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Applied Microbiology and Biotechnology

Eduard Puente-Massaguer, Martí Lecina, Francesc Gòdia
Polyethylenimine (PEI)-based transient gene expression (TGE) is nowadays a well-established methodology for rapid protein production in mammalian cells, but it has been used to a much lower extent in insect cell lines. A fast and robust TGE methodology for suspension Hi5 (Trichoplusia ni) cells is presented. Significant differences in size and morphology of DNA:PEI polyplexes were observed in the different incubation solutions tested. Moreover, minimal complexing time (< 1 min) between DNA and PEI in 150 mM NaCl solution provided the highest transfection efficiency...
October 13, 2018: Applied Microbiology and Biotechnology
Chao Liu, Jing Zhao, Jiao Liu, Xuan Guo, Deming Rao, Haiping Liu, Ping Zheng, Jibin Sun, Yanhe Ma
trans-Proline 4-hydroxylases (trans-P4Hs) hydroxylate free L-proline to trans-4-hydroxy-L-proline (trans-4-Hyp) is a valuable chiral synthon for important pharmaceuticals such as carbapenem antibiotics. However, merely few microbial trans-P4Hs have been identified, and trans-4-Hyp fermentations using engineered Escherichia coli strains expressing trans-P4Hs are usually performed at temperatures below 37 °C, which is likely due to poor stability and low activities. In the present study, a new trans-P4H from uncultured bacterium esnapd13 (UbP4H) with potential in the fermentative production of trans-4-Hyp at 37 °C was reported...
October 12, 2018: Applied Microbiology and Biotechnology
Matthew Chekwube Enebe, Olubukola Oluranti Babalola
The struggle for survival is a natural and a continuous process. Microbes are struggling to survive by depending on plants for their nutrition while plants on the other hand are resisting the attack of microbes in order to survive. This interaction is a tug of war and the knowledge of microbe-plant relationship will enable farmers/agriculturists improve crop health, yield, sustain regular food supply, and minimize the use of agrochemicals such as fungicides and pesticides in the fight against plant pathogens...
October 12, 2018: Applied Microbiology and Biotechnology
Gaspar Banfalvi
The in vitro susceptibility of gentamicin fractions against Fusarium growth was the subject of this retrospective study. Fusariosis was earlier an exceptionally rare human disease and an unrealistic idea to treat soil saprophytes and plant pathogens with expensive antibiotics such as gentamicins or their minor components. Disseminated fusariosis is now the second most frequent lethal fungal infection after aspergillosis especially in neutropenic patients with hematologic malignancy. Results of this study obtained between May and November 1973 were interesting but not practicable and remained unpublished...
October 12, 2018: Applied Microbiology and Biotechnology
Miye Kwon, Adrian Ho, Sukhwan Yoon
The recent drop in the price of natural gas has rekindled the interests in methanotrophs, the organisms capable of utilizing methane as the sole electron donor and carbon source, as biocatalysts for various industrial applications. As heterologous expression of the methane monooxygenases in more amenable hosts has been proven to be nearly impossible, future success in methanotroph biotechnology largely depends on securing phylogenetically and phenotypically diverse methanotrophs with relatively high growth rates...
October 12, 2018: Applied Microbiology and Biotechnology
Hirokazu Suzuki
The genus Geobacillus comprises thermophilic bacilli capable of endospore formation. The members of this genus provide thermostable proteins and can be used in whole cell applications at elevated temperatures; therefore, these organisms are of biotechnological importance. While these applications have been described in previous reviews, the present paper highlights the environmental adaptations and genome diversifications of Geobacillus spp. and their applications in evolutionary-protein engineering. Despite their obligate thermophilic properties, Geobacillus spp...
October 11, 2018: Applied Microbiology and Biotechnology
Shuaihua Gao, Yingxiu Lu, Yuanyuan Li, Rong Huang, Guojun Zheng
(-)-γ-Lactam ((-)-2-azabicyclo[2.2.1]hept-5-en-3-one) has attracted increasing attention as the chiral intermediate of carbocyclic nucleosides most of which serve as pharmaceutical agents such as anti-HIV/HBV drugs abacavir and carbovir. So far, developing in vitro (+)-γ-lactamase-mediated biotransformation has been one of the most efficient approaches for the production of (-)-γ-lactam. In this study, the catalytic activity of the (+)-γ-lactamase from Sulfolobus solfataricus P2 was engineered by semi-rational design...
October 11, 2018: Applied Microbiology and Biotechnology
Keita Takahashi, Ayumu Yano, Shiori Watanabe, Philippe Langella, Luis G Bermúdez-Humarán, Naoki Inoue
Efficient delivery of antigens to the gut-associated lymphoid tissue (GALT) is the most critical step for the induction of mucosal immunity by oral vaccines. As M cells are the main portal for luminal antigens into the GALT, the M cell-targeting of antigens affords a promising strategy toward the development of effective oral vaccines. Lactococcus lactis is a fascinating recombinant host for oral vaccines, as they survive and produce antigens in the gut and have a particularly safe profile for human use. In this study, we developed and evaluated an M cell-targeting oral immunization system using recombinant L...
October 11, 2018: Applied Microbiology and Biotechnology
Masarra M Sakr, Khaled M Aboshanab, Walid F Elkhatib, Mahmoud A Yassien, Nadia A Hassouna
The increasing occurrence of resistance among Pseudomonas aeruginosa clinical isolates necessitates finding alternatives to antibiotics for controlling the infection of such pathogenic bacteria. In this study, lactonase gene ahl-1 from Bacillus weihenstephanensis isolate-P65 was successfully cloned and expressed in Escherichia coli BL21 (DE3) under the control of T7 promoter for utilizing its quorum quenching activity against three multidrug-resistant (MDR) P. aeruginosa clinical isolates. The biological activity of the overexpressed lactonase enzyme (Ahl-1), tested using a synthetic signal and Chromobacterium violaceum CV026 as a biosensor, displayed good catalytic activity using hexanoyl homoserine lactone (HHL) as a substrate and Chromobacterium violaceum (CV026) as a biosensor (77...
October 11, 2018: Applied Microbiology and Biotechnology
Guizhen Wang, Yawen Gao, Hongsu Wang, Jianfeng Wang, Xiaodi Niu
Sortase B (SrtB) is a vital virulence factor that plays a critical role in Staphylococcus aureus (S. aureus) infections, indicating that it could be a latent target for S. aureus infections. In this study, phloretin, a natural compound that primarily exists in the pericarp and velamen of apples and pears, shows little anti-S. aureus activity, but significantly inhibited SrtB activity in vitro. The results of lactate dehydrogenase release and live/dead cell assays suggested that phloretin reduced human alveolar epithelial cell damage caused by S...
October 11, 2018: Applied Microbiology and Biotechnology
Muireann Egan, Francesca Bottacini, Mary O'Connell Motherway, Patrick G Casey, Ruth Morrissey, Silvia Melgar, Jean-Michel Faurie, Christian Chervaux, Tamara Smokvina, Douwe van Sinderen
Members of the Bifidobacterium genus are widely used as probiotics in fermented milk products. Bifidobacterium animalis subsp. animalis CNCM I-4602 grows and survives poorly in reconstituted skimmed milk (RSM). Availing of genome and transcriptome information, this poor growth and survival phenotype in milk was substantially improved by the addition of certain compounds, such as yeast extract, uric acid, glutathione, cysteine, ferrous sulfate, and a combination of magnesium sulfate and manganese sulfate. Carbohydrate utilization of CNCM I-4602 was also investigated, allowing the identification of several carbohydrate utilization gene clusters, and highlighting this strain's inability to utilize lactose, unlike the type strain of this subspecies, B...
October 10, 2018: Applied Microbiology and Biotechnology
Hanyu Wang, Qian Li, Xiaolin Kuang, Difan Xiao, Xuebing Han, Xiangdong Hu, Xi Li, Menggen Ma
Bioconversion of lignocellulosic biomass to high-value bioproducts by fermentative microorganisms has drawn extensive attentions worldwide. Lignocellulosic biomass cannot be efficiently utilized by microorganisms, such as Saccharomyces cerevisiae, but has to be pretreated prior to fermentation. Aldehyde compounds, as the by-products generated in the pretreatment process of lignocellulosic biomass, are considered as the most important toxic inhibitors to S. cerevisiae cells for their growth and fermentation...
October 10, 2018: Applied Microbiology and Biotechnology
Asmaa Mawad, Yosra A Helmy, Abdel-Gawad Shalkami, Dipak Kathayat, Gireesh Rajashekara
Microencapsulation enhances the oral delivery of probiotic bacteria. In this study, the probiotic Escherichia coli Nissle 1917 (EcN) was microencapsulated using alginate and chitosan nanoparticles. The result showed 90% encapsulation yield of EcN, and the encapsulated EcN displayed significantly (P < 0.05) increased survival in low pH (1.5), high bile salt concentration (4%), and high temperature (70 °C). The most effective cryopreservatives of EcN during freezing and thawing was skim milk and sucrose...
October 9, 2018: Applied Microbiology and Biotechnology
Hong-Duo Bao, Mao-da Pang, Ademola Olaniran, Xu-Hui Zhang, Hui Zhang, Yan Zhou, Li-Chang Sun, Stefan Schmidt, Ran Wang
Phages, the most abundant species in the mammalian gut, have numerous advantages as biocontrol agent over antibiotics. In this study, mice were orally treated with the lytic gut phage PA13076 (group B), the temperate phage BP96115 (group C), no phage (group A), or streptomycin (group D) over 31 days. At the end of the experiment, fecal microbiota diversity and composition was determined and compared using high-throughput sequencing of the V3-V4 hyper-variable region of the 16S rRNA gene and virus-like particles (VLPs) were quantified in feces...
October 9, 2018: Applied Microbiology and Biotechnology
Guihua Shen, Shuting Zhang, Xiaojiao Liu, Qipeng Jiang, Wei Ding
Application of soil amendments has been wildly used to increase soil pH and control bacterial wilt. However, little is known about causal shifts in the rhizosphere microbial community of crops, especially when the field naturally harbors the disease of bacterial wilt to tobacco for many years due to long-term continuous cropping and soil acidification. In this study, biochar (CP), lime (LM), oyster shell powder (OS) and no soil amendment additions (Control; CK) were assessed for their abilities to improve the soil acidification, change the composition of rhizosphere soil bacterial communities and thus control tobacco bacterial wilt...
October 9, 2018: Applied Microbiology and Biotechnology
Elisa Vignali, Fabio Tonin, Loredano Pollegioni, Elena Rosini
Peroxidases are well-known biocatalysts produced by all organisms, especially microorganisms, and used in a number of biotechnological applications. The enzyme DypB from the lignin-degrading bacterium Rhodococcus jostii was recently shown to degrade solvent-obtained fractions of a Kraft lignin. In order to promote the practical use, the N246A variant of DypB, named Rh_DypB, was overexpressed in E. coli using a designed synthetic gene: by employing optimized conditions, the enzyme was fully produced as folded holoenzyme, thus avoiding the need for a further time-consuming and expensive reconstitution step...
October 9, 2018: Applied Microbiology and Biotechnology
Manuela Oliveira, António Amorim
Recent advances in genetic data generation, through massive parallel sequencing (MPS), storage and analysis have fostered significant progresses in microbial forensics (or forensic microbiology). Initial applications in circumstances of biocrime, bioterrorism and epidemiology are now accompanied by the prospect of using microorganisms (i) as ancillary evidence in criminal cases; (ii) to clarify causes of death (e.g., drownings, toxicology, hospital-acquired infections, sudden infant death and shaken baby syndromes); (iii) to assist human identification (skin, hair and body fluid microbiomes); (iv) for geolocation (soil microbiome); and (v) to estimate postmortem interval (thanatomicrobiome and epinecrotic microbial community)...
October 9, 2018: Applied Microbiology and Biotechnology
Adivitiya, Babbal, Shilpa Mohanty, Yogender Pal Khasa
Streptokinase, a therapeutically important thrombolytic agent, is prone to C-terminal degradation and plasmin-mediated proteolytic processing. Since the protein was glycosylated during secretion from Pichia pastoris, therefore, the role of carbohydrate moieties on its stability was analyzed via in vivo blocking of N-glycosylation using tunicamycin where an increased degradation of streptokinase was observed. Further, the in vitro site-directed mutagenesis of the three putative N-glycosylation sites at asparagine residues 14, 265, and 377 to alanine revealed the essentiality of glycosylation of the 14th amino acid residue in its post-translational proteolytic stability without significantly affecting its biological activity...
October 8, 2018: Applied Microbiology and Biotechnology
Ana C Reis, Monika Čvančarová, Ying Liu, Markus Lenz, Timm Hettich, Boris A Kolvenbach, Philippe F-X Corvini, Olga C Nunes
In the last decade, biological degradation and mineralization of antibiotics have been increasingly reported feats of environmental bacteria. The most extensively described example is that of sulfonamides that can be degraded by several members of Actinobacteria and Proteobacteria. Previously, we reported sulfamethoxazole (SMX) degradation and partial mineralization by Achromobacter denitrificans strain PR1, isolated from activated sludge. However, further studies revealed an apparent instability of this metabolic trait in this strain...
October 8, 2018: Applied Microbiology and Biotechnology
Valeria Prigione, Federica Spina, Valeria Tigini, Samuele Giovando, Giovanna Cristina Varese
Tannins are secondary metabolites that are widely distributed in the plant kingdom. They act as growth inhibitors for many microorganisms: they are released upon microbial attack, helping to fight infection in plant tissues. Extraction of tannins from plants is an active industrial sector with several applications, including oenology, animal feeding, mining, the chemical industry, and, in particular, the tanning industry. However, tannins are also considered very recalcitrant pollutants in wastewater of diverse origin...
October 6, 2018: Applied Microbiology and Biotechnology
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