piRNAs in the human retina and retinal pigment epithelium reveal a potential role in intracellular trafficking and oxidative stress.

Long considered active only in the germline, the PIWI/piRNA pathway is now known to play a significant role in somatic cells, especially neurons. In this study, piRNAs were profiled in the human retina and retinal pigment epithelium (RPE). Furthermore, RNA immunoprecipitation with HIWI2 (PIWIL4) in ARPE19 cells yielded 261 piRNAs, and the expression of selective piRNAs in donor eyes was assessed by qRT-PCR. Intriguingly, computational analysis revealed complete and partial seed sequence similarity between piR-hsa-26131 and the sensory organ specific miR-183/96/182 cluster. Furthermore, the expression of retina-enriched piR-hsa-26131 was positively correlated with miR-182 in HIWI2-silenced Y79 cells. In addition, the lnc-ZNF169 sequence matched with two miRNAs of the let-7 family, and piRNAs, piR-hsa-11361 and piR-hsa-11360, which could modulate the regulatory network of retinal differentiation. Interestingly, we annotated four enriched motifs among the piRNAs and found that the piRNAs containing CACAATG and CTCATCAKYG motifs were snoRNA-derived piRNAs, which are significantly associated with developmental functions. However, piRNAs consisting of ACCACTANACCAC and AKCACGYTCSC motifs were mainly tRNA-derived fragments linked to stress response and sensory perception. Additionally, co-expression network analysis revealed cell cycle control, intracellular transport and stress response as the important biological functions regulated by piRNAs in the retina. Moreover, loss of piRNAs in HIWI2 knockdown ARPE19 confirmed altered expression of targets implicated in intracellular transport, circadian clock, and retinal degeneration. Moreover, piRNAs were dysregulated under oxidative stress conditions, indicating their potential role in retinal pathology. Therefore, we postulate that piRNAs, miRNAs, and lncRNAs might have a functional interplay during retinal development and functions to regulate retinal homeostasis.

Phosphorus recovery from the sludge generated from a continuous bipolar mode electrocoagulation (CBME) system.

Phosphorus is known to be a limited non-renewable resource. Phosphorus is obtained from phosphate rock, which is likely to be depleted in the next few decades. Therefore, it is very important to find alternate sources of phosphorus from which phosphorus can be recycled and recovered. This study focuses on the recovery of phosphorus from the sludge generated from a continuous bipolar mode electrocoagulation (CBME) system, used for treating a palm oil mill effluent (POME). The sludge generated from the CBME system is leached with oxalic acid and sulphuric acid for phosphorus recovery with and without thermal treatment. Acid leaching was carried out at various time intervals using various liquid/solid (L/S) ratios of acids and sludge. The CBME system caused a 73% removal of phosphorus from POME, where phosphorus is precipitated in sludge as iron phosphates or adsorbed as phosphates depending on the pH in the system. Acid leaching resulted in nearly 85% recovery of phosphorus with both sulphuric acid and oxalic acid for sludge combusted at 900 °C. Statistical analysis was carried out to find the significance of the operational conditions on the phosphorus yield. Acid leaching results in the formation of orthophosphates, which can be used as a raw material for synthesis of chemical fertilizers.

Biosynthesis of gold nanoparticles.

Synthesis of nanoparticles with interesting physico-chemical properties using efficient as well as eco-friendly technology is one of the main objectives of nanotechnology. Biological systems have been reported to synthesize inorganic materials under certain circumstances. Exploiting the biosynthetic potential of different organisms, nanoparticles of varying morphologies and sizes have been synthesized. Among the nanomaterials, gold has received considerable attention owing to its varied applications in the fields of nano-medicine, catalysis, electronics, and optics. This review gives an account on the biosynthesis of gold nanoparticles from microorganisms, plants, and other biological sources, with particular emphasis on the probable mechanisms leading to the formation of gold nanoparticles and the extent of control over nanoparticle properties that has been achieved so far in the biosynthetic protocols. It has been speculated that enzymes and/or proteins secreted by the organisms are involved in the bio-reduction and stabilization of the nanoparticles. The biosynthetic procedures could compete with existing solvent-based chemical synthetic procedures in order to achieve stable and monodisperse gold nanoparticles in large scale.

Critical analysis of application of generalized distance function for optimization of important variables for esterase synthesis by Saccharomyces cerevisiae.

The generalized distance function approach is employed to obtain a suitable near optimal conditions of variables. The optimal values of variables (medium constituents, microbiological parameters, and process parameters) have been evaluated separately using single responses (either specific esterase activity or cell mass) as per central-composite-design and multi-responses following generalized distance function approach. The optimal conditions (medium composition (gl(-1)): dextrose, 13.43; peptone, 7.285; yeast extract, 2.55; and malt extract, 1.695; microbiological parameters: slant age, 39.9h; inoculum age, 9.6 h; and number of cells, 1.49 x 10(8) numbers ml(-1); process conditions: temperature, 29.9 degrees C; and pH, 6.2) obtained by generalized distance approach can be considered as a 'near optimal' solution of interactive multi-response systems of intracellular esterase synthesis by Saccharomyces cerevisiae.

Production and applications of esterases.

Esterase plays a major role in the degradation of natural materials and industrial pollutants, viz., cereal wastes, plastics, and other toxic chemicals. It is useful in the synthesis of optically pure compounds, perfumes, and antioxidants. The potential applications of esterase with reference to agriculture, food, and pharmaceutical industries, are discussed in this review. Promising applications in this avenue can be supported by appropriate production strategies.

Regulation and degradation of HMGCo-A reductase.

The enzyme, 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) controls the biosynthesis of cholesterol. Hypercholesterolemia and atherosclerosis are critical health risk factors. One way of controlling these risk factors is to manipulate regulation as well as degradation of HMGR. At present, a class of compounds called statins, which are HMGR inhibitors, are used for the treatment of hypercholesterolemia. However, statins suffer major setbacks as their use produces more adverse reactions than the desirable one of inhibiting the enzyme. Genetically engineered forms of HMGR are also studied in primitive life forms like bacteria, but detailed investigation of this enzyme in human systems is certainly required. Extensive studies have been made on the regulatory aspects of this enzyme, but no breakthrough is conspicuous in the clinical background to find an alternative treatment for hypercholesterolemia. The immediate need is to find an alternate way of regulating degradation of the enzyme. This review presents the importance of regulation and degradation of the HMGR enzyme in different systems to gain possible insight into alternative schemes for regulating this enzyme and, if these exist, the feasibility of extending them same to studies in mammalian systems. A high degree of similarity exists between mammalian and yeast HMGR. Detailed studies reported on the regulation and degradation of the yeast enzyme also throw more light on the mammalian system, leading to a better understanding of ways of controlling hypercholesterolemia.

Extractive fermentation for improved production of endoglucanase by an intergeneric fusant of Trichoderma reesei/Saccharomyces cerevisiae using aqueous two-phase system.

Extractive aqueous two-phase fermentation of endoglucanase, a key enzyme for the conversion of cellulosic substances to fermentable sugars, from an intergeneric fusant of Trichoderma reesei/Saccharomyces cerevisiae is a meaningful approach for better production and simple recovery of this enzyme. A phase composition of 6.5% (w/w) dextran and 7.5% (w/w) polyethylene glycol 6000, having a partition coefficient of 2.89 and 1.31 for endoglucanase from an intergeneric fusant of T. reesei/S. cerevisiae and T. reesei (WT) (being a control in this study), respectively, was chosen for extractive fermentation of the enzyme. Endoglucanase production is higher in medium containing polyethylene glycol (PEG) 6000 than in medium without PEG 6000. Comparative analysis of endoglucanase fermentation by fusant and T. reesei was carried out in shake culture and environment-controlled bioreactor conditions. The fusant produced 0.43U of endoglucanase (overall production: 0.34U) in the top phase of an aqueous two-phase system (ATPS), compared to 0.3U in medium without the phase system in shake culture. In a batch reactor, the endoglucanase level for the fusant in the top phase of ATPS was 0.49U (overall production: 0.40U), compared to 0.38U produced in medium without aqueous two-phase components. To corroborate this study, T. reesei produced 8.41U of endoglucanase (overall production: 5.96U) in the top phase of ATPS, compared to 7.18U in the medium without the phase system in shake culture. On the other hand, in a batch bioreactor, T. reesei produced 10.13U of endoglucanase (overall production: 6.90U) in the top phase of ATPS, compared to 8.56U of the enzyme in medium without aqueous two-phase components. The lower overall enzyme production by T. reesei in the two-phase system might be due to limitation in oxygen transfer to the dispersed phase where the enzyme is produced. A higher cell concentration and a reduced lag phase was obtained in ATPS, compared to a similar medium without phase forming polymers for both the intergeneric fusant of T. reesei/S. cerevisiae and T. reesei.

Aqueous two-phase: the system of choice for extractive fermentation.

Extractive fermentation in aqueous two-phase systems is a meaningful approach to overcome low product yield in a conventional fermentation process, and by proper design of the two-phase system it is possible to obtain the product in a cell-free stream. The characteristics of an aqueous two-phase system, various polymers used for forming an aqueous two-phase system, the physicochemical parameters of the aqueous two-phase system, partitioning of biomolecules and cell mass and the effect of individual phase forming polymers on cell growth and product formation are reviewed in this article. The various extractive fermentation processes are also summarised here. At the end, the economic viability and scope of aqueous two-phase fermentation are briefly discussed in relation to the wider application of this topic.

Regulation and cloning of microbial chitinase genes.

A range of chitinase genes from microorganisms have been cloned and the potential uses of these genetically manipulated organisms are being investigated by various researchers. Fungi and yeast are better producers of chitinase than bacteria. Since fungi grow at a slower rate, there have been efforts to clone the fungal chitinase genes into fast-growing bacteria. This review gives a brief survey of recent progress in the regulation and cloning of microbial chitinase genes. Emphasis is placed on the post-translational modification and localization of the recombinant protein in the host. Various amino acid domains are present in this protein. The mode of catalytic activity of the recombinant protein in comparison to the wild-type protein is discussed in the available literature. The different mechanisms involved in the regulation of chitinase genes from various microorganisms is discussed by the researchers. The scope of future research and conclusions yet to be obtained in this particular area are also outlined in this review.

Effect of replacing helical glycine residues with alanines on reversible and irreversible stability and production of Aspergillus awamori glucoamylase.

To decrease irreversible thermoinactivation of Aspergillus awamori glucoamylase, five Gly residues causing helix flexibility were replaced with Ala residues. Mutation of Gly57 did not affect thermostability. Mutation of Gly137 doubled it at pHs 3.5 and 4.5 but barely changed it at pH 5.5. The Gly139-->Ala mutation did not change thermostability at pH 3.5, improved it at pH 4.5 and worsened it at pH 5.5. The Gly 137/Gly139-->Ala/Ala mutation gave 1.5-2-fold increased thermostabilities at pHs 3.5-5.5. Mutations of Gly251 and Gly383 decreased it at all pHs. Gly137-->Ala and Gly137/Gly139-->Ala/Ala glucoamylases are the most stable yet produced by mutation. Guanidine treatment at pH 4.5 decreased the reversible stabilities of Gly137-->Ala, Gly139-->Ala and Gly137/Gly139-->Ala/Ala glucoamylases at infinite dilution while not changing those of Gly251-->Ala and Gly383-->Ala glucoamylases, which is, in general, opposite to what occurred with thermoinactivation. Mutation of Gly57 greatly improved the extracellular glucoamylase production by yeast, that of Gly137 barely affected it and those of Gly139 and of both Gly137 and Gly139 strongly impeded it. These observations suggest that alpha-helix rigidity can affect reversible and irreversible glucoamylase stability differently, that the effects of multiple mutations within one alpha-helix to improve stability are not always additive and that even single mutations can strongly affect extracellular enzyme production.

Studies on improved techniques for immobilizing and stabilizing penicillin amidase associated with E. coli cells.

A method for catalyst development has been suggested for immobilizing whole E. coli cells containing penicillin amidase. Conventional methods have limitations, such as permeation of substrate and product through cellular membranes, leaching of protein and other cellular components into the reaction phase, lower specific activity compared to immobilized enzyme system, etc. The whole cell immobilization technique has been optimized for different process parameters. The most suitable conditions for this process were pH, 4.25; cell concentration, 3.75%; concentration of glutaraldehyde, 1.5%; level of bovine serum albumin as additional support, 2 mg ml-1. The reaction was continued for 2 h. The granular catalyst has good mechanical strength, low protein leachability, and high retention of penicillin amidase activity.

Method To Estimate Growth of Trichoderma reesei and Aspergillus wentii in Mixed Culture on Cellulosic Substrates.

A simple differential method based on measurement of an intracellular pigment of Aspergillus wentii was developed for estimation of the individual growths of two fungi, Trichoderma reesei and A. wentii, in mixed-culture fermentation of an insoluble cellulosic substrate.

Effect of culture phasing and a polysaccharide on production of xylanase by mixed culture of Trichoderma reesei D1-6 and Aspergillus wentii Pt 2804.

A significant increase in extracellular xylanase activity was observed in the mixed culture fermentation of Trichoderma reesei D1-6 and Aspergillus wentii Pt 2804 when A. wentii inoculation was phased by 15 h. A. wentii produced a polysaccharide, chiefly consisting of glucose monomeric units, which was required for expression of maximum xylanase activity. Expression of high activity of xylanase in the A. wentii phased mixed culture compared to that in either T. reesei or A. wentii single cultures appeared to be controlled by the combined action of a polysaccharide produced by A. wentii and the relative growth of the two fungi in the mixed culture.

Effect of culture phasing and mannanase on production of cellulase and hemicellulase by mixed culture of Trichoderma reesei D 1-6 and Aspergillus wentii pt 2804.

Significant increase in extracellular cellulase and hemicellulase activities was observed in the biosynthesis of cellulase enzyme in mixed culture fermentation of Trichoderma reesei D 1-6 and Aspergillus wentii Pt 2804 when the A. wentii inoculation was phased by 15 h. The optimal conditions of fermentation by the mixed culture have been established. Presence of mannanase has been found to affect the release as well as activity of cellulase enzyme produced in mixed culture.

Pretreatment of indian cane molasses for increased production of citric acid.

Surface culture citric acid fermentation was carried out by Aspergillus niger T55, a strain isolated from its natural source, using cane molasses, either untreated or treated by various methods. Citric acid biosynthesis is seriously impaired by both organic and inorganic inhibitors. A combined treatment of molasses with tricalcium phosphate, hydrochloric acid, and Sephadex fractionation minimizes the level of inorganic and organic inhibitors in molasses and increases the production of citric acid (65% weight yield based on total reducing sugar). The optimum level of individual metal ions for citric acid production depends on the concentration of other metals in the medium.