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Applied Microbiology and Biotechnology

Peilin Chen, Meilin Zhao, Feng Tang, Yanmin Hu, Xianjun Peng, Shihua Shen
Few studies have investigated the effect of environment on the root-associated microbiome, especially for woody plants in their native environment. The roots and rhizosphere soils of a native woody species (Broussonetia papyrifera) sampled across four different climate types in China were used to elucidate the influence of environment on the root-associated microbiome. Our results showed that the B. papyrifera root-associated microbiome contained abundant Proteobacteria and Basidiomycota, especially Pseudomonas and Rhizobium...
March 22, 2019: Applied Microbiology and Biotechnology
Anissa Dieudonné, David Pignol, Sandra Prévéral
The scientific community's interest in magnetotactic bacteria has increased substantially in recent decades. These prokaryotes have the particularity of synthesizing nanomagnets, called magnetosomes. The majority of research is based on several scientific questions. Where do magnetotactic bacteria live, what are their characteristics, and why are they magnetic? What are the molecular phenomena of magnetosome biomineralization and what are the physical characteristics of magnetosomes? In addition to scientific curiosity to better understand these stunning organisms, there are biotechnological opportunities to consider...
March 22, 2019: Applied Microbiology and Biotechnology
Tomo Kondo, Shigehiko Yumura
Methods for heterologous protein production in Escherichia coli have revolutionized biotechnology and the bioindustry. It is ultimately important to increase the amount of protein product from bacteria. To this end, a variety of tools, such as effective promoters, have been developed. Here, we present a versatile molecular tool based on a phenomenon termed "translation enhancement by a Dictyostelium gene sequence" ("TED") in E. coli. We found that protein expression was increased when a gene sequence of Dictyostelium discoideum was placed upstream of the Shine-Dalgarno sequence located between the promoter and the initiation codon of a target gene...
March 22, 2019: Applied Microbiology and Biotechnology
Alei Geng, Jian Wu, Rongrong Xie, Hongcheng Wang, Yanfang Wu, Xia Li, Fuxiang Chang, Jianzhong Sun
Arabinofuranosidase plays an essential role in the process of hydrolysis of arabinoxylan (AX). Thermostable, versatile, and efficient arabinofuranosidase is thus of great interest for the biorefinery industry. A GH51 arabinofuranosidase, Abf51, from Hungateiclostridium clariflavum DSM 19732 was heterogeneously expressed in Escherichia coli. Abf51 was found to have an optimal pH and temperature of 6.5 and 60 °C, respectively, with very high thermostability. At the optimal working temperature (60 °C), Abf51 retained over 90% activity after a 2-day incubation and over 60% activity after a 6-day incubation...
March 22, 2019: Applied Microbiology and Biotechnology
Anayeli Guadalupe Beltran-Aguilar, Santy Peraza-Echeverria, Luisa Alhucema López-Ochoa, Ileana Cecilia Borges-Argáez, Virginia Aurora Herrera-Valencia
The expression of transgenes in the nucleus is an attractive alternative for the expression of recombinant proteins in the green microalga Chlamydomonas reinhardtii. For this purpose, a strong inducible promoter that allows protein accumulation without possible negative effects on cell multiplication and biomass accumulation is desirable. A previous study at our laboratory identified that the CrGPDH3 gene from C. reinhardtii was inducible under NaCl treatments. In this study, we cloned and characterized a 3012 bp sequence upstream of the start codon of the CrGPDH3 gene, including the 285 bp 5' untranslated region...
March 21, 2019: Applied Microbiology and Biotechnology
Thanh-Tam Ho, Hosakatte Niranjana Murthy, Dayanand Dalawai, Medha A Bhat, Kee-Yoeup Paek, So-Young Park
A vast array of plant-based compounds has enriched red biotechnology to serve the human health and food. A peculiar medicinal plant which was an element of traditional Chinese medicine for centuries as a liver and kidney tonic, for life longevity and hair blackening, is Polygonum multiflorum Thunb. (PM) which is popularly known as "He shou wu" or "Fo-ti" and is rich in chemical components like stilbenes, quinones, and flavonoids which have been used as anti-aging, anti-alopecia, anti-cancer, anti-oxidative, anti-bacterial, anti-hyperlipidemia, anti-atherosclerosis, and immunomodulating and hepatoprotective agents in the modern medicine...
March 20, 2019: Applied Microbiology and Biotechnology
Baofeng Cui, Xinsheng Liu, Peng Zhou, Yuzhen Fang, Donghong Zhao, Yongguang Zhang, Yonglu Wang
Many recent studies have shown that flagellin fused to heterologous antigens can induce significantly enhanced humoral and cellular immune responses through its adjuvant activity. Therefore, in this study, two key B cell epitopes and a truncated VP1 (ΔVP1) protein from foot-and-mouth disease virus (FMDV) were expressed as flagellin fusion proteins in different patterns. Specifically, ΔVP1 and two duplicates of two key B cell epitopes (2×B1B2) were fused separately to the C-terminus of flagellin with a universal exogenous T cell epitope to construct FT (Flagellin-Truncated VP1) and FME (Flagellin-Multiple Epitopes)...
March 19, 2019: Applied Microbiology and Biotechnology
Kathrin Pommerehne, Jana Walisko, Anna Ebersbach, Rainer Krull
Rebeccamycin is an antibiotic and antitumor substance isolated from the filamentous bacterium Lentzea aerocolonigenes. After its discovery, investigations of rebeccamycin focused on elucidating its structure, biological activity, and biosynthetic pathway. For potential medical application, a sufficient drug supply has to be ensured, meaning that the production process of rebeccamycin plays a major role. In addition to the natural production of rebeccamycin in L. aerocolonigenes, where the complex cell morphology is an important factor for a sufficient production, rebeccamycin can also be heterologously produced or chemically synthesized...
March 19, 2019: Applied Microbiology and Biotechnology
Manyu Luo, Meiqi Zhao, Cedric Cagliero, Hua Jiang, Yueqing Xie, Jianwei Zhu, Hui Yang, Mengxiao Zhang, Ying Zheng, Yunsheng Yuan, Zixiu Du, Huili Lu
Antigen-binding fragments (Fabs) are an important part of monoclonal antibody (mAb) therapeutics and can be cost-effectively produced using an Escherichia coli (E. coli) expression system. However, Fabs tend to form undesirable aggregates when expressed in the cytoplasm of E. coli, substantially reducing the yield of correctly folded proteins. To solve this problem, in this study, we used five Fab fragments targeting IGF1R, Her2, VEGF, RANKL, and PD-1 to develop a novel system employing the alkaline phosphatase (phoA) promoter and the heat-stable enterotoxin II (STII) leader sequence to facilitate the efficient expression and extracellular secretion of Fabs...
March 18, 2019: Applied Microbiology and Biotechnology
Gyu Bi Lee, Yun Jae Kim, Jae Kyu Lim, Tae Wan Kim, Sung Gyun Kang, Hyun Sook Lee, Jung-Hyun Lee
The biosynthetic pathway of 2,3-butanediol (2,3-BDO) production from pyruvate under anaerobic conditions includes three enzymes: acetolactate synthase (ALS), acetolactate decarboxylase (ALDC), and acetoin reductase (AR). Recently, in anaerobic hyperthermophilic Pyrococcus furiosus, it has been reported that acetoin, a precursor of 2,3-BDO, is produced from pyruvate by ALS through a temperature-dependent metabolic switch. In this study, we first attempted to produce 2,3-BDO from Thermococcus onnurineus NA1 using a simple biosynthetic pathway by two enzymes (ALS and AR) at a high temperature...
March 18, 2019: Applied Microbiology and Biotechnology
José Alberto Martínez-Fernández, Daniel Bravo, Ángela Peirotén, Juan Luis Arqués, José María Landete
Bioengineering of probiotics allows the improvement of their beneficial characteristics. In this work, we develop a molecular tool that would allow the activation of desirable traits in probiotics once they reach the intestine. The activity of upstream regions of bile-inducible genes of Lactobacillus casei BL23 and Lactobacillus plantarum WCFS1 was analyzed using plasmids encoding an anaerobic fluorescent protein as reporter. The promoter P16090 from Lb. casei BL23 was selected and its bile induction confirmed in Lb...
March 18, 2019: Applied Microbiology and Biotechnology
Jing Li, Jian Sun, Xinyue Gao, Zhixin Wu, Guangdong Shang
Homologous recombination-based recombineering is a widely used DNA cloning and modification technique; recombineering efficiency improvement would be helpful for high-throughput DNA manipulation. Escherichia coli primase DnaG variants, such as DnaG Q576A and DnaG K580A, increase the recombineering efficiency via impairment of the interaction between primase and the replisome and boost the loading of more ssDNA on the replication fork. Bacterial adaptive immunity origin CRISPR-Cas9 is emerging as a powerful genome editing strategy...
March 16, 2019: Applied Microbiology and Biotechnology
Joanna Zebrowska, Joanna Jezewska-Frackowiak, Ewa Wieczerzak, Franciszek Kasprzykowski, Agnieszka Zylicz-Stachula, Piotr M Skowron
Over 470 prototype Type II restriction endonucleases (REases) are currently known. Most recognise specific DNA sequences 4-8 bp long, with very few exceptions cleaving DNA more frequently. TsoI is a thermostable Type IIC enzyme that recognises the DNA sequence TARCCA (R = A or G) and cleaves downstream at N11/N9. The enzyme exhibits extensive top-strand nicking of the supercoiled single-site DNA substrate. The second DNA strand of such substrate is specifically cleaved only in the presence of duplex oligonucleotides containing a cognate site...
March 16, 2019: Applied Microbiology and Biotechnology
Yota Tsuge, Naoto Kato, Shogo Yamamoto, Masako Suda, Toru Jojima, Masayuki Inui
Strain development is critical for microbial production of bio-based chemicals. The stereo-complex form of polylactic acid, a complex of poly-L- and poly-D-lactic acid, is a promising polymer candidate due to its high thermotolerance. Here, we developed Corynebacterium glutamicum strains producing high amounts of L- and D-lactic acid through intensive metabolic engineering. Chromosomal overexpression of genes encoding the glycolytic enzymes, glucokinase, glyceraldehyde-3-phosphate dehydrogenase, phosphofructokinase, triosephosphate isomerase, and enolase, increased L- and D-lactic acid concentration by 146% and 56%, respectively...
March 15, 2019: Applied Microbiology and Biotechnology
Yu-Ting Zhang, Jia-Yi Jiang, Tian-Qiong Shi, Xiao-Man Sun, Quan-Yu Zhao, He Huang, Lu-Jing Ren
Microalgae are arguably the most abundant single-celled eukaryotes and are widely distributed in oceans and freshwater lakes. Moreover, microalgae are widely used in biotechnology to produce bioenergy and high-value products such as polyunsaturated fatty acids (PUFAs), bioactive peptides, proteins, antioxidants and so on. In general, genetic editing techniques were adapted to increase the production of microalgal metabolites. The main genome editing tools available today include zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas nuclease system...
March 15, 2019: Applied Microbiology and Biotechnology
Wenting Yang, Wenwen Chen, Jingling Huang, Li Jin, Yawei Zhou, Jianing Chen, Na Zhang, Donglai Wu, Encheng Sun, Guangliang Liu
The variant strains of porcine epidemic diarrhea virus (PEDV) severely threaten the pig industry worldwide and cause up to 100% mortality in suckling piglets. It is critically important and urgent to develop tools for detection of PEDV infection. In this study, we developed six monoclonal antibodies (mAbs) targeting N protein of PEDV and analyzed their applications on enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), western blot assay, and flow cytometry assay. The results demonstrated that all these six mAbs were IgG1 isotype and κ chain...
March 15, 2019: Applied Microbiology and Biotechnology
R U Haque, F Paradisi, T Allers
Enzyme-mediated synthesis of pharmaceutical compounds is a 'green' alternative to traditional synthetic chemistry, and microbial engineering opens up the possibility of using whole cells as mini-factories. Whole-cell biocatalysis reduces cost by eliminating expensive enzyme purification and cofactor addition steps, as well as resulting in increased enzyme stability. Haloferax volcanii is a model halophilic archaeon encoding highly salt and organic solvent tolerant enzymes such as alcohol dehydrogenase (HvADH2), which catalyses the reduction of aldehydes and ketone in the presence of NADPH/NADH cofactor...
March 15, 2019: Applied Microbiology and Biotechnology
Nabbi Emamipour, Manouchehr Vossoughi, Fereidoun Mahboudi, Majid Golkar, Pezhman Fard-Esfahani
Production of insulin-like growth factor 1 (IGF1) in Escherichia coli mostly results in the formation of inclusion bodies. In the present study, IGF1 was fused to disulfide bond oxidoreductase A (DsbA) and expressed in SHuffle™ T7 strain, in order to obtain correctly folded protein. Soluble expression and IMAC purification of DsbA-IGF1 were optimized by applying the Box-Behnken design of response surface methodology. The optimization greatly increased concentration of soluble protein from 317 to 2600 mg/L, and IMAC yield from 400 to 1900 mg/L...
March 14, 2019: Applied Microbiology and Biotechnology
Huimin Shi, Xianqing Huang, Zheng Wang, Yejun Guan, Xuehong Zhang
Pyoluteorin (Plt) is a PKS-NRPS hybrid antibiotic that is produced by Pseudomonas spp. and shows strong antifungal and antibacterial activities. Pseudomonas protegens H78, which was isolated from the rape rhizosphere in Shanghai, can produce a large array of secondary metabolites, including antibiotics and siderophores. Plt is produced at low levels in the H78 wild-type strain. This study aimed to improve Plt production through combinatory genetic engineered strategies. Plt production was significantly enhanced (by14...
March 13, 2019: Applied Microbiology and Biotechnology
Eva Bermúdez-García, Carolina Peña-Montes, Isabel Martins, Joana Pais, Cristina Silva Pereira, Sergio Sánchez, Amelia Farrés
Four cutinase genes are encoded in the genome of the saprophytic fungus Aspergillus nidulans, but only two of them have proven to codify for active cutinases. However, their overall roles in cutin degradation are unknown, and there is scarce information on the regulatory effectors of their expression. In this work, the expression of the cutinase genes was assayed by multiplex qRT-PCR in cultures grown in media containing both inducer and repressor carbon sources. The genes ancut1 and ancut2 were induced by cutin and its monomers, while ancut3 was constitutively expressed...
March 13, 2019: Applied Microbiology and Biotechnology
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