Hydrogen sulfide (H2S) is a novel type signaling molecule in plants Seed germination is a key stage of life cycle of plants, which is vulnerable to environmental stress including high temperature However, under high temperature stress, whether pre-soaking of maize seeds with NaHS (a H2S donor) could improve seed germination and seedling growth and the possible mechanisms are not completely clear In this study, maize seeds pre-soaked with NaHS enhanced germination percentage, sprout length, root length, and fresh weight compared with the control without NaHS treatment, illustrating that H2S could improve maize seed germination and seedling growth under high temperature In addition, in comparison to the control, NaHS pre-soaking stimulated antioxidant enzymes [ascorbate peroxidase (APX), glutathione reductase (GR), guaiacol peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT)] activities and the contents of water soluble non-enzymatic antioxidants [ascorbic acid (AsA) and glutathione (GSH)], as well as the ratio of reduced antioxidant to oxidized antioxidant Moreover, pre-soaking with NaHS activated Δ1-pyrroline-5-carboxylate synthetase (P5CS) and ornithine aminotransferase [OAT; both are rate-limiting enzymes in proline (Pro) synthesis], betaine aldehyde dehydrogenase [BADH; a key enzyme in glycine betaine (GB)], and trehalose (Tre)-6-phosphate phosphatase (a key step in Tre synthesis), which in turn accumulated Pro, GB, and Tre in germinating seeds compared with the control Also, an improved germination by NaHS under high temperature was reinforced by the above osmotic adjustment substances (osmolytes) alone, while deteriorated by the inhibitors of osmolyte biosynthesis [gabaculine (GAB), disulfiram (DSF), and sodium citrate (SC)] These results imply that H2S could improve maize seed germination and seedling growth under high temperature by inducing antioxidant system and osmolyte biosynthesis

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Signaling Molecule Hydrogen Sulfide Improves Seed Germination and Seedling Growth of Maize (Zea mays L.) Under High Temperature by Inducing Antioxidant System and Osmolyte Biosynthesis.

Over the last two decades the recombinant production of therapeutic and industrially important proteins has gained more attention due to a dramatically increased demand. This has driven the effectual bioprocess development to maximize the yield. Thus several microbial, mammalian, and insect expression hosts have been improved. Among the microbial hosts, Escherichia coli and yeast systems are widely used for the production of most of the therapeutic proteins. Further, advances in media and process optimization technologies, such as scale-out technology, process analytical technology, high throughput process devices, and modern bioreactors have enabled a many fold improvement in the final yield. In addition to this, several computational tools and simulation models have been developed to achieve higher product yield. This chapter basically outlines the different important expression systems, their advantages and disadvantages, and the modern bioprocess technologies used for protein production.

Effectual Bioprocess Development for Protein Production

The zebrafish (Danio rerio) is a versatile model organism that has been used in biomedical research for several decades to study a wide range of biological phenomena. There are many technical advantages of using zebrafish over other vertebrate models. They are readily available, hardy, easy, and inexpensive to maintain in the laboratory, have a short life cycle, and have excellent fecundity. Due to its optical clarity and reproducible capabilities, it has become one of the predominant models of human genetic diseases. Zebrafish research has made rapid strides in the United States and Europe, but in India the field is at an early stage and many researchers still remain unaware of the full research potential of this tiny fish. The zebrafish model system was introduced into India in the early 2000s. Up to now, more than 200 scientific referred articles have been published by Indian researchers. This review gives an overview of the current state of knowledge for zebrafish research in India, with the aim of promoting wider utilization of zebrafish for high level biological studies.

Zebrafish: A Premier Vertebrate Model for Biomedical Research in Indian Scenario

Rapid identification of osmolytes in tropical microalgae and cyanobacteria by 1H HR-MAS NMR spectroscopy

The formation of inclusion bodies (IBs) is considered as an Achilles heel of heterologous protein expression in bacterial hosts. Wide array of techniques has been developed to recover biochemically challenging proteins from IBs. However, acquiring the active state even from the same protein family was found to be an independent of single established method. Here, we present a new strategy for the recovery of wide sub-classes of recombinant protein from harsh IBs. We found that numerous methods and their combinations for reducing IB formation and producing soluble proteins were not effective, if the inclusion bodies were harsh in nature. On the other hand, different practices with mild solubilization buffers were able to solubilize IBs completely, yet the recovery of active protein requires large screening of refolding buffers. With the integration of previously reported mild solubilization techniques, we proposed an improved method, which comprised low sarkosyl concentration, ranging from 0.05 to 0.1% coupled with slow freezing (− 1 °C/min) and fast thaw (room temperature), resulting in greater solubility and the integrity of solubilized protein. Dilution method was employed with single buffer to restore activity for every sub-class of recombinant protein. Results showed that the recovered protein’s activity was significantly higher compared with traditional solubilization/refolding approach. Solubilization of IBs by the described method was proved milder in nature, which restored native-like conformation of proteins within IBs.

Comparative study to develop a single method for retrieving wide class of recombinant proteins from classical inclusion bodies

In vivo study of Zebrafish dihydrofolate reductase (zDHFR), a 23 kDa monomeric protein was studied to check the expression level in the presence of sorbitol, glycerol, proline, glycine, glucose, sucrose and betaine. Different osmolytes, also known as chemical chaperones, assist in expression enhancement of protein. In the present study, it was observed that a few osmolytes effectively enhance the expression and functionality of zDHFR and hence provide substantial stability to the protein with glucose, sucrose and betaine being the exception. A general strategy to improve the activity of recombinant proteins would be to increase the cellular concentration of viscous organic compounds, termed osmolytes, or of molecular chaperones that can prevent aggregation and convert it into native active species of commercial and pharmacological importance.

Effect of various osmolytes on the expression and functionality of zebrafish dihydrofolate reductase: an in vivo study

Investigation of folding unfolding process of a new variant of dihydrofolate reductase protein from Zebrafish.

Quantification of differential efficacy of chemical chaperones in ameliorating solubilization and folding of zebrafish dihydrofolate reductase.

The proper functioning of organisms in stress conditions is an axiological exercise Proteins are vulnerable to denaturation and misfolding under hiked temperatures, increased hydrostatic pressure, the presence of chaotropic agents, etc Osmolytes are one of the most important groups of molecules that are employed by the cell as an adaptation to these harsh conditions These small molecules maintain cell volume, osmotic equilibrium, redox states, and energy quanta of the cell The current chapter reviews the versatility of the osmolytes in various metabolic functions and how widely they are distributed across the different classes of organisms (plants, animals, insects, marine animals, etc) This chapter discusses their diversity and the exact mechanism by virtue of which these osmolytes are able to impart stability to the proteins It also elucidates on the application of these osmolytes in treatment of various diseases and as possible drugs from the pharmaceutical point of view

Charu Thapliyal

Papers

Effect of various osmolytes on the expression and functionality of zebrafish dihydrofolate reductase: an in vivo study

Investigation of folding unfolding process of a new variant of dihydrofolate reductase protein from Zebrafish.

Quantification of differential efficacy of chemical chaperones in ameliorating solubilization and folding of zebrafish dihydrofolate reductase.

Osmolyte System and Its Biological Significance

Kinetics and thermodynamics of the thermal inactivation and chaperone assisted folding of zebrafish dihydrofolate reductase