, Brazil, precision 0.002 mm), and the average of five measurements for each film was used to calculate the tensile properties. For water vapour transmission (WVT) calculations, the average of three thickness measurements of each sample was used (Kechichian, Ditchfield, Veiga-Santos, & Tadini, 2010). The mechanical properties of the films were determined by the tensile test using a Universal Testing Machine (Instron, model 3367, USA) with the following parameters: a load cell of 1 kN and a speed of 50 mm min−1. For each film, five samples with dimensions of 50 mm × 150 mm

XL184 were analysed. The tensile strength (TS, MPa) and elongation at break (E, %) values were measured. TS was calculated by dividing the maximum Dinaciclib clinical trial load by the cross-sectional area of the film, and E was calculated by dividing the extension at the moment of rupture of the specimen by the initial length of the specimen and multiplying the result by 100 ( ASTM, 2008). Mechanical analysis were performed at 0, 10, 20 and 30 days of storage. The water vapour permeability (WVP) of the films was determined according to ASTM Standard Method 96-00 (ASTM, 2000), method E96, with some

modifications. The test film was sealed in a permeation cell containing anhydrous calcium chloride. The permeation cell was then placed in a controlled temperature–humidity chamber maintained at 75% relative humidity (RH) and 25 °C to maintain a 75% RH gradient across the film. Because the

RH inside of the cell was always lower than the outside, water vapour transport could be determined based on the amount of mass gained by the permeation cell. The samples were weighed until a constant weight was reached, and the weight values were plotted as a function of time. The slope of each line was calculated by linear regression (r2 > 0.99), and the water vapour transmission rate (WVTR, g/h/m2) was calculated from the slope of the straight line divided by the exposed film area (m2). The WVP (g/(m s Pa)) of the film was calculated Isotretinoin as follows: WVP=(WVTR·x)/3600(P1−P2)WVP=(WVTR·x)/3600(P1−P2)where x is the film thickness, and P1 − P2 represents the vapour pressure differential across the film. The WVP of the films was measured at day 0. Colour was measured using the Color Quest XE colorimeter (Huber Lab) and CIELab system with a D65 light source and an observation angle of 10°. The following parameters were used: opacity, Y=(Yb/Yw)·100Y=(Yb/Yw)·100, according the relationship between the opacity of the film superposed on the black standard (Yb) and opacity of the film superposed on the white standard (Yw), and b* (yellowness). Colour analysis were performed at 0, 10, 20 and 30 days of storage. The product was assessed for sensory acceptability at a central location.

Moreover, a transcriptomic analysis of B.

granulifera was included to reveal new peptide sequence present in this sea anemone species. This is the first peptidomic and transcriptomic study of the neurotoxic fractions of these sea anemones, and the first report that compares the overall peptide composition of sea anemones species belonging to two distinct families (Stichodactylidae vs. Actiniidae). We found that the neurotoxic fraction of B. granulifera has richer peptide diversity in relation to S. helianthus, as judging by the more complex reversed-phase profile and the resulting higher number of separated peptide components (156 vs. 113 peptides) and toxic fractions (17 vs. 6). However a similar study of B. cangicum yielded a considerable smaller number of peptide components (81) than B. granulifera, despite both sea anemone species belong to the same genus and their chromatographic profiles share a similar complexity and several similarities, therefore see more such difference does not seem to arise from the use of different selleck screening library mucus extraction methods (immersion in distilled

water vs. electrical stimulation). Our study expanded to 156 the estimated maximal number of peptides in the neurotoxic fraction of sea anemones. We emphasize the term “maximal number” as we showed that venom peptide diversity varies among sea anemone species. Moreover, likewise the previous study [85] we found some apparent venom composition overlaps. Structural studies will confirm whether a single neurotoxic peptide is present in two or more sea anemone species. Peptide toxins previously isolated and characterized from S. helianthus and B. granulifera were identified in the present study, with the exception of ShK [14] and ShPI-1 [22]. These toxins seem to be poorly represented in the S. helianthus exudate so it was not possible to detect them by mass spectrometry. ShK occurs in very low amounts either in freeze-dried mucus or in whole Paclitaxel ic50 body extract [14], so its purification included a precipitation step by heating the sample at low pH, prior to the chromatographic protocol. Likewise, the isolation of

ShPI-1 comprised a precipitation step (trichloroacetic acid treatment) before the chromatographic separation which included affinity chromatography [22], utilized in many instances as a powerful purification method when the protein of interest is a minor component of a complex mixture [13]. Our study confirmed the presence of a very distinguishable feature among sea anemone species of the genus Bunodosoma, a group of abundant and hydrophobic 4–5 kDa peptides that elute in the last reversed-phase fractions ( Fig. 2 and Fig. 3), so far comprising type 1 sodium channels toxins and APETx-like peptides. The sodium channel toxins are BcIII from B. caissarum [55], Bcg 28.19 and Bcg 30.24 from B. cangicum, BgII and BgIII from B. granulifera. The APETx-like peptides are BcIV from B. caissarum [64], Bcg 31.16, Bcg 28.78, Bcg 25.

The size of the nodes corresponds to the number of genes of the gene set, and the thickness of the connecting lines indicates the degree of overlap between the gene sets. The color of the nodes corresponds to the gene set collection from which the gene sets were taken. Green: lymphocyte signature database; yellow: TOX TFS target genes; purple: gene ontology; light blue: cell cycle; dark blue: tissue-specific blood cell types. The authors thank Hakan Baykus, Jenneke Riethoff-Poortman, and Norbert de Ruijter Palbociclib solubility dmso for their technical support and Wilma Blauw and Bert Weijers of the Small Animal Center of Wageningen University (Wageningen, The

Netherlands). Sandra W.M van Kol is recipient of grant MFA6809 from the Dutch technology foundation STW. “
“The authors click here regret that in the Abstract, Materials and methods, and Results sections, the unit of PCB126 concentration was incorrect. This has now been corrected below. 1. In the abstract, the PCB126 concentration should read nM and not pM. The authors

deeply regret any inconvenience this mistake may have caused and would like the readers to have the correct information. “
“Organophosphorus (OP) compounds, including pesticides and chemical warfare nerve agents (CWNAs), represent a threat to the general population, not only as possible weapons of terrorism (Okumura et al., 2005, Zurer, 1998, Hubbard et al., 2013, Baker, 2013 and Dolgin, 2013), but also as chemicals that could be released from transportation and storage facilities during industrial accidents. Given the rapid onset of symptoms and toxicity of OP nerve agents, a quick-acting therapeutic regimen that is efficacious over the broad spectrum of OPs is needed. To provide the most effective therapy,

medical countermeasures must be administered as soon as possible post-exposure. The current U.S. therapy regimen includes the administration of atropine in combination with the oxime acetylcholinesterase (AChE) reactivator pralidoxime chloride (2-PAM Cl) (Inchem.org, Calpain 1989, 1999), followed by the anticonvulsant diazepam depending on whether convulsive symptoms are observed. This approach is accomplished with the use of the DuoDote® autoinjector kit (Meridian Medical Technologies™, Columbia, MD; https://www.duodote.com/meridian.aspx#) by trained emergency medical services personnel. The DuoDote® is a two-chambered, self-propelled syringe used for the intramuscular (IM) injection of atropine (2.1 mg free base) and 2-PAM Cl (600 mg) through the same needle. Although the current treatment approach does protect against some OP toxicities, this protection does not extend across all OP CWNAs, i.e., it is not a broad-spectrum antidote (Worek and Thiermann, 2013 and Thiermann et al., 2013). Unfortunately, when OP pesticides are included as potential intoxicants, the spectrum of therapeutic effectiveness is even less.

4). This passive effect in nest thermoregulation is considerably higher in wasps than in honeybees (see insert of Fig. 4; compare also Kovac et al., 2007). A wasp RQ below 1 would shift the curve of wasp metabolism in terms of O2 consumption to even higher values, and this way increase the difference in energy turnover between bees and wasps. In phases of regulated nest temperature, therefore, a certain number of ectothermic wasps produce a higher amount of heat than the same number of ectothermic individuals in honeybee colonies at a certain ambient temperature.

This has also the consequence that fewer wasps are needed for active (endothermic) click here heat production. Relatively few thermally active wasps may take away much burden from other individuals which can stay

passive. At the upper range of Cyclopamine in vitro experimental temperatures (from ∼35 °C upwards) the wasps showed rest only sparsely. Both, number and duration of resting periods decreased with rising Ta and agitated movement predominated. Furthermore, many individuals showed cooling behavior, an indication that the individuals were not comfortable under these circumstances, and mainly wanted to escape the hostile environment. From 39.7 °C onwards only 37.5% (3 of 8 individuals) of the wasps could be measured in a true resting state ( Fig. 4, crossed boxes), all other individuals were measured during “rest” in their “deleterious range” ( Klok et al., 2004) or heat stupor ( Fleurat-Lessard and Dupuis, 2010), right after cyclic respiration had ceased (see Fig. 6, after stage 4). Other individuals tested did not show rest at all at these high temperatures clonidine and therefore were not included in this study. As a consequence, one could reason that Vespula generally does not show resting behavior at ambient temperatures above Ta ≈ 40 °C ( Fig. 4, dashed line). In any case occasional rest (observed only for one or two minutes) at these temperatures is at a very high energetic level. With rising ambient temperatures, an increasing number of individuals did not survive the experiments (see Fig. 4, mortality

in %) in spite of Ta being way under their CTmax (see Table 1). The time of exposure obviously plays a considerable role in the wasps’ thermal tolerance when Ta reaches the upper edge of viability (compare e.g. Terblanche et al., 2011 and Willmer et al., 2004). Activity CTmax (“knockdown temperature” as defined by Klok et al., 2004) and respiratory CTmax (“mortal fall”, ( Lighton and Turner, 2004)) of V. vulgaris were proved to be within narrow thermal margins (average 0.4 °C, Table 1). This has to be expected under normobaric conditions ( Stevens et al., 2010). The use of the residual of the absolute difference sum of CO2 production (rADS residual, see Fig. 6) proved eligible in determining the end point of cyclic respiration and respiratory CTmax.

0 array, and scanned as described previously (Steiner et al., 2004 and Wodicka et al., 1997). Gene expression profiles of the LPS- and LPS/Dex-treated human 3D co-cultures were compared to the ones of vehicle-treated

cells. Genes were considered changed, if they showed an at least 2 fold change (p ≤ 0.05, Student’s Selleck Venetoclax t-test) compared to vehicle treated cells upon LPS or LPS/Dex treatment. All treatments were performed in biological triplicates and each RNA sample was hybridized on a separate microarray. The original gene array data are available in Supplementary Fig. 3. For immunochistochemistry, 30-days-old human or 20-days-old rat 3D liver cells were washed in PBS and fixed in 4% paraformaldehyde (PAF)

for 30 min and washed three times with PBS. Cell permeabilization and blocking of the non-specific antibody-binding was performed in PBS containing 1% bovine serum albumin (BSA, Sigma) and 0.1% Triton X-100 (Fluka) for 1 h at RT. Permeabilized cells were incubated with primary antibodies against mouse albumin (1:100; cat #: A6684; Sigma), rat F4/80 (1:10; cat #: ab16911; Abcam), rabbit vimentin (1:50, cat #: 3932, Cell signaling), rabbit intercellular adhesion molecule 1 (ICAM-1) (1:100; cat #: HPA002126; Sigma) and goat dipeptidyl peptidase IV (DPPIV) (1:10; cat #: AF1180; R&D systems) overnight at 4 °C. The cells were then washed three times with PBS and incubated at RT for 1–2 h in the dark with the secondary antibodies: donkey anti-mouse CHIR-99021 chemical structure IgG (H + L) AlexaFluor 568 (1:200; cat #: A10037; Invitrogen, Molecular probes), Sheep F(ab′)2 anti-rabbit IgG (H + L) (Cy3 ®) (1:50; cat #: ab50503; Abcam), rabbit anti-rat IgG AlexaFluor 594 (H + L) (1:200; cat #: A21211; Invitrogen, Molecular probes) and rabbit anti-goat AlexaFluor 568 (1:200;

cat #: A11079; Invitrogen, Molecular probes). After an additional washing with PBS, the nuclei were counterstained with 300 nM DAPI for 1 h. This was followed by transfer of the screens containing the cells into microscopic glass slides and mounting the cells with Vectashield medium (Vector laboratories, Inc.). The specimens were examined using a confocal microscope (Leica DMI 4000B). To label Kupffer cells in human 3D co-culture, they were incubated with 4 μl fluorescence-labeled latex beads (cat #: 17154, Polysciences, Inc.) in 1 ml PJ34 HCl serum containing media for 1 h at 37 °C and subsequently washed extensively with PBS. To quantify the number of hepatocytes and Kupffer cells, flow cytometry analysis was performed from human 3D liver co-cultures. First, cells were washed three times with PBS and then detached from the scaffolds by 20 min incubation with Accutase (PAA Laboratories, GmbH) at 37 °C. Dissociated cells were centrifuged for 5 min at 1200 rpm, fixed with 2% PAF for 15 min at RT, and then permeabilized and blocked in 10% normal goat serum (NGS)/0.1% saponin in PBS for 1 h at RT.

cruzi antibody (produced in our laboratory, LBI/IOC-Fiocruz, Brazil), as previously described ( Silva et al., 1999). For confocal microscopy, parasite antigens were revealed with the same anti-T. cruzi antibody except that the secondary antibody goat anti-rabbit immunoglobulin

was labeled with FITC or TRITC (Amersham, England). Astrocytes and microglial cells were revealed with purified anti-glial fibrillary acidic protein (GFAP) antibody (Amersham, England) and purified anti-F4/80 rat antibody (Caltag, USA), respectively. Secondary anti-rat immunoglobulins labeled with FITC or TRITC (Amersham, England) were used to reveal glial cells. For positive controls, heart tissue sections from T. cruzi-infected mice at 30 dpi were used. For negative controls, brain tissue sections from infected mice were subjected to all the steps of the reaction excluding the addition selleck chemicals of the primary antibodies. The images were analyzed with a confocal microscope (LSM 410, Zeiss, Germany). The presence of T. cruzi antigens in brain tissue sections was also evaluated with a digital morphometric apparatus. The images were analyzed

with the AnaliSYS Program and the areas containing parasite molecules were identified as amastigote nests in microscopic fields. Three whole sections were analyzed per brain. TNF was assayed with the ELISA sandwich development kit assay from R&D (catalog # 900-k57 lot # 0104054) with rat anti-TNF mAb and a biotin-labeled polyclonal rabbit serum specific for the cytokine. TNF levels were calculated by reference to a standard curve selleckchem constructed with recombinant cytokine. The sensitivity of DCLK1 this method was 10 pg/mL. The assay was developed using the 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) substrate (Sigma, USA) and the reaction was stopped with 20 μL of 20% sulfuric acid solution. The optical density (OD) was read with a microplate reader set to 405 nm. For reverse transcriptase PCR (RT-PCR), mRNA was isolated from the whole encephalon and heart tissue of the C57BL/6 mice by acid guanidinium thiocyanate–phenol–chloroform

extraction. The RNA STAT-60 reverse transcriptase-PCR conditions, primer sequences used for the detection of TNF, housekeeping gene hypoxanthine–guanine phosphoribosyltransferase (HPRT) and PCR product sizes have been published elsewhere (dos Santos et al., 2001). The PCR products and a molecular weight marker were electrophoresed in 6% polyacrylamide gel and stained with silver nitrate. The densitometry analysis of the gels was conducted on a Densitometer CS-9301PC (Shimadzu, Japan). The PCR data were standardized using mRNA of the housekeeping gene HPRT and fold increases were determined by a comparison with NI controls. For real-time quantitative RT-PCR (RT-qPCR), total RNA from heart and whole brain samples was extracted using TRI Reagent (Sigma–Aldrich, USA).

Vicinal dithiols, which are likely to form intraprotein disulfides because of their proximity, can be identified on the basis of a selective labeling and reduction strategy. Protein dithiols in reduced protein samples can be selectively blocked with the dithiol specific reagent phenylarsine oxide (PAO) and then all other thiols alkylated with

NEM. Subsequently, PAO-blocked dithiols are selectively reduced using the PAO-specific reducing agent 2,3-dimercaptopropanesulfonic acid (DMPS) and labeled with an alkylating probe [19, 46 and 47]. Identification of novel proteins that undergo inter-protein disulfide formation is also possible using diagonal electrophoresis [48]. Protein samples are first resolved by non-reducing SDS-PAGE so that all thiol modifications remain intact. Then samples are resolved in the second dimension with DTT incorporated into the running medium. By incorporating the reduction Veliparib step at this point, proteins involved in inter-protein disulfide linkages will migrate off the diagonal and can be subsequently identified by peptide mass fingerprinting or with an antibody on a western blot if candidate proteins are suspected. The reliance of this technique on electrophoresis limits the potential resolving power for complex protein mixtures. This lack of sensitivity can be addressed to some extent if a thiol specific fluorescent probe

is incorporated during the reduction step. Although this would focus on the cysteine residues, NVP-BEZ235 chemical structure in this case other thiol modifications in addition to inter-protein disulfides would also be labeled. As both the glutathione and thioredoxin systems are critical for the maintenance of protein thiol redox homeostasis, techniques Nintedanib (BIBF 1120) have been developed to identify the protein targets of these interactions.

Lind et al. used a mutant glutaredoxin from E. coli to selectively reduce glutathionylated proteins following the general scheme described in Figure 3b [ 49•]. Although this strategy may identify constitutively glutathionylated proteins it is unclear if the mutant glutaredoxin is capable of reducing all glutathionylated proteins. Sensitive strategies for the identification of thioredoxin-conjugated proteins have relied on the blocking of unmodified thiols, followed by the treatment of oxidized thiols ± thioredoxin and blocking of thioredoxin-reduced thiols. Finally, oxidized thiols not affected by thioredoxin treatment are reduced and labeled resulting in a signal [ 50•]. Decreased signal probe intensity in thioredoxin treated samples is indicative of a target cysteine residue. Recently, Benhar and colleagues used a combined strategy of selective reduction of protein S-nitrosothiols and thioredoxin conjugation to specifically determine S-nitrosated targets of thioredoxin action [ 51]. Using stable isotope labeling by amino acids in cell culture (SILAC), entire proteomes can be differentially labeled with light or heavy lysine.

Today, however, post distemper, Baltic Sea and Kattegat populations are increasing by ∼12% per year. Although hunting

is illegal in the United Kingdom and in Norway and Canada (but not culling), it is legal to kill seals perceived to threaten fisheries. More of that subject later. In the United Kingdom, moreover, seals are protected by the 1970 Conservation of Seals Act, which prohibits most other forms of killing. Seals are also protected BMS-354825 datasheet throughout the European Union (Council Directive 83/129/EEC of 28 March 1983, and subsequent amendments). Similarly, in the United States the 1972 Marine Mammal Protection Act prohibits the killing

of any marine mammals. In the Western North Atlantic, the grey seal occurs typically in large numbers in the coastal waters off Canada and south to about New Jersey in the United States. Here, harbour seal numbers are increasing as, also post-distemper, they reclaim parts of their range, which naturally extends south to North Carolina. The largest colony is on Sable Island in Nova Scotia. Numbers of the spotted seal (Phoca largha) population in the Wadden Sea have, similarly, increased to a level 50% larger than find more just before the last distemper outbreak of 2002. In the Baltic, Kattegat and Limfjorden and Jutland (Denmark), the spotted seal population increased by 80% between 2010 and 2012. Etoposide ic50 The intrinsic rate of increase in this species is 12% per year, largely, it is believed, through immigration from the North Sea. As with the harbour and spotted

seals in Europe, grey seal numbers are increasing rapidly in the United States. The Marine Mammal Protection Act rescued their dwindling stocks and, today, there is a large breeding colony near Cape Cod, Massachusetts, where pups have rebounded from a handful in 1980 to more than 2,000 in 2008. By 2009, thousands of grey seals had taken up residence there either on or near popular swimming beaches and great white sharks, Carcharodon carcharias, are said to have started hunting them close to shore, as they must have done in years past. In recent years too, the number of grey seals in Canadian waters has been increasing and, predictably, there have been fisheries calls for a cull. Colonies also exist off the islands of Sylt and Amrum and on Heligoland in the German Bight of the eastern North Sea. Similarly, the grey seal population in the Baltic Sea grew fast (>10% per year) between the early 1990’s and the mid-2000’s. Subsequently, the rate decreased a few years ago to ∼6%, but the population has begun to increase again and numbers recorded in 2012 were the highest ever.

On the other hand, vit E (low dose) and Se (Low dose) elicited slight significant decrease in GPx activity

as compared to normal control group. Meanwhile, MSG (high dose) followed by administration of either vit E (high or low dose) or followed by Se at high or low dose elicited slight decrease in GPx activity with respect to normal control group. At the end of the 30 days of treatment, the spermatogenic cells in the seminiferous tubules appeared to have normal histological structure in control group. Testis of the male control treated rats appears to be oval in shape and showed normal seminiferous tubules – surrounded by few edematous stroma containing www.selleckchem.com/products/ch5424802.html small groups of leydig cells (Fig. 1A). In the MSG-treated group with high dose, seminiferous tubules were observed

filled by spermatogenic cells with few sperm formation and showing seminiferous tubules filled by spermatogonia only with few sperm formation (Fig. 1B and1 C). Meanwhile, in group 3 (vit E treated group ZD1839 ic50 high dose) there were testicular tissues showed normal seminiferous tubules surrounded by small groups of leydig cells (Fig. 1D). On the other hand, in group 4 (Se-treated group high dose), testicular tissues showed seminiferous tubules lined by layers of spermatogenic cells up to sperm formation (Fig. 1E). While, group 5 treated with (MSG + vit E at high dose) showed seminiferous tubules lined by few layers of spermetrogenic cells and few sperms (Fig. 1F). (-)-p-Bromotetramisole Oxalate While group 6 treated with (MSG + Se at high dose) showed seminiferous tubules lined by few layers of spermetrogenic cells and moderate number sperms (Fig. 1G). Fig. 2, Fig. 3, Fig. 4 and Fig. 5 Lipid peroxidation is one of the main processes of oxidative damage, which plays a critical role in the toxicity of many xenobiotic (Ognjanović et al., 2010). It was evaluated by assessment of TBARS [36]. In the present study, TBARS levels also

increased in the MSG treated rats. It is known that MSG produces reactive oxygen species. Therefore, antioxidant enzymes could play a crucial role on MSG toxicity [37]. Our results was in harmony with Tezcan et al. [38] who declared that MDA is one of the final decomposition of lipid peroxidation and it is also formed as a product of the cyclooxygenase reaction in prostaglandin metabolism and this assure our finding who conclude the presence of oxidative stress in rats treated with MSG in which there was high level of MDA. In agreement with previous study, the susceptibility of spermatozoa to oxidative stress as a consequence of the abundance of unsaturated fatty acids in the sperm plasma membrane and a very low concentration of cytoplasmic antioxidants is well known [39]. We demonstrated that the major reason for damage of testicular tissues is the increasing level of lipid peroxidation and these findings was in parallel with Aitken et al.

This is widely known. Well established journals seem to accept structural work if the SDS-PAGE (with Coomassie Blue stain) show >95% purity. There is another disturbing practice which is occasionally

seen that the band of the protein is shown at far end of the lane. This rules out detecting the presence of any proteolytic fragments or contaminating proteins Venetoclax mouse of lower molecular weight. Not all applications of the proteins require the same level of purity. This is an important point since there is a three way trade-off between purity vs. number of steps vs. cost of production (Figure 1). Industrial enzymes used in many industries do not require high purity. Reasonable level of specific activity Dabrafenib solubility dmso is sufficient. Proteins used for pharmaceutical applications (e.g. monoclonal antibodies or clot busters, hormones, etc.) not only require extremely high purity; regulatory agencies require that these preparations are specifically free of certain contaminants (Anicetti and Hancock, 1994 and Walsh and Headon, 1994) (Table 2). There is also a fairly widespread practice of measuring Km, Vmax and stability of proteins which are fairly impure. Unless, the preparation is standardized with respect to contaminants (like in commercially available industrial enzymes), such data actually cannot be relied upon (the reason for this is explained

later on). Finally, as may be clear from the above discussion, protein purity is a relative term. One of the most well characterized enzymes is bovine pancreatic RNase A (Richards and Wyckoff, 1971). Most of the work, including X-ray crystallography, has been carried out with a “pure” preparation obtained by many a final ion-exchange chromatographic step (Richards and Wyckoff, 1971). However, this preparation shows multiple proteins when subjected to multiple counter-current distribution process (Richards and Wyckoff, 1971)! In general, crystallization can be both a purification strategy (Przybycien et al., 2004) as well as a criterion of reasonable purity (Dixon et al., 1979). Precipitation,

both with and without an interface with affinity interactions is another efficient, simple and scalable approach (Mondal et al., 2006, Mondal and Gupta, 2006 and Niederauer and Glatz, 1992). Most of the industrial enzymes these days are produced by recombinant methods wherein overexpression leads to a considerably less heterogeneous protein preparation. Many proteins upon overexpression in Escherichia coli as host end up as inclusion bodies. In recent years, in many cases these inclusion bodies are being considered as carrier-free immobilized preparation of fairly pure enzymes ( Garcia-Fruitos et al., 2012). One of the key parameters in biocatalysis is the amount of protein present in the biocatalyst preparation.