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Applied Microbiology and Biotechnology

Ruben Hohnholz, Kim Julia Pohlmann, Tilman Achstetter
Multicopy episomal plasmids in yeast, used whenever elevated levels of foreign or homologous gene expression are necessary, are known to be less stable compared to the endogenous 2-μm plasmid they are based on, at least without selective pressure. Considering that rich medium favors growth rate and, simultaneously, is less expensive than selective medium, enhancing stability in non-selective medium is extremely desirable. In this study, we changed the architecture of a multicopy model expression plasmid, creating six isoforms (same size, same DNA content but different positions and orientations of the expression block) and studied mitotic stability, copy number, as well as reporter yEGFP3 expression between isoforms...
October 20, 2017: Applied Microbiology and Biotechnology
Botond Hegedüs, Péter B Kós, Gábor Bende, Naila Bounedjoum, Gergely Maróti, Krisztián Laczi, Márk Szuhaj, Katalin Perei, Gábor Rákhely
Novosphingobium resinovorum SA1 was the first single isolate capable of degrading sulfanilic acid, a widely used representative of sulfonated aromatic compounds. The genome of the strain was recently sequenced, and here, we present whole-cell transcriptome analyses of cells exposed to sulfanilic acid as compared to cells grown on glucose. The comparison of the transcript profiles suggested that the primary impact of sulfanilic acid on the cell transcriptome was a starvation-like effect. The genes of the peripheral, central, and common pathways of sulfanilic acid biodegradation had distinct transcript profiles...
October 19, 2017: Applied Microbiology and Biotechnology
Irnayuli R Sitepu, Luis A Garay, Lauren Enriquez, Russell Fry, John H Butler, Julian M Lopez, Atit Kanti, Sarah A Faulina, Agustinus J Nugroho, Blake A Simmons, Steven W Singer, Christopher W Simmons, Kyria Boundy-Mills
Pretreatment with ionic liquids (IL) such as 1-ethyl-3-methylimidazolium chloride or acetate is an effective method for aiding deconstruction of lignocellulosic biomass; however, the residual IL remaining in hydrolysates can be inhibitory to growth of ethanologenic or oleaginous yeasts that have been examined in the literature. The aim of this study was to identify oleaginous yeasts that are tolerant of the IL [C2C1Im][OAc] and [C2C1Im]Cl using 45 strains belonging to 38 taxonomically diverse species within phyla Ascomycota and Basidiomycota...
October 19, 2017: Applied Microbiology and Biotechnology
Lijuan Hou, Yu Li, Mingjie Chen, Zhengpeng Li
Compared with controls, treatment of the cultivation substrate for Volvariella volvacea with 0.02% NaAc at the "spraying water" stage increased the number of fruiting body primordia by 280, the mushroom yield by 16.25%, the number of fruiting bodies by 35.57%, and the biological efficiency by 16.28%. The average single mushroom weight increased by 19.33%, but there was no significant difference between treatments and controls. A correlation analysis revealed a significant (p < 0.05) positive correlation between the total yield and the number of fruiting bodies...
October 19, 2017: Applied Microbiology and Biotechnology
Scott J Funston, Konstantina Tsaousi, Thomas J Smyth, Matthew S Twigg, Roger Marchant, Ibrahim M Banat
Microbially produced rhamnolipids have significant commercial potential; however, the main bacterial producer, Pseudomonas aeruginosa, is an opportunistic human pathogen, which limits biotechnological exploitation. The non-pathogenic species Burkholderia thailandensis produces rhamnolipids; however, yield is relatively low. The aim of this study was to determine whether rhamnolipid production could be increased in Burkholderia thailandensis through mutation of genes responsible for the synthesis of the storage material polyhydroxyalkanoate (PHA), thereby increasing cellular resources for the production of rhamnolipids...
October 17, 2017: Applied Microbiology and Biotechnology
Gui-Ping Wen, Zi-Min Tang, Si-Ling Wang, Wen-Fang Ji, Wei Cai, Xu Zhang, Shou-Jie Huang, Ting Wu, Jun Zhang, Zi-Zheng Zheng, Ning-Shao Xia
Hepatitis E virus (HEV) is one of the major pathogens that cause acute viral hepatitis. The human (genotypes 1 and 2) and zoonotic (genotypes 3 and 4) groups of HEV present different epidemiology and clinical features. In this study, we developed a classification method for rapidly classifying HEV into human or zoonotic groups that combines a general antigen test with a zoonotic group-specific antigen test. Evaluation of serial samples from HEV-infected rhesus monkeys indicated that HEV antigen-positive samples can be classified using the antigen-based classification method...
October 16, 2017: Applied Microbiology and Biotechnology
Kinuko Ueno, Yuta Sakai, Chika Shono, Ippei Sakamoto, Kaori Tsukakoshi, Yukako Hihara, Koji Sode, Kazunori Ikebukuro
Cyanobacteria are one of the most attractive hosts for biofuel production; however, genetic approaches to regulate specific chromosomal genes in cyanobacteria remain limited. With the aim of developing a novel method to regulate chromosomal gene expression in cyanobacteria, we focused on riboregulatory technology. Riboregulators are composed of two RNA fragments whose interaction leads to target gene regulation with high specificity. In this study, we inserted a riboregulator sequence upstream of the chromosomal gene encoding AbrB-like transcriptional regulator, cyAbrB2, to investigate the utility of this tool...
October 16, 2017: Applied Microbiology and Biotechnology
Kateřina Svobodová, Čeněk Novotný
White-rot fungi are renowned for their remarkable potential to degrade a wide range of organic pollutants. They are applicable in standard bioreactors offering both the use of the continuous mode of action and easy upscaling of the biodegradation process. The recent advance in this field consisted in the use of various fungi and different types of reactors in the treatment of real wastewaters. Most degradation studies involving white-rot fungi carried out so far used controlled, aseptic conditions. However, during bioremediation of real wastewaters, the degradation capacity of the fungi would be significantly affected by autochthonous microorganisms...
October 16, 2017: Applied Microbiology and Biotechnology
Laura Salusjärvi, Mervi Toivari, Maija-Leena Vehkomäki, Outi Koivistoinen, Dominik Mojzita, Klaus Niemelä, Merja Penttilä, Laura Ruohonen
The important platform chemicals ethylene glycol and glycolic acid were produced via the oxidative D-xylose pathway in the yeast Saccharomyces cerevisiae. The expression of genes encoding D-xylose dehydrogenase (XylB) and D-xylonate dehydratase (XylD) from Caulobacter crescentus and YagE or YjhH aldolase and aldehyde dehydrogenase AldA from Escherichia coli enabled glycolic acid production from D-xylose up to 150 mg/L. In strains expressing only xylB and xylD, 29 mg/L 2-keto-3-deoxyxylonic acid [(S)-4,5-dihydroxy-2-oxopentanoic acid] (2K3DXA) was produced and D-xylonic acid accumulated to ca...
October 16, 2017: Applied Microbiology and Biotechnology
Xiao Xiao, Miaochun Fan, Entao Wang, Weimin Chen, Gehong Wei
Although the rhizomicrobiome has been extensively studied, little is known about the interactions between soil properties and the assemblage of plant growth-promoting microbes in the rhizosphere. Herein, we analysed the composition and structure of rhizomicrobiomes associated with soybean and alfalfa plants growing in different soil types using deep Illumina 16S rRNA sequencing. Soil pH, P and K significantly affected the composition of the soybean rhizomicrobiome, whereas soil pH and N had a significant effect on the alfalfa rhizomicrobiome...
October 16, 2017: Applied Microbiology and Biotechnology
Yong-Jie Zhang, Xiao-Qing Yang, Shu Zhang, Richard A Humber, Jianping Xu
Mitochondrial DNA is generally regarded to evolve faster than nuclear DNA in animals, whereas if this is also true in fungi remains unclear. Herein, we annotate the first complete mitochondrial genome (mitogenome) of the cyclosporin-producing fungus Tolypocladium inflatum and report the genome-wide sequence variations among five isolates originating from distantly separated localities. We found that T. inflatum has among the most compact of fungal mitogenomes; its 25 kb DNA molecule encodes all standard fungal mitochondrial genes and harbors only one intron...
October 15, 2017: Applied Microbiology and Biotechnology
B Vijayalakshmi Ayyar, M Zouhair Atassi
Myasthenia gravis (MG) is an autoimmune disease caused by sensitization of the immune system to self-antigens. We have previously shown that targeting MG-susceptible alleles can significantly inhibit proliferation of disease-specific T cells. In this work, we humanized a murine monoclonal antibody (mAb) LG11, capable of blocking MG-associated DQ beta 1 (DQB1) allele and reformatted it into single-chain fragment variable (scFv). A fully functional humanized scFv was obtained by optimizing variable domain orientations and linker lengths, along with the optimization of expression conditions and codons to suit Escherichia coli expression machinery...
October 15, 2017: Applied Microbiology and Biotechnology
Hanyu Wang, Yidan Ouyang, Chang Zhou, Difan Xiao, Yaping Guo, Lan Wu, Xi Li, Yunfu Gu, Quanju Xiang, Ke Zhao, Xiumei Yu, Likou Zou, Menggen Ma
Aldehydes generated as by-products during the pretreatment of lignocellulose are the key inhibitors to Saccharomyces cerevisiae, which is considered as the most promising microorganism for industrial production of biofuel, xylitol as well as other special chemicals from lignocellulose. S. cerevisiae has the inherent ability to in situ detoxify aldehydes to corresponding alcohols by multiple aldehyde reductases. Herein, we report that an uncharacterized open reading frame YKL071W from S. cerevisiae encodes a novel "classical" short-chain dehydrogenase/reductase (SDR) protein with NADH-dependent enzymatic activities for reduction of furfural (FF), glycolaldehyde (GA), formaldehyde (FA), and benzaldehyde (BZA)...
October 15, 2017: Applied Microbiology and Biotechnology
An-Dong Li, Liping Ma, Xiao-Tao Jiang, Tong Zhang
During the past years, antibiotic-resistant bacteria (ARB) leading for the spreading of antibiotic resistance genes (ARGs) became a global problem, especially multidrug-resistant (MDR) bacteria are considered the prime culprit of antibiotic resistance. However, the correlation between the antibiotic-resistant phenotype and the ARG profiles remains poorly understood. In the present study, metagenomic functional screening and metagenomic analysis of coliforms were combined to explore the phenotype and genotype of the ARBs from municipal sewage...
October 15, 2017: Applied Microbiology and Biotechnology
Mario Cezar Rodrigues Mano, Iramaia Angélica Neri-Numa, Juliana Bueno da Silva, Bruno Nicolau Paulino, Marina Gabriel Pessoa, Gláucia Maria Pastore
Oligosaccharides are polymers with two to ten monosaccharide residues which have sweetener functions and sensory characteristics, in addition to exerting physiological effects on human health. The ones called nondigestible exhibit a prebiotic behavior being fermented by colonic microflora or stimulating the growth of beneficial bacteria, playing roles in the immune system, protecting against cancer, and preventing cardiovascular and metabolic issues. The global prebiotics market is expected to grow around 12...
October 14, 2017: Applied Microbiology and Biotechnology
Teruyo Ojima-Kato, Naomi Yamamoto, Satomi Nagai, Keisuke Shima, Yumi Akiyama, Junji Ota, Hiroto Tamura
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based microbial identification is a popular analytical method. Strain Solution proteotyping software available for MALDI-TOF MS has great potential for the precise and detailed discrimination of microorganisms at serotype- or strain-level, beyond the conventional mass fingerprinting approaches. Here, we constructed a theoretically calculated mass database of Salmonella enterica subspecies enterica consisting of 12 biomarker proteins: ribosomal proteins S8, L15, L17, L21, L25, and S7, Mn-cofactor-containing superoxide dismutase (SodA), peptidyl-prolyl cis-trans isomerase C (PPIase C), and protein Gns, and uncharacterized proteins YibT, YaiA, and YciF, that can allow serotyping of Salmonella...
October 14, 2017: Applied Microbiology and Biotechnology
Wenyu Gu, Jeremy D Semrau
In aerobic methanotrophs, copper and cerium control the expression and activity of different forms of methane monooxygenase and methanol dehydrogenase, respectively. To exploit methanotrophy for the valorization of methane, it is crucial to determine if these metals exert more global control on gene expression in methanotrophs. Using RNA-Seq analysis we compared the transcriptome of Methylosinus trichosporium OB3b grown in the presence of varying amounts of copper and cerium. When copper was added in the absence of cerium, expression of genes encoding for both soluble and particulate methane monooxygenases varied as expected...
October 14, 2017: Applied Microbiology and Biotechnology
Dora Allegra Carbone, Giuseppe Olivieri, Antonino Pollio, Gabriele, Michael Melkonian
Scenedesmus is a genus of microalgae employed for several industrial uses. Industrial cultivations are performed in open ponds or in closed photobioreactors (PBRs). In the last years, a novel type of PBR based on immobilized microalgae has been developed termed porous substrate photobioreactors (PSBR) to achieve significant higher biomass density during cultivation in comparison to classical PBRs. This work presents a study of the growth of Scenedesmus vacuolatus in a Twin Layer System PSBR at different light intensities (600 μmol photons m(-2) s(-1) or 1000 μmol photons m(-2) s(-1)), different types and concentrations of the nitrogen sources (nitrate or urea), and at two CO2 levels in the gas phase (2% or 0...
October 14, 2017: Applied Microbiology and Biotechnology
Junfeng Dou, Wei Qin, Aizhong Ding, Xiang Liu, Yi Zhu
This study focused on the protein expression of a Microbacterium sp. strain that utilized various concentrations of benzo(a)pyrene (BaP) as the sole source of carbon and energy under anaerobic conditions. A total of 1539 protein species were quantified by isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC-MS/MS. GO, COG, and pathway enrichment analysis showed that most proteins demonstrated catalytic and binding functions and were mainly involved in metabolic processes, cellular processes, and single-organism processes...
October 14, 2017: Applied Microbiology and Biotechnology
Zijun Guo, Yongyue Lu, Fan Yang, Ling Zeng, Guangwen Liang, Yijuan Xu
Symbionts are associated with many insects and play several multifunctional roles in insect-microorganism mutualistic relationships. The trichlorphon-degrading symbiont Citrobacter freundii (CF-BD) of the oriental fruit fly Bactrocera dorsalis was recently discovered; however, its intraspecies transmission pathway among flies remains unknown. Here, we use fluorescence in situ hybridization (FISH), PCR detection, and a series of ingenious experiments to reveal that CF-BD was aggregated in rectal pads associated with the female ovipositor, and the CF-BD symbiont was vertically transmitted via egg surface contamination...
October 13, 2017: Applied Microbiology and Biotechnology
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