Restriction patterns revealed a diverse plasmid pool present in these strains. Plasmids were assigned to FrepB (Aeromonas salmonicida,Aeromonas veronii,Aeromonas sp., E. coli,Enterobacter sp.), FIC (A. salmonicida,Aeromonas sp.), FIA (Shigella sp.), I1 (A. veronii,Aeromonas sp., E. coli), HI1 (E. coli) and U (Aeromonas media) replicons. Nevertheless, 50% of the plasmids could not be assigned to any replicon type. Among integron-positive transconjugants, FrepB, I1 and HI1 replicons

were detected. Results showed that wastewaters enclose a rich plasmid pool associated with integron-carrying bacteria, capable of conjugating to different bacterial hosts. HDAC inhibitors cancer Moreover, replicons detected in this study in Aeromonas strains expand our current knowledge of plasmid diversity in this genus. Identification and classification of plasmids has been an important issue in recent decades to trace plasmid evolutionary origins and to elucidate their role in environmental processes and microbial adaptation (Johnson & Nolan, 2009a). Classification is usually based on genetic traits CX-5461 ic50 related to plasmid maintenance and replication control. Plasmids that use the same replication system belong to the same incompatibility group and compete for stable maintenance. Therefore,

plasmids belonging to the same incompatibility group cannot stably coexist in the same cell, although their accessory genes may be different (Couturier et al., 1988). The importance of plasmids in bacterial adaptation has been reported in several environments, such as soil (Lee et al., 2006), rivers (Shintani et al., RG7420 ic50 2008) and wastewaters (Verma et al., 2002). Despite the energetic burden, plasmids provide a fitness advantage to their hosts which allow them to persist across bacterial generations (Dionísio et al., 2005). The genetic traits harboured on plasmids may include genes involved in mechanisms such as resistance, energy metabolism, virulence, pathogenicity, symbiosis and/or dissemination, favouring the survival of bacterial hosts under selective pressures (Dionísio et al., 2002). Conjugation is considered a major pathway for horizontal gene transfer (HGT) among bacteria (Sørensen

et al., 2005). It involves direct cell-to-cell contact and DNA exchange usually mediated by a conjugative plasmid. Conjugative plasmids can be highly promiscuous and transfer may occur between different genera or even domains (Ochman et al., 2000). Antibiotic resistance plasmids are found in several bacterial genera, both Gram-negative and Gram-positive. Because of their wide distribution and because they may confer multiple resistance phenotypes, resistance plasmids are of both clinical and environmental concern. Several plasmid families carrying multiple antibiotic resistance determinants have been reported in Aeromonas spp. (Sørum et al., 2003; Picão et al., 2008; Fricke et al., 2009) and Enterobacteriaceae isolates (Carattoli, 2009).

The typical pharmacy-based EC consumer in this study had a tertiary education and worked either Quizartinib part-time or full-time.

Our findings are consistent with those of an international systematic review in which Anderson and Blenkinsopp established that women who request EC from pharmacies are generally better educated, working, possibly of a higher socioeconomic background and prefer to use a pharmacy on the basis of ease of access.[12] Ease of access is of particular importance in relation to EC, where the time elapsed between sexual intercourse and obtaining EC is a critical factor. We believe that almost all the women in our study said that they found the pharmacy very easy/easy to access for EC because most pharmacies have a high street presence, are open long evening and weekend hours and do not require them to make an appointment for an EC consultation. To determine whether EC should be dispensed, pharmacists are required to conduct a sexual health consultation to identify the women’s risk of pregnancy. We found that the majority of women said they felt very comfortable/comfortable discussing sexual health and EC with the pharmacist. Most women in our study also said that this

was not their first experience of obtaining EC and the majority said they previously got EC from a pharmacy, possibly indicating a preference for an accessible venue that is not outside their daily routine. However, we also found that nearly 30% of the women BTK inhibitor said they were very concerned/concerned about privacy in the pharmacy. This, and the fact that EC consultations are often in-depth and of a personal nature, suggest that it may be necessary to improve the level of anonymity and privacy in community pharmacies. Most pharmacy-based EC consumers said that they would be willing to accept a chlamydia test from the pharmacy; however, the

proportion was significantly higher for women attending rural, regional and remote WA pharmacies when compared to the Perth metropolitan region (P < 0.05). This could be an indication that women in rural and remote areas may have fewer options for sexual health services and prefer pharmacies on the basis of ease of access and longer opening hours. As discussed above, most had also indicated that they felt comfortable discussing sexual Isoconazole health-related issues with pharmacists, making pharmacies an obvious choice for chlamydia screening. Evidence also suggests that GPs and pharmacists think offering a chlamydia test with a sexual health consultation is highly appropriate.[18-20, 30, 31] A recent study found that Australian GPs believed chlamydia screening should be opportunistically offered during a sexual health consultation.[31] Similarly, four different pharmacy-based chlamydia screening studies also found that pharmacists preferred offering women a chlamydia test during an EC consultation.

The typical pharmacy-based EC consumer in this study had a tertiary education and worked either find more part-time or full-time.

Our findings are consistent with those of an international systematic review in which Anderson and Blenkinsopp established that women who request EC from pharmacies are generally better educated, working, possibly of a higher socioeconomic background and prefer to use a pharmacy on the basis of ease of access.[12] Ease of access is of particular importance in relation to EC, where the time elapsed between sexual intercourse and obtaining EC is a critical factor. We believe that almost all the women in our study said that they found the pharmacy very easy/easy to access for EC because most pharmacies have a high street presence, are open long evening and weekend hours and do not require them to make an appointment for an EC consultation. To determine whether EC should be dispensed, pharmacists are required to conduct a sexual health consultation to identify the women’s risk of pregnancy. We found that the majority of women said they felt very comfortable/comfortable discussing sexual health and EC with the pharmacist. Most women in our study also said that this

was not their first experience of obtaining EC and the majority said they previously got EC from a pharmacy, possibly indicating a preference for an accessible venue that is not outside their daily routine. However, we also found that nearly 30% of the women selleck products said they were very concerned/concerned about privacy in the pharmacy. This, and the fact that EC consultations are often in-depth and of a personal nature, suggest that it may be necessary to improve the level of anonymity and privacy in community pharmacies. Most pharmacy-based EC consumers said that they would be willing to accept a chlamydia test from the pharmacy; however, the

proportion was significantly higher for women attending rural, regional and remote WA pharmacies when compared to the Perth metropolitan region (P < 0.05). This could be an indication that women in rural and remote areas may have fewer options for sexual health services and prefer pharmacies on the basis of ease of access and longer opening hours. As discussed above, most had also indicated that they felt comfortable discussing sexual Demeclocycline health-related issues with pharmacists, making pharmacies an obvious choice for chlamydia screening. Evidence also suggests that GPs and pharmacists think offering a chlamydia test with a sexual health consultation is highly appropriate.[18-20, 30, 31] A recent study found that Australian GPs believed chlamydia screening should be opportunistically offered during a sexual health consultation.[31] Similarly, four different pharmacy-based chlamydia screening studies also found that pharmacists preferred offering women a chlamydia test during an EC consultation.

Escherichia coli strains were grown in Luria–Bertani (LB) medium. Listeria monocytogenes was grown in brain heart infusion (BHI) broth. Erythromycin (Ery) and chloramphenicol (Cm) were made up as concentrated stocks, and then added to media at the required levels. Where solid media was required, agar was added at 1.5%. Zinc limiting media was achieved via the addition of 0.5 M EDTA at the required concentration

followed by agitated overnight incubation to allow complete chelation. Where metals were VE-821 chemical structure used, they were prepared as 1 M stocks according to the manufacturer’s instructions and were added to media at the required concentrations. Gel extractions were performed using the Qiagen gel extraction kit (Qiagen). Plasmid DNA isolation was achieved utilizing the Qiagen QIAprep spin miniprep kit (Qiagen). Ligations were carried out using T4 DNA Ligase from Roche MAPK inhibitor Diagnostics GmbH (Mannheim, Germany) according to the manufacturer’s instructions. Restriction enzymes were purchased

from New England Biolabs and were used according to the manufacturer’s recommendations. The splicing by overlap extension (SOEing) PCR procedure described by Horton et al. (1990) was used to create an isogenic nonpolar deletion mutant of zurR. Primers SOE AB and SOE CD were used to amplify regions of similar size flanking the sequence to be deleted. The resulting products were mixed in a 1 : 1 ration and were amplified using SOE A and D primers. The resulting product was digested with the appropriate restriction enzymes (XbaI and EcoRI), cloned into the temperature sensitive shuttle vector pKSV7, and transformed into E. coli DH5α.

The plasmid was subsequently transformed into L. monocytogenes EGDe. Chromosomal integration of the plasmid at 42 °C was selected by serial passage of a single colony in prewarmed BHI/Cm broth and streaking onto prewarmed BHI/Cm agar. Plasmid excision and curing was brought about by continuous passaging in BHI at 30 °C followed by spread plating onto BHI agar at 30 °C. Replica plating onto BHI and BHI/Cm was used to select for loss of plasmid and appropriate deletion events. The mutation was ioxilan confirmed using primers upstream of SOE A (SOE X) and downstream of SOE D (SOE Y) (listed in Table 1). The pORI19 insertional mutant was constructed as outlined previously (Law et al., 1995; Rea et al., 2004). Briefly, a central portion of the zurR gene was amplified by PCR and cloned into the multiple cloning site of pORI19 (RepA−) and maintained in E. coli EC101. The plasmid was transformed by electroporation into L. monocytogenes EGDe containing the temperature sensitive RepA+ plasmid pVE6007. Loss of pVE6007 and integration of pORI19 to create zurR::pORI19 was carried out as described previously (Rea et al., 2004). No obvious specific ribosome-binding site was determined within zurM and zurR, which suggests a coupled translation mechanism (Dalet et al., 1999).

3% of patients (95% CI = 43.5 – 83.7%) (19 medications) with at least one moderately severe discrepancy and 45.5% of patients (95% CI = 24.6 – 66.3%) (10 medications) with a minor discrepancy. The results shows that discrepancies occur between the hospital discharge prescription and the patient’s further supply of medication as reported by the parent. The results indicate that 36.8% of patients experienced discrepancies post hospital discharge which is higher Selleckchem BTK inhibitor than the 14.1% of post discharge discrepancies experienced in older adults aged 65 from an adult study[2]. The results indicate that 7.6% of

patients (95% CI = 1.1% – 16.0%) who are discharged will experience an unintended moderately severe medication discrepancy post discharge. 1. Dean BD, Barber N. A validated reliable method of scoring the severity of medication

errors. selleckchem American Journal of Health-System Pharmacists. 1999; 56: 57–62. 2. Coleman EA, Smith JD, Raha D, Min S-J. Post hospital medication discrepancies. Prevalence and contributing factors. Archives of internal medicine. 2005; 165: 1842–1847. Rowan Yemm1, Debi Bhattacharya1, David Wright1, Anne Regan2, David Green2, Lorna Hollister2 1University of East Anglia, Norwich, Norfolk, UK, 2Colchester Hospital University NHS Foundation Trust, Colchester, Essex, UK In recent guidance, RPS emphasises the importance of communicating medication changes at discharge Charts were amended to allow better annotation of changes, which it was hoped would translate onto Electronic Discharge summaries (EDS) Whilst changes on charts increased from 51.7% to 62.8%, no improvement was seen on EDS or the general practitioner’s (GP) list after discharge More work is needed to establish how, if not from charts, doctors source information for preparing EDS. In 2011, RPS published guidance to support the transfer of care1, which emphasises the importance of communicating changes in medication across the interface. Colchester hospital volunteered as an early adopter, participating in a 6-month programme to assess the effect of amending inpatient medication charts during to include additional fields for annotating medication changes. This

study aimed to investigate the effect of these amendments on the annotation of new medicines, dose changes and discontinuations on charts, and whether this improves the quality of information provided on EDS, and GP-held information after discharge. Data were collected over 7-day periods in November 2011 (pre implementation), March and May (2 & 4 months post implementation) from 2 medical wards purposively selected for high patient turnover. Charts and EDS for patients discharged from study wards during data collection were reviewed. Researchers identified where medication changes had occurred, and whether these were annotated in new chart fields and explicitly stated on EDS. Fisher’s exact test was used to compare proportions. Short-term changes (e.g.

, 2002). However, altered fixation behavior has also been observed using entirely non-social materials (Joseph et al., 2009), suggesting a more general difference in visuo-motor processing in ASD. In the few published examples of eye-traces in ASD participants, fixations are often only slightly away from targeted

regions of interest (Pelphrey et al., 2002; Rice et al., 2012). Higher variability of saccadic amplitude has also been observed within (Takarae et al., 2004) and between autistic participants http://www.selleckchem.com/products/AZD8055.html (Goldberg et al., 2002; Takarae et al., 2004; Stanley-Cary et al., 2011). Collectively, these studies suggest that the precision of eye movements is less reliable in ASD. Much of the early visual cortical hierarchy is organized into a series of precise retinotopically mapped regions. A key aspect of these maps is that they display the so-called cortical magnification factor, which describes the fact that the region of visual space that we foveate on has considerably more cortex devoted to its processing than have more peripheral regions (Daniel & Whitteridge, 1961; Tootell et al., 1982). This non-uniformity of cortical representation is very dramatic indeed, such that in the squirrel

monkey, a 1° stimulus presented at fixation would activate approximately 42 mm2 of primary visual cortex (V1), whereas the same stimulus presented GSK126 purchase at 6° eccentricity would activate only about 1.5 mm2 (Adams & Horton, 2003), a near 30-fold difference in representation. The implications for macroscopic recordings of visual activity at the scalp surface are clear. Presentation of a stimulus at fixation will result in activation of a very large patch of early visual cortex relative to when it is presented at more peripheral Meloxicam locations, which is clearly borne out as a sharp amplitude decrease in the visual evoked potential (VEP)

under such circumstances (Schlykowa et al., 1993; Jedynak & Skrandies, 1998). Early visuo-spatial maps are developmentally driven, as is abundantly evident in patients with amblyopia, where functional imaging has shown that V1 receptive fields are shifted to represent more parafoveal locations for the strabismic eye (Conner et al., 2007). Even in neurotypical adults, rapid changes in the mapping of perceptual space can be induced by experimentally altering the relationship between eye movements and visual stimulation (Awater et al., 2005). It seems a reasonable proposition then that the titration of cortical space representation in early visual regions develops during infancy and early childhood and that this development is strongly influenced by the fidelity of the eye-gaze system.

The two major calpain isozymes are μ-calpain

and m-calpain, activated by micromolar and millimolar Ca2+, respectively. Activated calpain causes a limited degradation of a variety of proteins (cytoskeletal proteins, membrane integral proteins, certain enzymes, transcription factors, components in cell adhesion and Bioactive Compound Library signaling pathways). The ratio of calpastatin to calpain varies among tissues and species, and is an important factor in the control of calpain activity within the cell. The calpain–calpastatin system has been implicated in a variety of cellular physiological and pathological processes such as cell motility, myoblast fusion, signal transduction pathways, neurotoxicity, apoptosis and necrosis (Barnoy et al.,

1998; Goll et al., 2003; Nixon, 2003; Orrenius et al., 2003; Das et al., 2006; Liu et al., 2008). SH-SY5Y cells have been widely studied in connection with neuronal development and differentiation, and have been used as a cellular model for investigations on the calpain system in neuroblastoma and in neurodegenerative disorders (Grynspan et al., 1997; Hoerndli et al., 2004; Das et al., 2006). During a preliminary study on the effects of Bortezomib clinical trial amyloid-β-peptide (Aβ) on the calpain–calpastatin system in SH-SY5Y neuroblastoma cells, we unexpectedly found that calpastatin protein levels were increased in some samples of the cultured cells, as compared with the levels in other samples (E. Elkind, T. Vaisid, S. Barnoy & N.S. Kosower, unpublished data). The elevated calpastatin levels could not be explained by the culture conditions per

se. We were unaware of the fact that some of the cell culture samples we had then were contaminated with Olopatadine mycoplasma. Subsequently, when the diagnosis of mycoplasma contamination of these cells was established, we carried out a study of the calpain–calpastatin system in mycoplasma-contaminated SH-SY5Y cells. Mycoplasmas (class Mollicutes) are the smallest self-replicating, wall-less prokaryotes widely distributed in nature. They have limited biosynthetic abilities and most are parasites, exhibiting host and tissue specificities. Almost all of the mycoplasmas adhere to the surface of eukaryotic cells. Adherence of these organisms to the cells is essential for tissue colonization and the subsequent development of disease (Rottem, 2003). Some species may invade the cell (Rottem, 2003; Yavlovich et al., 2004). Mycoplasmas contaminate cultured cells, leading to a variety of alterations in the cells, including alterations in gene expression, protein synthesis, cell membrane composition and changes in signal transduction (Drexler & Uphoff, 2002; Rottem, 2003). Mycoplasma hyorhinis, first isolated from the respiratory tract of young pigs, was implicated in various swine diseases, and has also been detected in humans (Huang et al., 2001).

For spleens, livers and caecal contents, significant differences of loads were determined using the Cobimetinib research buy Mann–Whitney U-test. Samples with no detectable Salmonella were placed in the lowest rank, those with bacteria detected only following enrichment were placed in the next rank, and further samples were ranked according to the number of CFU. Differences were considered significant at the 5% level. HD11 avian macrophage-like cells (Beug et al., 1979) were seeded into 24-well plates at

a density of 2 × 105 cells per well in RPMI 1640 medium (Invitrogen, UK) supplemented with 10% foetal bovine serum (PAA laboratories Ltd, UK), 10% chicken serum (Sigma, UK), 2 mM L-glutamine, 100 U mL−1 penicillin/streptomycin, 2.5 μg mL−1 fungizone, hereafter referred to as HD11 medium. Cells were incubated for 48 h at 41 °C under 5% CO2. Twenty-four hours prior to assay, the cells were washed with 1× PBS and HD11 medium without fungizone and penicillin/streptomycin (HD11-Ab-free medium) was added. Salmonella strains were grown in L-broth statically at 37 °C overnight. HD11 cells were inoculated with 20 μL bacterial culture in triplicate and plates centrifuged at 30 g for 5 min. Bacteria in the inocula were enumerated by serial decimal dilution, plating onto CBA and an overnight incubation at 37 °C. Infected HD11 cells were incubated at 41 °C for 30 min to allow the uptake of bacteria.

Extracellular bacteria were removed by washing with PBS, and HD11 medium containing 100 μg mL−1 gentamicin was added to each well AG-014699 in vitro followed by incubation for 2 h at 41 °C under 5% CO2. Cells were then washed with PBS and the medium was replaced with HD11 containing 20 μg mL−1 gentamicin. At 2, 4 and 6 h post Salmonella addition, cells were washed with PBS and lysed by incubation in 1 mL 0.1% (v/v) Triton X-100 in PBS for 10 min. Numbers of viable bacteria per well were determined by serial dilution, plating on CBA and an overnight incubation at 37 °C. The assay was performed in triplicate. Macrophage survival was examined

using a Cytotox lactate dehydrogenase assay (Promega, UK). Five genomic islands (R1, R3, R4, R5 and R6) present in the sequenced SEn strain P125109, but absent from Typhimurium LT2, and Typhi CT18 were identified by Davidson (2008). Rucaparib These loci were chosen for deletion. Comparative genome analysis and PCR screening showed that all these loci were also present in the avian-adapted serotype Gallinarum (Davidson, 2008; Thomson et al., 2008), although for this serotype R5 did not contain a ST64B phage-like sequence found in SEn. In previous analysis (Thomson et al., 2008), these loci were termed as regions of difference (ROD) and spanned slightly different genes to those in Davidson (2008) (Table 1). The genomic sequence of SEn Thirsk flanking the islands was determined by sequencing PCR products and shown to be identical to the published P125109 sequence (Thomson et al.

, 2011). This biosynthetic pathway may therefore be evolutionarily distinct from other reported DFO pathways. blastp analysis revealed a putative DmdR repressor in S. arenicola CNS-205 (Sare_1414) and S. tropica CNB-440 (Strop_1456), with 62/63% identity and 72/73% similarity to DmdR1 in S. coelicolor A3(2) (Flores & Martín, 2004). blastn and EMBOSS Palindrome analyses identified four putative DmdR-binding sites

(iron boxes) in each of the Salinispora genomes. Two of the iron boxes are upstream of desE and desF in both species (Fig. 1). The des gene cluster organization is conserved in Streptomyces (Barona-Gómez et al., 2006) with all six genes in one locus whereas, in Salinispora, desF is 13- to 21-kb upstream Seliciclib manufacturer of desEABCD. Despite these differences, iron repression of des is consistent in both genera, as confirmed in Salinispora by transcript analysis (Fig. 2). The remaining two iron boxes are upstream of a periplasmic binding protein similar to ferric-enterobactin transporters, and a putative siderophore utilization protein in StBac1/SaBac2, which may encode a Class

I bacteriocin (Penn et al., 2009). No iron boxes were identified near sid2–sid4. Alignment of the eight putative iron box sequences enabled the prediction of a DmdR-binding consensus sequence for Salinispora: TAGGTTArCCT (Fig. 4). Although sid2 from S. tropica Protein Tyrosine Kinase inhibitor CNB-440 was transcribed in iron-limited cultures, the lack of detectable Rho siderophores in the des mutants, and their poor growth without iron, suggests that the sid2 compound was either not produced in detectable quantities or that it is unable to chelate iron. As iron supplementation increased the growth of the des mutant, another iron chelator may be produced at very low levels or with a lower affinity for iron than CAS, which would

not be detected by our methods. Because of the differential transcriptional response of sid2 to iron in the des mutants, however, it is unlikely that this additional iron chelator is associated with sid2. Sid2 possesses similarity to ybt (Bearden et al., 1997; Pelludat et al., 1998); however, there are several differences between the two gene clusters (Fig. 1b). The three methyltransferases in sid2 are not integrated into the NRPS/PKS genes, and several essential ybt genes are absent in sid2, namely the reductase ybtU, salicylate synthase ybtS and regulator ybtT (Fig. 1b) (Geoffroy et al., 2000; Miller et al., 2002), which may explain the lack of yersiniabactin-like siderophore production. Sid2 in S. arenicola CNS-205 is transcribed under iron-replete rather than iron-limited conditions, although no chemotypic difference was detected between the wild-type and sid2 mutant in iron-sufficient conditions. The altered transcriptional regulation may be due to mobilization of the sid2 cluster 846-kb downstream on a separate genomic island. The putative CoA ligase remains in the original locus with respect to the S. tropica CNB-440 sid2 gene cluster (Fig.

It is also plausible that the concentration of protease inhibitors in a given cocktail may be sufficient to prevent protein degradation, but insufficient to inhibit or kill bacteria in the samples. Accordingly, several investigators have reported that a concentration-dependent relationship exists between protease inhibitor and bacterial growth (Labbe et al., 2001). Grenier et al. (2001a) showed that there was no inhibition of bacteria with bestatin at 0.02 μg mL−1, but when the concentration of bestatin approached 10 μg mL−1, the bactericidal function Akt inhibitor reached a maximum. In the presence

of aprotinin, the growth of S. alboniger was also partially or completely inhibited, depending on the concentration of the protease inhibitor (Lopes et al., 1999). Clearly, then, a higher dose of protease inhibitor has the potential to interfere with the proliferation of bacteria, resulting in an alteration of bacterial composition. Because massive degradation of proteins caused by proteases has been

observed in proteomic studies, it was suggested that PI should be added in the preparation of samples. Here, we have presented results indicating that saliva samples with and without PI showed similar protein diversity in fractions both with high-molecular-weight proteins and low-molecular-weight species as judged by both 1D SDS-PAGE and LC-MS/MS analysis. Addition of protease inhibitors seemed to have no significant effect on the integrity of salivary samples. Alternatively keeping the samples on ice and processing them in <1 h may have been sufficient to preserve protein integrity. In summary, our study Alectinib datasheet lends considerable

evidence that a protease cocktail containing AEBSF, aprotinin, bestatin, E64, leupeptin, and pepstatin A has no effect on oral bacterial growth or total bacterial composition. These findings suggest that the addition of protease inhibitors in the preparation of saliva samples for protein research will not interfere with microbial DNA analysis. The study was supported by the National Institute of Dental and Craniofacial Research (NIDCR) Grant no. U19 DE018385. Program Officer: Dr Isaac R. Rodriguez-Chavez. “
“A halophilic isolate Salimicrobium halophilum strain LY20 producing extracellular amylase DOK2 and protease was isolated from Yuncheng, China. Production of both enzymes was synchronized with bacterial growth and reached a maximum level during the early-stationary phase. The amylase and protease were purified to homogeneity with molecular weights of 81 and 30 kDa, respectively. Optimal amylase activity was observed at 70 °C, pH 10.0% and 10% NaCl. Complete inhibition by EDTA, diethyl pyrocarbonate (DEPC), and phenylarsine oxide (PAO) indicated that the amylase was a metalloenzyme with histidine and cysteine residues essential for its catalysis. Maltose was the main product of starch hydrolysis, indicating an β-amylase activity. The purified protease from LY20 showed highest activity at 80 °C, pH 10.0% and 12.5% NaCl.