scholarly journals Applications of Bacillus subtilis Spores in Biotechnology and Advanced Materials

2020 ◽  
Vol 86 (17) ◽  
Author(s):  
Xiaopei Zhang ◽  
Amal Al-Dossary ◽  
Myer Hussain ◽  
Peter Setlow ◽  
Jiahe Li
Keyword(s):  
Bacillus Subtilis ◽  
Large Scale ◽  
Genetic Manipulation ◽  
Industrial Applications ◽  
Advanced Materials ◽  
Content Type ◽  
Interdisciplinary Approaches ◽  
Food Shortages ◽  
Material Sciences ◽  
Basic Studies

ABSTRACT The bacterium Bacillus subtilis has long been an important subject for basic studies. However, this organism has also had industrial applications due to its easy genetic manipulation, favorable culturing characteristics for large‐scale fermentation, superior capacity for protein secretion, and generally recognized as safe (GRAS) status. In addition, as the metabolically dormant form of B. subtilis, its spores have attracted great interest due to their extreme resistance to many environmental stresses, which makes spores a novel platform for a variety of applications. In this review, we summarize both conventional and emerging applications of B. subtilis spores, with a focus on how their unique characteristics have led to innovative applications in many areas of technology, including generation of stable and recyclable enzymes, synthetic biology, drug delivery, and material sciences. Ultimately, this review hopes to inspire the scientific community to leverage interdisciplinary approaches using spores to address global concerns about food shortages, environmental protection, and health care.

Even if politicians deal, Lebanon faces a bleak future

2021 ◽  
Keyword(s):  
Foreign Aid ◽  
Rural Areas ◽  
Large Scale ◽  
Economic Reforms ◽  
Financial Strain ◽  
Economic Conditions ◽  
Medium Term ◽  
Content Type ◽  
Global Pandemic ◽  
Food Shortages

Significance The audit and wider structural economic reforms are preconditions for urgently needed foreign aid. Economic conditions in Lebanon are still worsening, with power cuts, food shortages and rising poverty. Impacts A new government would allow reform planning to resume and temporarily stall the decline of the currency. The easing of the global pandemic will somewhat reduce the financial strain, as Lebanon reopens its economy. Soaring poverty rates could provoke large-scale ‘bread riots’ in the coming months. Further devaluation of the currency will make poor Lebanese more dependent on sectarian protection and strengthen patronage. If the situation worsens, sectarian rural areas could revert to warlordism in the medium term.

Low friction bio-inspired polydopamine/polytetrafluoroethylene coating performance in hydrodynamic bearings

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Marius Rutkevičius ◽  
Jimmy Dong ◽  
Darren Tremelling ◽  
Julia Viertel ◽  
Samuel Beckford
Keyword(s):  
Large Scale ◽  
Journal Bearing ◽  
Low Cost ◽  
Optical Microscope ◽  
Polymer Coatings ◽  
Industrial Applications ◽  
Low Friction ◽  
Coating Performance ◽  
Content Type ◽  
Coating Delamination

Purpose Low friction polymer coatings able to withstand high loadings and many years of continuous operation are difficult to formulate at low cost, but could find many applications in industry. This study aims to analyze and compare friction and wear performance of novel polydopamine/polytetrafluoroethylene (PDA/PTFE) and traditional tin Babbitt coatings applied to an industrial journal bearing. Design/methodology/approach This paper tested PTFE based coating, co-deposited with PDA, a biopolymer allowing sea mussels to adhere to ocean rocks. This coating was deposited on flat steel substrates and on a curved cast iron hydrodynamic journal bearing surface. The flat substrates were analyzed with a tribometer and an optical microscope, while the coated bearing liners were tested in an industrial laboratory setting at different speeds and different radial loads. Findings PDA/PTFE coating showed 2-3 times lower friction compared to traditional tin Babbitt for flat substrates, but higher friction in the bearing liners. PDA/PTFE also showed considerable wear through coating delamination and abrasion in the bearing liners. Research limitations/implications Five future modifications to mitigate coating flaws are provided, which include modifications to coating thickness and its surface finish. Originality/value While the novel coating showed excellent results on flat substrates, coating performance in a large scale bearing was found to be poor. This study shows that coating preparation needs to be improved to avoid frictional losses and unwanted damage to bearings. We provide several routes that could improve coating performance in industrial applications.

scholarly journals Engineering the Genome of Thermus thermophilus Using a Counterselectable Marker

2015 ◽  
Vol 197 (6) ◽  
pp. 1135-1144 ◽  
Author(s):  
Jennifer F. Carr ◽  
Michael E. Danziger ◽  
Athena L. Huang ◽  
Albert E. Dahlberg ◽  
Steven T. Gregory
Keyword(s):  
Resistance Gene ◽  
Large Scale ◽  
Genetic Manipulation ◽  
Thermus Thermophilus ◽  
Point Mutations ◽  
Trna Synthetase ◽  
Biological Research ◽  
Lethal Mutant ◽  
Α Subunit ◽  
Content Type

ABSTRACTThermus thermophilusis an extremely thermophilic bacterium that is widely used as a model thermophile, in large part due to its amenability to genetic manipulation. Here we describe a system for the introduction of genomic point mutations or deletions using a counterselectable marker consisting of a conditionally lethal mutant allele ofpheSencoding the phenylalanyl-tRNA synthetase α-subunit. Mutant PheS with an A294G amino acid substitution renders cells sensitive to the phenylalanine analogp-chlorophenylalanine. Insertion of the mutantpheSallele via a linked kanamycin resistance gene into a chromosomal locus provides a gene replacement intermediate that can be removed by homologous recombination usingp-chlorophenylalanine as a counterselective agent. This selection is suitable for the sequential introduction of multiple mutations to produce a final strain unmarked by an antibiotic resistance gene. We demonstrated the utility of this method by constructing strains bearing either a point mutation in or a precise deletion of therrsBgene encoding 16S rRNA. We also used this selection to identify spontaneous, large-scale deletions in the pTT27 megaplasmid, apparently mediated by either of theT. thermophilusinsertion elements ISTth7and ISTth8. One such deletion removed 121 kb, including 118 genes, or over half of pTT27, including multiple sugar hydrolase genes, and facilitated the development of a plasmid-encoded reporter system based on β-galactosidase. The ability to introduce mutations ranging from single base substitutions to large-scale deletions provides a potentially powerful tool for engineering the genome ofT. thermophilusand possibly other thermophiles as well.IMPORTANCEThermus thermophilusis an extreme thermophile that has played an important part in the development of both biotechnology and basic biological research. Its suitability as a genetic model system is established by its natural competence for transformation, but the scarcity of genetic tools limits the kinds of manipulations that can currently be performed. We have developed a counterselectable marker that allows the introduction of unmarked deletions and point mutations into theT. thermophilusgenome. We find that this marker can also be used to select large chromosomal deletions apparently resulting from aberrant transposition of endogenous insertion sequences. This system has the potential to advance the genetic manipulation of this important model organism.

scholarly journals Exploiting the Diversity of Saccharomycotina Yeasts To Engineer Biotin-Independent Growth of Saccharomyces cerevisiae

2020 ◽  
Vol 86 (12) ◽  
Author(s):  
Anna K. Wronska ◽  
Meinske P. Haak ◽  
Ellen Geraats ◽  
Eva Bruins Slot ◽  
Marcel van den Broek ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Large Scale ◽  
De Novo ◽  
Laboratory Strain ◽  
Optimal Growth ◽  
Industrial Applications ◽  
Fast Growth ◽  
Growth Media ◽  
Free Medium ◽  
Content Type

ABSTRACT Biotin, an important cofactor for carboxylases, is essential for all kingdoms of life. Since native biotin synthesis does not always suffice for fast growth and product formation, microbial cultivation in research and industry often requires supplementation of biotin. De novo biotin biosynthesis in yeasts is not fully understood, which hinders attempts to optimize the pathway in these industrially relevant microorganisms. Previous work based on laboratory evolution of Saccharomyces cerevisiae for biotin prototrophy identified Bio1, whose catalytic function remains unresolved, as a bottleneck in biotin synthesis. This study aimed at eliminating this bottleneck in the S. cerevisiae laboratory strain CEN.PK113-7D. A screening of 35 Saccharomycotina yeasts identified six species that grew fast without biotin supplementation. Overexpression of the S. cerevisiae BIO1 (ScBIO1) ortholog isolated from one of these biotin prototrophs, Cyberlindnera fabianii, enabled fast growth of strain CEN.PK113-7D in biotin-free medium. Similar results were obtained by single overexpression of C. fabianii BIO1 (CfBIO1) in other laboratory and industrial S. cerevisiae strains. However, biotin prototrophy was restricted to aerobic conditions, probably reflecting the involvement of oxygen in the reaction catalyzed by the putative oxidoreductase CfBio1. In aerobic cultures on biotin-free medium, S. cerevisiae strains expressing CfBio1 showed a decreased susceptibility to contamination by biotin-auxotrophic S. cerevisiae. This study illustrates how the vast Saccharomycotina genomic resources may be used to improve physiological characteristics of industrially relevant S. cerevisiae. IMPORTANCE The reported metabolic engineering strategy to enable optimal growth in the absence of biotin is of direct relevance for large-scale industrial applications of S. cerevisiae. Important benefits of biotin prototrophy include cost reduction during the preparation of chemically defined industrial growth media as well as a lower susceptibility of biotin-prototrophic strains to contamination by auxotrophic microorganisms. The observed oxygen dependency of biotin synthesis by the engineered strains is relevant for further studies on the elucidation of fungal biotin biosynthesis pathways.

scholarly journals Complementation of Cobalamin Auxotrophy in Synechococcus sp. Strain PCC 7002 and Validation of a Putative Cobalamin RiboswitchIn Vivo

2016 ◽  
Vol 198 (19) ◽  
pp. 2743-2752 ◽  
Author(s):  
Adam A. Pérez ◽  
Zhenfeng Liu ◽  
Dmitry A. Rodionov ◽  
Zhongkui Li ◽  
Donald A. Bryant
Keyword(s):  
Large Scale ◽  
De Novo ◽  
Expression System ◽  
Industrial Applications ◽  
Methionine Synthase ◽  
Methionine Biosynthesis ◽  
Content Type ◽  
Methyl Donor ◽  
Synechococcus Sp

ABSTRACTThe euryhaline cyanobacteriumSynechococcussp. strain PCC 7002 has an obligate requirement for exogenous vitamin B12(cobalamin), but little is known about the roles of this compound in cyanobacteria. Bioinformatic analyses suggest that only the terminal enzyme in methionine biosynthesis, methionine synthase, requires cobalamin as a coenzyme inSynechococcussp. strain PCC 7002. Methionine synthase (MetH) catalyzes the transfer of a methyl group fromN5-methyl-5,6,7,8-tetrahydrofolate tol-homocysteine duringl-methionine synthesis and uses methylcobalamin as an intermediate methyl donor. Numerous bacteria and plants alternatively employ a cobalamin-independent methionine synthase isozyme, MetE, that catalyzes the same methyl transfer reaction as MetH but usesN5-methyl-5,6,7,8-tetrahydrofolate directly as the methyl donor. The cobalamin auxotrophy ofSynechococcussp. strain PCC 7002 was complemented by using themetEgene from the closely related cyanobacteriumSynechococcussp. strain PCC 73109, which possesses genes for both methionine synthases. This result suggests that methionine biosynthesis is probably the sole use of cobalamin inSynechococcussp. strain PCC 7002. Furthermore, a cobalamin-repressible gene expression system was developed inSynechococcussp. strain PCC 7002 that was used to validate the presence of a cobalamin riboswitch in the promoter region ofmetEfromSynechococcussp. strain PCC 73109. This riboswitch acts as a cobalamin-dependent transcriptional attenuator formetEin that organism.IMPORTANCESynechococcussp. strain PCC 7002 is a cobalamin auxotroph because, like eukaryotic marine algae, it uses a cobalamin-dependent methionine synthase (MetH) for the final step ofl-methionine biosynthesis but cannot synthesize cobalaminde novo. Heterologous expression ofmetE, encoding cobalamin-independent methionine synthase, fromSynechococcussp. strain PCC 73109, relieved this auxotrophy and enabled the construction of a truly autotrophicSynechococcussp. strain PCC 7002 more suitable for large-scale industrial applications. Characterization of a cobalamin riboswitch expands the genetic toolbox forSynechococcussp. strain PCC 7002 by providing a cobalamin-repressible expression system.

scholarly journals Proteome Analyses of Strains ATCC 51142 and PCC 7822 of the Diazotrophic Cyanobacterium Cyanothece sp. under Culture Conditions Resulting in Enhanced H2Production

2012 ◽  
Vol 79 (4) ◽  
pp. 1070-1077 ◽  
Author(s):  
Uma K. Aryal ◽  
Stephen J. Callister ◽  
Sujata Mishra ◽  
Xiaohui Zhang ◽  
Janani I. Shutthanandan ◽  
...  
Keyword(s):  
Large Scale ◽  
Genetic Manipulation ◽  
Nitrogenase Activity ◽  
Calvin Cycle ◽  
Proteome Analysis ◽  
Culture Conditions ◽  
Pentose Phosphate ◽  
Diurnal Changes ◽  
Content Type ◽  
Diurnal Cycles

ABSTRACTCultures of the cyanobacterial genusCyanothecehave been shown to produce high levels of biohydrogen. These strains are diazotrophic and undergo pronounced diurnal cycles when grown under N2-fixing conditions in light-dark cycles. We seek to better understand the way in which proteins respond to these diurnal changes, and we performed quantitative proteome analysis ofCyanothecesp. strains ATCC 51142 and PCC 7822 grown under 8 different nutritional conditions. Nitrogenase expression was limited to N2-fixing conditions, and in the absence of glycerol, nitrogenase gene expression was linked to the dark period. However, glycerol induced expression of nitrogenase during part of the light period, together with cytochromecoxidase (Cox), glycogen phosphorylase (Glp), and glycolytic and pentose phosphate pathway (PPP) enzymes. This indicated that nitrogenase expression in the light was facilitated via higher levels of respiration and glycogen breakdown. Key enzymes of the Calvin cycle were inhibited inCyanotheceATCC 51142 in the presence of glycerol under H2-producing conditions, suggesting a competition between these sources of carbon. However, inCyanothecePCC 7822, the Calvin cycle still played a role in cofactor recycling during H2production. Our data comprise the first comprehensive profiling of proteome changes inCyanothecePCC 7822 and allow an in-depth comparative analysis of major physiological and biochemical processes that influence H2production in both strains. Our results revealed many previously uncharacterized proteins that may play a role in nitrogenase activity and in other metabolic pathways and may provide suitable targets for genetic manipulation that would lead to improvement of large-scale H2production.

scholarly journals BSGatlas: a unified Bacillus subtilis genome and transcriptome annotation atlas with enhanced information access

2021 ◽  
Vol 7 (2) ◽  
Author(s):  
Adrian Sven Geissler ◽  
Christian Anthon ◽  
Ferhat Alkan ◽  
Enrique González-Tortuero ◽  
Line Dahl Poulsen ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Type Species ◽  
Large Scale ◽  
Information Access ◽  
Model Systems ◽  
Transcription Start ◽  
Content Type ◽  
Link Type ◽  
Genome Wide ◽  
Transcript Map

A large part of our current understanding of gene regulation in Gram-positive bacteria is based on Bacillus subtilis , as it is one of the most well studied bacterial model systems. The rapid growth in data concerning its molecular and genomic biology is distributed across multiple annotation resources. Consequently, the interpretation of data from further B. subtilis experiments becomes increasingly challenging in both low- and large-scale analyses. Additionally, B. subtilis annotation of structured RNA and non-coding RNA (ncRNA), as well as the operon structure, is still lagging behind the annotation of the coding sequences. To address these challenges, we created the B. subtilis genome atlas, BSGatlas, which integrates and unifies multiple existing annotation resources. Compared to any of the individual resources, the BSGatlas contains twice as many ncRNAs, while improving the positional annotation for 70 % of the ncRNAs. Furthermore, we combined known transcription start and termination sites with lists of known co-transcribed gene sets to create a comprehensive transcript map. The combination with transcription start/termination site annotations resulted in 717 new sets of co-transcribed genes and 5335 untranslated regions (UTRs). In comparison to existing resources, the number of 5′ and 3′ UTRs increased nearly fivefold, and the number of internal UTRs doubled. The transcript map is organized in 2266 operons, which provides transcriptional annotation for 92 % of all genes in the genome compared to the at most 82 % by previous resources. We predicted an off-target-aware genome-wide library of CRISPR–Cas9 guide RNAs, which we also linked to polycistronic operons. We provide the BSGatlas in multiple forms: as a website (https://rth.dk/resources/bsgatlas/), an annotation hub for display in the UCSC genome browser, supplementary tables and standardized GFF3 format, which can be used in large scale -omics studies. By complementing existing resources, the BSGatlas supports analyses of the B. subtilis genome and its molecular biology with respect to not only non-coding genes but also genome-wide transcriptional relationships of all genes.

scholarly journals Purification and characterisation of thermostable α-amylases from microbial sources

BioResources ◽  
2019 ◽  
Vol 15 (1) ◽  
pp. 2005-2029
Author(s):  
Si J. Lim ◽  
Siti Nur Hazwani-Oslan ◽  
Siti N. Oslan
Keyword(s):  
Column Chromatography ◽  
Large Scale ◽  
Genetic Manipulation ◽  
Promising Result ◽  
Potato Starch ◽  
Industrial Applications ◽  
Wheat Starch ◽  
Starch Degradation ◽  
Affinity Column ◽  
Microbial Sources

α-Amylases (E.C 3.2.1.1) hydrolyse starch into smaller moieties such as maltose and glucose by breaking α-1,4-glycosidic linkages. The application of α-amylases in various industries has made the large-scale productions of these enzymes crucial. Thermostable α-amylase that catalyses starch degradation at the temperatures higher than 50 °C is favourable in harsh industrial applications. Due to ease in genetic manipulation and bulk production, this enzyme is most preferably produced by microorganisms. Bacillus sp. and Escherichia coli are commonly used microbial expression hosts for α-amylases (30 to 205 kDa in molecular weight). These amylases can be purified using ultrafiltration, salt precipitation, dialysis, and column chromatography. Recently, affinity column chromatography has shown the most promising result where the recovery rate was 38 to 60% and purification up to 13.2-fold. Microbial thermostable α-amylases have the optimum temperature and pH ranging from 50 °C to 100 °C and 5.0 to 10.5, respectively. These enzymes have high specificity towards potato starch, wheat starch, amylose, and amylopectin. EDTA (1 mM) gave the highest inhibitory effect (79%), but Ca2+ (5 mM) was the most effective co-factor with 155%. This review provides insight regarding thermostable α-amylases obtained from microbial sources for industrial applications.

scholarly journals Toward Improvement of Erythromycin A Production in an Industrial Saccharopolyspora erythraea Strain via Facilitation of Genetic Manipulation with an ArtificialattBSite for Specific Recombination

2011 ◽  
Vol 77 (21) ◽  
pp. 7508-7516 ◽  
Author(s):  
Jiequn Wu ◽  
Qinglin Zhang ◽  
Wei Deng ◽  
Jiangchao Qian ◽  
Siliang Zhang ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Large Scale ◽  
Genetic Manipulation ◽  
Saccharopolyspora Erythraea ◽  
Scale Production ◽  
Content Type ◽  
Large Scale Production ◽  
Endogenous Genes ◽  
Erythromycin A ◽  
Adenosylmethionine Synthetase

ABSTRACTLarge-scale production of erythromycin A (Er-A) relies on the organismSaccharopolyspora erythraea, in which lack of a typicalattBsite largely impedes the application of phage ΦC31 integrase-mediated recombination into site-specific engineering. We herein report construction of an artificialattBsite in an industrialS. erythraeastrain, HL3168 E3, in an effort to break the bottleneck previously encountered during genetic manipulation mainly from homologous or unpredictable nonspecific integration. Replacement of a cryptic gene,nrps1-1, with a cassette containing eightattBDNA sequences did not affect the high Er-producing ability, setting the stage for precisely engineering the industrial Er-producing strain for foreign DNA introduction with a reliable conjugation frequency. Transfer of either exogenous or endogenous genes of importance to Er-A biosynthesis, including theS-adenosylmethionine synthetase gene for positive regulation,vhbfor increasing the oxygen supply, and two tailoring genes,eryKanderyG, for optimizing the biotransformation at the late stage, was achieved by taking advantage of this facility, allowing systematic improvement of Er-A production as well as elimination of the by-products Er-B and Er-C in fermentation. The strategy developed here can generally be applicable to other strains that lack theattBsite.

scholarly journals Recent Molecular Tools for the Genetic Manipulation of Highly Industrially Important Mucoromycota Fungi

2021 ◽  
Vol 7 (12) ◽  
pp. 1061
Author(s):  
Hassan Mohamed ◽  
Tahira Naz ◽  
Junhuan Yang ◽  
Aabid Manzoor Shah ◽  
Yusuf Nazir ◽  
...  
Keyword(s):  
Large Scale ◽  
Genetic Manipulation ◽  
Value Added ◽  
Industrial Applications ◽  
Fungal Species ◽  
Model Organisms ◽  
Molecular Tools ◽  
Genetic Tools ◽  
Efficient Gene ◽  
Genetic Modifications

Mucorales is the largest and most well-studied order of the phylum Mucormycota and is known for its rapid growth rate and various industrial applications. The Mucorales fungi are a fascinating group of filamentous organisms with many uses in research and the industrial and medical fields. They are widely used biotechnological producers of various secondary metabolites and other value-added products. Certain members of Mucorales are extensively used as model organisms for genetic and molecular investigation and have extended our understanding of the metabolisms of other members of this order as well. Compared with other fungal species, our understanding of Mucoralean fungi is still in its infancy, which could be linked to their lack of effective genetic tools. However, recent advancements in molecular tools and approaches, such as the construction of recyclable markers, silencing vectors, and the CRISPR-Cas9-based gene-editing system, have helped us to modify the genomes of these model organisms. Multiple genetic modifications have been shown to generate valuable products on a large scale and helped us to understand the morphogenesis, basic biology, pathogenesis, and host–pathogen interactions of Mucoralean fungi. In this review, we discuss various conventional and modern genetic tools and approaches used for efficient gene modification in industrially important members of Mucorales.

Sign in / Sign up

Export Citation Format

Share Document