Taxonomy of mycelial actinobacteria isolated from Saharan soils and their efficiency to reduce aflatoxin B1 content in a solid-based medium

Aflatoxin B1 (AFB1) is a carcinogenic compound produced by filamentous fungi. In order to reduce AFB1 occurrence in foodstuffs, 13 strains of mycelial actinobacteria were tested in vitro for the efficacy to reduce AFB1 content; all were isolated from the Saharan soils of Algeria. Firstly, morphological study and molecular analysis, based on the 16S rRNA gene, indicated that these strains belong to Actinomadura, Nocardiopsis, Nonomuraea, Saccharothrix and Streptomyces genera. Secondly, each strain’s efficacy to reduce pure AFB1 content was studied in ISP2-medium. After a 4-day incubation at 30°C on AFB1-supplemented medium (5 ppm of AFB1), AFB1 was extracted and quantified. AFB1 content was reduced by all strains (42.9–97.6%). The three most efficient reducers (94.9–97.6%) were two strains belonging to the genus Streptomyces and one to the genus Saccharothrix. Among the latter, strains ACD6 and ABH19 showed no adsorption mechanism involved, suggesting a potential degradation mechanism. These findings led us to suggest that these actinobacterial strains could be used as decontamination treatments for the reduction of AFB1 content.     

Identification of the di-<Emphasis Type="Italic">n</Emphasis>-butyl phthalate-biodegrading strains and the biodegradation pathway in strain LMB-1

DNA isolated from a greenhouse soil (Nanjing, Jiangsu Province, China) was suitable for PCR amplification of gene segment coding for the 16S rRNA. Diverse PCR products were characterized by cloning and sequencing, and analysis of bacterial colonies showed the presence over 26 phyla. The most bacteria belonged to Proteobacteria, Actinobacteria, Gemmatimonadetes, Acidobacteria and Planctomycetes. Furthermore, after the enrichment procedure of DBP-degrading microorganisms, 4 strains were isolated from the soil sample with di-n-butyl phthalate (DBP) biodegradability, and they were identified to be Rhizobium sp., Streptomyces sp., Pseudomonas sp. and Acinetobacter sp. Analysis of the degradation products by LC-MS led to identification of metabolites of DBP in strain LMB-1 (identified as Rhizobium sp.) which suggests that DBP was degraded through β-oxidation, demethylation, de-esterification and cleavage of aromatic ring.     

Antimicrobial potential and taxonomic investigation of piezotolerant <Emphasis Type="Italic">Streptomyces</Emphasis> sp. NIOT-Ch-40 isolated from deep-sea sediment

Microbial-derived natural products from extreme niches such as deepsea are known to possess structural and functional novelty. With this background, the present study was designed to investigate the bioprospecting potential and systematics of a deep-sea derived piezotolerant bacterial strain NIOT-Ch-40, showing affiliation to the genus Streptomyces based on 16S RNA gene similarity. Preliminary screening for the presence of biosynthetic genes like polyketide synthase I, polyketide synthase II, non ribosomal peptide synthase, 3-amino-5-hydroxybenzoic acid synthase and spiroindimicin followed by antibacterial activity testing confirmed the presence of potent bioactivity. The secondary metabolites produced during fermentation in Streptomyces broth at 28?°C for 7 days were extracted with ethyl acetate. The extract exhibited a specific inhibitory activity against Gram-positive bacteria and was significantly effective (p?<?0.0001) against methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentration and minimum bactericidal concentration against MRSA was 1.5 µg/mL, which was statistically significant in comparison with erythromycin. A multifaceted analysis of the Streptomyces spp. was carried out to delineate the strain NIOT-Ch-40 at a higher resolution which includes morphological, biochemical and molecular studies. Piezotolerance studies and comparison of fatty acid profiles at high pressures revealed that it could be considered as one of the taxonomic markers, especially for the strains isolated from the deep sea environments. In conclusion, the observation of comparative studies with reference strains indicated towards the strain NIOT-Ch-40 as an indigenous marine piezotolerant Streptomyces sp. with a higher probability of obtaining novel bioactive metabolites.     

Diversity of Culturable Plant Growth-Promoting Bacterial Endophytes Associated with Sugarcane Roots and Their Effect of Growth by Co-Inoculation of Diazotrophs and Actinomycetes

Application of environmentally friendly agents to reduce the use of chemicals and to enhance growth of plants is an ultimate goal of sustainable agriculture. The use of plant growth-promoting endophytes has become of great interest as a way to enhance plant growth and additionally protect plants from phytopathogens. In this study, 135 isolates of endophytic bacteria including actinomycetes were isolated from roots of commercial sugarcane plants cultivated in Thailand and were characterized for plant growth-promoting (PGP) traits. Based on morphological and 16S rRNA sequence analysis, the endophytes were distributed into 14 genera of which the most dominant species belong to Bacillus, Enterobacter, Microbispora, and Streptomyces. Two strains of endophytic diazotrophs, Bacillus sp. EN-24 and Enterobacter sp. EN-21; and two strains of actinomycetes, Microbispora sp. GKU 823 and Streptomyces sp. GKU 895, were selected based on their PGP traits including 1-aminocyclopropane-1-decarboxylate deaminase, indole-3-acetic acid, nitrogen fixation, phosphate solubilization, and siderophore production for evaluation of sugarcane growth enhancement by individual and co-inoculation. Sixty days after co-inoculation by endophytic diazotrophs and actinomycetes, the growth parameters of sugarcane plants were significantly greater than that of individual and un-inoculated plants. The results indicated that these endophytes have high potential as PGP agents that could be applied to promote sugarcane growth and could be developed as active added value biofertilizers in the future.     

Production and characterization of cellulase from <Emphasis Type="Italic">E. coli</Emphasis> EgRK2 recombinant based oil palm empty fruit bunch

Oil Palm Empty Fruit Bunch (OPEFB) is an abundant biomass resource in Indonesia, which contains 41.3 ~ 46.5% (w/w) of cellulose. This research examined the production of cellulase by the E. coli EgRK2 recombinant strain using an OPEFB substrate. The production of the enzyme was initially examined to identify optimum growth conditions, by observing the growth and activity of E. coli EgRK2 compared to its wild type. Our results showed that the optimum production time, pH and temperature of the recombinant growth and cellulase activity were achieved at 24 h, and at 7 and 40°C, respectively. Using these optimum conditions, the enzyme was produced, and experiments were carried out to examine the enzyme characteristics, produced from both strains, on hydrolysis of cellulose from OPEFB. Our results showed that the activity of the enzyme produced by the recombinant almost doubled compared to that of the wild type, although the optimum pH for both strains was pH 6. Higher activity was achieved by the recombinant compared to the wild type strain, and values were 1.905 and 1.366 U/mL, respectively. The optimum temperature for hydrolysis by cellulase occurred at 50°C for Bacillus sp. RK2, and 60°C for Bacillus sp. EgRK2. The Michaelis-Menten constant (Km) and maximum velocity (Vmax) for OPEFB degradation by E. coli EgRK2 were 0.26% and 1.750 μmol/mL/sec, which were significantly better values than those of the wild type. Control experiments for the degradation test using CMC also showed a better Vmax value for E. coli EgRK2 compared to the wild type, which is 2.543 and 1.605 μmol/mL/sec, respectively.     

Identification and characterization of a new agar-degrading strain with the novel properties of saccharides inhibition and nitrogen fixation

In this study, a new agar-degrading strain was isolated from soil with agar as a sole carbon source and energy. Based on its morphological, physiological, biochemical characterization and 16S rDNA sequence, the strain was identified as Streptomyces lavendulae UN-8. The extracellular agarase activity reached 0.03 U/ml after fermentation in shake flask (250 ml), which was close to other reported non-marine microorganisms. Furthermore, it is interesting that the growth of UN-8 would be inhibited by glucose (40 g/L) and maltose (40 g/L) with the inhibitory rate of 100% and 70%, respectively. Besides, UN-8 could be grown on the solid medium without any nitrogen sources, then the possible nitrogen fixation gene nifU was cloned from its genomic DNA. The deduced amino acid sequence of nifU has high similarity (98%) with nitrogen fixation protein NifU from Streptomyces sp. NRRL S-104 (KJY22454.1) and Streptomyces sp. NRRL F-4428 (KJK52526.1) based on NCBI blast. It is suggested that the nifU gene of UN-8 also encoded nitrogen fixation protein NifU. These results provided some new information for the further understanding of agar-degrading strain.     

<Emphasis Type="Italic">Streptomyces gamaensis</Emphasis> sp. nov., a novel actinomycete with antifungal activity isolated from soil in Gama,Chad

During an investigation exploring potential sources of novel species and natural products, a novel actinomycete with antifungal activity, designated strain NEAU-Gz11^T, was isolated from a soil sample, which was collected from Gama, Chad. The isolate was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain NEAU-Gz11^T belongs to the genus Streptomyces with high sequence similarity to Streptomyces hiroshimensis JCM 4098^T (98.0 %). Similarities to other type strains of the genus Streptomyces were lower than 98.0 %. However, the physiological and biochemical characteristics and low levels of DNA–DNA relatedness could differentiate the isolate genotypically and phenotypically from S. hiroshimensis JCM 4098^T. Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces gamaensis sp. nov. is proposed. The type strain is NEAU-Gz11^T (=CGMCC 4.7304^T=DSM 101531^T).     

Cost-effective production of cellulose hydrolysing enzymes from <Emphasis Type="Italic">Trichoderma</Emphasis> sp. RCK65 under SSF and its evaluation in saccharification of cellulosic substrates

Trichoderma sp. is a potential cellulase producing mesophilic fungi which grow under mild acidic condition. In this study, growth and nutritional conditions were manipulated for the maximum and cost-effective production of cellulase using lab strain Trichoderma sp. RCK65 and checked for its efficiency in hydrolysis of Prosopis juliflora (a woody substrate). Preliminary studies suggested that when 48 h old secondary fungal culture (20 % v/w) was inoculated in wheat bran moistened with mineral salt solution (pH 4.5 and 1:3 solid to moisture ratio), incubated at 30 °C and after 72 h, it produced maximum cellulase (CMCase 145 U/gds, FPase 38 U/gds and β-glucosidase 105 U/gds). However, using statistical approach a S:L ratio (1:1) was surprisingly found to be optimum that improved cellulase that is CMCase activity by 6.21 %, FPase activity by 23.68 % and β-glucosidase activity by 37.28 %. The estimated cost of crude enzyme (Rs. 5.311/1000 FPase units) seems to be economically feasible which may be due to high enzyme titre, less cultivation time and low media cost. Moreover, when the crude enzyme was used to saccharify pretreated Prosopis juliflora (a woody substrate), it resulted up to 83 % (w/w) saccharification.     

Microbial secondary metabolites ameliorate growth, <Emphasis Type="Italic">in planta</Emphasis> contents and lignification in <Emphasis Type="Italic">Withania somnifera</Emphasis> (L.) Dunal

In the present investigation, metabolites of Streptomyces sp. MTN14 and Trichoderma harzianum ThU significantly enhanced biomass yield (3.58 and 3.48 fold respectively) in comparison to the control plants. The secondary metabolites treatments also showed significant augmentation (0.75–2.25 fold) in withanolide A, a plant secondary metabolite. Lignin deposition, total phenolic and flavonoid content in W. somnifera were maximally induced in treatment having T. harzianum metabolites. Also, Trichoderma and Streptomyces metabolites were found much better in invoking in planta contents and antioxidants compared with their live culture treatments. Therefore, identification of new molecular effectors from metabolites of efficient microbes may be used as biopesticide and biofertilizer for commercial production of W. somnifera globally.     

Characterization of phosphate-solubilizing bacteria exhibiting the potential for growth promotion and phosphorus nutrition improvement in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) in calcareous soils of Sinaloa,Mexico

Greenhouse bioassays were used to examine the ability of selected strains of the rhizobacteria Sinorhizobium meliloti, Bacillus flexus and B. megaterium to solubilize phosphorus (P) and to affect growth promotion and phosphorus nutrition in maize. These bacterial strains were found to decrease the pH and solubilize some forms of insoluble P, such as tricalcium phosphate and hydroxyapatite, as well as to exhibit acid and alkaline phosphatase enzymatic activities in culture medium, properties that are possibly involved in P solubilization. Inoculation of the strains separately and as a consortium of the three bacteria (S. meliloti, B. flexus and B. megaterium) in P-deficient soil (4.33 w/v P) fertilized without P improved plant height, shoot and root dry weight, as well as P nutrition in the maize plants. Use of the B. flexus and B. megaterium strains separately and in a consortium positively affected several growth parameters and P nutrition in plants supplemented with insoluble P. No effect was observed when pots in which the seedlings were growing were supplied with soluble fertilizer. A second assay using a P-deficient soil (6.64 w/v P) showed that inoculation with the consortium of B. flexus and B. megaterium significantly increased growth and total P content in maize plants. A dose–response P fertilization experiment using sterile P-deficient soil led us to conclude that inoculation to soil of the mixture of B. flexus and B. megaterium may improve P nutrition and growth to a level previously attained by the addition of soluble P-fertilizer at 40 w/v P. A non-sterile experiment showed a beneficial response with B. megaterium but not with B. flexus. We propose utilizing these bacteria in P-deficient alkaline soils in future field trials in order to evaluate their potential as biofertilizers.     

Plant growth promotion and suppression of <Emphasis Type="Italic">Phytophthora drechsleri</Emphasis> damping-off in cucumber by cellulase-producing <Emphasis Type="Italic">Streptomyces</Emphasis>

Phytophthora drechsleri damping-off is one of the most important diseases of cucumber (Cucumis sativus). Salinity is a serious problem for crop production and affects diversity and activity of soil microorganisms. Application of salt-tolerant biocontrol agents may be beneficial in order to protect plants against pathogenic fungi in saline soils. In this study, a total of 717 Streptomyces isolates were isolated from the rhizosphere of cucumber, out of which two isolates showed more than 70% inhibitory effect against P. drechsleri and had cellulase activity in the presence and absence of NaCl. In a greenhouse experiment, two Streptomyces isolates with the highest antagonistic activity, strains C 201 and C 801, reduced seedling damping-off of cucumber caused by P. drechsleri by 77 and 80%, respectively, in artificially infested soils. Strain C 201 increased dry weight of seedlings up to 21% in greenhouse experiments. Phylogenetic analyses of 16S rRNA gene sequence reveals that strains C 201 and C 801 are closely related to S. rimosus and S. monomycini respectively. Increased activity of polyphenol oxidase (PPO) and peroxidase (POX) enzymes in Streptomyces-treated plants proved the biocontrol-induced systemic resistance (ISR) in cucumber plants against P. drechsleri.     

Laboratory and simulated-field bioassays for assessing mixed cultures of <Emphasis Type="Italic">Lysinibacillus sphaericus</Emphasis> against <Emphasis Type="Italic">Aedes aegypti</Emphasis> (Diptera: Culicidae) larvae resistant to temephos

Aedes aegypti (L.) is the main vector of tropical diseases such as dengue, chikungunya and Zika. Due to the overuse of insecticides, Ae. aegypti resistant populations have increased. Biological control with Lysinibacillus sphaericus (Ahmed) has been used against Culex sp. and Anopheles sp. Although Ae. aegypti is refractory to the binary toxin of L. sphaericus spores, vegetative cells have been shown to be effective against Ae. aegypti larvae. In this work, the effect of L. sphaericus vegetative cells on Ae. aegypti temephos-resistant larvae was assessed under lab and simulated field conditions. L. sphaericus caused about 90% mortality of insecticide-resistant Ae. aegypti larvae under simulated field conditions. Likewise, Ae. aegypti larvae were more sensitive to mixed cultures of L. sphaericus than to individual strains; then, the most effective mixed culture exhibited an LC₅₀ of 1.21 × 10⁵ CFU/mL with Rockefeller larvae and 8.04 × 10⁴ CFU/mL with field-collected larvae. Additionally, we found that mixed cultures composed of two L. sphaericus strains were more effective than a culture formed by the three strains. Our results suggest that mixed cultures comprising L. sphaericus vegetative cells could be useful for controlling temephos-resistant populations of Ae. aegypti, as evidenced by the effectiveness demonstrated under laboratory and simulated field conditions.     

Isolation and Characterization of Toxic Cyanobacteria from Different Natural Sources

We isolated seven algologically and five bacteriologically pure cultures of toxin-producing cyanobacteria from Turgen gorge (Kazakhstan), Karlovy Vary (Czech Republic), and Shar-Nuur Lake, Bayan Ulgiiregion (Mongolia) springs. According to the Daphnia magna test, Desertifilum sp. and Nostoc sp. strains were the most toxic in the test of isolated strains (complete death of all test organisms was detected after 48 h). These strains possessed the highest inhibitory effect on proliferation of the HeLa cancer cell line. The Anabaena sp. 35 and Nostoc sp. 4 strains were also high toxic. Model strains Synechocystis PCC 6803 and Synechococcus elongates PCC 7942, as well as the strain isolated in the present work, Synechococcus sp. 55, were less toxic. Mass spectrometry made it possible to assign cyanobacterial toxins to cyclic depsipeptides. Two cyclic depsipeptides, micropeptin T and oscillapeptin, were detected in Desertifilum sp. extracts. Cryptophycin and small amounts of cyclic depsipeptide micropeptin SD were detected in Nostoc sp. extract.     

<Emphasis Type="Italic">Streptomyces urticae</Emphasis> sp. nov., isolated from rhizosphere soil of <Emphasis Type="Italic">Urtica urens</Emphasis> L.

Two novel Gram-stain positive, spore-forming, aerobic actinomycetes, designated NEAU-PCY-1^T and NEAU-PCY-2, were isolated from rhizosphere soil of Urtica urens L. collected from Anshan, Liaoning Province, northeast China. The 16S rRNA gene sequence analysis showed that strains NEAU-PCY-1^T and NEAU-PCY-2 exhibited 99.8% similarity with each other and are closely related to Streptomyces abietis DSM 42080^T (98.2, 98.3%) and Streptomyces fildesensis DSM 41987^T (98.0, 98.1%). Phylogenetic analysis based on the 16S rRNA gene sequences indicated that the two strains formed a cluster with these two closely related species. Moreover, DNA–DNA hybridization results and some phenotypic, physiological and biochemical properties differentiated the two strains from their close relatives in the genus Streptomyces. Based on a polyphasic taxonomy study, strains NEAU-PCY-1^T and NEAU-PCY-2 are considered to represent a novel species of the genus Streptomyces, for which the name Streptomyces urticae sp. nov. is proposed, with NEAU-PCY-1^T (=?DSM 105115^T?=?CCTCC AA 2017015^T) as the type strain.     

Characterization and enzymatic hydrolysis of hydrothermally treated β-1,3–1,6-glucan from <Emphasis Type="Italic">Aureobasidium pullulans</Emphasis>

The chemical structure of hydrothermally treated β-1,3–1,6-glucan from Aureobasidium pullulans was characterized using techniques such as gas chromatography/mass spectrometry (GC/MS) and nuclear magnetic resonance (NMR). The chemical shifts of anomeric carbons observed in the ¹³C-NMR spectra suggested the presence of single flexible chains of polysaccharide in the sample. β-1,3–1,6-Glucan from A. pullulans became water-soluble, with an average molecular weight of 128,000 Da after hydrothermal treatment, and the solubility in water was approximately 10% (w/w). Sample (3% w/v) was completely hydrolyzed to glucose by enzymatic reaction with Lysing enzymes from Trichoderma harzianum. Gentiobiose (Glcβ1 → 6Glc) and glucose were released as products during the reaction, and the maximum yield of gentiobiose was approximately 70% (w/w). The molar ratio of gentiobiose to glucose after 1 h reaction suggested that the sample is likely highly branched. Sample (3% w/v) was also hydrolyzed to glucose by Uskizyme from Trichoderma sp., indicating that it is very sensitive to enzymatic hydrolysis.     

Unraveling the cellulolytic and hemicellulolytic potential of two novel <Emphasis Type="Italic">Streptomyces</Emphasis> strains

The Streptomyces spp. are notorious plant biomass decomposers in soil environments, but only few strains were biochemically and genetically characterized. Here, we employed functional screening along with genomic sequencing for identification of novel lignocellulolytic Streptomyces strains. Streptomyces strains isolated from soil were functional screened based on their cellulolytic and hemicellulolytic capacities by enzymatic plate assays containing carboxymethylcellulose (CMC) and beechwood xylan as sole carbon source. Subsequently, genomes of Streptomyces strains were sequenced, annotated, and interpreted to correlate their genetic contents with biochemical properties. Among the 80 bacterial isolates that were screened for enzymatic activity, two Streptomyces strains (named as F1 and F7) exhiting higher endoglucanase and endoxylanase activities were selected for biochemical and genomic characterization. After cultivation on steam-pretreated sugarcane bagasse-based medium, the supernatant of the strains F1 and F7 exhibited enzymatic activity against different substrates, such as arabinan, rye arabinoxylan, β-glucan, starch, CMC, xylan, and chitin. Furthermore, strain F7 was able to degrade pectin, mannan, and lichenan. The genomic analysis of both strains revealed a diversity of carbohydrate-active enzymes. The F1 and F7 genomes encode 33 and 44 different types of glycosyl hydrolases families, respectively. Moreover, the genomic analysis also identified genes related to degradation of lignin-derived aromatic compounds. Collectively, the study revealed two novel Streptomyces strains and further insights on the degradation capability of lignocellulolytic bacteria, from which a number of technologies can arise, such as saccharification processes.     

Combined use of <Emphasis Type="Italic">GAP</Emphasis> and <Emphasis Type="Italic">AOX1</Emphasis> promoters and optimization of culture conditions to enhance expression of <Emphasis Type="Italic">Rhizomucor miehei</Emphasis> lipase

Rhizo mucor miehei lipase (RML) is an industrially important enzyme, but its application is limited due to its high cost. In this study, a series of measures such as codon optimization, propeptide addition, combined use of GAP and AOX1 promoters, and optimization of culture conditions were employed to increase the expression of RML. Three transformants of the constitutive-inducible combined Pichia pastoris strains were generated by transforming the pGAPZαA-rml vector into the pPIC9K-rml/GS115 strain, which resulted in high-expression yields of RML. Using the shake flask method, highest enzyme activity corresponding to 140 U/mL was observed in the strain 3-17, which was about sixfold higher than that of pPIC9K-rml/GS115 or pGAPZαA-rml/GS115. After optimization of culture conditions by response surface methodology, the lipolytic activity of strain 3-17 reached 175 U/mL in shake flasks. An increase in the copy number simultaneously with the synergistic effect provided by two promoters led to enhanced degree of protein expression.     

<Emphasis Type="Italic">Streptomyces xiangluensis</Emphasis> sp. nov., a novel actinomycete isolated from soil

A novel actinomycete, designated strain NEAU-LA29^T, was isolated from soil collected from Xianglu Mountain and subjected to a polyphasic taxonomic study. Based on a polyphasic taxonomic approach comprising chemotaxonomic, phylogenetic, morphological and physiological characterisation, the isolate has been affiliated to the genus Streptomyces. 16S rRNA gene sequence analysis showed that the isolate is closely related to Streptomyces vastus JCM4524^T (98.8% identity) and Streptomyces cinereus DSM43033^T (97.9%). However, multilocus sequence analysis based on five other house-keeping genes (atpD, gyrB, rpoB, recA and trpB) and low DNA–DNA relatedness values enabled the strain to be differentiated from these closely related species of the genus Streptomyces. Thus, strain NEAU-LA29^T is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces xiangluensis sp. nov. is proposed. The type strain is NEAU-LA29^T (=?CGMCC 4.7466^T?=?DSM 105786^T).     

Endophytic <Emphasis Type="Italic">Streptomyces</Emphasis> sp. AC35, a producer of bioactive isoflavone aglycones and antimycins

In this research, a microbial endophytic strain obtained from the rhizosphere of the conifer Taxus baccata and designated as Streptomyces sp. AC35 (FJ001754.1 Streptomyces, GenBank) was investigated. High 16S rDNA gene sequence similarity suggests that this strain is closely related to S. odorifer. The major fatty acid profile of intracellular lipids was also carried out to further identify this strain. Atomic force microscopy and scanning acoustic microscopy were used to image our strain. Its major excreted substances were extracted, evaluated for antimicrobial activity, purified, and identified by ultraviolet–visible spectroscopy (UV–vis), liquid chromatography–mass spectrometry (LC–MS/MS) and nuclear magnetic resonance as the bioactive isoflavone aglycones—daidzein, glycitein and genistein. Batch cultivation, performed under different pH conditions, revealed enhanced production of antimycin components when the pH was stable at 7.0. Antimycins were detected by HPLC and identified by UV–vis and LC–MS/MS combined with the multiple reaction monitoring. Our results demonstrate that Streptomyces sp. AC35 might be used as a potential source of effective, pharmaceutically active compounds.     

Expression,purification and characterization of a thermostable leucine dehydrogenase from the halophilic thermophile <Emphasis Type="Italic">Laceyella sacchari</Emphasis>

Objective

A potential thermotolerant l-leucine dehydrogenase from Laceyella sacchari (Ls-LeuDH) was over-expressed in E. coli, purified and characterized.

Results

Ls-LeuDH had excellent thermostability with a specific activity of 183 U/mg at pH 10.5 and 25 °C. It retained a high activity in 200 mM carbonate buffer from pH 9.5 to 11. The optimal temperature for Ls-LeuDH was 60 °C.

Conclusion

It is the first time that a thermostable and highly active LeuDH originating from L. sacchari has been characterized. It may be useful for medical and pharmaceutical applications.