BioTech

Open AccessReview

DNA Data Storage

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Tomasz Buko

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Nella Tuczko

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Takao Ishikawa

BioTech 2023, 12(2), 44; https://doi.org/10.3390/biotech12020044 - 01 Jun 2023

Abstract

The demand for data storage is growing at an unprecedented rate, and current methods are not sufficient to accommodate such rapid growth due to their cost, space requirements, and energy consumption. Therefore, there is a need for a new, long-lasting data storage medium [...] Read more.

The demand for data storage is growing at an unprecedented rate, and current methods are not sufficient to accommodate such rapid growth due to their cost, space requirements, and energy consumption. Therefore, there is a need for a new, long-lasting data storage medium with high capacity, high data density, and high durability against extreme conditions. DNA is one of the most promising next-generation data carriers, with a storage density of 10¹⁹ bits of data per cubic centimeter, and its three-dimensional structure makes it about eight orders of magnitude denser than other storage media. DNA amplification during PCR or replication during cell proliferation enables the quick and inexpensive copying of vast amounts of data. In addition, DNA can possibly endure millions of years if stored in optimal conditions and dehydrated, making it useful for data storage. Numerous space experiments on microorganisms have also proven their extraordinary durability in extreme conditions, which suggests that DNA could be a durable storage medium for data. Despite some remaining challenges, such as the need to refine methods for the fast and error-free synthesis of oligonucleotides, DNA is a promising candidate for future data storage. Full article

(This article belongs to the Topic Computational Intelligence and Bioinformatics (CIB))

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Open AccessArticle

Influence of Growth Medium Composition on Physiological Responses of Escherichia coli to the Action of Chloramphenicol and Ciprofloxacin

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BioTech 2023, 12(2), 43; https://doi.org/10.3390/biotech12020043 - 01 Jun 2023

Abstract

The ability of hydrogen sulfide (H₂S) to protect bacteria from bactericidal antibiotics has previously been described. The main source of H₂S is the desulfurization of cysteine, which is either synthesized by cells from sulfate or transported from the medium, [...] Read more.

The ability of hydrogen sulfide (H₂S) to protect bacteria from bactericidal antibiotics has previously been described. The main source of H₂S is the desulfurization of cysteine, which is either synthesized by cells from sulfate or transported from the medium, depending on its composition. Applying electrochemical sensors and a complex of biochemical and microbiological methods, changes in growth, respiration, membrane potential, SOS response, H₂S production and bacterial survival under the action of bactericidal ciprofloxacin and bacteriostatic chloramphenicol in commonly used media were studied. Chloramphenicol caused a sharp inhibition of metabolism in all studied media. The physiological response of bacteria to ciprofloxacin strongly depended on its dose. In rich LB medium, cells retained metabolic activity at higher concentrations of ciprofloxacin than in minimal M9 medium. This decreased number of surviving cells (CFU) by 2–3 orders of magnitude in LB compared to M9 medium, and shifted optimal bactericidal concentration (OBC) from 0.3 µg/mL in M9 to 3 µg/mL in LB. Both drugs induced transient production of H₂S in M9 medium. In media containing cystine, H₂S was produced independently of antibiotics. Thus, medium composition significantly modifies physiological response of E. coli to bactericidal antibiotic, which should be taken into account when interpreting data and developing drugs. Full article

(This article belongs to the Section Medical Biotechnology)

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Graphical abstract

Open AccessArticle

Application of Chlorella vulgaris Beijerinck as a Biostimulant for Growing Cucumber Seedlings in Hydroponics

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Galiya I. Vildanova

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Rezeda Z. Allaguvatova

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Dina F. Kunsbaeva

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Natalia V. Sukhanova

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Lira A. Gaysina

BioTech 2023, 12(2), 42; https://doi.org/10.3390/biotech12020042 - 22 May 2023

Abstract

Hydroponics is a promising method for growing agricultural plants and is especially relevant in the context of global climate change. Microscopic algae, including Chlorella vulgaris, have great potential for use in hydroponic systems as natural growth stimulators. The effect of the suspension [...] Read more.

Hydroponics is a promising method for growing agricultural plants and is especially relevant in the context of global climate change. Microscopic algae, including Chlorella vulgaris, have great potential for use in hydroponic systems as natural growth stimulators. The effect of the suspension of an authentic strain of Chlorella vulgaris Beijerinck on the length of cucumber shoots and roots, as well as its dry biomass, was studied. During cultivation in a Knop medium with the addition of Chlorella suspension, the length of the shoots was shortened from 11.30 to 8.15 cm, while the length of the roots also decreased from 16.41 to 10.59 cm. At the same time, the biomass of the roots increased from 0.04 to 0.05 g. The data obtained indicate the positive effect of the suspension of the Chlorella vulgaris authentic strain on the dry biomass of cucumber plants in hydroponic conditions and make it possible to recommend this strain for use when growing plants in hydroponic systems. Full article

(This article belongs to the Special Issue New Technologies in Agricultural and Plant Biotechnology)

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Open AccessReview

Trends in Biological Ammonia Production

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BioTech 2023, 12(2), 41; https://doi.org/10.3390/biotech12020041 - 19 May 2023

Abstract

Food production heavily depends on ammonia-containing fertilizers to improve crop yield and profitability. However, ammonia production is challenged by huge energy demands and the release of ~2% of global CO₂. To mitigate this challenge, many research efforts have been made to [...] Read more.

Food production heavily depends on ammonia-containing fertilizers to improve crop yield and profitability. However, ammonia production is challenged by huge energy demands and the release of ~2% of global CO₂. To mitigate this challenge, many research efforts have been made to develop bioprocessing technologies to make biological ammonia. This review presents three different biological approaches that drive the biochemical mechanisms to convert nitrogen gas, bioresources, or waste to bio-ammonia. The use of advanced technologies—enzyme immobilization and microbial bioengineering—enhanced bio-ammonia production. This review also highlighted some challenges and research gaps that require researchers’ attention for bio-ammonia to be industrially pragmatic. Full article

(This article belongs to the Section Industrial Biotechnology)

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Open AccessArticle

Mass Cultivation of Microalgae: II. A Large Species Pulsing Blue Light Concept

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Hans Chr. Eilertsen

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Jo Strømholt

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John-Steinar Bergum

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Gunilla Kristina Eriksen

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Richard Ingebrigtsen

BioTech 2023, 12(2), 40; https://doi.org/10.3390/biotech12020040 - 17 May 2023

Abstract

If mass cultivation of photoautotrophic microalgae is to gain momentum and find its place in the new “green future”, exceptional optimizations to reduce production costs must be implemented. Issues related to illumination should therefore constitute the main focus, since it is the availability [...] Read more.

If mass cultivation of photoautotrophic microalgae is to gain momentum and find its place in the new “green future”, exceptional optimizations to reduce production costs must be implemented. Issues related to illumination should therefore constitute the main focus, since it is the availability of photons in time and space that drives synthesis of biomass. Further, artificial illumination (e.g., LEDs) is needed to transport enough photons into dense algae cultures contained in large photobioreactors. In the present research project, we employed short-term O₂ production and 7-day batch cultivation experiments to evaluate the potential to reduce illumination light energy by applying blue flashing light to cultures of large and small diatoms. Our results show that large diatom cells allow more light penetration for growth compared to smaller cells. PAR (400–700 nm) scans yielded twice as much biovolume-specific absorbance for small biovolume (avg. 7070 μm³) than for large biovolume (avg. 18,703 μm³) cells. The dry weight (DW) to biovolume ratio was 17% lower for large than small cells, resulting in a DW specific absorbance that was 1.75 times higher for small cells compared to large cells. Blue 100 Hz square flashing light yielded the same biovolume production as blue linear light in both the O₂ production and batch experiments at the same maximum light intensities. We therefore suggest that, in the future, more focus should be placed on researching optical issues in photobioreactors, and that cell size and flashing blue light should be central in this. Full article

(This article belongs to the Special Issue Interaction between Microalgae and Light: Biotechnological Insights)

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Open AccessArticle

Metabolites Potentially Determine the High Antioxidant Properties of Limosilactobacillus fermentum U-21

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BioTech 2023, 12(2), 39; https://doi.org/10.3390/biotech12020039 - 17 May 2023

Abstract

Many kinds of Lactobacillus are common occupants of humans’ digestive tract that support the preservation of a balanced microbial environment that benefits host health. In this study, the unique lactic acid bacterium strain Limosilactobacillus fermentum U-21, which was isolated from the feces of [...] Read more.

Many kinds of Lactobacillus are common occupants of humans’ digestive tract that support the preservation of a balanced microbial environment that benefits host health. In this study, the unique lactic acid bacterium strain Limosilactobacillus fermentum U-21, which was isolated from the feces of a healthy human, was examined for its metabolite profile in order to compare it to that of the strain L. fermentum 279, which does not have antioxidant (AO) capabilities. By using GC × GC−MS, the metabolite fingerprint of each strain was identified, and the data were then subjected to multivariate bioinformatics analysis. The L. fermentum U-21 strain has previously been shown to possess distinctive antioxidant properties in in vivo and in vitro studies, positioning it as a drug candidate for the treatment of Parkinsonism. The production of multiple distinct compounds is shown by the metabolite analysis, demonstrating the unique characteristics of the L. fermentum U-21 strain. According to reports, some of the L. fermentum U-21 metabolites found in this study have health-promoting properties. The GC × GC−MS-based metabolomic tests defined strain L. fermentum U-21 as a potential postbiotic with significant antioxidant potential. Full article

(This article belongs to the Special Issue Natural Antioxidants: Determination in Food and Nutraceuticals and Implications on Human Health)

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Graphical abstract

Open AccessReview

Recent Developments in Protein Lactylation in PTSD and CVD: Novel Strategies and Targets

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BioTech 2023, 12(2), 38; https://doi.org/10.3390/biotech12020038 - 15 May 2023

Abstract

In 1938, Corneille Heymans received the Nobel Prize in physiology for discovering that oxygen sensing in the aortic arch and carotid sinus was mediated by the nervous system. The genetics of this process remained unclear until 1991 when Gregg Semenza while studying erythropoietin, [...] Read more.

In 1938, Corneille Heymans received the Nobel Prize in physiology for discovering that oxygen sensing in the aortic arch and carotid sinus was mediated by the nervous system. The genetics of this process remained unclear until 1991 when Gregg Semenza while studying erythropoietin, came upon hypoxia-inducible factor 1, for which he obtained the Nobel Prize in 2019. The same year, Yingming Zhao found protein lactylation, a posttranslational modification that can alter the function of hypoxia-inducible factor 1, the master regulator of cellular senescence, a pathology implicated in both post-traumatic stress disorder (PTSD) and cardiovascular disease (CVD). The genetic correlation between PTSD and CVD has been demonstrated by many studies, of which the most recent one utilizes large-scale genetics to estimate the risk factors for these conditions. This study focuses on the role of hypertension and dysfunctional interleukin 7 in PTSD and CVD, the former caused by stress-induced sympathetic arousal and elevated angiotensin II, while the latter links stress to premature endothelial cell senescence and early vascular aging. This review summarizes the recent developments and highlights several novel PTSD and CVD pharmacological targets. They include lactylation of histone and non-histone proteins, along with the related biomolecular actors such as hypoxia-inducible factor 1α, erythropoietin, acid-sensing ion channels, basigin, and Interleukin 7, as well as strategies to delay premature cellular senescence by telomere lengthening and resetting the epigenetic clock. Full article

(This article belongs to the Section Medical Biotechnology)

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Open AccessReview

Recent Genome-Editing Approaches toward Post-Implanted Fetuses in Mice

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BioTech 2023, 12(2), 37; https://doi.org/10.3390/biotech12020037 - 11 May 2023

Abstract

Genome editing, as exemplified by the CRISPR/Cas9 system, has recently been employed to effectively generate genetically modified animals and cells for the purpose of gene function analysis and disease model creation. There are at least four ways to induce genome editing in individuals: [...] Read more.

Genome editing, as exemplified by the CRISPR/Cas9 system, has recently been employed to effectively generate genetically modified animals and cells for the purpose of gene function analysis and disease model creation. There are at least four ways to induce genome editing in individuals: the first is to perform genome editing at the early preimplantation stage, such as fertilized eggs (zygotes), for the creation of whole genetically modified animals; the second is at post-implanted stages, as exemplified by the mid-gestational stages (E9 to E15), for targeting specific cell populations through in utero injection of viral vectors carrying genome-editing components or that of nonviral vectors carrying genome-editing components and subsequent in utero electroporation; the third is at the mid-gestational stages, as exemplified by tail-vein injection of genome-editing components into the pregnant females through which the genome-editing components can be transmitted to fetal cells via a placenta-blood barrier; and the last is at the newborn or adult stage, as exemplified by facial or tail-vein injection of genome-editing components. Here, we focus on the second and third approaches and will review the latest techniques for various methods concerning gene editing in developing fetuses. Full article

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Open AccessReview

Bioremediation of Organic Pollutants in Soil–Water System: A Review

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Pankaj Kumar Gupta

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Manvi Gandhi

BioTech 2023, 12(2), 36; https://doi.org/10.3390/biotech12020036 - 11 May 2023

Abstract

Soil–water pollution is of serious concern worldwide. There is a public outcry against the continually rising problems of pollution to ensure the safest and healthiest subsurface environment for living beings. A variety of organic pollutants causes serious soil–water pollution, toxicity and, therefore, the [...] Read more.

Soil–water pollution is of serious concern worldwide. There is a public outcry against the continually rising problems of pollution to ensure the safest and healthiest subsurface environment for living beings. A variety of organic pollutants causes serious soil–water pollution, toxicity and, therefore, the removal of a wide range of organic pollutants from contaminated matrix through the biological process rather than physico-chemical methods is an urgent need to protect the environment and public health. Being an ecofriendly technology, bioremediation can solve the problems of soil–water pollution due to hydrocarbons as it is a low-cost and self-driven process that utilises microorganisms and plants or their enzymes to degrade and detoxify pollutants and thus, promote sustainable development. This paper describes the updates on the bioremediation and phytoremediation techniques which have been recently developed and demonstrated at the plot-scale. Further, this paper provides details of wetland-based treatment of BTEX contaminated soils and water. The knowledge acquired in our study contributes extensively towards understanding the impact of dynamic subsurface conditions on engineered bioremediation techniques. Full article

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Open AccessCommunication

Development of the Follow-Up Human 3D Oral Cancer Model in Cancer Treatment

by

Kazuyo Igawa

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Kenji Izumi

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Yoshinori Sakurai

BioTech 2023, 12(2), 35; https://doi.org/10.3390/biotech12020035 - 11 May 2023

Abstract

As function preservation cancer therapy, targeted radiation therapies have been developed for the quality of life of cancer patients. However, preclinical animal studies evaluating the safety and efficacy of targeted radiation therapy is challenging from the viewpoints of animal welfare and animal protection, [...] Read more.

As function preservation cancer therapy, targeted radiation therapies have been developed for the quality of life of cancer patients. However, preclinical animal studies evaluating the safety and efficacy of targeted radiation therapy is challenging from the viewpoints of animal welfare and animal protection, as well as the management of animal in radiation-controlled areas under the regulations. We fabricated the human 3D oral cancer model that considers the time axis of the follow up in cancer treatment. Therefore, in this study, the 3D model with human oral cancer cells and normal oral fibroblasts was treated based on clinical protocol. After cancer treatment, the histological findings of the 3D oral cancer model indicated the clinical correlation between tumor response and surrounding normal tissue. This 3D model has potential as a tool for preclinical studies alternative to animal studies. Full article

(This article belongs to the Special Issue Feature Papers in Medical Biotechnology)

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Open AccessReview

Nanotherapeutic Approaches to Treat COVID-19-Induced Pulmonary Fibrosis

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BioTech 2023, 12(2), 34; https://doi.org/10.3390/biotech12020034 - 05 May 2023

Abstract

There have been significant collaborative efforts over the past three years to develop therapies against COVID-19. During this journey, there has also been a lot of focus on understanding at-risk groups of patients who either have pre-existing conditions or have developed concomitant health [...] Read more.

There have been significant collaborative efforts over the past three years to develop therapies against COVID-19. During this journey, there has also been a lot of focus on understanding at-risk groups of patients who either have pre-existing conditions or have developed concomitant health conditions due to the impact of COVID-19 on the immune system. There was a high incidence of COVID-19-induced pulmonary fibrosis (PF) observed in patients. PF can cause significant morbidity and long-term disability and lead to death in the long run. Additionally, being a progressive disease, PF can also impact the patient for a long time after COVID infection and affect the overall quality of life. Although current therapies are being used as the mainstay for treating PF, there is no therapy specifically for COVID-induced PF. As observed in the treatment of other diseases, nanomedicine can show significant promise in overcoming the limitations of current anti-PF therapies. In this review, we summarize the efforts reported by various groups to develop nanomedicine therapeutics to treat COVID-induced PF. These therapies can potentially offer benefits in terms of targeted drug delivery to lungs, reduced toxicity, and ease of administration. Some of the nanotherapeutic approaches may provide benefits in terms of reduced immunogenicity owing to the tailored biological composition of the carrier as per the patient needs. In this review, we discuss cellular membrane-based nanodecoys, extracellular vesicles such as exosomes, and other nanoparticle-based approaches for potential treatment of COVID-induced PF. Full article

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Open AccessArticle

A Bacterial Myeloperoxidase with Antimicrobial Properties

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Claire Stines-Chaumeil

BioTech 2023, 12(2), 33; https://doi.org/10.3390/biotech12020033 - 05 May 2023

Abstract

The four mammalian peroxidases (myeloperoxidase, eosinophilperoxidase, lactoperoxidase, and thyroid peroxidase) are widely studied in the literature. They catalyze the formation of antimicrobial compounds and participate in innate immunity. Owing to their properties, they are used in many biomedical, biotechnological, and agro-food applications. We [...] Read more.

The four mammalian peroxidases (myeloperoxidase, eosinophilperoxidase, lactoperoxidase, and thyroid peroxidase) are widely studied in the literature. They catalyze the formation of antimicrobial compounds and participate in innate immunity. Owing to their properties, they are used in many biomedical, biotechnological, and agro-food applications. We decided to look for an enzyme that is easiest to produce and much more stable at 37 °C than mammalian peroxidases. To address this question, a peroxidase from Rhodopirellula baltica, identified by bioinformatics tools, was fully characterized in this study. In particular, a production and purification protocol including the study of heme reconstitution was developed. Several activity tests were also performed to validate the hypothesis that this peroxidase is a new homolog of mammalian myeloperoxidase. It has the same substrate specificities as the human one and accepts I⁻, SCN⁻, Br⁻, and Cl⁻ as (pseudo-) halides. It also exhibits other auxiliary activities such as catalase and classical peroxidase activities, and it is very stable at 37 °C. Finally, this bacterial myeloperoxidase can kill the Escherichia coli strain ATCC25922, which is usually used to perform antibiograms. Full article

(This article belongs to the Section Medical Biotechnology)

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Open AccessReview

Post-Harvest Prevention of Fusariotoxin Contamination of Agricultural Products by Irreversible Microbial Biotransformation: Current Status and Prospects

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Natalia V. Statsyuk

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Sophya B. Popletaeva

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Larisa A. Shcherbakova

BioTech 2023, 12(2), 32; https://doi.org/10.3390/biotech12020032 - 05 May 2023

Abstract

Biological degradation of mycotoxins is a promising environmentally-friendly alternative to chemical and physical detoxification methods. To date, a lot of microorganisms able to degrade them have been described; however, the number of studies determining degradation mechanisms and irreversibility of transformation, identifying resulting metabolites, [...] Read more.

Biological degradation of mycotoxins is a promising environmentally-friendly alternative to chemical and physical detoxification methods. To date, a lot of microorganisms able to degrade them have been described; however, the number of studies determining degradation mechanisms and irreversibility of transformation, identifying resulting metabolites, and evaluating in vivo efficiency and safety of such biodegradation is significantly lower. At the same time, these data are crucial for the evaluation of the potential of the practical application of such microorganisms as mycotoxin-decontaminating agents or sources of mycotoxin-degrading enzymes. To date, there are no published reviews, which would be focused only on mycotoxin-degrading microorganisms with the proved irreversible transformation of these compounds into less toxic compounds. In this review, the existing information about microorganisms able to efficiently transform the three most common fusariotoxins (zearalenone, deoxinyvalenol, and fumonisin B1) is presented with allowance for the data on the corresponding irreversible transformation pathways, produced metabolites, and/or toxicity reduction. The recent data on the enzymes responsible for the irreversible transformation of these fusariotoxins are also presented, and the promising future trends in the studies in this area are discussed. Full article

(This article belongs to the Section Agricultural and Food Biotechnology)

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Open AccessArticle

Efficient Purification of Polyhistidine-Tagged Recombinant Proteins Using Functionalized Corundum Particles

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BioTech 2023, 12(2), 31; https://doi.org/10.3390/biotech12020031 - 03 May 2023

Abstract

Immobilized metal affinity chromatography (IMAC) is a popular and valuable method for the affinity purification of polyhistidine-tagged recombinant proteins. However, it often shows practical limitations, which might require cumbersome optimizations, additional polishing, and enrichment steps. Here, we present functionalized corundum particles for the [...] Read more.

Immobilized metal affinity chromatography (IMAC) is a popular and valuable method for the affinity purification of polyhistidine-tagged recombinant proteins. However, it often shows practical limitations, which might require cumbersome optimizations, additional polishing, and enrichment steps. Here, we present functionalized corundum particles for the efficient, economical, and fast purification of recombinant proteins in a column-free format. The corundum surface is first derivatized with the amino silane APTES, then EDTA dianhydride, and subsequently loaded with nickel ions. The Kaiser test, well known in solid-phase peptide synthesis, was used to monitor amino silanization and the reaction with EDTA dianhydride. In addition, ICP-MS was performed to quantify the metal-binding capacity. His-tagged protein A/G (PAG), mixed with bovine serum albumin (BSA), was used as a test system. The PAG binding capacity was around 3 mg protein per gram of corundum or 2.4 mg per 1 mL of corundum suspension. Cytoplasm obtained from different E. coli strains was examined as examples of a complex matrix. The imidazole concentration was varied in the loading and washing buffers. As expected, higher imidazole concentrations during loading are usually beneficial when higher purities are desired. Even when higher sample volumes, such as one liter, were used, recombinant protein down to a concentration of 1 µg/mL could be isolated selectively. Comparing the corundum material with standard Ni–NTA agarose beads indicated higher purities of proteins isolated using corundum. His6-MBP-mSA2, a fusion protein consisting of monomeric streptavidin and maltose-binding protein in the cytoplasm of E. coli, was purified successfully. To show that this method is also suitable for mammalian cell culture supernatants, purification of the SARS-CoV-2-S-RBD-His8 expressed in human Expi293F cells was performed. The material cost of the nickel-loaded corundum material (without regeneration) is estimated to be less than 30 cents for 1 g of functionalized support or 10 cents per milligram of isolated protein. Another advantage of the novel system is the corundum particles’ extremely high physical and chemical stability. The new material should be applicable in small laboratories and large-scale industrial applications. In summary, we could show that this new material is an efficient, robust, and cost-effective purification platform for the purification of His-tagged proteins, even in challenging, complex matrices and large sample volumes of low product concentration. Full article

(This article belongs to the Section Industrial Biotechnology)

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Open AccessArticle

Impact of Biomass Drying Process on the Extraction Efficiency of C-Phycoerythrin

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Ariadna H. Vergel-Suarez

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Janet B. García-Martínez

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Germán L. López-Barrera

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Andrés F. Barajas-Solano

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Antonio Zuorro

BioTech 2023, 12(2), 30; https://doi.org/10.3390/biotech12020030 - 23 Apr 2023

Abstract

Drying the biomass produced is one of the critical steps to avoid cell degradation; however, its high energy cost is a significant technological barrier to improving this type of bioprocess’s technical and economic feasibility. This work explores the impact of the biomass drying [...] Read more.

Drying the biomass produced is one of the critical steps to avoid cell degradation; however, its high energy cost is a significant technological barrier to improving this type of bioprocess’s technical and economic feasibility. This work explores the impact of the biomass drying method of a strain of Potamosiphon sp. on the extraction efficiency of a phycoerythrin-rich protein extract. To achieve the above, the effect of time (12–24 h), temperature (40–70 °C), and drying method (convection oven and dehydrator) were determined using an I-best design with a response surface. According to the statistical results, the factors that most influence the extraction and purity of phycoerythrin are temperature and moisture removal by dehydration. The latter demonstrates that gentle drying of the biomass allows removing the most significant amount of moisture from the biomass without affecting the concentration or quality of temperature-sensitive proteins. Full article

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Open AccessReview

Cyanidiales-Based Bioremediation of Heavy Metals

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BioTech 2023, 12(2), 29; https://doi.org/10.3390/biotech12020029 - 18 Apr 2023

Abstract

With growing urbanization and ongoing development activities, the consumption of heavy metals has been increasing globally. Although heavy metals are vital for the survival of living beings, they can become hazardous when they surpass the permissible limit. The effect of heavy metals varies [...] Read more.

With growing urbanization and ongoing development activities, the consumption of heavy metals has been increasing globally. Although heavy metals are vital for the survival of living beings, they can become hazardous when they surpass the permissible limit. The effect of heavy metals varies from normal to acute depending on the individual, so it is necessary to treat the heavy metals before releasing them into the environment. Various conventional treatment technologies have been used based on physical, chemical, and biological methods. However, due to technical and economic constraints and poor sustainability towards the environment, the use of these technologies has been limited. Microalgal-based heavy metal removal has been explored for the past few decades and has been seen as an effective, environment-friendly, and inexpensive method compared to conventional treatment technology. Cyanidiales that belong to red algae have the potential for remediation of heavy metals as they can withstand and tolerate extreme stresses of heat, acid salts, and heavy metals. Cyanidiales are the only photosynthetic organisms that can survive and thrive in acidic mine drainage, where heavy metal contamination is often prevalent. This review focuses on the algal species belonging to three genera of Cyanidiales: Cyanidioschyzon, Cyanidium, and Galdieria. Papers published after 2015 were considered in order to examine these species’ efficiency in heavy metal removal. The result is summarized as maximum removal efficiency at the optimum experimental conditions and based on the parameters affecting the metal ion removal efficiency. This study finds that pH, initial metal concentration, initial algal biomass concentration, algal strains, and growth temperature are the major parameters that affect the heavy metal removal efficiency of Cyanidiales. Full article

Open AccessArticle

Protein Delivery to Insect Epithelial Cells In Vivo: Potential Application to Functional Molecular Analysis of Proteins in Butterfly Wing Development

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Yugo Nakazato

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Joji M. Otaki

BioTech 2023, 12(2), 28; https://doi.org/10.3390/biotech12020028 - 16 Apr 2023

Abstract

Protein delivery to cells in vivo has great potential for the functional analysis of proteins in nonmodel organisms. In this study, using the butterfly wing system, we investigated a method of protein delivery to insect epithelial cells that allows for easy access, treatment, [...] Read more.

Protein delivery to cells in vivo has great potential for the functional analysis of proteins in nonmodel organisms. In this study, using the butterfly wing system, we investigated a method of protein delivery to insect epithelial cells that allows for easy access, treatment, and observation in real time in vivo. Topical and systemic applications (called the sandwich and injection methods, respectively) were tested. In both methods, green/orange fluorescent proteins (GFP/OFP) were naturally incorporated into intracellular vesicles and occasionally into the cytosol from the apical surface without any delivery reagent. However, the antibodies were not delivered by the sandwich method at all, and were delivered only into vesicles by the injection method. A membrane-lytic peptide, L17E, appeared to slightly improve the delivery of GFP/OFP and antibodies. A novel peptide reagent, ProteoCarry, successfully promoted the delivery of both GFP/OFP and antibodies into the cytosol via both the sandwich and injection methods. These protein delivery results will provide opportunities for the functional molecular analysis of proteins in butterfly wing development, and may offer a new way to deliver proteins into target cells in vivo in nonmodel organisms. Full article

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Open AccessArticle

Molecular Characterization of Dehydrin in Azraq Saltbush among Related Atriplex Species

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Anas Musallam

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Saeid Abu-Romman

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Monther T. Sadder

BioTech 2023, 12(2), 27; https://doi.org/10.3390/biotech12020027 - 07 Apr 2023

Abstract

Atriplex spp. (saltbush) is known to survive extremely harsh environmental stresses such as salinity and drought. It mitigates such conditions based on specialized physiological and biochemical characteristics. Dehydrin genes (DHNs) are considered major players in this adaptation. In this study, a [...] Read more.

Atriplex spp. (saltbush) is known to survive extremely harsh environmental stresses such as salinity and drought. It mitigates such conditions based on specialized physiological and biochemical characteristics. Dehydrin genes (DHNs) are considered major players in this adaptation. In this study, a novel DHN gene from Azrak (Jordan) saltbush was characterized along with other Atriplex species from diverse habitats. Intronless DHN-expressed sequence tags (495–761 bp) were successfully cloned and sequenced. Saltbush dehydrins contain one S-segment followed by three K-segments: an arrangement called SK3-type. Two substantial insertions were detected including three copies of the K2-segemnet in A. canescens. New motif variants other than the six-serine standard were evident in the S-segment. AhaDHN1 (A. halimus) has a cysteine residue (SSCSSS), while AgaDHN1 (A. gardneri var. utahensis) has an isoleucine residue (SISSSS). In contrast to the conserved K1-segment, both the K2- and K3-segment showed several substitutions, particularly in AnuDHN1 (A. nummularia). In addition, a parsimony phylogenetic tree based on homologs from related genera was constructed. The phylogenetic tree resolved DHNs for all of the investigated Atriplex species in a superclade with an 85% bootstrap value. Nonetheless, the DHN isolated from Azraq saltbush was uniquely subclustred with a related genera Halimione portulacoides. The characterized DHNs revealed tremendous diversification among the Atriplex species, which opens a new venue for their functional analysis. Full article

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Open AccessReview

Biomolecular Liquid–Liquid Phase Separation for Biotechnology

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BioTech 2023, 12(2), 26; https://doi.org/10.3390/biotech12020026 - 01 Apr 2023

Abstract

The liquid–liquid phase separation (LLPS) of biomolecules induces condensed assemblies called liquid droplets or membrane-less organelles. In contrast to organelles with lipid membrane barriers, the liquid droplets induced by LLPS do not have distinct barriers (lipid bilayer). Biomolecular LLPS in cells has attracted [...] Read more.

The liquid–liquid phase separation (LLPS) of biomolecules induces condensed assemblies called liquid droplets or membrane-less organelles. In contrast to organelles with lipid membrane barriers, the liquid droplets induced by LLPS do not have distinct barriers (lipid bilayer). Biomolecular LLPS in cells has attracted considerable attention in broad research fields from cellular biology to soft matter physics. The physical and chemical properties of LLPS exert a variety of functions in living cells: activating and deactivating biomolecules involving enzymes; controlling the localization, condensation, and concentration of biomolecules; the filtration and purification of biomolecules; and sensing environmental factors for fast, adaptive, and reversible responses. The versatility of LLPS plays an essential role in various biological processes, such as controlling the central dogma and the onset mechanism of pathological diseases. Moreover, biomolecular LLPS could be critical for developing new biotechnologies such as the condensation, purification, and activation of a series of biomolecules. In this review article, we introduce some fundamental aspects and recent progress of biomolecular LLPS in living cells and test tubes. Then, we discuss applications of biomolecular LLPS toward biotechnologies. Full article

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