, 2006). Reaction mixture I (50 μL) contained 100 mM HEPES (pH 7.0), 10 mM α-ketoglutarate, 0.5 mM FeSO4·7H2O, 0.5 mM ascorbate, variable concentrations (0.3–40 mM) of l-leucine, l-threonine or l-methionine, and aliquots of purified dioxygenase. Reaction I was incubated at 30 °C for 30 min, at which point it was arrested by placement on ice. The amount of enzyme applied

was selected to ensure that the increase in synthesized succinate was linear AZD6244 order during the reaction. To determine the concentration of the synthesized succinate, 2.5 μL of reaction mixture I was added to reaction mixture II (up to final volume of 25 μL), which contained 100 mM Tris–HCl (pH 8.0), 1 mM phosphoenolpyruvate, 0.3 mM NADH, 10 mM MgCl2, 0.3 mM CoA, 0.3 mM ATP, 3 μg succinyl-coenzyme A synthetase Venetoclax purchase E. coli (purified by IMAC as his6-tag-fused protein) and 0.25 μL of a solution of pyruvate kinase (PK)/lactate dehydrogenase (LDH) from rabbit muscle (Sigma) (0.186 U of PK and 0.226 U of LDH). Reaction II was incubated at 30 °C for 1 h and halted by placement on ice. Subsequently, the absorbance at 340 nm was measured, and the concentration of synthesized succinate was deduced from a calibration curve obtained by performing reaction

II with succinate standards. Enzymatic activity was quantified by measuring the amount of succinate produced per minute and per milligram of enzyme. The KM and Vmax parameters with standard errors for l-leucine, l-threonine and l-methionine were deduced from Michaelis–Menten kinetic equation plots obtained from nonlinear regression analysis of experimental data using SigmaPlot (http://www.systat.com). The preparation and identification of l-methionine sulfoxide and hydroxylated l-leucine was performed as previously described (Hibi et al., 2011). A biomass sample of BL21(DE3) [pET-HT-BPE] strain from fresh-made LB-agar plates was inoculated into 400 mL of LB broth (2 × 200 mL) supplemented with Ap

(100 mg L−1) and cultivated at 37 °C until A555 nm = 1 was reached. Subsequently, IPTG was added to a final concentration of 1 mM, and the culture was incubated for an additional 2 h. The biomass was harvested by centrifugation and re-suspended in 5 mL of 50 mM HEPES (pH 7) and lysed by one pass through a French press (1000 psi). The five reaction mixtures (2 mL volume) Thiamine-diphosphate kinase then consisted of 25 mM l-threonine, 25 mM α-ketoglutarate, 100 mM HEPES (pH 7), 10 mM FeSO4·7H2O and 1 mL of cell lysate. The reactions were incubated at 37 °C for 15 h with vigorous shaking. Amino acid hydroxylation was monitored by TLC analysis using ninhydrin (2-propanol/acetone/ammonia/water = 25 : 25 : 6 : 4). A 10 mL volume of the resulting supernatant was passed through a 0.22 μm filter and applied to a preparative TLC plate. The hydroxylated l-Thr was collected, eluted with water, freeze-dried and analysed by ESI-MS as described in (Hibi et al., 2011).

A concordant approach to realising the recommendations may enhance pharmacy professionals’ engagement in CPD and pave the way for CPD in revalidation in due course. The authors declare that they have no conflicts of interest to disclose. This study was supported by the RPSGB with Department of Health

funding. We would like to thank the RPSGB research steering committee, which later became part of the GPhC, click here including Dr Andreas Hasman and in particular Professor Christine Bond and Dr Peter Wilson, for supporting our initial proposal and helping to bring it to fruition. MeSH is the National Library of Medicine’s controlled vocabulary thesaurus. It consists of sets of terms naming descriptors in a hierarchical structure that permits searching at various levels of specificity. The MeSH thesaurus is used by NLM for indexing articles from 5400 biomedical journals for the Medline/PubMED® database.

MeSH has a hierarchical structure in an extensive tree structure representing increasing levels of specificity. Mesh heading (MH) is the term used in the Medline database as the indexing term. The term reflects a meaning; its use indicates the topics discussed by the work cited. Entry terms are used as pointers to the MH. The presence of an entry term in the record is an indication that this topic should be indexed by the given MH. A variety of search terms was constructed for use within the databases using the following rationale. Searching for ‘Pharmacy’ within MeSH returns a tree structure but also a number of related Nutlin-3a terms, detailed below: Pharmacy (falls under All MeSH categories>Disciplines and Occupations Category>Health Occupations>Pharmacy) Pharmacist (falls under All MeSH categories>Persons Category>Occupational Groups>Health Personnel>Pharmacists AND All MeSH categories>Health Care Category>Health Care

Facilities, Manpower, and Services>Health Personnel>Pharmacists) Entry terms: Continuing pharmacy education (falls under All MeSH categories>Education>Education, Professional>Education, Pharmacy>Education, Pharmacy, Continuing) Entry terms: Searching cAMP for ‘Continuing professional development’ within the MeSH browser (‘Find terms with any fragment’ option) does not return a tree structure but a number of related terms, some already covered above, and the additional term detailed below: Education, Professional, Retraining (falls under All MeSH categories>Anthropology, Education, Sociology and Social Phenomena Category>Education>Education, Professional>Education, Continuing>Education, Professional, Retraining) Entry terms: The following search was conducted again in August 2010. The 395 papers from the Medline database, as shown below, were transported and saved in a unique file using Endnote software.

The residues vital for metal binding and catalysis (Q56, C106, H148, E149 and H152) were within 30 nm around the metal ion. MD simulations CSF-1R inhibitor of MtbPDF

and G151D structures revealed no significant differences in the positioning of metal-binding residues and their average distance from the Fe2+ ion. This supports the equal Fe content in MtbPDF and G151D, as seen from the AAS results. The side chains of residues lining the substrate-binding cavity (G49, V50, G51, E104, G105, C106, L107, R144 and M145) of G151D showed slight fluctuations in positioning compared with MtbPDF. The average distance between side chain atoms of M145 with L107 in G151D was increased by 20 nm compared with MtbPDF (Fig. S2). Similarly, the distance between side

chain atoms of G49, V50 and G51 with those of 104EGCL107 was increased by 5–10 nm in G151D (Fig. S3). These differences might have contributed to the increase in space within the peptide binding pocket of G151D. These differences were reported to be decreased in the R77-79K SB431542 supplier mutation of MtbPDF, leading to a reduction in size of the substrate binding site (Saxena et al., 2008). Three arginines in the insertion sequence (77RRR79) (Fig. 1a) of MtbPDF were reported to be responsible for the observed resistance to oxidative stress (Saxena et al., 2008). The higher sensitivity of the G151D mutant to oxidizing agents led us to look into the structural variations in the loop containing three arginines. During MD simulations, the side chain of R77 in G151D was displaced by 35 nm from Fe2+, losing its stabilization from hydrogen bonding with side chain atoms of D128 (Fig. 4c). This destabilizes the loop containing three arginines, which was reported to interact with the core helix in MtbPDF to provide oxidative stress stability. The predicted mechanism of this interaction was an ‘action-at-distance’, in which the R77-79 present

in the loop away from the active site modulates the thermostability and resistance to H2O2 in MtbPDF. Although the arginine side chains are reported to interact and scavenge oxygen (Saxena et al., 2008), the actual mechanism by which these residues prevent Fe2+ and/or metal-coordinating cystein from oxidizing is still not clear. In G151D, destabilization of the loop containing three arginines might have led to increased Selleckchem Etoposide oxidation of Fe2+ and/or metal-coordinating cystein. More systematic studies on this property would unveil the underlying mechanism of action. The free energy of binding of substrate N-formyl-Met-Ala-Ser into MtbPDF was −6.34 kcal mol−1 and for G151D was −7.25 kcal mol−1. Superimposition of the two docked structures indicated that the positioning of residues at the P′ and position of the substrate (formyl group and Met) was essentially the same in both cases. But residues at the and positions of the substrate (Ala and Ser) were better aligned in G151D than in MtbPDF (Fig. 4d).

2 and 1.3 kb, respectively) were

observed in the agarose gel (Fig. 2b). The difference in size indicated that TPMA0004 was the mycF disruption mutant. For genetic complementation for the mycF disruption mutant TPMA0004, pMG508 including mycF was transferred to TPMA0004 buy EPZ5676 by intergeneric conjugation. The transconjugant TPMA0009 isolated from the conjugation plate containing apramycin and nalidixic acid produced M-II (8.29 μg mL−1) (Fig. 3). The amount of M-II produced by TPMA0009 was approximately 55% of that produced by the wild strain A11725. PCR was performed with several primer pairs to confirm the genetic condition of TPMA0009. As shown in Fig. 2b, the transconjugant TPMA0009 producing M-II was the homogenous mycF complementation strain in which the mycF gene was inserted into the chromosome by a site-specific recombination between the artificial attB site on the chromosome and the attP site on Trichostatin A cost pMG508. The disruption cassette FRT-neo-oriT-FRT-attB

was used to obtain the mycE disruption mutant TPMA0003 and the mycF disruption mutant TPMA0004 of M. griseorubida. In particular, PCR targeting with the phage λ-Red recombinase was performed to isolate the mycF disruption mutant. Furthermore, from these mutants, the homogenous complementation strains TPMA0003 and TPMA0004 were isolated by a site-specific recombination between the artificial attB site on the mutant chromosomes and the attP on pSET152 used as a vector. Recently, a simple and highly efficient

PCR-targeting system was developed for the gene targeting of Streptomyces strains (Gust et al., 2003). However, genetic engineering cannot be performed for actinomycete strains lacking the bacteriophage φC31 attB attachment site using vectors possessing a φC31 int gene and an Ribonucleotide reductase attP site. In this study, gene disruption and complementation studies could be performed for M. griseorubida, which lacked the bacteriophage φC31 attB site on the chromosome, using the disruption cassette FRT-neo-oriT-FRT-attB. A multiple gene disruption and complementation scheme using the disruption cassette is shown in Fig. 4. In this study, the complementation plasmid pMG508 possessing the int gene encoding integrase, the attP site, and the resistant marker aac(3)IV was inserted into the φC31 attB attachment site, which was flanked by the resistant marker neo and oriT on the mycF disruption mutant. For additional gene disruption and complementation studies of the complementation strain TPMA0009, resistant markers other than neo and aac(3)IV should be used. However, if a gene disruption mutant with the resistant marker eliminated was obtained by in-frame disruption, additional gene disruption studies can be performed with the same resistant marker.

Faculty and/or preceptors converge their thoughtfulness on the preliminary understanding selleck chemicals llc of the reflective process by the student, boosting the student’s distinct nonverbal communication and ultimately providing well-thought-out facets to equipoise the flexible nature of reflective writing. The Authors declare that they have no conflicts of interest to disclose. “
“Objectives  The aims of this study were to determine the frequency of prescription compounding by community pharmacists, identify factors that influence pharmacists’ decisions to provide compounding services,

and evaluate physicians’ perspectives on prescribing medications that require compounding. Methods  The study was a cross-sectional survey administered via face-to-face structured interviews with randomly selected community pharmacists and physicians from different areas of the West Bank. Key findings  Of the 260 community pharmacists who were contacted, 212 agreed to participate in the survey, giving a response rate of 81.5%. Overall, 153 (72.2%) of respondent pharmacists provided compounding services. Compounded prescriptions accounted for 1973 (1.55%) of 126 840 prescriptions PARP activation dispensed in a typical month. Among the compounders, 112 (73.2%)

pharmacists reported that their goal in providing full pharmaceutical care to their patients was the most important motivator. The most frequently reported reason for not providing compounding was ‘I do not receive prescriptions that require compounding’ by 43 out of 59 (72.9%) pharmacists. A total of 179 out of 220 physicians consented to participate in this study giving a response oxyclozanide rate of 81.4%. The majority of physicians (142, 79.3%) did not prescribe compounded medicines. The most important reason for their decision to prescribe compounded medicines was the unavailability of the required dosage forms. The most commonly cited reason for

not prescribing them was a lack of trust in the quality of the compounded formulations. Conclusion  While most respondent pharmacists provide a compounding service this represents only a small percentage of the total volume of dispensed prescriptions. Most responding physicians do not prescribe medications that require compounding because they lack trust in the quality of the compounded formulations. “
“Objectives The aim of the study was to determine the public’s views on weight-management services, including pharmacies as a potential venue, and the extent of current pharmacy involvement in weight management. Methods Two questionnaires were developed for face-to-face interview in one Primary Care Trust area: one for the general public and one for community pharmacists. Key findings Interviews were conducted with 177 members of the public, 75% of whom had tried to lose weight. More had used over-the-counter weight-loss products than prescribed medicines. There was greater awareness of commercial weight-management clinics than of NHS-led initiatives.

Another potential limitation anti-PD-1 monoclonal antibody of this study is the different origins of the populations. While the AHC group mainly consisted of Central European individuals, who were infected by sexual

transmission, the majority of patients in the CHC group were Southern European injecting drug users (IDUs). In both groups, the HCV genotype distribution was in accordance with the results of the EuroSIDA cohort study [16], which reported a slightly lower prevalence of genotypes 1 and 2 relative to genotype 3 in the Southern European CHC population, as compared with Central Europe. In addition, the ethnicity of patients in the two cohorts, a factor strongly associated with the prevalence of different IL-28B genotypes [1,4], might have differed. However, most patients were Caucasian in this study, and accordingly the prevalence of the rs12979860 CC genotype was very similar in patients with AHC and CHC (47.5%vs. 45.2%). Furthermore, similar differences in HCV genotype ERK inhibitor chemical structure distribution in relation to the IL-28B genotype were found within the group of German patients with CHC. Therefore, it is unlikely that demographic differences had an impact on the study results. Relationships between rs12979860 genotype CC and a higher baseline HCV viral load [1,4] and between genotype CC and higher

transaminase levels [10] have previously been found in HCV-monoinfected patients. The IL-28B genotype CC is associated with lower expression of interferon-stimulated genes [17]. The presence of the IL-28B CC genotype may therefore lead to elevated HCV replication and higher levels of necrosis and inflammation, in response to higher activity of HCV. However, data on the impact of these SNPs on viral replication are contradictory [6,8,10]. Recently, Lindh et al. proposed that the higher viral load in CHC patients with the CC genotype may be attributable to a significantly Meloxicam higher clearance rate in CC carriers

with a low viral load, causing a higher proportion of those with the CC genotype and a higher viral load in the CHC population [18]. In our study, the plasma HCV viral load was higher in patients with the CC genotype and AHC, while in those with CHC there was no significant difference in this parameter according to IL-28B genotype. This may be attributable to the fact that HIV/HCV-coinfected patients show higher levels of viraemia than HCV-monoinfected subjects with CHC [19]. In this setting, a subtle effect of IL-28B genotype on HCV viral load may not be detected. Finally, significantly higher ALT levels were observed in patients with IL-28B CC, supporting the above theory. Most homosexual male patients with AHC carried HIV before becoming infected with HCV, whereas IDU patients with CHC are presumed to be infected with HCV before, or at the same time as, HIV. Because of this, the immunodeficiency in patients with AHC could have been more profound.

To illustrate, strain 12 to which the IMP–COL combination was synergistic was highly resistant to both IMP (MICIMP > 32 mg L−1) and COL (MICCOL = 128 mg L−1). Combining IMP and COL decreased MICIMP from 32 to 6 mg L−1 and MICCOL from 128 to 32 mg L−1. This result yielded an FIC index of 0.44, meeting the definition of synergy. However, as per CLSI breakpoint, MICIMP of 6 mg L−1 against A. baumannii indicates IMP non-susceptibility, while MICCOL of 32 mg L−1 against A. baumannii indicates Caspase activity COL resistance. Therefore, this combination was considered clinically insignificant. The same conclusion applies to the other synergistic combinations that were observed

in this study. We conclude that the effect of antibiotic combinations on our outbreak strains of MDR A. baumannii seemed highly strain-specific. The complex genetic background of each A. baumannii strain seems to exert differential effects on bacterial response to antibiotic combinations. The choice of antibiotic combinations should be dictated by results of susceptibility tests performed on each strain.

Further investigations are warranted to ascertain the molecular basis of the COL-DOX synergy. This project was supported by an investigator-initiated grant from Merck. We thank the Cedars-Sinai Microbiology Laboratory and Hospital Epidemiology Department staff for assistance in technical aspects STA-9090 ic50 and data collection, respectively. We thank Drs. Michael Jacobs, Andrea Endimiani, and Ms. Saralee Bajaksouzian of Case Western Reserve University for assistance with MICs. A portion of this manuscript was presented at the 45th Annual Meeting of the Infectious Disease Society of America (2007, San Diego, CA). Y.M. is partially supported by the Cedars-Sinai Clinical Scholars’ Funding Award. R.A.B. is supported by the VISN 10 Geriatric Research Education and Clinical Care Center (GRECC), Merit Review Program of the Veterans Administration, and the National Institute of Health (R01AI072219-05).

All other authors have no financial disclosures. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should Branched chain aminotransferase be directed to the corresponding author for the article. “
“Rapidly increasing bacterial resistance to existing therapies creates an urgent need for the development of new antibacterials. Tirapazamine (TPZ, 3-amino-1,2,4-benzotriazine 1,4 dioxide) is a prodrug undergoing clinical trials for various types of cancers. In this study, we showed that TPZ has antibacterial activity, particularly at low oxygen levels. With Escherichia coli, TPZ was bactericidal under both aerobic and anaerobic conditions. Escherichia coli mutants deficient in homologous recombination were hypersusceptible to TPZ, suggesting that drug toxicity may be due to DNA damage. Moreover, E.

CMV oesophagitis is treated with ganciclovir 5 mg/kg bd iv for 2–4 weeks, or until symptoms/signs have resolved (category III recommendation) [14,15]. Valganciclovir may be substituted for iv ganciclovir at 900 mg bd orally for some or all of the duration if symptoms are not severe enough to interfere with oral absorption on the basis of studies showing efficacy for CMV disease in transplant patients [16] but there is a paucity of data in HIV-related CMV disease of the gastrointestinal tract (category IV recommendation). Secondary CMV prophylaxis for oesophageal disease is

not routinely indicated, learn more unless there is concomitant ophthalmological disease. Herpes simplex oesophagitis is treated with aciclovir 5–10 mg/kg tid iv, followed by 400 mg five times a day orally for a total of 14 days (category III recommendation) [17] or oral valaciclovir Ku-0059436 ic50 1 g bd orally (see 6 Herpes viruses for a discussion of prophylaxis of HSV). Foscarnet 90 mg/kg bd iv has been used in cases

of ganciclovir-resistant CMV or 40 mg/kg bd or tid for aciclovir-resistant HSV [15]. After presentation with infectious oesophagitis, early initiation of HAART should be considered (category IV recommendation) [18]. As elsewhere in these guidelines, early initiation of HAART is favoured on the basis that improved survival without AIDS progression or death has been seen when HAART is initiated within the first two Tyrosine-protein kinase BLK weeks of treatment of the opportunistic infection [18]. This recommendation is extrapolated from a series in which most cases were not related to oesophageal opportunistic infection but is also supported by evidence of functional immunological benefits of antiretrovirals against organisms such as Candida spp. [19]. Diarrhoea is a common problem for people with HIV in both resource-poor and resource-rich settings, regardless of antiretroviral exposure. In the pre-HAART

era, 30–70% of HIV-seropositive individuals experienced diarrhoea, and among European patients with CD4 counts <50 cells/μL, 49% would expect to develop diarrhoea within 1 year and 96% within 3 years [20]. In resource-poor areas, incidence and severity continue to be higher. Early clinical observations confirmed that diarrhoeal illness was linked to reduced quality of life and poorer survival [21]. Diarrhoea may be the presenting symptom of lymphoma and Kaposi’s sarcoma, may affect up to 40–50% of those taking antiretroviral therapy (ART), can be induced by other medications and may be the result of an incompletely defined direct effect of HIV on the gut mucosa termed HIV-associated enteropathy [22–25].

One basic hypothesis states that either the PPIase activity or some chaperone activity of Mip and Mip-like proteins might be involved in the maturation and trafficking of proteins derived from pathogens. It goes on to add that these activities may also allow Mip and

Mip-like proteins to recognize host receptors and inhibit the host’s defense response. Despite studies performed in numerous laboratories, none of Mip’s substrates or molecular targets has yet been discovered. Xcc, a Gram-negative Gammaproteobacterium, is the causal agent of black rot disease in cruciferous crops worldwide (Hayward, 1993). Our own recent studies have shown that a mip-like gene (here called mipXcc) exists within Xcc and encodes a protein, MipXcc, which exhibits a PPIase activity specifically inhibited by FK-506 OSI-744 order (Zang et al., 2007). Mutagenesis analysis revealed that Xcc requires a functional MipXcc for full virulence and proliferation in host plants. Further study showed that, in mutants lacking a working mipXcc, Xcc was unable to produce its usual amounts of exopolysaccharide and its extracellular proteases were significantly less active (Zang et al., 2007). Although the mechanism by which MipXcc affects the activity of extracellular proteases remains unclear, we have made an effort to address

this issue. In this study, we provide evidence that Osimertinib in vivo MipXcc interacts with the major Xcc protease PrtA and assists its maturation in the periplasm. The bacterial strains and plasmids used in this project are listed

in Table 1. The primers used are listed in Table 2. Escherichia coli strains JM109 and M15 (Qiagen, Germany) were grown in LB medium at 37 °C. The bacterial two-hybrid Arachidonate 15-lipoxygenase reporter strain (here named BTHrst) (Stratagene, La Jolla, CA) was grown in M9 His-dropout medium at 30 °C. Xcc strains were grown in NYG medium at 28 °C. Antibiotics were used at the following final concentrations: rifampicin, 50 μg mL−1; kanamycin, 25 μg mL−1; ampicillin, 100 μg mL−1; chloramphenicol, 34 μg mL−1; gentamicin, 10 μg mL−1; streptomycin, 12.5 μg mL−1; and tetracycline, 15 μg mL−1 for E. coli and 5 μg mL−1 for Xcc. Standard DNA manipulation was performed as described by Sambrook & Russell (2001). The conjugation of Xcc to E. coli was performed as described by Turner et al. (1985). Restriction enzymes and DNA ligase were purchased from Promega (Madison, WI) and used in accordance with the manufacturer’s instructions. All clones were confirmed by sequencing. Fragment of prtA was PCR-amplified and cloned into pLAFR3. The resulting plasmid pR3PrtA was introduced into the mipXcc mutant NK2699 and the prtA mutant 001F10 by triparental conjugation. Fragment of mipXcc was conjugated at the 3′ end with 6xHis coding sequences, then PCR-amplified and cloned into pLAFR3. The derived plasmid pR3MipH6 was introduced into NK2699.

Cel5M was identified as a cold-active cellulase with an optimal temperature of 30 °C and it was active within a narrow pH range with an optimum at pH 4.5. Phylogenetic analysis showed that Cel5M represented a new subfamily of the glycosyl hydrolase family 5, representing an opportunity for research into and applications of novel cold-active cellulases. Glycoside hydrolases (GHs) have been classified into more than 100 families according to similarities in their amino acid sequence (Henrissat & Davies, 1997) and into clans according to their three-dimensional structures.

GH5, which belongs to glycoside hydrolase clan A, is a superfamily with a conserved overall structure and mechanism (Leggio & Larsen, 2002). Cold-active cellulases have gained considerable attention for both industrial applications and fundamental research because of their unique structural and catalytic characteristics (Zeng et al., 2006). Only www.selleckchem.com/products/ly2157299.html a few cold-active cellulases have been reported so far, CelG from Pseudoalteromonas haloplanktis (Violot et al., 2003)

and CelX from Pseudoalteromonas sp. DY3 (Zeng et al., 2006). Both CelG and CelX belong to GH5 and consist of a catalytic module (CM) and a carbohydrate-binding module (CBM), separated by a linker region selleck kinase inhibitor (LR) that plays a key role in cold adaptation of cold-active cellulases (Sonan et al., 2007). In the present study, a gene encoding a novel cold-active endo-β-1,4-glucanase (named Cel5M) from psychrophilic deep-sea bacteria Pseudomonas sp. MM15 was isolated. The deduced protein sequence lacked the typical cellulase domain structures of CBM and LR, providing an opportunity for investigating its novel cold-adaptation mechanism. Phylogenetic analysis showed that Cel5M represents a new subfamily in GH5. Carboxymethyl cellulase (CMCase) producing Pseudomonas sp. MM15, deposited in

the China Center of Industrial Culture Collection under strain collection number CICC 10441, was isolated from the deep-sea sediment of the Southern Okinawa Trough using the method described by Ibrahim & El-Diwany (2007). The in situ environment of the deep-sea sediments with a water depth of 1245 m was characterized by a strong terrestrial input of organic matters, thus favoring the activity of various ALOX15 extracellular enzyme-producing bacteria (Dang et al., 2009). A genomic library of Pseudomonas sp. MM15 was constructed using plasmid pUC19 (TaKaRa, Japan) and Escherichia coli DH 5α following the procedure described by Chen et al. (2011). After 14 h incubation at 37 °C, the colonies were transferred onto carboxymethyl cellulose (CMC; Sigma) plates (1 g L−1 KH2PO4; 5 g L−1 NaCl; 10 g L−1 yeast extract; 10 g L−1 peptone; 10 g L−1 CMC and 15 g L−1 agar). After another 14 h growth at 37 °C, the plates were stained with Congo red (1 g L−1) for 15 min and then washed with 1 M NaCl solution for 5 min.