Santi Lata Sahoo

Production of α-Amylase by Aspergillus terreus NCFT 4269.10 Using Pearl Millet and Its Structural Characterization.

In this investigation, Aspergillus terreus NCFT4269.10 was employed in liquid static surface (LSSF) and solid state (SSF) fermentation to assess the optimal conditions for α-amylase biosynthesis. One-variable-at-a-time approach (quasi-optimum protocol) was primarily used to investigate the effect of each parameter on production of amylase. The maximum amylase production was achieved using pearl millet (PM) as substrate by SSF (19.19 ± 0.9 Ug−1) and also in presence of 1 mM magnesium sulfate, 0.025% (w/v) gibberellic acid, and 30 mg/100 ml (w/v) of vitamin E (~60-fold higher production of amylase) with the initial medium pH of 7.0 and incubation at 30 °C for 96 h. In addition, maltose, gelatin and isoleucine also influenced the α-amylase production. Amylase was purified to homogeneity with molecular mass around 15.3 kDa. The enzyme comprised of a typical secondary structure containing α-helix (12.2%), β-pleated sheet (23.6%), and β-turn (27.4%). Exploitation of PM for α-amylase production with better downstream makes it the unique enzyme for various biotechnological applications.

Development of a simplified protocol for in vitro propagation of a valuable medicinal plant Plumbago zeylanica Linn. through nodal explants found in Odisha, India

An efficient and economic protocol for rapid in vitro propagation using nodal explants obtained from 2 year old, field grown medicinal plants of Plumbago zeylanica L. belonging to the family Plumbaginaceae was successfully achieved from the nodal segments. Shoot development was maximum (95%) on Murashige and Skoog’s (MS) basal medium supplemented with 6- Benzyl amino purine (BAP) (2.0 mg/l), indole-3-acetic acid (IAA) (1.5 mg/l) and indole-3-butyric acid (IBA) (1.0 mg/l) with (19.56±0.04) mean number of shoots per explants and the maximum shoot length was found to be (4.98±0.87). Rooting of the differentiated shoots was achieved in MS medium with triple auxins combination of α- naphthalene acetic acid (NAA) (1.5 mg/l), IAA (1.5 mg/l) and IBA (2.0 mg/l) with (18.54±0.09) mean number of roots per shoots and the mean root length was found to be (7.63±0.83). Regenerated plantlets were successfully acclimated in the green house and after a hardening period of 4 weeks 100% transplantation success was achieved under the natural condition. The plantlets derived through in vitro propagation mimic the morphological characteristics of the donor plants taken for the present investigation.

Role of auxins in the in vitro rooting and micropropagation of Holarrhena antidysenterica Wall., a woody aromatic medicinal plant, through nodal explants from mature trees

An economic and efficient procedure has been outlined for plant regeneration of an important medicinal shrub, Holarrhena antidysenterica W. using nodal explants obtained from about 20-year-old mature trees growing in the field, belonging to the family of Apocynaceae. Shoot development was maximum (90%) on Murashige and Skoog’s (MS) basal medium supplemented with α-naphthalene acetic acid (NAA, 2.0 mg/L), indole-3-acetic acid (IAA, 1.0 mg/L), and KIN (1.0 mg/L) with (10.06 ± 0.24) mean number of shoots per explants and the maximum shoot length was found to be 4.01 ± 0.37. The role of auxins were instrumental as rooting of the differentiated shoots was best in MS medium with combination of indole-3-butyric acid (IBA, 1.5 mg/L) and IAA (1.5 mg/L) with 13.14 ± 0.08 mean number of roots per shoots and the mean root length was found to be 5.61 ± 0.03. Regenerated plantlets were successfully acclimated in the green house and after a hardening period of 4 weeks, 90% transplantation success was achieved under the natural condition. The established in vitro propagated plants were identical and uniform on the basis of the morphology and growth characteristics to the donor plants used in the study.

Thermostable acidic protease production in Aspergillus terreus NCFT 4269.10 using chickling vetch peels

Abstract A newly isolated fungus, Aspergillus terreus NCFT4269.10, was employed in both solid state (SSF) and liquid static surface culture (LSSC) for the production of protease using different agro-residues. Among different substrates appraised, chickling vetch peels (CVP) supported the enhanced production of protease both at LSSC and SSF (499.99 ± 11 U/ml; 5266.8 ± 202.5 U/gds, respectively). In the presence of peptone (1%, w/v), leucine (5 mM/100 ml), Fe2+ (1 mM) and riboflavin (10 mg/100 ml) with a medium pH of 5.0 incubated at 30 °C for 96 h, 3-fold higher protease production was achieved in LSSC compared with control. Fermentation kinetics studies revealed that the highest specific growth rate of A. terreus was observed in fermentation medium supplemented with riboflavin (10 mg/100 ml), i.e., 256.45 mg l−1 h−1. The growth-associated coefficient of enzyme production (α) by A. terreus was maximal when protease was produced using Fe2+. Further, the protease was purified to electrophoretic homogeneity and its molecular mass was determined as 23.8 kDa. The present strain suggests the potential utilization of inexpensive agro-residues (CVP) as medium components for the efficient industrial production using LSSC.

Proton induced X-ray emission-based analysis of trace element composition of cotyledon derived in vitro callus culture of Abrus precatorius L.: a multimedicinal wild legume

Quantitative estimation of inorganic elements of cotyledon-derived callus tissues of a medicinal legume, Abrus precatorius L. was determined using Proton induced X-ray emission technique. Nine trace elements namely Cu, Cr, Co, Zn, Mn, Fe, Se, Br and Ti in addition to two macro-elements K and Ca were identified, quantified and compared with each culture passage to monitor the variation in trace element accumulation. Experimental findings revealed that, 10-week-old calli was more efficient than others in accumulating inorganic elements. These results through light on the implication of in vitro callus cultures as a viable, alternative and proliferating renewable resource of medicinally useful elements for designing of drugs and also helping in natural germplasm conservation.

Proteomic and genomic responses of plants to nutritional stress

Minerals or trace elements in small amount are essential nutrients for every plant, but when the internal concentration exceeds the threshold, these essential elements do create phytotoxicity. Plant responses to elemental stresses are very common due to different anthropogenic activities; however it is a complex phenomenon with individual characteristics for various species. To cope up with the situation, a plant produces a group of strategies both in proteomic and genomic level to overcome it. Controlling the metal stress is known to activate a multigene response resulting in the changes in various proteins, which directly affects almost all biological processes in a living cell. Therefore, proteomic and genomic approaches can be useful for elucidating the molecular responses under metal stress. For this, it is tried to provide the latest knowledge and techniques used in proteomic and genomic study during nutritional stress and is represented here in review form.