Balwinder Singh Sooch

Recent insights into microbial catalases: Isolation, production and purification

Catalase, an oxidoreductase enzyme, works as a detoxification system inside living cells against reactive oxygen species formed as a by-product of different metabolic reactions. The enzyme is found in a wide range of aerobic and anaerobic organisms. Catalase has also been employed in various analytical and diagnostic methods in the form of biosensors and biomarkers in addition to its other applications in textile, paper, food and pharmaceutical industries. New applications for catalases are constantly emerging thanks to their high turnover rate, distinct evolutionary origin, relatively simple and well-defined reaction mechanisms. The following review provides comprehensive information on isolation, production and purification of catalases with different techniques from various microbial sources along with their types, structure, mechanism of action and applications.

Differential proteomics of human seminal plasma: A potential target for searching male infertility marker proteins

The clinical fertility tests, available in the market, fail to define the exact cause of male infertility in almost half of the cases and point toward a crucial need of developing better ways of infertility investigations. The protein biomarkers may help us toward better understanding of unknown cases of male infertility that, in turn, can guide us to find better therapeutic solutions. Many clinical attempts have been made to identify biomarkers of male infertility in sperm proteome but only few studies have targeted seminal plasma. Human seminal plasma is a rich source of proteins that are essentially required for development of sperm and successful fertilization. This viewpoint article highlights the importance of human seminal plasma proteome in reproductive physiology and suggests that differential proteomics integrated with functional analysis may help us in searching potential biomarkers of male infertility.

Influence of multiple bioprocess parameters on production of lipase from Pseudomonas sp. BWS-5

The aim of the present work was to study the influence of multiple bioprocess parameters for the maximum production of lipase from Pseudomonas sp. BWS-5. The culture reached the stationary phase of growth after 36h of incubation when the maximum lipase production was obtained at flask level. The different media components such as carbon sources, nitrogen sources, trace elements and process parameters such as the pH of the medium, temperature and time of incubation, agitation/stationary conditions, etc. were optimized at flask level and at bioreactor level. The maximum enzyme production of 298 IU/mL was obtained with the use of simple medium with pH 6.5 containing glucose (1 %, w/v), peptone (3 %, w/v) and KCl (0.05 %, w/v) after 30h of incubation at 37°C under agitation (200 rpm) conditions with 0.75 vvm of air supply.

Differential proteomics of sperm: insights, challenges and future prospects

Male factors account for 40% of infertility cases and most are caused by low sperm count, poor sperm quality or both. Defects in sperm are directly linked to reproductive malfunctions, and these defects may be caused by genetic mutations, environmental factors and exposure to free radicals, for example. Almost half of the male infertility cases have no known cause, indicating the lack of sensitive tests for the diagnosis of infertility. Proteomics has evolved as a major research field in biology and medicine, to identify and validate potent targets, at the molecular level, for development of more sensitive diagnostic tools. The recent advances in this field focus on the identification of differentially expressed proteins and analyzing their functional aspects for better understanding of the biological pathways. It not only provides a platform to discover biomarkers of infertility, but may also help in the design of effective male contraceptives. This article discusses various insights of proteomics for exploring biomarkers of male infertility in sperm. It also discusses the enhanced understanding of reproductive physiology offered by data produced by proteomic studies of spermatozoa.

Interaction analysis identifies semenogelin I fragments as new binding partners of PIP in human seminal plasma.

Identification of protein-protein interactions is vital for complete understanding of a biological process and for functional characterization of a protein in related biochemical pathways. In this study, we performed analysis of prolactin inducible protein (PIP) interactions in human seminal plasma. PIP and its interacting partners were co-immunoprecipitated, analyzed by SDS-PAGE and identified by MALDI-TOF mass spectrometry. Three major interacting partners were identified, viz. human serum albumin, zinc-α-2 glycoprotein and semenogelin I fragments. This is the first report of interaction between PIP and semenogelin I fragments in human seminal plasma or elsewhere with a suggestive role in reproductive physiology which might be helpful for spermatozoa to acquire their motility.

Isolation and identification of Concanavalin A binding glycoproteins from human seminal plasma: A step towards identification of male infertility marker proteins

Human seminal plasma contains a large array of proteins of clinical importance which are essentially needed to maintain the reproductive physiology of spermatozoa and for successful fertilization. Thus, isolation and identification of seminal plasma proteins is of paramount significance for their biophysical characterization and functional analysis in reproductive physiological processes. In this study, we have isolated Concanavalin-A binding glycoproteins from human seminal plasma and subsequently identified them by MALDI-TOF/MS analysis. The major proteins, as identified in this study, are Aminopeptidase N, lactoferrin, prostatic acid phosphatase, zinc-alpha-2-glycoprotein, prostate specific antigen, progestagen-associated endometrial protein, Izumo sperm-egg fusion protein and prolactin inducible protein. This paper also reports preliminary studies to identify altered expression of these proteins in oligospermia and azoospermia in comparison to normospermia. In oligospermia, five proteins were found to be downregulated while in azoospermia, four proteins were downregulated and two proteins were upregulated. Thus, this study is of immense biomedical interest towards identification of potential male infertility marker proteins in seminal plasma.

Optimization of Cellulase Production from Newly Isolated Bacillus sp. Y3

Cellulose, a major constituent of plant cell wall, is the most abundant biological polymer on earth. The use of various cellulolytic microorganisms for the bioconversion of cellulose into value added products has attracted a worldwide attention. Hence the present work was aimed to isolate new cellulase producing microorganisms and further to investigate the effect of nutritional and process parameters on cellulase production from selected isolated culture. Out of 20 cellulase producing bacterial strains isolated during the study, Y3 isolate was found to be best for the production of cellulase enzyme. This isolate was then characterized for its morphological and biochemical characters and identified as Bacillus sp. Y3. The effect of different parameters like carbon sources, nitrogen sources, temperature, pH, inoculum concentration and incubation time was monitored with selected strain for cellulase production. The maximum FPase and CMCase activity of Bacillus sp. Y3 was 6.84 IU/mL and 7.82 IU/ mL, respectively, when the basal media of pH 7 containing CMC (1%, w/v) and peptone (1%, w/v) was inoculated with 2% (v/v) inoculum and incubated at 37°C for 96 hours at 120 rpm. The FPase (6.84 IU/mL) and CMCase activity (7.82 IU/mL) obtained after optimization was much higher than FPase (1.97 IU/mL) and CMCase activity (2.48 IU/ mL) before optimization.

A Study on the Antioxidant Activity of Pyridylselenium Compounds and their Slow Release from Poly(acrylamide) Hydrogels

Abstract The antioxidant activity of pyridylselenium compounds has been evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and nitric oxide (NO) scavenging methods. Pyridylselenium compounds have shown far superior (100–1000 times) antioxidant property than ebselen. The control release of bis(2-pyridyl) diselenide from poly(acrylamide) hydrogels has been studied in order to evaluate its release mechanism and diffusion coefficient. The later study also demonstrates that the pyridylselenium loading into the polymer matrix increases the magnitude and the rate of the radical scavenging activity of the poly(acrylamide) hydrogels. GRAPHICAL ABSTRACT

Quantification studies in human seminal plasma samples identify prolactin inducible protein as a plausible marker of azoospermia.

Background: Prolactin inducible protein (PIP) is a ~17 kDa protein, which is known to play vital roles in immunoregulation, fertility, antimicrobial activity, apoptosis and tumour progression. Objectives: This study reports quantification of PIP concentration in human seminal plasma (SP) samples. Methodology: PIP was purified by immunoprecipitation and its concentration in human SP samples was quantified by ELISA method. Results: Average concentration of PIP in normozoospermia, oligozoospermia and azoospermia was 290.3 ± 71.5 µg/mL, 306.4 ± 71.2 µg/mL and 60.5 ± 23.6 µg/mL respectively. Conclusion: There was no significant variation in PIP levels in normozoospermia and oligozoospermia while its expression was down-regulated in azoospermia, indicating that PIP may be a plausible marker of azoospermia.

Covalent linkage of alkalothermophilic catalase onto functionalized cellulose

Catalase from a thermophilic bacterium belonging to the genus Geobacillus was purified and covalently immobilized onto a functionalized polymer via a spacer with an objective to improve its kinetic and biochemical properties. This is the first report on the purification and immobilization of catalase from the genus Geobacillus. The tetrameric catalase of about 221 kDa was successfully purified using a multistep purification strategy. A shift in pH and temperature optima from 8.0 to 9.0 and 55 °C to 60 °C, respectively was recorded after covalent binding of catalase onto the functionalized matrix. The kinetic constants i.e. Km, Vmax, Kcat and Kcat/Km were found to be 1.2 mM, 4.43 × 106 IU, 6.3 × 105 s−1 and 5.25 × 108 s−1 M−1 for free, and 1.8 mM, 4.01 × 106 IU, 5.9 × 105 s−1 and 3.20 × 108 s−1 M−1 for immobilized catalase, respectively. The ease of binding of alkalothermophilic catalase from a novel isolated bacterium G. extremocatsoochus sp. nov., MTCC 5873 onto a low cost activated cellulose support demonstrated enhanced pH and thermal stability as compared to its free counterpart. This immobilized catalase preparation with improved characteristics has good potential for diverse applications. The present findings provide valuable information on how to tailor enzymes and supported polymer matrices to improve the performance of a biocatalyst.