@article{BongaertsBovenbergKraemeretal.2002, author = {Bongaerts, Johannes and Bovenberg, Roel and Kr{\"a}mer, Marco and M{\"u}ller, Ulrike and Raeven, Leon and Wubbolts, Marcel}, title = {Metabolic engineering to produce fine chemicals in Escherichia coli}, series = {Chemie - Ingenieur - Technik (CIT)}, volume = {Vol. 74}, journal = {Chemie - Ingenieur - Technik (CIT)}, number = {Iss. 5}, issn = {1522-2640 (E-Journal); 0009-286X (Print)}, pages = {694}, year = {2002}, language = {en} } @article{BongaertsEsserLorbachetal.2011, author = {Bongaerts, Johannes and Esser, Simon and Lorbach, Volker and Al-Momani, L{\´o}ay and M{\"u}ller, Michael A. and Franke, Dirk and Grondal, Christoph and Kurutsch, Anja and Bujnicki, Robert and Takors, Ralf and Raeven, Leon and Wubbolts, Marcel and Bovenberg, Roel and Nieger, Martin and Sch{\"u}rmann, Melanie and Trachtmann, Natalie and Kozak, Stefan and Sprenger, Georg A. and M{\"u}ller, Michael}, title = {Diversity-oriented production of metabolites derived from chorismate and their use in organic synthesis}, series = {Angewandte Chemie International Edition}, volume = {Vol. 50}, journal = {Angewandte Chemie International Edition}, number = {Iss. 34}, publisher = {Wiley}, address = {Weinheim}, issn = {1521-3773 (E-Journal); 0570-0833 (Print); 1433-7851 (Print)}, pages = {7781 -- 7786}, year = {2011}, language = {en} } @article{BongaertsKraemerMuelleretal.2001, author = {Bongaerts, Johannes and Kr{\"a}mer, Marco and M{\"u}ller, Ulrike and Raeven, Leon and Wubbolts, Marcel}, title = {Metabolic engineering for microbial production of aromatic amino acids and derived compounds}, series = {Metabolic engineering}, volume = {Vol. 3}, journal = {Metabolic engineering}, number = {Iss. 4}, issn = {1096-7184 (E-Journal); 1096-7176 (Print)}, pages = {289 -- 300}, year = {2001}, language = {en} } @article{BongaertsZoschkeWeidneretal.1995, author = {Bongaerts, Johannes and Zoschke, Sascha and Weidner, Uwe and Linden, Gottfried}, title = {Transcriptional regulation of the proton translocating NADH}, series = {Molecular microbiology}, volume = {Vol. 16}, journal = {Molecular microbiology}, number = {Iss. 3}, issn = {1365-2958 (E-Journal); 0950-382x (Print)}, pages = {521 -- 534}, year = {1995}, language = {en} } @article{BorgmeierBongaertsMeinhardt2012, author = {Borgmeier, Claudia and Bongaerts, Johannes and Meinhardt, Friedhelm}, title = {Genetic analysis of the Bacillus licheniformis degSU operon and the impact of regulatory mutations on protease production}, series = {Journal of biotechnology}, volume = {159}, journal = {Journal of biotechnology}, number = {1-2}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, doi = {10.1016/j.jbiotec.2012.02.011}, pages = {12 -- 20}, year = {2012}, abstract = {Disruption experiments targeted at the Bacillus licheniformis degSU operon and GFP-reporter analysis provided evidence for promoter activity immediately upstream of degU. pMutin mediated concomitant introduction of the degU32 allele - known to cause hypersecretion in Bacillus subtilis - resulted in a marked increase in protease activity. Application of 5-fluorouracil based counterselection through establishment of a phosphoribosyltransferase deficient Δupp strain eventually facilitated the marker-free introduction of degU32 leading to further protease enhancement achieving levels as for hypersecreting wild strains in which degU was overexpressed. Surprisingly, deletion of rapG - known to interfere with DegU DNA-binding in B. subtilis - did not enhance protease production neither in the wild type nor in the degU32 strain. The combination of degU32 and Δupp counterselection in the type strain is not only equally effective as in hypersecreting wild strains with respect to protease production but furthermore facilitates genetic strain improvement aiming at biological containment and effectiveness of biotechnological processes.}, language = {en} } @article{DegeringEggertPulsetal.2010, author = {Degering, Christian and Eggert, Thorsten and Puls, Michael and Bongaerts, Johannes and Evers, Stefan and Maurer, Karl-Heinz and Jaeger, Karl-Erich}, title = {Optimization of protease secretion in Bacillus subtilis and Bacillus licheniformis by screening of homologous and herologous signal peptides}, series = {Applied and environmental microbiology}, volume = {76}, journal = {Applied and environmental microbiology}, number = {19}, publisher = {American Society for Microbiology}, address = {Washington, DC}, issn = {1098-5336 (E-Journal); 0003-6919 (Print); 0099-2240 (Print)}, doi = {10.1128/AEM.01146-10}, pages = {6370 -- 6378}, year = {2010}, abstract = {Bacillus subtilis and Bacillus licheniformis are widely used for the large-scale industrial production of proteins. These strains can efficiently secrete proteins into the culture medium using the general secretion (Sec) pathway. A characteristic feature of all secreted proteins is their N-terminal signal peptides, which are recognized by the secretion machinery. Here, we have studied the production of an industrially important secreted protease, namely, subtilisin BPN′ from Bacillus amyloliquefaciens. One hundred seventy-three signal peptides originating from B. subtilis and 220 signal peptides from the B. licheniformis type strain were fused to this secretion target and expressed in B. subtilis, and the resulting library was analyzed by high-throughput screening for extracellular proteolytic activity. We have identified a number of signal peptides originating from both organisms which produced significantly increased yield of the secreted protease. Interestingly, we observed that levels of extracellular protease were improved not only in B. subtilis, which was used as the screening host, but also in two different B. licheniformis strains. To date, it is impossible to predict which signal peptide will result in better secretion and thus an improved yield of a given extracellular target protein. Our data show that screening a library consisting of homologous and heterologous signal peptides fused to a target protein can identify more-effective signal peptides, resulting in improved protein export not only in the original screening host but also in different production strains.}, language = {en} } @article{DeppeBongaertsO'Connelletal.2011, author = {Deppe, Veronika Maria and Bongaerts, Johannes and O'Connell, Timothy and Maurer, Karl-Heinz and Meinhardt, Friedhelm}, title = {Enzymatic deglycation of Amadori products in bacteria}, series = {Applied microbiology and biotechnology}, volume = {Vol. 90}, journal = {Applied microbiology and biotechnology}, number = {Iss. 2}, publisher = {Springer}, address = {Berlin}, issn = {1432-0614 (E-Journal); 0171-1741 (Print); 0175-7598 (Print); 0340-2118 (Print)}, pages = {399 -- 406}, year = {2011}, language = {en} } @article{DeppeKlatteBongaertsetal.2011, author = {Deppe, Veronika Maria and Klatte, Stephanie and Bongaerts, Johannes and Maurer, Karl-Heinz and O'Connell, Timothy and Meinhardt, Friedhelm}, title = {Genetic control of Amadori product degradation in Bacillus subtilis via regulation of frlBONMD expression by FrlR}, series = {Applied and environmental microbiology}, volume = {Vol. 77}, journal = {Applied and environmental microbiology}, number = {No. 9}, publisher = {American Society of Mechanical Engineers (ASME)}, address = {New York}, issn = {1098-5336 (E-Journal); 0003-6919 (Print); 0099-2240 (Print)}, pages = {2839 -- 2846}, year = {2011}, language = {en} } @article{FalkenbergBottBongaertsetal.2022, author = {Falkenberg, Fabian and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {Phylogenetic survey of the subtilase family and a data-mining-based search for new subtilisins from Bacillaceae}, series = {Frontiers in Microbiology}, volume = {2022}, journal = {Frontiers in Microbiology}, number = {13}, publisher = {Frontiers}, address = {Lausanne}, issn = {1664-302X}, doi = {10.3389/fmicb.2022.1017978}, pages = {Artikel 13:1017978}, year = {2022}, abstract = {The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family Bacillaceae using a data mining approach. The obtained 1,400 sequences were phylogenetically classified in the context of the subtilase family. This required an updated comprehensive overview of the different groups within this family. To fill this gap, we conducted a phylogenetic survey of the S8 family with characterised holotypes derived from the MEROPS database. The analysis revealed the presence of eight previously uncharacterised groups and 13 subgroups within the S8 family. The sequences that emerged from the data mining with the set filter parameters were mainly assigned to the subtilisin subgroups of true subtilisins, high-alkaline subtilisins, and phylogenetically intermediate subtilisins and represent an excellent source for new subtilisin candidates.}, 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{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{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{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} } @techreport{HaegerBongaertsSiegert2023, author = {Haeger, Gerrit and Bongaerts, Johannes and Siegert, Petra}, title = {Abschlussbericht Teil II: Eingehende Darstellung Neue biobasierte Lipopeptide aus nachhaltiger Produktion (LipoPep)}, pages = {17Seiten}, year = {2023}, language = {de} } @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{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{JossekBongaertsSprenger2001, author = {Jossek, Ralf and Bongaerts, Johannes and Sprenger, Georg A.}, title = {Characterization of a new feedback-resistant 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase AroF of Escherichia coli}, series = {FEMS microbiology letters}, volume = {Vol. 202}, journal = {FEMS microbiology letters}, number = {Iss. 1}, issn = {1574-6968}, pages = {145 -- 148}, year = {2001}, language = {en} } @article{KraemerBongaertsBovenbergetal.2003, author = {Kr{\"a}mer, Marco and Bongaerts, Johannes and Bovenberg, Roel and Kremer, Susanne and M{\"u}ller, Ulrike and Orf, Sonja and Wubbolts, Marcel and Raeven, Leon}, title = {Metabolic engineering for microbial production of shikimic acid}, series = {Metabolic engineering}, volume = {Vol. 5}, journal = {Metabolic engineering}, number = {Iss. 4}, issn = {1096-7184 (E-Journal); 1096-7176 (Print)}, pages = {277 -- 283}, year = {2003}, language = {en} } @article{KueppersSteffenHellmuthetal.2014, author = {K{\"u}ppers, Tobias and Steffen, Victoria and Hellmuth, Hendrik and O'Connell, Timothy and Bongaerts, Johannes and Maurer, Karl-Heinz and Wiechert, Wolfgang}, title = {Developing a new production host from a blueprint: Bacillus pumilus as an industrial enzyme producer}, series = {Microbial cell factories}, volume = {13}, journal = {Microbial cell factories}, publisher = {BioMed Central}, address = {London}, issn = {1475-2859 (E-Journal)}, doi = {10.1186/1475-2859-13-46}, pages = {Article No. 46}, year = {2014}, language = {en} } @article{MolinnusMuschallikGonzalezetal.2018, author = {Molinnus, Denise and Muschallik, Lukas and Gonzalez, Laura Osorio and Bongaerts, Johannes and Wagner, Torsten and Selmer, Thorsten and Siegert, Petra and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Development and characterization of a field-effect biosensor for the detection of acetoin}, series = {Biosensors and Bioelectronics}, volume = {115}, journal = {Biosensors and Bioelectronics}, publisher = {Elsevier}, address = {Amsterdam}, doi = {10.1016/j.bios.2018.05.023}, pages = {1 -- 6}, year = {2018}, abstract = {A capacitive electrolyte-insulator-semiconductor (EIS) field-effect biosensor for acetoin detection has been presented for the first time. The EIS sensor consists of a layer structure of Al/p-Si/SiO₂/Ta₂O₅/enzyme acetoin reductase. The enzyme, also referred to as butane-2,3-diol dehydrogenase from B. clausii DSM 8716T, has been recently characterized. The enzyme catalyzes the (R)-specific reduction of racemic acetoin to (R,R)- and meso-butane-2,3-diol, respectively. Two different enzyme immobilization strategies (cross-linking by using glutaraldehyde and adsorption) have been studied. Typical biosensor parameters such as optimal pH working range, sensitivity, hysteresis, linear concentration range and long-term stability have been examined by means of constant-capacitance (ConCap) mode measurements. Furthermore, preliminary experiments have been successfully carried out for the detection of acetoin in diluted white wine samples.}, language = {en} }