3 Biotech

The heterotrimeric guanine-nucleotide-binding proteins (G-proteins) play a crucial role in signal transduction and regulate plant responses against biotic and abiotic stresses. Necrotrophic pathogens trigger Gα subunit and, in contrast, sometimes Gβγ dimers. Beneficial microbes play a vital role in the activation of heterotrimeric G-proteins in plants against biotrophic and necrotrophic pathogens. The subunits of G-protein (α, β, and γ) are activated differentially against different kinds of pathogens which in turn regulates the entry of the pathogen in a plant cell. Defense mediated by G-proteins in plants imparts resistance against several pathogens. Activation of different G-protein subunits depends on the mode of nutrition of the pathogen. The current review discussed the role of the three subunits against various pathogens. It appeared to be specific in the individual host–pathogen system as well as the role of effectors in the induction of G-proteins. We also discussed the G-protein-mediated production of reactive oxygen species (ROS), including H2O2, activation of NADPH oxidases, hypersensitive response (HR), phospholipases, and ion channels in response to microorganisms.

Costus spicatus (spiked spiralflag ginger) is traditionally utilised for its advantages such as antidiabetic, antihyperlipidemic, diuretic, antimicrobial, and anticancer properties. However, there is no scientific evidence on the nephroprotective potential of this plant. Thus, this study tested the nephroprotective effect of the polyphenol-rich extract of Costus spicatus leaves (PCSL) using preclinical models, including the HeK cell line and Wistar albino rats against cisplatin-induced toxicity. It also determined the polyphenolic compounds using high-performance thin-layer chromatography (HPTLC). PCSL showed significant (p < 0.05) nephroprotective potential against cisplatin-induced nephrotoxicity in HeK cells. Moreover, in vivo studies revealed significant (p < 0.05) amelioration in serum biochemical markers and antioxidant enzymes against cisplatin-induced nephrotoxicity. PCSL significantly inhibited the level of inflammatory cytokines such as TNF-α, IL-6, and IL-1β. Moreover, PCSL restored the damage of the kidney tissues and ameliorated interstitial haemorrhage, congestion in capillaries, inflammatory cell infiltration, vacuolated cytoplasm, and tubular epithelial injury with widened Bowman's space. In addition, HPTLC analysis revealed that PCSL comprised polyphenolic compounds such as caffeic acid, quercetin, and ferulic acid. In conclusion, PCSL exerted nephroprotective potential by modulating the expression of inflammation, oxidative stress, and histological architecture of kidney tissues.

In this study, we developed an effective technology for the extraction of sericin from silk cocoons by deep sea water (DSW). We focused on extraction of sericin in the absence of chemical additives to obtain a safe, effective, inexpensive sericin powder. Sericin was extracted using a simple high-temperature process involving heating, condensation with Molus alba, filtering with cotton cloth, cold storage, and lyophilization. The results showed that the yield of sericin (26%) extracted by DSW was approximately 2% higher than that obtained using a chemical buffer (0.2 M Na2CO3, 24%). The marine mineral sericin M. alba (MSM) showed a size distribution of 15–250 kDa, with major peaks at 75–250 kDa with a galactose chain. Additionally, this MSM product had high antioxidant, whitening, and antibiosis effects and could be safely stored for a long time. Thus, our findings supported the use of a DSW extraction method, which was ecofriendly and yielded a proteinous, biodegradable biopolymer, for preparation of sericin.

Saccharomyces cerevisiae KCCM 51299, a potential probiotic yeast overproducing glutathione, has been isolated from among 272 yeast strains from the relatively safe Nuruk. The genome sequence of S. cerevisiae KCCM 51299 was analyzed and a near-complete genome (12 Mb) with 19 contigs was assembled after PacBio single-molecule real-time (SMRT) sequencing. The genome of S. cerevisiae KCCM 51299 was compared to the S. cerevisiae s288c reference genome. Additionally, genes involved in glutathione biosynthesis were identified, and the glutathione biosynthesis pathway was constructed in silico based on these genes. Furthermore, S. cerevisiae KCCM 51299 genes were compared with those in other yeast genomes. Finally, genome-scale in silico flux analysis was carried out, and a metabolic engineering strategy for glutathione biosynthesis was generated. These results provide useful information to further develop eukaryotic probiotics to overproduce glutathione.

The 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR; EC1.1.1.267), an NADPH-dependent reductase, plays a pivotal role in the methylerythritol 4-phosphate pathway (MEP), in the conversion of 1-deoxy-d-xylulose-5-phosphate (DXP) into MEP. Photochemical profiles, as well as pharmaceutical activities of Centella asiatica (L.), one of the most valuable medicinal plants, divulge the presence of secondary metabolites called Centellosides. Despite well-studied pharmaceutical activities, not much is known about the genes responsible for the synthesis of these compounds. In the present study, the full-length DXR gene sequence (JQ965955) of Centella submitted in NCBI was characterized using various bioinformatics tools and tissue specific differential expression studies were also carried out. The full-length CDNA of CaDXR contains an open reading frame (ORF) of 1425 bp which encodes a peptide of 474 amino acids. The molecular weight of this protein was found to be 51.5 kDa with isoelectric point of 6.33. The protein contains three conserved domain, namely NADPH (GSTGSIGT and LAAGSNV), substrate binding (LPADSEHSAI and NKGLEVIEAHY) and Cys-Ser-(Ala/Met/Val/Thr) cleavage-site domains. Phylogenetic studies of CaDXR sequence show close homology with DXR sequence of Angelica sinensis and Daucus carota subsp sativus as they all belong to Apiaceae family. In silico analysis predicted that CaDXR protein contains 21 α-helix and 11 β-sheets and further DXR protein model was validated by Ramachandran plot analysis. The results of molecular dynamics (MD) simulations unveil dynamic stability of the proposed model and docking studies suggest that the NDP cofactor tightly binds in the active site of the protein with a strong network of hydrogen and hydrophobic interactions. The expression studies by semi-RT followed by qRT—PCR suggests that CaDXR is differentially expressed in different tissues (with maximal expression in the node and lowest in the roots). Thus, characterization and structure–function analysis of DXR gene in Centella facilitate us to understand not only the functions of DXR gene but also regulatory mechanisms involved in the MEP pathway in C. asiatica plant at the molecular level.

Watercress (Nasturtium officinale R. Br.) is a cruciferous plant consumed by people worldwide. This vegetable contains numerous health-benefiting compounds; however, gene information and metabolomic profiling of individual parts for this plant species are scarce. In this study, we investigated the expression patterns of phenylpropanoid biosynthetic genes and the content of phenylpropanoids in different parts of watercress. We identified 11 genes encoding enzymes related to the phenylpropanoid biosynthetic pathway and analyzed the expression patterns of these genes in the leaves, stems, roots, flowers, and seeds of watercress. The result showed that most of the genes were expressed at the highest levels in the flowers. HPLC analysis performed in samples from these same parts revealed the presence of seven phenylpropanoid-derived compounds. The content of total phenylpropanoids was the highest in flowers, followed by that in the leaves, whereas the lowest level was generally detected in the stems. Rutin was the most abundant phenylpropanoid in all plant segments, while quercetin was detected only in the flowers and roots. This study provides useful information for further molecular and functional research involving N. officinale and closely related species.

The present studies were attempted to develop direct shoot organogenesis from in vitro grown leaf explants of Lysimachia laxa and comparative evaluation of phytochemical and antioxidant potential of in vitro raised and wild plants extracts. The fresh leaves of this species are used for deworming gastrointestinal worm infection in traditional medicine. Overexploitation of this species and poor regeneration has led to rapid decline in wild population, therefore, present investigation was attempted to develop an efficient rapid mass propagation protocol for this species. Our result showed significantly (P < 0.05) high adventitious shoot proliferation of 17.21 ± 0.24 number per leaf explants cultured in Murashige and Skoog medium fortified with 1.25 mg L−1 thiadizuron and 1.0 mg L−1 α-naphthalene acetic acid. Further enhancement was achieved through elongation medium fortified with 1.0 mg L−1 6-benzylaminopurine by average shoot number of 31.1 ± 0.80 and length of 5.96 ± 0.13 cm. Murashige and Skoog medium fortified with 0.50 mg L−1 Indole-3-acetic acid showed high rooting induction (100%) with average root number of 11.70 and length 7.35 cm. All rooted plants were successfully acclimatized in greenhouse and transferred to field condition with a survival rate of 97%. The contents of phenolic and flavonoid were higher in in vitro raised plant in compared to wild plant extracts. Antioxidants assay showed high radical scavenging activity of IC50 1.61 ± 0.07 mg dry material and reducing power of 49.79 ± 0.11 mg/g ascorbic acid equivalent by aqueous methanol extracts of in vitro raised 3-months-old plants in compare to the wild plants. The present protocol is a viable option for pharmaceutical or nutraceutical industries for sustainable utilization of L. laxa with enhanced of phytochemical and antioxidant potency which is not reported elsewhere.

In the present study, we aimed to produce CGP/PVA films containing entrapped anti-inflammatory drugs for wound dressing applications. Using a 33–1 fractional factorial design, the effect of each component was evaluated on the physicochemical and morphological properties of the produced materials. The best formulation for entrapment of diclofenac sodium and ketoprofen was also determined. The produced films presented high swelling capacity, with some formulations showing o porous structure. CGP/PVA films showed a maximum retention of 75.6% for diclofenac sodium and 32.2% for ketoprofen, and both drugs were released in a controlled manner for up to 7 h. The drug release kinetic was studied, and the data were fitted using a Korsmeyer–Peppas model, which suggested that the release mechanism is controlled by diffusion. These results indicate that CGP/PVA-based matrices have great potential to be used as drug–delivery systems for wound dressing applications, contributing to prolonging the drug’s action time and then improving their anti-inflammatory efficacy.

Among various problems faced by mankind, health-related concerns are prevailing since long which are commonly found in the form of infectious diseases and different metabolic disorders. The clinical cure and management of such abnormalities are greatly dependent on the availability of their diagnoses. The conventional diagnostics used for such purposes are extremely powerful; however, most of these are limited by time-consuming protocols and require higher volume of test sample, etc. A new evolving technology called “biosensor” in this context shows an enormous potential for an alternative diagnostic device, which constantly compliments the conventional diagnoses. In this review, we have summarized different kinds of biosensors and their fundamental understanding with various state-of-the-art examples. A critical examination of different types of biosensing mechanisms is also reported highlighting the advantages of electrochemical biosensors for its great potentials in next-generation commercially viable modules. In recent years, a number of nanomaterials are extensively used to enhance not only the performance of biosensing mechanism, but also obtain robust, cheap, and fabrication-friendly durable mechanism. Herein, we have summarized the importance of nanomaterials in biosensing mechanism, their syntheses as well as characterization techniques. Subsequently, we have discussed the probe fabrication processes along with various techniques for assessing its analytical performances and potentials for commercial viability.

Transcription and translation in eukaryotes are distinct processes of the molecular cascade leading to protein production from genetic material. However, establishing correlation between mRNA expression and protein abundance, the end results of the two processes of central dogma, remains a challenge. For transgenic plants, such correlation between mRNA and protein expression serves as a guide to design the transgene, in particular the choices of promoter and codon usage to ensure stable expression of the target protein in relevant tissues under various stress conditions. To elucidate level of mRNA–protein correlation in a commercial transgenic cotton plant Gossypium hirsutum, Bollgard II® (MON15985), we present the results of Cry1Ac protein expression correlating with corresponding mRNA levels. Protein was quantitated using a home-grown validated ELISA assay with a monoclonal–polyclonal antibody pair, whereas mRNA level was detected by a real-time quantitative PCR assay using standardized reference genes. Our results indicate that protein and mRNA levels are highly correlated in the leaves, but not in squares and stem. The correlations seem to be consistent between young and mature leaves and increase over time of harvesting of samples from months 1–3. These findings demonstrate that transcript level measurement could serve as a proxy to protein abundance for this commercially important cotton species, particularly for leaf tissues which are the most vulnerable organs to cotton bollworms and other pathogens.