation in engineering design, nutrients production that contributes towards a better and more sustainable future. This review intends to uncover a potential renewable source that could significantly contribute to various goals under the SDGs. The prospects of algae tackling the socio-ecological, economic, and environmental issues faced globally are discussed, along with approaches of algae that can be utilized to achieve many of the SDGs are reviewed and discussed. Moreover, the recent trends in terms of engineering application that co-relate to novel algae-based technology has also been included. Apart from that, algae have high oil content which is suitable for producing affordable and clean energy, which can be used for biofuels or electricity generation. The promising characteristics of algae will lead to its global acceptance and utilization for sustainability to help create a better world.

Toxic cyanobacterial blooms are recurrent for few decades throughout the globe, due to climate change, atmospheric warming and various anthropogenic activities with severe impacts of potential toxins on various ecosystems finally affecting the entire environment. These cyanobacteria are merely unexplored regarding their biochemical components except toxins. Variable influences and interactions of different factors including nitrogen, carbon, and availability of light are well known to crucially regulate cyanobacterial growth and metabolism. Thus, current research work is motivated for the evaluation and optimization of the effects of the aforementioned vital factors for improvement of biomass and lipid production of a freshwater, toxic strain of Anabaena circinalis. The modelling and optimization of factors such as nitrogen, light intensity and bicarbonate concentration (source of carbon) to maximize growth and lipid production were based on 20 design point experiments by Response Surface Methodology (RSM) and optimized values were further improved and validated by Particle Swarm Optimization (PSO) algorithm. The maximum optima were obtained 1.829 g L-1 and 39.64 % for biomass production and lipid content respectively from PSO optimization with two different sets of optimal values of factors. It shows 0.44 % and 2.77 % higher values of responses than that of RSM optimization. These asynchronous findings pioneered the enhanced lipid accumulation as well as the growth of a toxic cyanobacterium by optimizing interaction effects of culture conditions through various statistical and computational approaches.

Acid stress caused by the accumulation of acidic metabolites severely affects the fermentation performance of lactic acid bacteria. In this study, to overcome the impact of acid stress during growth, nine membrane transporters were introduced in Lactococcus lactis NZ9000 to study their effects on acid tolerance. The engineered strains that overexpressed the metal ATP-binding cassette (ABC) transporters zitP (metal ABC transporter permease) and zitQ (metal ABC transporter ATP-binding protein) exhibited 14.5 and 9.5-fold higher survival rates, respectively, at pH 4.0 for 4 h than the control strain. During acid stress, the two recombinant strains maintained relatively higher ATP concentrations, i.e., 7.7- and 11.7-fold higher, respectively, than the control strain at pH 4.0 for 3 h. Subsequently, transcriptome analysis revealed that genes associated with ABC transporters, metal ion transport, transcriptional regulation, and stress response exhibited differentially expressed. The transcriptional level of ecfA2 gene (energy-coupling factor transporter ATPase) was substantially higher in L. lactis (ZitQ) during acid stress, and the ecfA2 gene was overexpressed in L. lactis. This recombinant strain L. lactis (EcfA2) exhibited a 598.7-fold higher survival rate than the control strain at pH 4.0 for 4 h. This study showed that the membrane transporters ZitP and ZitQ could increase acid tolerance and provided a strategy for constructing robust strains that can be used in food industry.

In this study, the effect of ozonation process on the poly(ADP-ribose) polymerase 1 gene expression (PARP-1) and related the NADPH metabolism in strawberry fruit during storage was determined. Our results showed that ozonation with gas at both 10 and 100 ppm concentrations increased the expression of PARP-1 in the fruit during storage. Furthermore, the ozonation process initially increased the level of NAD+ and NADH in the fruit, which corresponds to a higher ATP level. The storage of the fruit in an ozone atmosphere also contributed to increased activity of the NAD+ kinase, leading to increased levels of NADP+ . In turn, the higher activity of glucose-6-phosphate dehydrogenase caused the ozonated fruit to show a higher level of NADPH. However, as the storage period extended and thus with increasing expression of PARP-1 in the ozonated fruit, the level of NAD+ decreased. In general, the ozonated fruit, which had a higher level of NADPH, showed a higher content of reduced glutathione, which in turn contributed to an increase in the antioxidant activity of the fruit and, ultimately, to a lower accumulation of reactive oxygen species.

Oleaginous microalgae represent potential feedstocks for the sustainable production of lipids thanks to their ability to accumulate triacylglycerols (TAGs). TAG accumulation in several algal species is strongly induced under specific conditions such as nutrient deprivation and high light which, however, also negatively impact growth. Genetic modification of lipogenic pathways can potentially enhance TAG accumulation without negatively affecting growth, avoiding the trade-off between biomass and lipid productivity. In this study, the phospholipid: diacylglycerol acyltransferase (PDAT), an enzyme involved in membrane lipid recycling, was overexpressed in the seawater alga Nannochloropsis gaditana. PDAT overexpression induced increased TAG content in actively growing algae cultures while no effects were observed in conditions naturally stimulating strong lipid accumulation such as high light and nitrogen starvation. The increase of TAG content was confirmed also in a strain cultivated in industrially relevant conditions even though PDAT overexpression, if too strong, the gene overexpression becomes detrimental for growth in the longer term. Results overall suggest that genetic modulation of the PDAT gene represents a promising strategy to increase microalgae lipid content by minimizing negative effects on biomass productivity.

In this study, we applied a tube-based reverse transcription loop-mediated isothermal amplification technique using preloaded amplification and detection reagents for simple screening of viable vancomycin-resistant Enterococcus in a cordless manner. We adopted an mRNA-based approach to detect live Enterococcus in vancomycin-treated cultures. We used agarose to preload and store all reagents for amplification and detection inside the tube, which could achieve on-site isothermal nucleic acid amplification and detection in less than 1 h without using sophisticated instruments. Moreover, the use of a portable insulated water tumbler eliminated the need for electricity, which is usually important in nucleic acid amplification-based assays. The water tumbler acted as a heat source to supply a stable heat required for the amplification reaction, which could last up to 45 min. In addition, colorimetric detection was realized using pH-based methods. The detection was triggered by shaking the tube so that the amplified solution was reacted with phenolphthalein embedded in the tube cap. The introduced one-pot strategy has many advantages such as easy and cordless operation, low cost, disposability, and less chance of contamination because the amplification and detection occur in a closed system. The system could have a great impact on nucleic acid analyses in instrument-free and low-resource areas.

Fusaric acid (FA) is a secondary fungal metabolite, which is widespread on corn and corn-based feed and food; FA has non-specific toxicity. Biosensor method is an express and easy-to-use method for quantitative and qualitative assessment of FA effect. Search for cultures has been performed for the formation of laboratory models of FA biosensor with the Clark-type oxygen electrode as transducer: respiration intensity of chosen cultures changed in the presence of FA. Resting cells of Fusarium oxysporum f. sp. vasinfectum and Bacillus subtilis were used as receptors of the amperometric biosensor for FA determination in aqueous solution. To enhance the sensitivity of detection, induction by substrate was performed for Bacillus subtilis. Response-concentration linear dependencies were obtained in a range of 0.5-500 FA mg/L. Biosensor models were applied to characterize influence of FA on microbial cells and investigate some features of FA transport. The dependences of the cells' response to FA on FA concentration were obtained; the kinetic parameters S0.5 and Vmax were determined for each culture. Inhibition-threshold FA (Sit) concentrations were similar for both studied cultures. At concentrations lower than Sit, the process of simple diffusion governed FA transport into cells and caused the cells' response to FA for non-induced culture.

The red palm weevil (RPW), Rhynchophorus ferrugineus, is considered a severe pest of palms. Usually, the early stages of infection are without visible signs. An attractive early sensing approach of non-visible infections is based on volatile organic compounds (VOCs). In this study, a whole-cell bacterial biosensor was used for the identification of RPW in date palm (Phoenix dactylifera). The cells are genetically modified to produce light in the presence of general stresses. The bioluminescent bacterial panel is based on three genetically engineered Escherichia coli strains that are sensitive to cytotoxicity (TV1061), genotoxicity (DPD2794), or quorum-sensing (K802NR). The bioluminescent bacterial panel detects the presence of VOCs and a change in the light signal is then generated, reflecting the health status of the date palm tree. The bioreporter bacteria cells are immobilized in calcium alginate tablets and placed in a sealed jar without direct contact with the tested sample, thereby exposing them only to the VOCs in the surrounding air. The immobilized bacteria cells were exposed to the air near infected by RPW or uninfected sugar canes, date palm tree pieces, and on date palm trees. Commercial plate reader was used for signal measurement. The findings show that quorum-sensing was induced by all the tested samples of infected sugar canes, date palm tree pieces, and date palm trees. While, cytotoxicity was induced only by infected date palm tree pieces, and genotoxicity was induced only by infected date palm trees. The bacterial monitoring results enable the identification of specific signatures that will allow a quick and accurate diagnosis.

Halomonas smyrnensis AAD6T is a moderately halophilic bacterium proven to be a powerful biotechnological tool with its ability to accumulate valuable biopolymers such as levan and poly(3-hydroxybutyrate) (PHB). Levan is a fructose homopolymer with β-2,6 fructofuranosidic linkages on the polymer backbone, and its distinctive applications in various industries such as food, pharmaceutical, medical, and chemical have been well-defined. On the other hand, PHB is a promising raw material to produce biodegradable plastics. Although it was shown in our previous studies that H. smyrnensis AAD6T exhibits one of the highest conversion yields of sucrose to levan reported to date, novel strategies are required to overcome high costs of levan production. In this study, we aimed at increasing levan productivity of H. smyrnensis AAD6T cultures using random mutagenesis techniques combined (i.e., ethyl methanesulfate treatment and/or ultraviolet irradiation). After several consecutive treatments, mutant strains BAE2, BAE5 and BAE6 were selected as efficient levan producers, as BAE2 standing out as the most efficient one not only in sucrose utilization and levan production rates, but also in final PHB concentrations. The mutants' whole genome sequences were analysed to determine the mutations occurred. Several mutations in genes related to central carbon metabolism and osmoregulation were found. Our results suggest that random mutagenesis can be a facile and efficient strategy to enhance the performance of extremophiles in adverse conditions.

Bacterial outer membrane vesicles (OMVs) are small unilamellar proteoliposomes, which are involved in various functions including cell to cell signaling and protein excretion. Here, we have engineered the OMVs of Escherichia coli to nano-scaled bioreactors for the degradation of β-lactam antibiotics. This was exploited by targeting a β-lactamase (i.e., CMY-10) into the OMVs of a hyper-vesiculating E. coli BL21(DE3) mutant. The CMY-10-containing OMVs, prepared from the E. coli mutant cultures, were able to hydrolyze β-lactam ring of nitrocefin and meropenem to a specific rate of 6.6 × 10-8 and 3.9 × 10-12 μmol/min/µm3 of OMV, which is approximately 100 and 600-fold greater than those of E. coli-based whole-cell biocatalsyts. Furthermore, CMY-10, which was encapsulated in the engineered OMVs, was much more stable against temperature and acid stresses, as compared to free enzymes in aqueous phase. The OMV-based nano-scaled reaction system would be useful for the remediation of a variety of antibiotics pollution for food and agricultural industry.

The effect of tungsten and selenium on cell growth and production of metabolites such as acetic acid and ethanol when fermenting syngas using "Clostridium autoethanogenum" was investigated to improve the process efficiency. General concentrations of selenium and tungsten in the medium are 0.01 µM during acetogenic syngas fermentation. We conducted culture experiments at concentrations of 0, 0.001, 0.01 and 0.1 µM for each heavy metal. The effect of selenium on cell growth and total metabolite production was greater than that of tungsten as the effect of selenium on formate dehydrogenase, an important enzyme of the Wood-Ljungdahl pathway, is greater than that of tungsten. Although an increase in tungsten had a marginal effect on total metabolite production, the ethanol/acetic acid production ratio increased significantly due to a decrease in acetic acid and an increase in ethanol production. Thus, tungsten plays a key role in activating aldehyde:ferredoxin oxidoreductase, a key enzyme in the reduction of acetate to ethanol. A specific ethanol productivity of 0.462 g ethanol/g DCW∙d was obtained in a culture using 0.01 µM selenium and 0.1 µM tungsten, which was 2.18 times higher than when using 0.01 µM of both selenium and tungsten.

The present study reports a strategy to produce polyhydroxyalkanoates (PHAs) by culturing the marine bacterium Rhodovulum sulfidophilum DSM-1374. The study was carried out by growing the bacterium anaerobically for 720 h under 16/8 light/dark cycle. Two analytical techniques such as proton magnetic nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy (FT-IR) were used to determine that the polyester produced was poly-3-hydroxybutirate-co-3-hydroxyvalerate (PHBV). This study showed that the excess of lactate and the limitation of N-P nutrients under a light-dark cycle enhanced PHBV synthesis and achieved a PHBV concentration of 330 mg/L in the R. sulfidophilum culture. During the 30 days of bacterial cultivation, the percentage of polymer in the six harvested dry biomasses gradually increased from 13.7% to 23.4%. In addition, the study showed that PHBV synthesis stopped during the 8-h dark phase and restarted in the light. The light-dark cycle study also showed that R. sulfidophilum DSM-1374 can be grown outdoors because the cells are exposed to the natural light-dark cycle.

The aim of this study was to establish a sustainable alternative callus culture of Cereus hildmannianus for the production and bioactive determination of phenolic compounds from this species. The conventional callus was cultivated using agar and Murashige and Skoog (MS) medium, while for the alternative culture the agar was replaced with a cotton support covered with filter paper and MS medium (incubated at 32 °C with photoperiod of 16 h), and the morphological characteristics and growth index were assessed (8 weeks). Extracts were obtained by maceration followed by partition, characterized by nuclear magnetic resonance - NMR and ultra-high performance liquid chromatography - UHPLC, quantified (phenolic compounds) by UV-Vis methods, and their antioxidant, antitumor activities, as well as cytotoxicity, were evaluated. The establishment of an alternative callus culture was carried out successfully. Characteristic signals of phenolic compounds were determined by NMR, and 46 compounds with fragment ions were identified using UHPLC analysis. The highest concentrations of phenolic compounds, and greatest antioxidant and antitumor activities, were obtained with the dichloromethane fractions of both callus tissue cultures, which were not cytotoxic. The callus culture from C. hildmannianus has shown promise as a source for the sustainable production of phenolic compounds with antioxidant and antiproliferative activities and thus, has potential use as a natural antitumor product.

This study evaluated the effects of the exogenous application of ethylenediaminetetraacetic acid (EDTA), indole-3-acetic acid (IAA) and iron sulfate (FeSO4) upon the phytochemical mechanisms of fenugreek grown under Pb-excess (2000 mg L-1 PbCl2). The results showed that chemical additives of EDTA and IAA as well as FeSO4 improved fenugreek germination parameters. The radicle length and the amylase activity were significantly improved under IAA treatment compared to EDTA and FeSO4. Exogenous FeSO4 was more effective to improving growth parameters. Moreover, the decrease in hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels was noted under all chemical additives especially under IAA application. In addition, it was more effective than EDTA and Fe in increasing catalase, glutathione (GSH), ascorbate peroxidase (APX), flavonoids and phenols while the increment superoxide dismutase (SOD) production was more pronounced under EDTA addition to Pb than other chelators. HPLC analysis revealed that the gallic was the major phenol produced under all chelators addition especially with IAA. In addition, the syringic acid was only produced with exogenous IAA while the quercetin was only detected under EDTA addition. Our results exhibited a higher IAA efficiency than EDTA and FeSO4 in mitigating Pb stress in fenugreek through up-regulated mechanisms of the antioxidant system for reducing reactive oxygen species (ROS) activities and enhancing special phenols.

This study aims to investigate the bioproduction and prospective biological applications of a natural red pigment from Talaromyces purpureogenus AUMC2603. Maximum pigment yield was achieved by a numerical optimization at pH 6, temperature 25 °C, and an 18-day incubation period on Yeast Malt Broth (YMB) media. The crude pigment was separated and purified into two pigment fractions via solid-phase extraction and then characterized as anthraquinone (dominant) and herquinone by LC/MS and 1HNMR analysis. The crude pigment extract and the two separated fractions displayed a potential antioxidant activity. Additionally, they showed a powerful anticancer activity towards cancer cell lines, MCF-7, HepG-2, and HCT116 with less cytotoxicity on normal cell lines, MCF12F and BJ-1T. The radioiodination efficiency of the radiosynthesized 99mTc-anthraquinone pigment complex was also investigated and optimized, obtaining a radiochemical yield of 92.70 % ± 0.89 %. An in vivo biodistribution study of the 99mTc-anthraquinone pigment complex demonstrated a high kidney uptake of 34 % injected dose per gram of organ tissue 60 min after intravenous injection, and the complex retention remained high up to 120 min. The current study is the first bioassay report on the efficacy of a purified anthraquinone from T. purpureogenus as a potent agent for kidney radio-imaging that could be applied in kidney cancer diagnosis.