@article{HentschkeHagerHojdis2014, author = {Hentschke, Reinhard and Hager, Jonathan and Hojdis, Nils}, title = {Molecular Modeling Approach to the Prediction of Mechanical Properties of Silica-Reinforced Rubbers}, series = {Journal of Applied Polymer Science}, volume = {131}, journal = {Journal of Applied Polymer Science}, number = {18}, publisher = {Wiley}, address = {New York, NY}, issn = {1097-4628}, doi = {10.1002/app.40806}, pages = {1 -- 9}, year = {2014}, abstract = {Recently, we have suggested a nanomechanical model for dissipative loss in filled elastomer networks in the context of the Payne effect. The mechanism is based on a total interfiller particle force exhibiting an intermittent loop, due to the combination of short-range repulsion and dispersion forces with a long-range elastic attraction. The sum of these forces leads, under external strain, to a spontaneous instability of "bonds" between the aggregates in a filler network and attendant energy dissipation. Here, we use molecular dynamics simulations to obtain chemically realistic forces between surface modified silica particles. The latter are combined with the above model to estimate the loss modulus and the low strain storage modulus in elastomers containing the aforementioned filler-compatibilizer systems. The model is compared to experimental dynamic moduli of silica filled rubbers. We find good agreement between the model predictions and the experiments as function of the compatibilizer's molecular structure and its bulk concentration.}, language = {en} } @article{HenkenOosterhuisOehlschlaegeretal.2012, author = {Henken, F. E. and Oosterhuis, K. and {\"O}hlschl{\"a}ger, Peter and Bosch, L. and Hooijberg, E. and Haanen, J. B. A. G. and Steenbergen, R. D. M.}, title = {Preclinical safety evaluation of DNA vaccines encoding modified HPV16 E6 and E7}, series = {Vaccine}, volume = {30}, journal = {Vaccine}, number = {28}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0264-410X}, doi = {10.1016/j.vaccine.2012.04.013}, pages = {4259 -- 4266}, year = {2012}, abstract = {Persistent infection with high-risk human papillomaviruses (hrHPV) can result in the formation of anogenital cancers. As hrHPV proteins E6 and E7 are required for cancer initiation and maintenance, they are ideal targets for immunotherapeutic interventions. Previously, we have described the development of DNA vaccines for the induction of HPV16 E6 and E7 specific T cell immunity. These vaccines consist of 'gene-shuffled' (SH) versions of HPV16 E6 and E7 that were fused to Tetanus Toxin Fragment C domain 1 (TTFC) and were named TTFC-E6SH and TTFC-E7SH. Gene-shuffling was performed to avoid the risk of inducing malignant transformation at the vaccination site. Here, we describe the preclinical safety evaluation of these candidate vaccines by analysis of their transforming capacity in vitro using established murine fibroblasts (NIH 3T3 cells) and primary human foreskin keratinocytes (HFKs). We demonstrate that neither ectopic expression of TTFC-E6SH and TTFC-E7SH alone or in combination enabled NIH 3T3 cells to form colonies in soft agar. In contrast, expression of HPV16 E6WT and E7WT alone or in combination resulted in effective transformation. Similarly, retroviral transduction of HFKs from three independent donors with both TTFC-E6SH and TTFC-E7SH alone or in combination did not show any signs of immortalization. In contrast, the combined expression of E6WT and E7WT induced immortalization in HFKs from all donors. Based on these results we consider it justified to proceed to clinical evaluation of DNA vaccines encoding TTFC-E6SH and TTFC-E7SH in patients with HPV16 associated (pre)malignancies.}, language = {en} } @article{HengsbachEngelCwienczeketal.2023, author = {Hengsbach, Jan-Niklas and Engel, Mareike and Cwienczek, Marcel and Stiefelmaier, Judith and Tippk{\"o}tter, Nils and Ulber, Roland}, title = {Scalable unseparated bioelectrochemical reactors by using a carbon fiber brush as stirrer and working electrode}, series = {ChemElectroChem}, volume = {10}, journal = {ChemElectroChem}, number = {21}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {2196-0216}, doi = {10.1002/celc.202300440}, pages = {9 Seiten}, year = {2023}, abstract = {The concept of energy conversion into platform chemicals using bioelectrochemical systems (BES) has gained increasing attention in recent years, as the technology simultaneously provides an opportunity for sustainable chemical production and tackles the challenge of Power-to-X technologies. There are many approaches to realize the industrial scale of BES. One concept is to equip standard bioreactors with static electrodes. However, large installations resulted in a negative influence on various reactor parameters. In this study, we present a new single-chamber BES based on a stirred tank reactor in which the stirrer was replaced by a carbon fiber brush, performing the functions of the working electrode and the stirrer. The reactor is characterized in abiotic studies and electro-fermentations with Clostridium acetobutylicum. Compared to standard reactors an increase in butanol production of 20.14±3.66 \% shows that the new BES can be efficiently used for bioelectrochemical processes.}, language = {en} } @article{HendersonScheerWolf2009, author = {Henderson, Colin J. and Scheer, Nico and Wolf, C. Roland}, title = {Advances in the generation of mouse models to elucidate the pathways of drug metabolism in rodents and man}, series = {Expert Review of Clinical Pharmacology}, volume = {2}, journal = {Expert Review of Clinical Pharmacology}, number = {2}, publisher = {Taylor \& Francis}, address = {London}, issn = {1751-2441}, doi = {10.1586/17512433.2.2.105}, pages = {105 -- 109}, year = {2009}, language = {en} } @article{HendersonMclaughlinScheeretal.2015, author = {Henderson, Colin J. and Mclaughlin, Lesley A. and Scheer, Nico and Stanley, Lesley A. and Wolf, C. Roland}, title = {Cytochrome b5 Is a Major Determinant of Human Cytochrome P450 CYP2D6 and CYP3A4 Activity In Vivo s}, series = {Molecular Pharmacology}, volume = {87}, journal = {Molecular Pharmacology}, number = {4}, publisher = {ASPET}, address = {Bethesda}, issn = {1521-0111}, doi = {10.1124/mol.114.097394}, pages = {733 -- 739}, year = {2015}, language = {en} } @article{HemmerlingMerschenzQuackWunderlich1988, author = {Hemmerling, H.-J. and Merschenz-Quack, Angelika and Wunderlich, H.}, title = {Crystal structures of indeno[1,2-d]imidazoles. XIth European Crystallographic Meeting, Vienna 1988}, series = {Zeitschrift f{\"u}r Kristallographie - Crystalline Materials}, volume = {185}, journal = {Zeitschrift f{\"u}r Kristallographie - Crystalline Materials}, number = {H. 1-4}, issn = {2196-7105 (E-Books); 2194-4946 (Print)}, pages = {256}, year = {1988}, language = {en} } @article{HeinzeMangPeteretal.2014, author = {Heinze, Daniel and Mang, Thomas and Peter, Karin and M{\"o}ller, Martin and Weichold, Oliver}, title = {Synthesis of low molecular weight poly(vinyl acetate) and its application as plasticizer}, series = {Journal of applied polymer science}, volume = {131}, journal = {Journal of applied polymer science}, number = {9}, publisher = {Wiley}, address = {New York}, issn = {1097-4628 (E-Journal); 0021-8995 (Print)}, doi = {10.1002/app.40226}, pages = {Article No. 40226}, year = {2014}, abstract = {Poly(vinyl acetate), PVAc, with a degree of polymerization Xn = 10 was prepared by chain-transfer radical polymerization using carbon tetrachloride and used as oligomeric plasticizer for commercial PVAc. However, the chlorinated chain ends cause a low thermal stability requiring mild Cl/H substitution. The product exhibits high thermal stability and excellent melt-compounding properties. Blends of oligomeric and commercial PVAc show single glass transition temperatures which decrease with higher oligomer content and exhibit small negative deviations from Fox' linear additivity rule. This indicates plasticization and miscibility being mainly due to entropic effects. Injection-moulded thick specimens show ductile behaviour at oligomer contents >10 wt \%, while sheets with a thickness of 0.2-0.5 mm appear flexible already at 7.5 wt \%. The oxygen permeability coefficients are an order of magnitude lower than those of low-density polyethylene. Due to the sum of their properties, the plasticized sheets present a promising alternative in the preparation of barrier materials.}, language = {en} } @article{HeinzeMangPopescuetal.2016, author = {Heinze, D. and Mang, Thomas and Popescu, C. and Weichold, O.}, title = {Effect of side chain length and degree of polymerization on the decomposition and crystallization behaviour of chlorinated poly(vinyl ester) oligomers}, series = {Thermochimica Acta}, volume = {637}, journal = {Thermochimica Acta}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0040-6031 (electronic)}, doi = {10.1016/j.tca.2016.05.015}, pages = {143 -- 153}, year = {2016}, abstract = {Four members of a homologous series of chlorinated poly(vinyl ester) oligomers CCl₃-(CH₂CH (OCO(CH₂)ₘCH₃))ₙ-Cl with degrees of polymerization of 10 and 20 were prepared by telomerisation using carbon tetrachloride. The number of side chain carbon atoms ranges from 2 (poly(vinyl acetate) to 18 (poly(vinyl stearate)). The effect of the n-alkyl side chain length and of the degree of polymerization on the thermal stability and crystallization behaviour of the synthesized compounds was investigated. All oligomers degrade in two major steps by first losing HCl and side chains with subsequent breakdown of the backbone. The members with short side chains, up to poly(vinyl octanoate), are amorphous and show internal plasticization, whereas those with high number of side chain carbon atoms are semi-crystalline due to side-chain crystallization. A better packing for poly(vinyl stearate) is also noticeable. The glass transition and melting temperatures as well as the onset temperature of decomposition are influenced to a larger extent by the side chain length than by the degree of polymerization. Thermal stability is improved if both the size and number of side chains increase, but only a long side chain causes a significant increase of the resistance to degradation. This results in a stabilization of PVAc so that oligomers from poly(vinyl octanoate) on are stable under atmospheric conditions. Thus, the way to design stable, chlorinated PVEs oligomers is to use a long n-alkyl side chain.}, language = {en} } @article{HeineHerrmannSelmeretal.2014, author = {Heine, A. and Herrmann, G. and Selmer, Thorsten and Terwesten, F. and Buckel, W. and Reuter, K.}, title = {High resolution crystal structure of clostridium propionicum β-Alanyl-CoA:Ammonia Lyase, a new member of the "Hot Dog Fold" protein superfamily}, series = {Proteins}, volume = {82}, journal = {Proteins}, number = {9}, publisher = {Wiley-Liss}, address = {New York}, issn = {1097-0134 (E-Journal); 0887-3585 (Print)}, doi = {10.1002/prot.24557}, pages = {2041 -- 2053}, year = {2014}, abstract = {Clostridium propionicum is the only organism known to ferment β-alanine, a constituent of coenzyme A (CoA) and the phosphopantetheinyl prosthetic group of holo-acyl carrier protein. The first step in the fermentation is a CoA-transfer to β-alanine. Subsequently, the resulting β-alanyl-CoA is deaminated by the enzyme β-alanyl-CoA:ammonia lyase (Acl) to reversibly form ammonia and acrylyl-CoA. We have determined the crystal structure of Acl in its apo-form at a resolution of 0.97 {\AA} as well as in complex with CoA at a resolution of 1.59 {\AA}. The structures reveal that the enyzme belongs to a superfamily of proteins exhibiting a so called "hot dog fold" which is characterized by a five-stranded antiparallel β-sheet with a long α-helix packed against it. The functional unit of all "hot dog fold" proteins is a homodimer containing two equivalent substrate binding sites which are established by the dimer interface. In the case of Acl, three functional dimers combine to a homohexamer strongly resembling the homohexamer formed by YciA-like acyl-CoA thioesterases. Here, we propose an enzymatic mechanism based on the crystal structure of the Acl·CoA complex and molecular docking. Proteins 2014; 82:2041-2053. © 2014 Wiley Periodicals, Inc.}, language = {en} } @article{HasegawaKapelyukhTaharaetal.2011, author = {Hasegawa, Maki and Kapelyukh, Yury and Tahara, Harunobu and Seibler, Jost and Rode, Anja and Krueger, Sylvia and Lee, Dongtao N. and Wolf, C. Roland and Scheer, Nico}, title = {Quantitative prediction of human pregnane X receptor and cytochrome P450 3A4 mediated drug-drug interaction in a novel multiple humanized mouse line}, series = {Molecular Pharmacology}, volume = {80}, journal = {Molecular Pharmacology}, number = {33}, publisher = {ASPET}, address = {Bethesda, Md.}, issn = {1521-0111}, doi = {10.1124/mol.111.071845}, pages = {518 -- 528}, year = {2011}, language = {en} } @article{HarishWriggersJungketal.2016, author = {Harish, Ajay B. and Wriggers, Peter and Jungk, Juliane and Hojdis, Nils and Recker, Carla}, title = {Mesoscale Constitutive Modeling of Non-Crystallizing Filled Elastomers}, series = {Computational Mechanics}, volume = {57}, journal = {Computational Mechanics}, publisher = {Springer}, address = {Berlin}, issn = {1432-0924}, doi = {10.1007/s00466-015-1251-1}, pages = {653 -- 677}, year = {2016}, abstract = {Elastomers are exceptional materials owing to their ability to undergo large deformations before failure. However, due to their very low stiffness, they are not always suitable for industrial applications. Addition of filler particles provides reinforcing effects and thus enhances the material properties that render them more versatile for applications like tyres etc. However, deformation behavior of filled polymers is accompanied by several nonlinear effects like Mullins and Payne effect. To this day, the physical and chemical changes resulting in such nonlinear effect remain an active area of research. In this work, we develop a heterogeneous (or multiphase) constitutive model at the mesoscale explicitly considering filler particle aggregates, elastomeric matrix and their mechanical interaction through an approximate interface layer. The developed constitutive model is used to demonstrate cluster breakage, also, as one of the possible sources for Mullins effect observed in non-crystallizing filled elastomers.}, language = {en} } @article{HansScheerRiedletal.2004, author = {Hans, Stefan and Scheer, Nico and Riedl, Iris and Weiz{\"a}cker, Elisabeth von and Blader, Patrick and Campos-Ortega, Jos{\´e} A.}, title = {her3, a zebrafish member of the hairy-E(spl) family, is repressed by Notch signalling}, series = {Development}, volume = {131}, journal = {Development}, number = {12}, issn = {1477-9129}, doi = {10.1242/dev.01167}, pages = {2957 -- 2969}, year = {2004}, language = {en} } @article{HandtkeVollandMethlingetal.2014, author = {Handtke, Stefan and Volland, Sonja and Methling, Karen and Albrecht, Dirk and Becher, D{\"o}rte and Nehls, Jenny and Bongaerts, Johannes and Maurer, Karl-Heinz and Lalk, Michael and Liesegang, Heiko and Voigt, Birgit and Daniel, Rolf and Hecker, Michael}, title = {Cell physiology of the biotechnological relevant bacterium Bacillus pumilus - An omics-based approach}, series = {Journal of Biotechnology}, journal = {Journal of Biotechnology}, number = {192(A)}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, doi = {10.1016/j.jbiotec.2014.08.028}, pages = {204 -- 214}, year = {2014}, abstract = {Members of the species Bacillus pumilus get more and more in focus of the biotechnological industry as potential new production strains. Based on exoproteome analysis, B. pumilus strain Jo2, possessing a high secretion capability, was chosen for an omics-based investigation. The proteome and metabolome of B. pumilus cells growing either in minimal or complex medium was analyzed. In total, 1542 proteins were identified in growing B. pumilus cells, among them 1182 cytosolic proteins, 297 membrane and lipoproteins and 63 secreted proteins. This accounts for about 43\% of the 3616 proteins encoded in the B. pumilus Jo2 genome sequence. By using GC-MS, IP-LC/MS and H NMR methods numerous metabolites were analyzed and assigned to reconstructed metabolic pathways. In the genome sequence a functional secretion system including the components of the Sec- and Tat-secretion machinery was found. Analysis of the exoproteome revealed secretion of about 70 proteins with predicted secretion signals. In addition, selected production-relevant genome features such as restriction modification systems and NRPS clusters of B. pumilus Jo2 are discussed.}, language = {en} } @article{HandtkeSchroeterJuergenetal.2014, author = {Handtke, Stefan and Schroeter, Rebecca and J{\"u}rgen, Britta and Methling, Karen and Schl{\"u}ter, Rabea and Albrecht, Dirk and Hijum, Sacha A. F. T. van and Bongaerts, Johannes and Maurer, Karl-Heinz and Lalk, Michael and Schweder, Thomas and Hecker, Michael and Voigt, Birgit}, title = {Bacillus pumilus reveals a remarkably high resistance to hydrogen peroxide provoked oxidative stress}, series = {PLOS one}, volume = {9}, journal = {PLOS one}, number = {1}, publisher = {PLOS}, address = {San Francisco}, issn = {1932-6203}, doi = {10.1371/journal.pone.0085625}, pages = {e85625}, year = {2014}, abstract = {Bacillus pumilus is characterized by a higher oxidative stress resistance than other comparable industrially relevant Bacilli such as B. subtilis or B. licheniformis. In this study the response of B. pumilus to oxidative stress was investigated during a treatment with high concentrations of hydrogen peroxide at the proteome, transcriptome and metabolome level. Genes/proteins belonging to regulons, which are known to have important functions in the oxidative stress response of other organisms, were found to be upregulated, such as the Fur, Spx, SOS or CtsR regulon. Strikingly, parts of the fundamental PerR regulon responding to peroxide stress in B. subtilis are not encoded in the B. pumilus genome. Thus, B. pumilus misses the catalase KatA, the DNA-protection protein MrgA or the alkyl hydroperoxide reductase AhpCF. Data of this study suggests that the catalase KatX2 takes over the function of the missing KatA in the oxidative stress response of B. pumilus. The genome-wide expression analysis revealed an induction of bacillithiol (Cys-GlcN-malate, BSH) relevant genes. An analysis of the intracellular metabolites detected high intracellular levels of this protective metabolite, which indicates the importance of bacillithiol in the peroxide stress resistance of B. pumilus.}, language = {en} } @article{HalbachScheer2000, author = {Halbach, Thorsten and Scheer, Nico}, title = {Transcriptional activation by the PHD finger is inhibited through an adjacent leucine zipper that binds 14-3-3 proteins}, series = {Nucleic Acids Research}, volume = {28}, journal = {Nucleic Acids Research}, number = {18}, issn = {1362-4962}, doi = {10.1093/nar/28.18.3542}, pages = {3542 -- 3550}, year = {2000}, language = {en} } @article{HagerHentschkeHojdisetal.2015, author = {Hager, Jonathan and Hentschke, Reinhard and Hojdis, Nils and Karimi-Varzaneh, Hossein Ali}, title = {Computer Simulation of Particle-Particle Interaction in a Model Polymer Nanocomposite}, series = {Macromolecules}, volume = {48}, journal = {Macromolecules}, number = {24}, issn = {1520-5835}, doi = {10.1021/acs.macromol.5b01864}, pages = {9039 -- 9049}, year = {2015}, language = {en} } @article{HafidiElHatkaSchmitzetal.2024, author = {Hafidi, Youssef and El Hatka, Hicham and Schmitz, Dominik and Krauss, Manuel and Pettrak, J{\"u}rgen and Biel, Markus and Ittobane, Najim}, title = {Sustainable soil additives for water and micronutrient supply: swelling and chelating properties of polyaspartic acid hydrogels utilizing newly developed crosslinkers}, series = {Gels}, volume = {10}, journal = {Gels}, number = {3}, publisher = {MDPI}, address = {Basel}, issn = {2310-2861}, doi = {10.3390/gels10030170}, pages = {Artikel 170}, year = {2024}, abstract = {Drought and water shortage are serious problems in many arid and semi-arid regions. This problem is getting worse and even continues in temperate climatic regions due to climate change. To address this problem, the use of biodegradable hydrogels is increasingly important for the application as water-retaining additives in soil. Furthermore, efficient (micro-)nutrient supply can be provided by the use of tailored hydrogels. Biodegradable polyaspartic acid (PASP) hydrogels with different available (1,6-hexamethylene diamine (HMD) and L-lysine (LYS)) and newly developed crosslinkers based on diesters of glycine (GLY) and (di-)ethylene glycol (DEG and EG, respectively) were synthesized and characterized using Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) and regarding their swelling properties (kinetic, absorbency under load (AUL)) as well as biodegradability of PASP hydrogel. Copper (II) and zinc (II), respectively, were loaded as micronutrients in two different approaches: in situ with crosslinking and subsequent loading of prepared hydrogels. The results showed successful syntheses of di-glycine-ester-based crosslinkers. Hydrogels with good water-absorbing properties were formed. Moreover, the developed crosslinking agents in combination with the specific reaction conditions resulted in higher water absorbency with increased crosslinker content used in synthesis (10\% vs. 20\%). The prepared hydrogels are candidates for water-storing soil additives due to the biodegradability of PASP, which is shown in an exemple. The incorporation of Cu(II) and Zn(II) ions can provide these micronutrients for plant growth.}, language = {en} } @article{HaegerProbstJaegeretal.2023, author = {Haeger, Gerrit and Probst, Johanna and Jaeger, Karl-Erich and Bongaerts, Johannes and Siegert, Petra}, title = {Novel aminoacylases from Streptomyces griseus DSM 40236 and their recombinant production in Streptomyces lividans}, series = {FEBS Open Bio}, volume = {13}, journal = {FEBS Open Bio}, number = {12}, publisher = {Wiley}, address = {Hoboken, NJ}, issn = {2211-5463}, doi = {10.1002/2211-5463.13723}, pages = {2224 -- 2238}, year = {2023}, abstract = {Amino acid-based surfactants are valuable compounds for cosmetic formulations. The chemical synthesis of acyl-amino acids is conventionally performed by the Schotten-Baumann reaction using fatty acyl chlorides, but aminoacylases have also been investigated for use in biocatalytic synthesis with free fatty acids. Aminoacylases and their properties are diverse; they belong to different peptidase families and show differences in substrate specificity and biocatalytic potential. Bacterial aminoacylases capable of synthesis have been isolated from Burkholderia, Mycolicibacterium, and Streptomyces. Although several proteases and peptidases from S. griseus have been described, no aminoacylases from this species have been identified yet. In this study, we investigated two novel enzymes produced by S. griseus DSM 40236ᵀ . We identified and cloned the respective genes and recombinantly expressed an α-aminoacylase (EC 3.5.1.14), designated SgAA, and an ε-lysine acylase (EC 3.5.1.17), designated SgELA, in S. lividans TK23. The purified aminoacylase SgAA was biochemically characterized, focusing on its hydrolytic activity to determine temperature- and pH optima and stabilities. The aminoacylase could hydrolyze various acetyl-amino acids at the Nα -position with a broad specificity regarding the sidechain. Substrates with longer acyl chains, like lauroyl-amino acids, were hydrolyzed to a lesser extent. Purified aminoacylase SgELA specific for the hydrolysis of Nε -acetyl-L-lysine was unstable and lost its enzymatic activity upon storage for a longer period but could initially be characterized. The pH optimum of SgELA was pH 8.0. While synthesis of acyl-amino acids was not observed with SgELA, SgAA catalyzed the synthesis of lauroyl-methionine.}, language = {en} } @article{HaegerGrankinWagner2023, author = {Haeger, Gerrit and Grankin, Alina and Wagner, Michaela}, title = {Construction of an Aspergillus oryzae triple amylase deletion mutant as a chassis to evaluate industrially relevant amylases using multiplex CRISPR/Cas9 editing technology}, series = {Applied Research}, journal = {Applied Research}, number = {Early View}, publisher = {Wiley-VCH}, issn = {2702-4288}, doi = {10.1002/appl.202200106}, pages = {1 -- 15}, year = {2023}, abstract = {Aspergillus oryzae is an industrially relevant organism for the secretory production of heterologous enzymes, especially amylases. The activities of potential heterologous amylases, however, cannot be quantified directly from the supernatant due to the high background activity of native α-amylase. This activity is caused by the gene products of amyA, amyB, and amyC. In this study, an in vitro CRISPR/Cas9 system was established in A. oryzae to delete these genes simultaneously. First, pyrG of A. oryzae NSAR1 was mutated by exploiting NHEJ to generate a counter-selection marker. Next, all amylase genes were deleted simultaneously by co-transforming a repair template carrying pyrG of Aspergillus nidulans and flanking sequences of amylase gene loci. The rate of obtained triple knock-outs was 47\%. We showed that triple knockouts do not retain any amylase activity in the supernatant. The established in vitro CRISPR/Cas9 system was used to achieve sequence-specific knock-in of target genes. The system was intended to incorporate a single copy of the gene of interest into the desired host for the development of screening methods. Therefore, an integration cassette for the heterologous Fpi amylase was designed to specifically target the amyB locus. The site-specific integration rate of the plasmid was 78\%, with exceptional additional integrations. Integration frequency was assessed via qPCR and directly correlated with heterologous amylase activity. Hence, we could compare the efficiency between two different signal peptides. In summary, we present a strategy to exploit CRISPR/Cas9 for gene mutation, multiplex knock-out, and the targeted knock-in of an expression cassette in A. oryzae. Our system provides straightforward strain engineering and paves the way for development of fungal screening systems.}, language = {en} } @article{HaegerBongaertsSiegert2022, author = {Haeger, Gerrit and Bongaerts, Johannes and Siegert, Petra}, title = {A convenient ninhydrin assay in 96-well format for amino acid-releasing enzymes using an air-stable reagent}, series = {Analytical Biochemistry}, journal = {Analytical Biochemistry}, number = {624}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1096-0309}, doi = {10.1016/j.ab.2022.114819}, pages = {Artikel 114819}, year = {2022}, abstract = {An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP). Furthermore, we found that the use of a basic, aqueous buffer enhances stability of RP. An acidic protocol for the quantification of lysine was developed by addition of glacial acetic acid. The assay allows for parallel processing in a 96-well format with measurements microtiter plates.}, language = {en} } @article{GuoMiyamotoWagneretal.2014, author = {Guo, Yuanyuan and Miyamoto, Ko-ichiro and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Theoretical study and simulation of light-addressable potentiometric sensors}, series = {Physica status solidi (A) : applications and materials}, volume = {211}, journal = {Physica status solidi (A) : applications and materials}, number = {6}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0031-8965}, doi = {10.1002/pssa.201330354}, pages = {1467 -- 1472}, year = {2014}, abstract = {The light-addressable potentiometric sensor (LAPS) is a semiconductor-based potentiometric sensor using a light probe with an ability of detecting the concentration of biochemical species in a spatially resolved manner. As an important biomedical sensor, research has been conducted to improve its performance, for instance, to realize high-speed measurement. In this work, the idea of facilitating the device-level simulation, instead of using an equivalent-circuit model, is presented for detailed analysis and optimization of the performance of the LAPS. Both carrier distribution and photocurrent response have been simulated to provide new insight into both amplitude-mode and phase-mode operations of the LAPS. Various device parameters can be examined to effectively design and optimize the LAPS structures and setups for enhanced performance.}, language = {en} } @article{GoetzeBaerWinkelmannetal.2005, author = {Goetze, Sandra and Baer, Alexandra and Winkelmann, Silke and Nehlsen, Kristina and Seibler, Jost and Maass, Karin and Bode, J{\"u}rgen}, title = {Performance of genomic bordering elements at predefined genomic loci}, series = {Molecular and Cellular Biology}, volume = {25}, journal = {Molecular and Cellular Biology}, number = {6}, issn = {1098-5549}, doi = {10.1128/MCB.25.6.2260-2272.2005}, pages = {2260 -- 2272}, year = {2005}, language = {en} } @article{GlaserLubitzLovelandetal.2009, author = {Glaser, Stefan and Lubitz, Sandra and Loveland, Kate L. and Ohbo, Kazu and Robb, Lorraine and Schwenk, Frieder and Seibler, Jost and Roellig, Daniela and Kranz, Andrea and Anastassiadis, Konstantinos and Stewart, A. Francis}, title = {The histone 3 lysine 4 methyltransferase, Mll2, is only required briefly in development and spermatogenesis}, series = {Epigenetics \& Chromatin}, volume = {2}, journal = {Epigenetics \& Chromatin}, number = {5}, issn = {1756-8935}, doi = {10.1186/1756-8935-2-5}, pages = {1 -- 16}, year = {2009}, language = {en} } @article{GielenWolfBerndtetal.1979, author = {Gielen, Hans-G{\"u}nther and Wolf, G{\"u}nter and Berndt, Heinz and Zahn, Helmut}, title = {Synthese der Fragmente A1-8, A9-15 und A16-21 der Schafinsulin-A-Kette unter Verwendung des S-tert-Butylmercaptorestes als Thiolschutzgruppe}, series = {Hoppe-Seyler's Zeitschrift f{\"u}r physiologische Chemie}, volume = {360}, journal = {Hoppe-Seyler's Zeitschrift f{\"u}r physiologische Chemie}, number = {2}, issn = {1437-4315}, doi = {10.1515/bchm2.1979.360.2.1535}, pages = {1535 -- 1548}, year = {1979}, language = {de} } @article{GhoschBaierSchuetzetal.2016, author = {Ghosch, S. and Baier, M. and Sch{\"u}tz, J. and Schneider, Felix and Scherer, Ulrich W.}, title = {Analysis of electronic autoradiographs by mathematical post-processing}, series = {Radiation Effects and Defects in Solids: Incorporating plasma science and plasma technology}, volume = {171}, journal = {Radiation Effects and Defects in Solids: Incorporating plasma science and plasma technology}, number = {1-2}, publisher = {Taylor \& Francis}, address = {London}, issn = {1029-4953}, doi = {10.1080/10420150.2016.1155587}, pages = {161 -- 172}, year = {2016}, abstract = {Autoradiography is a well-established method of nuclear imaging. When different radionuclides are present simultaneously, additional processing is needed to distinguish distributions of radionuclides. In this work, a method is presented where aluminium absorbers of different thickness are used to produce images with different cut-off energies. By subtracting images pixel-by-pixel one can generate images representing certain ranges of β-particle energies. The method is applied to the measurement of irradiated reactor graphite samples containing several radionuclides to determine the spatial distribution of these radionuclides within pre-defined energy windows. The process was repeated under fixed parameters after thermal treatment of the samples. The greyscale images of the distribution after treatment were subtracted from the corresponding pre-treatment images. Significant changes in the intensity and distribution of radionuclides could be observed in some samples. Due to the thermal treatment parameters the most significant differences were observed in the ³H and ¹⁴C inventory and distribution.}, language = {en} } @article{GerigkMaassTakorsetal.2000, author = {Gerigk, M. and Maaß, D. and Takors, Ralf and Kreutzer, A. and Wandrey, Christian and Bongaerts, Johannes and Wubbolts, Marcel}, title = {Fermentative Herstellung von L-Phenylalanin im Fed-Batch Verfahren mit E. coli unter Einbindung eines integrierten Aufarbeitungsverfahrens}, series = {Chemie-Ingenieur-Technik (CIT)}, volume = {Vol. 72}, journal = {Chemie-Ingenieur-Technik (CIT)}, number = {Iss. 9}, issn = {1522-2640 (E-Journal); 0009-286X (Print)}, pages = {926 -- 927}, year = {2000}, language = {de} } @article{GerigkMaassKreutzeretal.2002, author = {Gerigk, M. and Maaß, D. and Kreutzer, A. and Sprenger, G. and Bongaerts, Johannes and Wubbolts, Marcel and Takors, Ralf}, title = {Enhanced pilot-scale fed-batch L-phenylalanine production with recombinant Escherichia coli by fully integrated reactive extraction}, series = {Bioprocess and biosystems engineering}, volume = {Vol. 25}, journal = {Bioprocess and biosystems engineering}, number = {Iss. 1}, issn = {1432-0797 (E-Journal); 1615-7605 (E-Journal); 0178-515X (Print); 1615-7591 (Print)}, pages = {43 -- 52}, year = {2002}, language = {en} } @article{GerigkBujnickiGanpoNkwenkwaetal.2002, author = {Gerigk, M. and Bujnicki, R. and Ganpo-Nkwenkwa, E. and Bongaerts, Johannes and Sprenger, G. and Takors, Ralf}, title = {Process control for enhanced L-phenylalanine production using different recombinant Escherichia coli strains}, series = {Biotechnology and bioengineering}, volume = {Vol. 80}, journal = {Biotechnology and bioengineering}, number = {Iss. 7}, issn = {1097-0290 (E-Journal); 0006-3592 (Print)}, pages = {746 -- 754}, year = {2002}, language = {en} } @article{GebeshuberKornauthDongetal.2013, author = {Gebeshuber, Christoph A. and Kornauth, Christoph and Dong, Lihua and Sierig, Ralph and Seibler, Jost and Reiss, Martina and Tauber, Stefanie and Bilban, Martin and Wang, Shijun and Kain, Renate and B{\"o}hmig, Georg A. and Moeller, Marcus J. and Gr{\"o}ne, Hermann-Josef and Englert, Christoph and Martinez, Javier and Kerjaschki, Dontscho}, title = {Focal segmental glomerulosclerosis is induced by microRNA-193a and its downregulation of WT1}, series = {Nature Medicine}, volume = {19}, journal = {Nature Medicine}, number = {4}, issn = {1078-8956}, doi = {10.1038/nm.3142}, pages = {481 -- 487}, year = {2013}, language = {en} } @article{FeuerriegelStahlbergWeisenburgeretal.1995, author = {Feuerriegel, Uwe and Stahlberg, R. and Weisenburger, P. and Steiger, F.}, title = {Massen- und Energiebilanzen unter Einbeziehung der Untersuchungen an der Thermoselect-Anlage Fondotoce (Italien). Stahlberg, R.; Feuerriegel, U.; Weisenburger, P.; Steiger, F.}, series = {Thermoselect : der neue Weg, Restm{\"u}ll umweltgerecht zu behandeln ; [Tagungsband zur Fachtagung Thermoselect am 19. Januar 1995 in Rastatt/Baden] / G{\"u}nther H{\"a}ßler. 2. durchges. und erw. Aufl.}, journal = {Thermoselect : der neue Weg, Restm{\"u}ll umweltgerecht zu behandeln ; [Tagungsband zur Fachtagung Thermoselect am 19. Januar 1995 in Rastatt/Baden] / G{\"u}nther H{\"a}ßler. 2. durchges. und erw. Aufl.}, publisher = {Goerner}, address = {Karlsruhe}, isbn = {3-9803665-4-5}, pages = {55 -- 84}, year = {1995}, language = {de} } @article{FeuerriegelStahlbergRiegeletal.1995, author = {Feuerriegel, Uwe and Stahlberg, R. and Riegel, J. and Steiger, F.}, title = {Thermoselect - Energie- und Rohstoffgewinnung: Teil 3. Massen- und Energiebilanz unter Ber{\"u}cksichtigung der Wirtschaftlichkeit der Synthesegasnutzung. Stahlberg, R.; Riegel, J.; Feuerriegel, U.; Steiger, F.}, series = {Chemische Technik . 47 (1995), H. 3}, journal = {Chemische Technik . 47 (1995), H. 3}, isbn = {0045-6519}, pages = {113 -- 121}, year = {1995}, language = {de} } @article{FeuerriegelStahlberg1996, author = {Feuerriegel, Uwe and Stahlberg, R.}, title = {Performance of the Thermoselect Demonstration Plant at Fondotoce, Italy: Product Quality and Distribution of Chemical Elements in the Material Flow. Stahlberg, R. ; Feuerriegel, U.}, series = {Solid waste management: thermal treatment \& waste-to-energy technologies : proceedings of an international specialty conference sponsored by the Air \& Waste Management Association, [Washington, DC, April 18 - 21, 1995] / comp. by James D. Kilgroe}, journal = {Solid waste management: thermal treatment \& waste-to-energy technologies : proceedings of an international specialty conference sponsored by the Air \& Waste Management Association, [Washington, DC, April 18 - 21, 1995] / comp. by James D. Kilgroe}, publisher = {Air \& Waste Management Assoc.}, address = {Pittsburgh, Pa.}, pages = {XVI, 965 S : graph. Darst.}, year = {1996}, language = {en} } @article{FeuerriegelStahlberg1994, author = {Feuerriegel, Uwe and Stahlberg, R.}, title = {Thermoselect - Energie- und Rohstoffgewinnung: Teil 1. Verfahrensgrundlagen zur unterbrechungslosen Verwertung von Restabf{\"a}llen. Stahlberg, R. ; Feuerriegel, U.}, series = {Chemische Technik . 46 (1994), H. 5}, journal = {Chemische Technik . 46 (1994), H. 5}, isbn = {0045-6519}, pages = {257 -- 266}, year = {1994}, language = {en} } @article{FeuerriegelStahlberg1995, author = {Feuerriegel, Uwe and Stahlberg, R.}, title = {Thermoselect - Energie- und Rohstoffgewinnung: Teil 2. Untersuchungen an der Thermoselect-Anlage Fondotoce (Italien), Qualit{\"a}ten der Produkte und Nachweis der Verteilung der chemischen Elemente in den Stoffstr{\"o}men. Stahlberg, R. ; Feuerriegel, U.}, series = {Chemische Technik . 47 (1995), H. 1}, journal = {Chemische Technik . 47 (1995), H. 1}, isbn = {0045-6519}, pages = {1 -- 10}, year = {1995}, language = {de} } @article{FeuerriegelStahlberg1995, author = {Feuerriegel, Uwe and Stahlberg, R.}, title = {Das Thermoselect-Verfahren zur Energie- und Rohstoffgewinnung - Konzepte, Verfahren, Kosten}, series = {Thermische Abfallentsorgung - Konzepte, Kosten, Verfahren. Tagung Veitsh{\"o}chheim, 27./28.06.1995, VDI-Gesellschaft Energietechnik}, journal = {Thermische Abfallentsorgung - Konzepte, Kosten, Verfahren. Tagung Veitsh{\"o}chheim, 27./28.06.1995, VDI-Gesellschaft Energietechnik}, publisher = {VDI-Verl.}, address = {D{\"u}sseldorf}, isbn = {0083-5560}, pages = {319 -- 319}, year = {1995}, language = {de} } @article{FeuerriegelStahlberg1995, author = {Feuerriegel, Uwe and Stahlberg, R.}, title = {Thermoselect - Energie- und Rohstoffgewinnung, Verfahrensgrundlagen zur unterbrechungslosen Verwertung von Restabf{\"a}llen. Stahlberg, R.; Feuerriegel, U.}, series = {Thermoselect : der neue Weg, Restm{\"u}ll umweltgerecht zu behandeln ; [Tagungsband zur Fachtagung Thermoselect am 19. Januar 1995 in Rastatt/Baden] / G{\"u}nther H{\"a}ßler. 2. durchges. und erw. Aufl.}, journal = {Thermoselect : der neue Weg, Restm{\"u}ll umweltgerecht zu behandeln ; [Tagungsband zur Fachtagung Thermoselect am 19. Januar 1995 in Rastatt/Baden] / G{\"u}nther H{\"a}ßler. 2. durchges. und erw. Aufl.}, publisher = {Goerner}, address = {Karlsruhe}, isbn = {3-9803665-4-5}, pages = {13 -- 32}, year = {1995}, language = {de} } @article{FeuerriegelStahlberg1995, author = {Feuerriegel, Uwe and Stahlberg, R.}, title = {Untersuchungen an der Thermoselect-Anlage Fondotoce (Italien), Qualit{\"a}ten der Produkte und Nachweis der Verteilung der chemischen Elemente in den Stoffstr{\"o}men. Stahlberg, R.; Feuerriegel, U.}, series = {Thermoselect : der neue Weg, Restm{\"u}ll umweltgerecht zu behandeln ; [Tagungsband zur Fachtagung Thermoselect am 19. Januar 1995 in Rastatt/Baden] / G{\"u}nther H{\"a}ßler. 2. durchges. und erw. Aufl.}, journal = {Thermoselect : der neue Weg, Restm{\"u}ll umweltgerecht zu behandeln ; [Tagungsband zur Fachtagung Thermoselect am 19. Januar 1995 in Rastatt/Baden] / G{\"u}nther H{\"a}ßler. 2. durchges. und erw. Aufl.}, publisher = {Goerner}, address = {Karlsruhe}, isbn = {3-9803665-4-5}, pages = {33 -- 54}, year = {1995}, language = {de} } @article{FeuerriegelStahlberg1995, author = {Feuerriegel, Uwe and Stahlberg, R.}, title = {Thermoselect - Energie- und Rohstoffgewinnung. Stahlberg, R. ; Feuerriegel, U.}, series = {Abfallwirtschaft Stoffkreisl{\"a}ufe / Terratec '95, Kongress West-Ost-Transfer Umwelt vom 1. bis 3. M{\"a}rz 1995. Leipziger Messe. Hrsg. von Hasso Scheffler ...}, journal = {Abfallwirtschaft Stoffkreisl{\"a}ufe / Terratec '95, Kongress West-Ost-Transfer Umwelt vom 1. bis 3. M{\"a}rz 1995. Leipziger Messe. Hrsg. von Hasso Scheffler ...}, publisher = {Teubner}, address = {Stuttgart [u.a.]}, isbn = {3-8154-3513-7}, pages = {179 -- 194}, year = {1995}, language = {de} } @article{FeuerriegelMaHoffmann1990, author = {Feuerriegel, Uwe and Ma, J. and Hoffmann, U.}, title = {Untersuchungen zur Rußfilterregeneration mit Hilfe von Z{\"u}ndhilfen. Ma, J.; Hoffmann, U.; Feuerriegel, U}, series = {Erd{\"o}l und Kohle - Erdgas, Brennstoff-Chemie. 43 (1990)}, journal = {Erd{\"o}l und Kohle - Erdgas, Brennstoff-Chemie. 43 (1990)}, isbn = {0014-0058}, pages = {150 -- 157}, year = {1990}, language = {de} } @article{FeuerriegelKuenschStahlbergetal.1994, author = {Feuerriegel, Uwe and K{\"u}nsch, M. and Stahlberg, R. and Steiger, F.}, title = {Massen- und Energiebilanz des Thermoselect-Verfahrens}, series = {Thermoselect-Verfahren zur Ent- und Vergasung von Abf{\"a}llen / Franz J. Schweitzer [Hrsg.]}, journal = {Thermoselect-Verfahren zur Ent- und Vergasung von Abf{\"a}llen / Franz J. Schweitzer [Hrsg.]}, publisher = {EF-Verl. f{\"u}r Energie- u. Umwelttechnik}, address = {Berlin}, isbn = {3-924511-47-0}, pages = {207 S. : graph. Darst.}, year = {1994}, language = {de} } @article{FeuerriegelKloseSloboshaninetal.1994, author = {Feuerriegel, Uwe and Klose, W. and Sloboshanin, S. and Goebel, H. [u.a.]}, title = {Deactivation of a palladium-supported alumina catalyst by hydrogen sulfide during the oxidation of methane}, series = {Langmuir: the ACS journal of surfaces and colloids. 10 (1994), H. 10}, journal = {Langmuir: the ACS journal of surfaces and colloids. 10 (1994), H. 10}, isbn = {0743-74363}, pages = {3567 -- 3570}, year = {1994}, language = {en} } @article{FeuerriegelKlose1993, author = {Feuerriegel, Uwe and Klose, W.}, title = {Reaktionskinetik der katalytischen Methan-Oxidation und experimentelle Untersuchungen zur Desaktivierung eines Palladium-Tr{\"a}gerkatalysators durch Schwefelwasserstoff. Klose, W. ; Feuerriegel, U.}, series = {Chemie - Ingenieur - Technik. 65 (1993), H. 10}, journal = {Chemie - Ingenieur - Technik. 65 (1993), H. 10}, isbn = {0009-286X}, pages = {1242 -- 1243}, year = {1993}, language = {de} } @article{FeuerriegelKlose1998, author = {Feuerriegel, Uwe and Klose, W.}, title = {Untersuchungen zum Mechanismus der Desaktivierung eines Palladium-Tr{\"a}gerkatalysators durch Vergiftung mit Schwefelwasserstoff}, series = {Chemie - Ingenieur - Technik. 70 (1998), H. 4}, journal = {Chemie - Ingenieur - Technik. 70 (1998), H. 4}, isbn = {0009-286X}, pages = {449 -- 452}, year = {1998}, language = {de} } @article{FeuerriegelKlose1998, author = {Feuerriegel, Uwe and Klose, W.}, title = {Investigations into the Mechanism of the Deactivation of a Palladium-Supported Catalyst by Hydrogen Sulfide Poisoning. Klose, W. ; Feuerriegel, U.}, series = {Chemical engineering \& technology. 21 (1998), H. 7}, journal = {Chemical engineering \& technology. 21 (1998), H. 7}, isbn = {0930-7516}, pages = {562 -- 564}, year = {1998}, language = {en} } @article{FeuerriegelHoffmannWalenziketal.2001, author = {Feuerriegel, Uwe and Hoffmann, Ulrich and Walenzik, C. and K{\"o}gel, J.}, title = {F{\"u}r Scale-up´s und Betriebsmuster, Projekt Trocknungstechnik an der FH Aachen}, series = {CITplus : das Praxismagazin f{\"u}r Verfahrenstechnik und Apparatebau. 4 (2001)}, journal = {CITplus : das Praxismagazin f{\"u}r Verfahrenstechnik und Apparatebau. 4 (2001)}, isbn = {1436-2597}, pages = {24 -- 25}, year = {2001}, language = {de} } @article{Feuerriegel1998, author = {Feuerriegel, Uwe}, title = {Energetische Verwertung von Kunststoffabf{\"a}llen}, series = {Chemie und Technologie makromolekularer Stoffe :, 16. Kolloquium. / Fachhochschule / Fachbereich Chemieingenieurwesen. 20. November 1998 an der FH Aachen, Fachbereich Chemieingenieurwesen}, journal = {Chemie und Technologie makromolekularer Stoffe :, 16. Kolloquium. / Fachhochschule / Fachbereich Chemieingenieurwesen. 20. November 1998 an der FH Aachen, Fachbereich Chemieingenieurwesen}, publisher = {Fachhochschule Aachen}, address = {Aachen}, pages = {169 -- 189}, year = {1998}, language = {de} } @article{Feuerriegel2001, author = {Feuerriegel, Uwe}, title = {Trennen und Trocknen. FH Aachen verf{\"u}gt {\"u}ber einen Zentrifugentrockner im Technikumsmaßstab}, series = {Lebensmitteltechnik. 33 (2001), H. 7-8}, journal = {Lebensmitteltechnik. 33 (2001), H. 7-8}, isbn = {0047-4290}, pages = {22 -- 23}, year = {2001}, language = {de} } @article{FengSeiblerAlamietal.1999, author = {Feng, Yong-Qing and Seibler, Jost and Alami, Raouf and Eisen, Andrew and Westerman, Karen A. and Leboulch, Philippe and Fiering, Steven and Bouhassira, Eric E.}, title = {Site-specific chromosomal integration in mammalian cells: highly efficient CRE recombinase-mediated cassette exchange}, series = {Journal of Molecular Biology}, volume = {292}, journal = {Journal of Molecular Biology}, number = {4}, issn = {0022-2836}, doi = {10.1006/jmbi.1999.3113}, pages = {779 -- 785}, year = {1999}, language = {en} } @article{FalkenbergVossBottetal.2023, author = {Falkenberg, Fabian and Voß, Leonie and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {New robust subtilisins from halotolerant and halophilic Bacillaceae}, series = {Applied Microbiology and Biotechnology}, volume = {107}, journal = {Applied Microbiology and Biotechnology}, publisher = {Springer Nature}, address = {Berlin}, issn = {1432-0614}, doi = {10.1007/s00253-023-12553-w}, pages = {3939 -- 3954}, year = {2023}, abstract = {The aim of the present study was the characterisation of three true subtilisins and one phylogenetically intermediate subtilisin from halotolerant and halophilic microorganisms. Considering the currently growing enzyme market for efficient and novel biocatalysts, data mining is a promising source for novel, as yet uncharacterised enzymes, especially from halophilic or halotolerant Bacillaceae, which offer great potential to meet industrial needs. Both halophilic bacteria Pontibacillus marinus DSM 16465ᵀ and Alkalibacillus haloalkaliphilus DSM 5271ᵀ and both halotolerant bacteria Metabacillus indicus DSM 16189 and Litchfieldia alkalitelluris DSM 16976ᵀ served as a source for the four new subtilisins SPPM, SPAH, SPMI and SPLA. The protease genes were cloned and expressed in Bacillus subtilis DB104. Purification to apparent homogeneity was achieved by ethanol precipitation, desalting and ion-exchange chromatography. Enzyme activity could be observed between pH 5.0-12.0 with an optimum for SPPM, SPMI and SPLA around pH 9.0 and for SPAH at pH 10.0. The optimal temperature for SPMI and SPLA was 70 °C and for SPPM and SPAH 55 °C and 50 °C, respectively. All proteases showed high stability towards 5\% (w/v) SDS and were active even at NaCl concentrations of 5 M. The four proteases demonstrate potential for future biotechnological applications.}, language = {en} } @article{FalkenbergRahbaFischeretal.2022, author = {Falkenberg, Fabian and Rahba, Jade and Fischer, David and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T}, series = {FEBS Open Bio}, volume = {12}, journal = {FEBS Open Bio}, number = {10}, publisher = {Wiley}, address = {Hoboken, NJ}, issn = {2211-5463}, doi = {10.1002/2211-5463.13457}, pages = {1729 -- 1746}, year = {2022}, abstract = {Halophilic and halotolerant microorganisms represent a promising source of salt-tolerant enzymes suitable for various biotechnological applications where high salt concentrations would otherwise limit enzymatic activity. Considering the current growing enzyme market and the need for more efficient and new biocatalysts, the present study aimed at the characterization of a high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T. The protease gene was cloned and expressed in Bacillus subtilis DB104. The recombinant protease SPAO with 269 amino acids belongs to the subfamily of high-alkaline subtilisins. The biochemical characteristics of purified SPAO were analyzed in comparison with subtilisin Carlsberg, Savinase, and BPN'. SPAO, a monomer with a molecular mass of 27.1 kDa, was active over a wide range of pH 6.0-12.0 and temperature 20-80 °C, optimally at pH 9.0-9.5 and 55 °C. The protease is highly oxidatively stable to hydrogen peroxide and retained 58\% of residual activity when incubated at 10 °C with 5\% (v/v) H2O2 for 1 h while stimulated at 1\% (v/v) H2O2. Furthermore, SPAO was very stable and active at NaCl concentrations up to 5.0 m. This study demonstrates the potential of SPAO for biotechnological applications in the future.}, language = {en} }