Partial purification and characterization of limonoate dehydrogenase from Rhodococcus fascians for the degradation of limonin

An extracellular limonoate dehydrogenase was purified 10-fold from a cell-free extract of Rhodococcus fascians by ammonium sulfate precipitation, dialysis, and ultrafiltration. This purified dehydrogenase catalyzed the
conversion of limonoate to 17-dehydrolimonoate. The enzyme showed optimum activity at pH 8.0 and 40oC, with K_m value of 0.9 µM, and requires Zn ions and sulfhydryl groups for catalytic action. The enzyme activity was inhibited by Hg²⁺ and NaN₃ ions. The degradation of limonin (66%) in Kinnow mandarin juice was successfully demonstrated with partially
purified limonoate dehydrogenase. With scale-up preparation of limonoate dehydrogenase, a successful debittering operation of fruit juices appears feasible.

Examining the development of the Victorian Certificate of Applied Learning and its implications for schools and teacher education in Australia

The Victorian Certificate of Applied Learning (VCAL) is a very successful senior secondary school qualification introduced in the Australian state of Victoria in 2002. Applied learning in the VCAL engages senior students in a combination of work-based learning, service-learning, and project-based learning and aims to provide them with the skills, knowledge, and attitudes to make informed choices regarding pathways to work and further education. The program has enjoyed rapid growth and its system-wide adoption by Victorian secondary schools, Technical and Further Education (TAFE) institutions, Registered Training Organizations (RTOs), and Adult and Community Education (ACE) providers has broadened significantly the range of senior schooling pathway options for young people. This paper will examine reasons for developing an applied learning senior secondary certificate and its rapid growth in Victoria since 2002. The authors draw on a number of case studies to profile the unique nature of applied learning in the VCAL, including its dimensions of service learning, work-based learning, and project-based learning. These case studies are also used to discuss a number of implications that have emerged from the use of applied learning in the VCAL, including approaches to teaching and assessment that will support applied learning and the development of new partnerships between VCAL providers and community partners. Finally, the paper considers significant implications the VCAL has created for teacher education in Victoria by discussing the new Graduate Diploma of Education (Applied Learning) developed by Deakin University.

Youth literacy development through applied learning and the national curriculum

The introduction of a national curriculum into Australian schools coincides with an increasing body of research emphasising the importance of completing Year 12 or its equivalent if young people are to achieve successful transitions beyond school. This research is reflected in Australian government policy targets to achieve secondary school completion rates to at least 90% by 2015. State governments across the country have also embraced these targets, with some states already achieving above this level thanks to innovative curriculum and pedagogical initiatives that are more attuned to applied ways of learning.

In this chapter we examine the challenges faced by school leaders and teachers as they re-conceptualise approaches to youth literacy development through applied learning pedagogy. We begin by exploring definitions and applications of applied learning and examining the relatively recent growth of applied learning in secondary schools. We discuss the impetus for using applied learning approaches to literacy development and consider the challenges manifest in the day-to-day professional practices of secondary school teachers who use applied learning to engage young people who may otherwise leave school early. We conclude by exploring tensions emerging from the use of applied learning approaches to youth literacy development in Australia and the new demands created by a national curriculum.

Nature, extent and antecedents of risk management in accounting, law and biotechnology firms in Australia

This paper provides empirical evidence on the nature and the extent of risks faced by small and medium-sized biotechnology and professional service firms (accounting and law) in Australia, as well as on the style of their adopted risk management methods and approaches. The findings of the study indicate that the top three risks faced by these firms are related to reputation, recruiting and retaining skilled staff, and cost management. The study also finds that more than half of the respondent firms manage risk in an integrated manner. The results of this study provide useful insights into the nature, extent and driving forces of risk management practices in these firms.

Applied econometrics and implications for energy economics research

Developments in applied econometrics, particularly with regard to unit root tests and cointegration tests, have motivated a rich empirical literature on energy economics over the last decade. This study reviews recent developments in time series econometrics applications in the energy economics literature. We first consider the literature on the integration properties of energy variables. We begin with a discussion of the implications of whether energy variables contain a unit root and proceed to examine how results differ according to the specific unit root or stationarity test employed. We then proceed to examine recent developments in the literature on cointegration, Granger causality and long-run estimates between (disaggregated) energy consumption and economic growth. We review both single country and panel studies and pay particular attention to studies which have expanded the literature through adding variables such as financial development and trade, in addition to energy consumption to the augmented production function, as well as studies which have extended the literature through examining disaggregated energy consumption by type. In each case we highlight best practice in the literature, point to limitations in the literature, including econometric modeling challenges, and suggest recommendations for future research. A key message of our survey is that the profession needs to guard against 'overload' of research in these areas as most applied studies are no longer adding anything more to what is already known. © 2014 Elsevier B.V. All rights reserved.

Evaluation of glucose dehydrogenase and pyrroloquinoline quinine (pqq) mutagenesis that renders functional inadequacies in host plants

The rhizospheric zone abutting plant roots usually clutches a wealth of microbes. In the recent past, enormous genetic resources have been excavated with potential applications in host plant interaction and ancillary aspects. Two Pseudomonas strains were isolated and identified through 16S rRNA and rpoD sequence analyses as P. fluorescens QAU67 and P. putida QAU90. Initial biochemical characterization and their root-colonizing traits indicated their potential role in plant growth promotion. Such aerobic systems, involved in gluconic acid production and phosphate solubilization, essentially require the pyrroloquinoline quinine (PQQ)- dependent glucose dehydrogenase (GDH) in the genome. The PCR screening and amplification of GDH and PQQ and subsequent induction of mutagenesis characterized their possible role as antioxidants as well as in growth promotion, as probed in vitro in lettuce and in vivo in rice, bean, and tomato plants. The results showed significant differences (p ≤ 0.05) in parameters of plant height, fresh weight, and dry weight, etc., deciphering a clear and in fact complementary role of GDH and PQQ in plant growth promotion. Our study not only provides direct evidence of the in vivo role of GDH and PQQ in host plants but also reveals their functional inadequacy in the event of mutation at either of these loci.

ADAMTS4 and ADAMTS5 knockout mice are protected from versican but not aggrecan or brevican proteolysis during spinal cord injury

The chondroitin sulfate proteoglycans (CSPGs) aggrecan, versican, and brevican are large aggregating extracellular matrix molecules that inhibit axonal growth of the mature central nervous system (CNS). ADAMTS proteoglycanases, including ADAMTS4 and ADAMTS5, degrade CSPGs, representing potential targets for ameliorating axonal growth-inhibition by CSPG accumulation after CNS injury. We investigated the proteolysis of CSPGs in mice homozygous for Adamts4 or Adamts5 null alleles after spinal cord injury (SCI). ADAMTS-derived 50-60 kDa aggrecan and 50 kDa brevican fragments were observed in Adamts4-/-, Adamts5-/-, and wt mice but not in the sham-operated group. By contrast Adamts4-/- and Adamts5-/- mice were both protected from versican proteolysis with an ADAMTS-generated 70 kDa versican fragment predominately observed in WT mice. ADAMTS1, ADAMTS9, and ADAMTS15 were detected by Western blot in Adamts4-/- mice' spinal cords after SCI. Immunohistochemistry showed astrocyte accumulation at the injury site. These data indicate that aggrecan and brevican proteolysis is compensated in Adamts4-/- or Adamts5-/- mice by ADAMTS proteoglycanase family members but a threshold of versican proteolysis is sensitive to the loss of a single ADAMTS proteoglycanase during SCI. We show robust ADAMTS activity after SCI and exemplify the requirement for collective proteolysis for effective CSPG clearance during SCI.

Problems in seawater industrial desalination processes and potential sustainable solutions: a review

Seawater desalination has significantly developed towards membrane technology than phase change process during last decade. Seawater reverse osmosis (SWRO) in general is the most familiar process due to higher water recovery and lower energy consumption compared to other available desalination processes. Despite major advancements in SWRO technology, desalination industry is still facing significant amount of practical issues. Therefore, the potentials and problems faced by current SWRO industries and essential study areas are discussed in this review for the benefit of desalination industry. It is important to consider all the following five components in SWRO process i.e. (1) intake (2) pre-treatment (3) high pressure pumping (4) membrane separation (performance of membranes and brine disposal) and (5) product quality. Development of higher corrosion resistant piping materials or coating materials, valves, and pumps is believed to be in higher research demand. Furthermore, brine management, that includes brine disposal and resource recovery need further attention. Pre-treatment sludge management and reduced cleaning in place flush volume will reduce the capital costs associated with evaporation ponds and the maintenance costs associated with disposal and transportation reducing the unit cost of water. © 2013 Springer Science+Business Media Dordrecht.

Fe-bLf nanoformulation targets survivin to kill colon cancer stem cells and maintains absorption of iron, calcium and zinc

To validate the anticancer efficacy of alginate-enclosed, chitosan-conjugated, calcium phosphate, iron-saturated bovine lactoferrin (Fe-bLf) nanocarriers/nanocapsules (NCs) with improved sustained release and ability to induce apoptosis by downregulating survivin, as well as cancer stem cells.

Expression and purification of soluble recombinant full length HIV-1 Pr55(Gag) protein in Escherichia coli

The HIV-1 Gag precursor protein, Pr55(Gag), is a multi-domain polyprotein that drives HIV-1 assembly. The morphological features of HIV-1 suggested Pr55(Gag) assumes a variety of different conformations during virion assembly and maturation, yet structural determination of HIV-1 Pr55(Gag) has not been possible due to an inability to express and to isolate large amounts of full-length recombinant Pr55(Gag) for biophysical and biochemical analyses. This challenge is further complicated by HIV-1 Gag's natural propensity to multimerize for the formation of viral particle (with ∼2500 Gag molecules per virion), and this has led Pr55(Gag) to aggregate and be expressed as inclusion bodies in a number of in vitro protein expression systems. This study reported the production of a recombinant form of HIV-1 Pr55(Gag) using a bacterial heterologous expression system. Recombinant HIV-1 Pr55(Gag) was expressed with a C-terminal His×6 tag, and purified using a combination of immobilized metal affinity chromatography and size exclusion chromatography. This procedure resulted in the production of milligram quantities of high purity HIV-1 Pr55(Gag) that has a mobility that resembles a trimer in solution using size exclusion chromatography analysis. The high quantity and purity of the full length HIV Gag will be suitable for structural and functional studies to further understand the process of viral assembly, maturation and the development of inhibitors to interfere with the process.

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.

Characterization of the extracellular polymeric substances and microbial community of aerobic granulation sludge exposed to cefalexin

This study characterizes the extracellular polymeric substances and bacterial community composition of aerobic granules exposed to cefalexin (CLX). The presence of CLX potentially decreases granular stabilities, resulting in a lowered granule diameter. Chemical oxygen demand and NH4+-N removal efficiencies were slightly decreased and the denitrification process was inhibited with CLX addition. Extracellular polymeric substance contents were significantly increased in aerobic granules exposed to CLX. The shifts of fluorescence intensities and peak locations in 3D-EEM fluorescence spectra indicated changes of EPS components. High-throughput sequencing analysis showed aerobic granules with CLX addition in synthetic wastewater had superior diversity of microbial species, and this was the reason that the level and components of EPS changed. The species richness for bacteria was increased from 341 to 352, which was revealed by Chao1. The Shannon index of diversity rose slightly from 3.59 to 3.73 with CLX addition. The abundance of Proteobacteria significantly decreased, while the abundance of Bacteroidetes and Chloroflexi underwent a highly significant increase in aerobic granules exposed to CLX.