Promising Biotechnological Tool Research Articles

Nanoparticle elicitation: A promising strategy to modulate the production of bioactive compounds in hairy roots

Hairy root culture is one of the promising biotechnological tools to obtain the stable and sustainable production of specialized metabolites from plants under controlled environment conditions. Various strategies have been adopted to enhance the accumulation of bioactive compounds in hairy roots yet their utilization at the commercial scale is restricted to only a few products. Recently, nanotechnology has been emerged as an active technique that has revolutionized the many sectors in an advantageous way. Elicitation using nanoparticles has been recognized as an effective strategy for enhancing the bioactive compounds of interest in plants. Nanoparticles elicit the activity of defense-related compounds through activation of the specific transcription factors involved in specialized metabolites production. This review discusses the recent progress in using nanoparticles to enhance specialized metabolite biosynthesis using hairy root culture system and the significant achievements in this area of research. Biotic and abiotic elicitors to improve the production of bioactive compounds in hairy roots, different types of nanoparticles as eliciting agents, their properties as dependent on shape, most widely used nanoparticles in plant hairy root systems are described in detail. Further challenges involved in application of nanoparticles, their toxicity in plant cells and risks associated to human health are also envisaged. No doubt, nanoparticle elicitation is a remarkable approach to obtain phytochemicals from hairy roots to be utilized in various sectors including food, medicines, cosmetics or agriculture but it is quite essential to understand the inter-relationships between the nanoparticles and the plant systems in terms of specifics such as type, dosage and time of exposure as well as other important parameters.

Bothrops snake venom L-amino acid oxidases impair biofilm formation of clinically relevant bacteria

The present work addressed the abilities of two L-amino acid oxidases isolated from Bothrops moojeni (BmooLAAO-I) and Bothrops jararacussu (BjussuLAAO-II) snake venoms to control the growth and prevent the biofilm formation of clinically relevant bacterial pathogens. Upon S. aureus (ATCC BAA44) and S. aureus (clinical isolates), BmooLAAO-I (MIC = 0.12 and 0.24 μg/mL, respectively) and BjussuLAAO-II (MIC = 0.15 μg/mL) showed a potent bacteriostatic effect. Against E. coli (ATCC BAA198) and E. coli (clinical isolates), BmooLAAO-I (MIC = 15.6 and 62.5 μg/mL, respectively) and BjussuLAAO-II (MIC = 4.88 and 9.76 μg/mL, respectively) presented a lower extent effect. Also, BmooLAAO-I (MICB50 = 0.195 μg/mL) and BjussuLAAO-II (MICB50 = 0.39 μg/mL) inhibited the biofilm formation of S. aureus (clinical isolates) in 88% and 89%, respectively, and in 89% and 53% of E. coli (clinical isolates). Moreover, scanning electron microscopy confirmed that the toxins affected bacterial morphology by increasing the roughness of the cell surface and inhibited the biofilm formation. Furthermore, analysis of the tridimensional structures of the toxins showed that the surface-charge distribution presents a remarkable positive region close to the glycosylation motif, which is more pronounced in BmooLAAO-I than BjussuLAAO-II. This region may assist the interaction with bacterial and biofilm surfaces. Collectively, our findings propose that venom-derived antibiofilm agents are promising biotechnological tools which could provide novel strategies for biofilm-associated infections.

The light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibits cortical reaction in mouse oocytes.

Introduction: Cortical reaction is a secretory process that occurs after a spermatozoon fuses with the oocyte, avoiding the fusion of additional sperm. During this exocytic event, the cortical granule membrane fuses with the oocyte plasma membrane. We have identified several molecular components involved in this process and confirmed that SNARE proteins regulate membrane fusion during cortical reaction in mouse oocytes. In those studies, we microinjected different nonpermeable reagents to demonstrate the participation of a specific protein in the cortical reaction. However, the microinjection technique has several limitations. In this work, we aimed to assess the potential of cell-penetrating peptides (CPP) as biotechnological tools for delivering molecules into oocytes, and to evaluate the functionality of the permeable tetanus toxin (bound to CPP sequence) during cortical reaction. Methods: Arginine-rich cell-penetrating peptides have demonstrated the optimal internalization of small molecules in mammalian cells. Two arginine-rich CPP were used in the present study. One, labeled with 5-carboxyfluorescein, to characterize the factors that can modulate its internalization, and the other, the permeable light chain of tetanus toxin, that cleaves the SNAREs VAMP1 and VAMP3 expressed in mouse oocytes. Results: Results showed that fluorescent CPP was internalized into the oocyte cytoplasm and that internalization was dependent on the concentration, time, temperature, and maturation stage of the oocyte. Using our functional assay to study cortical reaction, the light chain of tetanus toxin bound to arginine-rich cell-penetrating peptide inhibited cortical granules exocytosis. Discussion: Results obtained from the use of permeable peptides demonstrate that this CPP is a promising biotechnological tool to study functional macromolecules in mouse oocytes.

RETRACTED: Biochemical characterization of a cold-adapted κ-carrageenase from Colwellia echini: A biotechnological tool for efficient preparation of carrageenan oligosaccharides

κ-Carrageenase (EC 3.2.1.83) is a glycoside hydrolase that efficiently breaks down κ-carrageenan into κ-carrageenan oligosaccharides (κ-COS). In this study, a novel GH16 family κ-carrageenase called CeCgkB was cloned from the genome of Colwellia echini. Escherichia coli BL21 (DE3) was used to express CeCgkB. CeCgkB has a complete length of 987 bp, encoding 329 amino acid residues, and a molecular weight of 37.44 kDa. Biochemical characterization indicated that CeCgkB exhibits cold-adapted properties and was able to reach a maximum activity of 1530 U/mg at pH 8.0 and 35 °C. Kinetic parameters showed that CeCgkB has a low Km value (2 mg/mL), indicating high substrate affinity. As an endo-type κ-carrageenase, CeCgkB can degrade κ-carrageenan into products of varying DP, with a preference for DP4, DP6, and DP10. ESI-MS analysis revealed that the major products of CeCgkB consist of κ-carratetraose, κ-carrahexaose, and κ-carradecaose. As a result of CeCgkB's high activity, cold-adapted, and principal products, it is a promising biotechnological tool for the efficient preparation of κ-carrageenan oligosaccharides.

Cyanoremediation and phyconanotechnology: cyanobacteria for metal biosorption toward a circular economy.

Cyanobacteria are widespread phototrophic microorganisms that represent a promising biotechnological tool to satisfy current sustainability and circularity requirements. They are potential bio-factories of a wide range of compounds that can be exploited in several fields including bioremediation and nanotechnology sectors. This article aims to illustrate the most recent trends in the use of cyanobacteria for the bioremoval (i.e., cyanoremediation) of heavy metals and metal recovery and reuse. Heavy metal biosorption by cyanobacteria can be combined with the consecutive valorization of the obtained metal-organic materials to get added-value compounds, including metal nanoparticles, opening the field of phyconanotechnology. It is thus possible that the use of combined approaches could increase the environmental and economic feasibility of cyanobacteria-based processes, promoting the transition toward a circular economy.

Structure-guided engineering of a receptor-agonist pair for inducible activation of the ABA adaptive response to drought

Strategies to activate abscisic acid (ABA) receptors and boost ABA signaling by small molecules that act as ABA receptor agonists are promising biotechnological tools to enhance plant drought tolerance. Protein structures of crop ABA receptors might require modifications to improve recognition of chemical ligands, which in turn can be optimized by structural information. Through structure-based targeted design, we have combined chemical and genetic approaches to generate an ABA receptor agonist molecule (iSB09) and engineer a CsPYL1 ABA receptor, named CsPYL15m, which efficiently binds iSB09. This optimized receptor-agonist pair leads to activation of ABA signaling and marked drought tolerance. No constitutive activation of ABA signaling and hence growth penalty was observed in transformed Arabidopsis thaliana plants. Therefore, conditional and efficient activation of ABA signaling was achieved through a chemical-genetic orthogonal approach based on iterative cycles of ligand and receptor optimization driven by the structure of ternary receptor-ligand-phosphatase complexes.

Bacterial volatile organic compounds (VOCs) promote growth and induce metabolic changes in rice.

Plant growth-promoting bacteria (PGPB) represent an eco-friendly alternative to reduce the use of chemical products while increasing the productivity of economically important crops. The emission of small gaseous signaling molecules from PGPB named volatile organic compounds (VOCs) has emerged as a promising biotechnological tool to promote biomass accumulation in model plants (especially Arabidopsis thaliana) and a few crops, such as tomato, lettuce, and cucumber. Rice (Oryza sativa) is the most essential food crop for more than half of the world's population. However, the use of VOCs to improve this crop performance has not yet been investigated. Here, we evaluated the composition and effects of bacterial VOCs on the growth and metabolism of rice. First, we selected bacterial isolates (IAT P4F9 and E.1b) that increased rice dry shoot biomass by up to 83% in co-cultivation assays performed with different durations of time (7 and 12 days). Metabolic profiles of the plants co-cultivated with these isolates and controls (without bacteria and non-promoter bacteria-1003-S-C1) were investigated via 1H nuclear magnetic resonance. The analysis identified metabolites (e.g., amino acids, sugars, and others) with differential abundance between treatments that might play a role in metabolic pathways, such as protein synthesis, signaling, photosynthesis, energy metabolism, and nitrogen assimilation, involved in rice growth promotion. Interestingly, VOCs from IAT P4F9 displayed a more consistent promotion activity and were also able to increase rice dry shoot biomass in vivo. Molecular identification by sequencing the 16S rRNA gene of the isolates IAT P4F9 and E.1b showed a higher identity with Serratia and Achromobacter species, respectively. Lastly, volatilomes of these and two other non-promoter bacteria (1003-S-C1 and Escherichia coli DH5α) were evaluated through headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. Compounds belonging to different chemical classes, such as benzenoids, ketones, alcohols, sulfide, alkanes, and pyrazines, were identified. One of these VOCs, nonan-2-one, was validated in vitro as a bioactive compound capable of promoting rice growth. Although further analyses are necessary to properly elucidate the molecular mechanisms, our results suggest that these two bacterial isolates are potential candidates as sources for bioproducts, contributing to a more sustainable agriculture.

Somatic embryogenesis in Abies nebrodensis, an endangered Sicilian fir

Somatic embryogenesis, as a promising biotechnological tool for many conifer trees, has never been applied for the Abies nebrodensis species. Although all the encouraging results previously obtained by the EU LIFE (European LIFE program) funded projects in over ten years, the critically endangered Sicilian fir remains alarmingly close to extinction. In this study, we reported the first protocol of somatic embryogenesis obtained from mature zygotic embryos of the Abies nebrodensis. Seeds from Abies adult trees with specific identification numbers (IN) were collected and full seeds were identified by X-ray. Different experiments were carried out for callus initiation, from both zygotic immature and mature embryos, testing different culture media. The immature embryos did not give embryogenic tissue (ET). Embryogenic callus (EC) was successfully induced from mature embryos with variable frequencies (0–40%). Schenk and Hilderbrandt (SH) was the most suitable initiation medium where the obtained callus initiation rate reached up to 40% for IN7 (first experiment). 6-benzylaminopurine (BAP) showed to be essential to induce EC (second experiment). IN8 presented the highest callus initiation rate (40%) among all tested donor trees, whereas IN13 recorded the lowest rate with 4% (third experiment). ET maturation from each singular embryo of IN7, IN8, IN10 and IN21 was successfully achieved in SH medium containing 37,83 µM abscisic acid (ABA), 8% of polyethylene glycol (PEG-4000) and 4% maltose. The encapsulation technology was assessed on the obtained ET and its proliferation was observed after encapsulation.

Fungal protein (Mycoprotein) what to know about

Microbes in general and fungi especially are promising biotechnological tools that are used for green synthesis of numerous products. Fungi in particular are potent producers of many important compounds used in different applications. Hence, this review aims to explore fungi and their potential as a source of various bioactive compounds like mycoprotein.

Fungi between Immunostimulatory and Immunosuppressive

Microbes in general and fungi especially are promising biotechnological tools that are used for green synthesis of numerous products. Fungi in particular are potent producers of many industrially important compounds. β-Glucans are a group of biologically-active fibres or polysaccharides from natural sources with proven medical significance. β-Glucans are known to have antitumor, anti-inflammatory, anti-obesity, anti-allergic, anti-osteoporotic, and immunomodulating activities. On the other hand, cyclosporin is a cyclic undecapeptide with a variety of biological activities including immunosuppressive, antiinflammatory, antifungal, and antiparasitic properties. It is an extremely powerful immunosuppressant and is approved for the use in organ transplantation to prevent graft rejection in kidney, liver, heart, lung, and combined heart–lung transplants. Hence, this review aims to focus on β-glucan as immunostimulatory and Cyclosporine A as Immunosuppressants

Peptide functionalized Dynabeads for the magnetic carrier separation of rare-earth fluorescent lamp phosphors

Novel separation processes for end of life fluorescent lamp phosphors could greatly contribute towards a more sustainable rare-earth element market. Furthermore, surface-binding peptides bound to magnetic carriers are a promising biotechnological tool for selective particle separation processes. In this work, we investigate the magnetic carrier separation of the three most common rare-earth fluorescent lamp phosphors, facilitated by Dynabeads functionalized with previously identified selectively surface-binding peptides. We present an active ester-mediated coupling to chemically immobilize the peptides on amine and carboxylic acid coated beads. We report on the impact of the peptide functionalization on the surface properties of the beads, based on zeta potential investigations of variously functionalized beads and a Raman spectroscopic structural study of the investigated peptides. Fluorometrically, we show that the phosphor removal strongly depends on the medium and the surface coating of the beads. Furthermore, the Raman spectroscopic evidence of various simultaneously present disulfide bond conformations indicates an equilibrium of multiple peptide conformations and/or the presence of intermolecular disulfide bonds. Moreover, we found that carboxylic acid coated Dynabeads have a high affinity for the red phosphor Y2O3:Eu3+and based on the determined isoelectric points we hypothesize that this is driven by electrostatic surface interactions. This work can contribute towards novel rare-earth phosphor separation processes and towards a better understanding of magnetic carrier separation processes facilitated by surface-binding peptides.

Abstract P2055: Secondary Structure And Antisense Oligonucleotides Regulate Cardiac Messenger RNA Translation And Modulates Cardiomyocyte Hypertrophy

Translation of upstream open reading frames (uORFs) typically abrogates translation for the main ORF (mORF) and regulates protein expression. uORFs are present in nearly 50% of messenger RNA (mRNA) transcripts, but their impact and regulation on biological processes are understudied. Mining of ribosome profiling data from next-generation sequencing analysis of human and mouse hearts reveals that most translated uORFs reside within mRNAs of transcription factors, which play essential roles in cardiomyocyte (CM) development and growth. We show that the uORF start codon synergizes with an immediate downstream double-stranded RNA (dsRNA) element to activate uORF and inhibit mORF translation. Biochemical and genetic evidence support that this mechanism is utilized by mRNAs encoding multiple transcription factors such as GATA4 (GATA binding protein 4), encoding a master transcription regulator of CM differentiation and pathological hypertrophy. Intriguingly, a trans-acting regulatory factor, DEAD-box RNA helicase DDX3X, is involved in unwinding the dsRNA structure and promoting mORF translation. We develop two types of antisense oligonucleotides (ASOs) that mimic the unwinding or stabilizing of the dsRNA secondary structure, thereby suppressing or enhancing the translation of this uORF, respectively. Genetic or chemical inactivation of this GATA4 uORF using CRISPR-Cas9 or ASOs in human embryonic stem cells lead to enhanced differentiation into CMs and cellular hypertrophy. At the organismal level, short-term treatment of isoproterenol and surgery-induced cardiac hypertrophy mouse models with uORF-enhancing ASOs reduces GATA4 mRNA translation and antagonizes cardiac hypertrophy and remodeling. As a summary, this uORF-dsRNA element is a new tunable regulator of cardiac transcription factor mRNA translation and can be targeted to alter cellular and organismal phenotypes. Moreover, mechanism-based translation-manipulating ASOs can serve as promising biotechnological tools to regulate gene expression in vitro and in vivo and can be adapted to create novel RNA-based therapeutics.

EDITING GENOMES VIA CRISPR/CAS9 AND APPLICATIONS IN MAIZE IMPROVEMENT

The CRISPR/Cas9 system is a gene editing method capable of accurately locating and altering a specific sequence of a genome. Currently, this technique stands out as a promising biotechnological tool for the improvement of different crops, overcoming the limitations of conventional breeding methods and accelerating the development of plants’ agronomic characteristics. The basic principle of genome editing via CRISPR is to cause a double-stranded DNA break at a predetermined location by a sgRNA (single guide RNA) and through internal repair mechanisms cause mutations and, consequently, genetic variability. In prokaryotic organisms, it is an immune system that naturally protects against viral infection and has been modified to create different applications in higher organisms in the field of medicine and agriculture. In agriculture, it has been used in a large number of mono and dicotyledonous plants, including those of great commercial value such as soybean, cotton and maize. In maize, this technique has been used as a biotechnological tool for increasing production, quality, nutritional value, tolerance to biotic and abiotic stresses. CRISPR-Cas9 technology has enormous potential to contribute to increased food production with great benefit to the environment. This review explores the bases for the use of CRISPR/Cas9 technology to generate genetic variability and its use in the development of maize cultivars with better agronomic characteristics.

Agri-biotechnology of coriander (Coriandrum sativum L.): an inclusive appraisal.

Bountiful expression of bioactivity of phytochemicals obtained from spice crops like coriander gifts them the label of being natural antioxidants. It is well-accepted and time-tested towards contributing to human wellbeing. The accomplishment of coriander production is fundamentally influenced by genetic, agroclimatic, and agronomic factors. Despite the fact that there are very restricted options to manage the first two factors, the third one is apparently imperative to arbitrate as far as the elevated yield and enhanced quality are concerned. On the other hand, an indomitable, object-oriented, controlled agrotechnological and biotechnological intervention can also contribute towards better yield and quality of coriander. There are several accounts of the successful use of such technologies in order to genetically improve the qualitative and quantitative indicators of coriander. However, often these areas are not comprehensively explored and utilized. In that context, the present review highlights the botanical features, origin and distribution, multi-dimensional importance, pre- and post-harvest crop management, phytochemical production, and germplasm conservation, including the in vitro-based regeneration methods along with molecular marker-based biotechnological and omics approaches attempted in coriander until date. In addition, the possibility of the yet-to-be-explored agri-biotechnological methods and their potential for genetic improvement of this crop has also been reviewed in this appraisal. KEY POINTS: • Coriander, used both as an herb and spice, is popular in the pharmaceutical and culinary industries. • The current review provides insight into agrotechnological and biotechnological interventions for better yield and quality. • Provides novel ideas to harness the comprehensive qualitative and quantitative genetic improvement based on the potential use of promising biotechnological tools and techniques.

Fungal Protein (Mycoprotein) What to Know About

Microbes in general and fungi especially are promising biotechnological tools that are used for green synthesis of numerous products. Fungi in particular are potent producers of many important compounds used in different applications. Hence, this review aims to explore fungi and their potential as a source of various bioactive compounds like mycoprotein.

Bioactive Potential of some Fascinating Edible Mushrooms Macrolepiota, Russula, Amanita, Vovariella and Grifola as a Treasure of Multipurpose Therapeutic Natural Product

Mushrooms are macro fungi that exist everywhere around us. They have significant roles in human health-life as source of nutrition and bioactive compounds. Many edible mushrooms have been reported as promising biotechnological tools for production of secondary metabolites of various biological activities. The genus Macrolepiota; Russula; Amanita; Vovariella and Grifola are a group of edible mushrooms that are distributed all over the world. The studies on different species of these edible mushrooms have revealed their nutritional medicinal potentials. This review aims to present the importance of the genera Macrolepiota; Russula; Amanita; Vovariella and Grifola as both food and medicine, and they offer new insights to researchers to develop new drugs and nutraceuticals

Molecular characterization of the Rpv3 locus towards the development of KASP markers for downy mildew resistance in grapevine (Vitis spp.)

Plasmopara viticola is the oomycete that causes downy mildew in grapevines. Varying levels of resistance to P. viticola across grape cultivars allowed quantitative trait loci to be identified. The Rpv3 locus is located at chromosome 18, in a region enriched in TIR-NBS-LRR genes, and the phenotype associated is a high hypersensitive response. In this work, we aimed to identify candidate genes associated with resistance to downy mildew on the Rpv3 locus and to evaluate their transcriptional profiles in a susceptible and a resistant grapevine cultivar after challenging with P. viticola. Candidate genes were identified by in silico functional enrichment tests. Many predicted genes associated with resistance to diseases were found at the Rpv3 locus. In total, seventeen genes were evaluated by RT-qPCR. Differences in the steady-state expression of these genes were observed between the two cultivars. Four genes were found to be expressed only in Villard Blanc, suggesting their association to the hypersensitivity reaction. Aiming to assist marker assisted-selection for downy mildew resistance, we show the efficient use of a set of SSR markers. Furthermore, from on a set of forty-one Rpv3-located SNPs, whose segregation was tested in the populations studied, the two segregating markers, Rpv3_15 and Rpv3_33, were considered efficient for downy mildew resistance identification. This study presents a genomic characterization of the Rpv3 locus, confirms its involvement in resistance against P. viticola infection and presents promising biotechnological tools for the selection of young resistant individuals.

Sphingobacterium prati sp. nov., isolated from agricultural soil and involved in lignocellulose deconstruction.

A bacterial strain, arapr2T, was isolated from agricultural soil sampled in Reims, France. Based on its 16S rRNA gene sequence, the strain was affiliated to the family Sphingobacteriaceae and more specifically to the genus Sphingobacterium . The strain had 98.31 % 16S rRNA gene sequence similarity to its closest relative Sphingobacterium canadense CR11T and 98.25 % to Sphingobacterium pakistanensis NCCP-246T. Genome relatedness indexes revealed that the average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values between arapr2T and its closest relative ( S. canadense CR11T) were 92.97 % and 52.00 %, respectively; for S. pakistanensis NCCP-246T, the ANI and dDDH values were 82.46 and 27.6%, respectively. The genomic DNA of strain arapr2T was 6.02 Mbp long, had a DNA G+C content of 40.4 mol% and had 5504 protein-coding genes. The results obtained in this study suggests that strain arapr2T (CIP 111872T=LMG 31848T) represents a new species for which the name Sphingobacterium prati sp. nov. is proposed. Due to the fact that this strain has been isolated using wheat straw as carbon source, this novel bacterial strain represents a promising biotechnological tool for the fractionation of lignocellulosic biomass in the context of biorefinery development.

RNAi of a Putative Grapevine Susceptibility Gene as a Possible Downy Mildew Control Strategy.

Downy mildew, caused by the oomycete Plasmopara viticola, is one of the diseases causing the most severe economic losses to grapevine (Vitis vinifera) production. To date, the application of fungicides is the most efficient method to control the pathogen and the implementation of novel and sustainable disease control methods is a major challenge. RNA interference (RNAi) represents a novel biotechnological tool with a great potential for controlling fungal pathogens. Recently, a candidate susceptibility gene (VviLBDIf7) to downy mildew has been identified in V. vinifera. In this work, the efficacy of RNAi triggered by exogenous double-stranded RNA (dsRNA) in controlling P. viticola infections has been assessed in a highly susceptible grapevine cultivar (Pinot noir) by knocking down VviLBDIf7 gene. The effects of dsRNA treatment on this target gene were assessed by evaluating gene expression, disease severity, and development of vegetative and reproductive structures of P. viticola in the leaf tissues. Furthermore, the effects of dsRNA treatment on off-target (EF1α, GAPDH, PEPC, and PEPCK) and jasmonic acid metabolism (COI1) genes have been evaluated. Exogenous application of dsRNA led to significant reductions both in VviLBDIf7 gene expression, 5 days after the treatment, and in the disease severity when artificial inoculation was carried out 7 days after dsRNA treatments. The pathogen showed clear alterations to both vegetative (hyphae and haustoria) and reproductive structures (sporangiophores) that resulted in stunted growth and reduced sporulation. Treatment with dsRNA showed signatures of systemic activity and no deleterious off-target effects. These results demonstrated the potential of RNAi for silencing susceptibility factors in grapevine as a sustainable strategy for pathogen control, underlying the possibility to adopt this promising biotechnological tool in disease management strategies.

Overproduction of Soyasaponin I by Tipuana speciosa (Benth.) cell culture

Plant cell and tissue cultures are promising biotechnological tools for the production of a myriad of bioactive secondary metabolites on demand. Discoveries of cell cultures capable of producing specific medicinal compounds at a rate similar or superior to that of intact plants have been accelerated in the last few years. Soyasaponins have recently gained more attention due to their biological activities and health-promoting functions. There are many reports relating the bioactive benefits of soyasaponins to their chemical structure mainly Soyasaponin I which possess unique pharmacological actions. Our previous study indicated that Tipuana speciosa (Benth.) leaves extract showed promising antimalarial and nephroprotective activities. This study aimed to isolate Soyasaponin I, a major compound in the leaves extract and the use of cell suspension culture as a biotechnological route for high-yield production of this bioactive molecule. The compound was identified by LC-MS/MS analysis and other NMR techniques. To the best of our knowledge, there is no previous studies have been undertaken for the isolation and the production of Soyasaponin I from T. speciosa leaves using plant tissue culture technology. Our data showed that the callus and cell aggregate of T. speciosa represent an over producer cell lines (220 folds) compared to the whole plant.