High-throughput strategies for the discovery and engineering of enzymes for biocatalysis

Innovations in novel enzyme discoveries impact upon a wide range of industries for which biocatalysis and biotransformations represent a great challenge, i.e., food industry, polymers and chemical industry. Key tools and technologies, such as bioinformatics tools to guide mutant library design, molecular biology tools to create mutants library, microfluidics/microplates, parallel miniscale bioreactors and mass spectrometry technologies to create high-throughput screening methods and experimental design tools for screening and optimization, allow to evolve the discovery, development and implementation of enzymes and whole cells in (bio)processes. These technological innovations are also accompanied by the development and implementation of clean and sustainable integrated processes to meet the growing needs of chemical, pharmaceutical, environmental and biorefinery industries. This review gives an overview of the benefits of high-throughput screening approach from the discovery and engineering of biocatalysts to cell culture for optimizing their production in integrated processes and their extraction/purification.

[1]  Ralph A. Cacho,et al.  Next-generation sequencing approach for connecting secondary metabolites to biosynthetic gene clusters in fungi , 2015, Front. Microbiol..

[2]  Ludovic Duponchel,et al.  Highly sensitive terahertz spectroscopy in microsystem , 2012 .

[3]  V. Lattanzio,et al.  Experimental design for in-house validation of a screening immunoassay kit. The case of a multiplex dipstick for Fusarium mycotoxins in cereals , 2013, Analytical and Bioanalytical Chemistry.

[4]  Andrew N. Holding,et al.  XL-MS: Protein cross-linking coupled with mass spectrometry. , 2015, Methods.

[5]  Andreas Vogel,et al.  Single-cell high-throughput screening to identify enantioselective hydrolytic enzymes. , 2008, Angewandte Chemie.

[6]  Valérie Leclère,et al.  Norine, the knowledgebase dedicated to non-ribosomal peptides, is now open to crowdsourcing , 2015, Nucleic Acids Res..

[7]  Hong Wang,et al.  Rapid screening and identification of compounds with DNA-binding activity from Folium Citri Reticulatae using on-line HPLC-DAD-MS(n) coupled with a post column fluorescence detection system. , 2016, Food chemistry.

[8]  Richard M Caprioli,et al.  Absolute Quantitative MALDI Imaging Mass Spectrometry: A Case of Rifampicin in Liver Tissues. , 2016, Analytical chemistry.

[9]  Rodney Rothstein,et al.  Quality control of DNA break metabolism: in the ‘end’, it's a good thing , 2008, The EMBO journal.

[10]  D. Figeys,et al.  Bottom-Up Proteomics (2013-2015): Keeping up in the Era of Systems Biology. , 2016, Analytical chemistry.

[11]  Feng Zhang,et al.  CRISPR-assisted editing of bacterial genomes , 2013, Nature Biotechnology.

[12]  L. G. de la Torre,et al.  Microfluidic tools toward industrial biotechnology , 2016, Biotechnology progress.

[13]  P. Dhulster,et al.  An improvement of surfactin production by B. subtilis BBG131 using design of experiments in microbioreactors and continuous process in bubbleless membrane bioreactor. , 2016, Bioresource technology.

[14]  Ludovic C. Gillet,et al.  Mass Spectrometry Applied to Bottom-Up Proteomics: Entering the High-Throughput Era for Hypothesis Testing. , 2016, Annual review of analytical chemistry.

[15]  Wilfried Mokwa,et al.  Microfluidic biolector—microfluidic bioprocess control in microtiter plates , 2010, Biotechnology and bioengineering.

[16]  S. Sørensen,et al.  Green fluorescent protein (GFP) leakage from microbial biosensors provides useful information for the evaluation of the scale-down effect. , 2011, Biotechnology journal.

[17]  Peter Neubauer,et al.  Direct and indirect use of GFP whole cell biosensors for the assessment of bioprocess performances: Design of milliliter scale‐down bioreactors , 2013, Biotechnology progress.

[18]  Nicolas Szita,et al.  Membrane‐aerated microbioreactor for high‐throughput bioprocessing , 2004, Biotechnology and bioengineering.

[19]  Fabienne Courtois,et al.  Picoliter cell lysate assays in microfluidic droplet compartments for directed enzyme evolution. , 2012, Chemistry & biology.

[20]  N. Bourzgui,et al.  Cold plasma functionalized TeraHertz BioMEMS for enzyme reaction analysis. , 2009, Biosensors & bioelectronics.

[21]  R. Huber,et al.  Screening for enzyme activity in turbid suspensions with scattered light , 2011, Biotechnology progress.

[22]  T. Vernet,et al.  Automated high-throughput process for site-directed mutagenesis, production, purification, and kinetic characterization of enzymes. , 2006, Analytical biochemistry.

[23]  Scott L Diamond,et al.  Printing chemical libraries on microarrays for fluid phase nanoliter reactions , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Le Cong,et al.  Multiplex Genome Engineering Using CRISPR/Cas Systems , 2013, Science.

[25]  W. Quax,et al.  Selection strategies for improved biocatalysts , 2007, The FEBS journal.

[26]  Alois Jungbauer,et al.  Continuous downstream processing of biopharmaceuticals. , 2013, Trends in biotechnology.

[27]  Jürgen Hubbuch,et al.  Rational and systematic protein purification process development: the next generation. , 2009, Trends in biotechnology.

[28]  K. Lewis,et al.  Uncultured microorganisms as a source of secondary metabolites , 2010, The Journal of Antibiotics.

[29]  M. Blackwell.  The fungi: 1, 2, 3 ... 5.1 million species? , 2011, American journal of botany.

[30]  R. Barrangou,et al.  A CRISPR design for next-generation antimicrobials , 2014, Genome Biology.

[31]  J. Kur,et al.  Cloning and characterization of a novel cold-active glycoside hydrolase family 1 enzyme with β-glucosidase, β-fucosidase and β-galactosidase activities , 2013, BMC Biotechnology.

[32]  N. Kelleher,et al.  Top Down proteomics: facts and perspectives. , 2014, Biochemical and biophysical research communications.

[33]  D. Weuster‐Botz,et al.  New miniature stirred-tank bioreactors for parallel study of enzymatic biomass hydrolysis. , 2012, Bioresource technology.

[34]  R. Huber,et al.  Utilizing high-throughput experimentation to enhance specific productivity of an E.coli T7 expression system by phosphate limitation , 2011, BMC biotechnology.

[35]  Markus Michael Müller,et al.  Regulatory and metabolic network of rhamnolipid biosynthesis: Traditional and advanced engineering towards biotechnological production , 2011, Applied Microbiology and Biotechnology.

[36]  Rainer Cramer,et al.  Advances in MALDI and Laser-Induced Soft Ionization Mass Spectrometry , 2016, Springer International Publishing.

[37]  Uwe T Bornscheuer,et al.  Strategies for the discovery and engineering of enzymes for biocatalysis. , 2013, Current opinion in chemical biology.

[38]  Marcelo Abel Soria,et al.  Improvement of a two-stage fermentation process for docosahexaenoic acid production by Aurantiochytrium limacinum SR21 applying statistical experimental designs and data analysis. , 2010, Bioresource technology.

[39]  Zhong Sheng Sun,et al.  Recent Progress in CRISPR/Cas9 Technology. , 2016, Journal of genetics and genomics = Yi chuan xue bao.

[40]  Gabrielle Chataigné,et al.  Burkholderia genome mining for nonribosomal peptide synthetases reveals a great potential for novel siderophores and lipopeptides synthesis , 2016, MicrobiologyOpen.

[41]  Radka Snajdrova,et al.  A combination of in vivo selection and cell sorting for the identification of enantioselective biocatalysts. , 2011, Angewandte Chemie.

[42]  Krist V. Gernaey,et al.  A continuous membrane microbioreactor system for development of integrated pectin modification and separation processes , 2011 .

[43]  Yanbing Zhu,et al.  Characterization of a new and thermostable esterase from a metagenomic library. , 2013, Microbiological research.

[44]  Chad W. Euler,et al.  Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials , 2014, Nature Biotechnology.

[45]  D. Swaney,et al.  Proteomic Analysis of Protein Posttranslational Modifications by Mass Spectrometry. , 2016, Cold Spring Harbor protocols.

[46]  M. Mrksich,et al.  Self-assembled monolayers that transduce enzymatic activities to electrical signals. , 2003, Angewandte Chemie.

[47]  Ronny Martínez,et al.  A fluorescent hydrogel-based flow cytometry high-throughput screening platform for hydrolytic enzymes. , 2014, Chemistry & biology.

[48]  In-Won Kim,et al.  Computational approaches for rational design of proteins with novel functionalities , 2012, Computational and structural biotechnology journal.

[49]  M Wessling,et al.  Membranes and microfluidics: a review. , 2006, Lab on a chip.

[50]  David J. Beebe,et al.  Microbial metabolomics in open microscale platforms , 2016, Nature Communications.

[51]  C. Orrego,et al.  Application of response surface design to solvent, temperature and lipase selection for optimal monoglyceride production , 2011 .

[52]  John E. Scott,et al.  Development and Validation of a High-Throughput Intrinsic ATPase Activity Assay for the Discovery of MEKK2 Inhibitors , 2013, Journal of biomolecular screening.

[53]  Želimir Kurtanjek,et al.  Modeling and kinetic parameter estimation of alcohol dehydrogenase‐catalyzed hexanol oxidation in a microreactor , 2012 .

[54]  J. Edel,et al.  Compartmentalization of electrophoretically separated analytes in a multiphase microfluidic platform. , 2012, Analytical chemistry.

[55]  Carla S. Jones,et al.  Minimum Information about a Biosynthetic Gene cluster. , 2015, Nature chemical biology.

[56]  Jianxiu Wang,et al.  Sensitive and continuous screening of inhibitors of β-site amyloid precursor protein cleaving enzyme 1 (BACE1) at single SPR chips. , 2013, Analytical chemistry.

[57]  J Büchs,et al.  Introduction to advantages and problems of shaken cultures. , 2001, Biochemical engineering journal.

[58]  Eduardo J. S. Brás,et al.  A microfluidic platform for physical entrapment of yeast cells with continuous production of invertase , 2017 .

[59]  Yaojun Tong,et al.  CRISPR-Cas9 Based Engineering of Actinomycetal Genomes. , 2015, ACS synthetic biology.

[60]  Robert Mistrik,et al.  Applications of Fourier Transform Ion Cyclotron Resonance (FT-ICR) and Orbitrap Based High Resolution Mass Spectrometry in Metabolomics and Lipidomics , 2016, International journal of molecular sciences.

[61]  Chulhwan Park,et al.  β-Galactosidase-immobilised microreactor fabricated using a novel technique for enzyme immobilisation and its application for continuous synthesis of lactulose , 2012 .

[62]  Kenneth D Greis,et al.  Mass spectrometry for enzyme assays and inhibitor screening: an emerging application in pharmaceutical research. , 2007, Mass spectrometry reviews.

[63]  P. Dhulster,et al.  New integrated bioprocess for the continuous production, extraction and purification of lipopeptides produced by Bacillus subtilis in membrane bioreactor. , 2013 .

[64]  Kaushal Kumar,et al.  High-throughput process development for biopharmaceutical drug substances. , 2011, Trends in biotechnology.

[65]  Marcel Ottens,et al.  Purifying biopharmaceuticals: knowledge-based chromatographic process development. , 2014, Trends in biotechnology.

[66]  D. Haft,et al.  SMURF: Genomic mapping of fungal secondary metabolite clusters. , 2010, Fungal genetics and biology : FG & B.

[67]  W. Duetz,et al.  Microtiter plates as mini-bioreactors: miniaturization of fermentation methods. , 2007, Trends in microbiology.

[68]  Krist V Gernaey,et al.  Application of microbioreactors in fermentation process development: a review , 2009, Analytical and bioanalytical chemistry.

[69]  A. H. Smits,et al.  Characterizing Protein-Protein Interactions Using Mass Spectrometry: Challenges and Opportunities. , 2016, Trends in biotechnology.

[70]  Won-Gun Koh,et al.  Fabrication of microfluidic devices incorporating bead-based reaction and microarray-based detection system for enzymatic assay , 2009 .

[71]  K. Lewis,et al.  A new antibiotic kills pathogens without detectable resistance , 2015, Nature.

[72]  Tim Stakenborg,et al.  Review of microfluidic microbioreactor technology for high-throughput submerged microbiological cultivation. , 2013, Biomicrofluidics.

[73]  Tilmann Weber,et al.  Bioinformatics Tools for the Discovery of New Nonribosomal Peptides. , 2016, Methods in molecular biology.

[74]  Huimin Zhao,et al.  High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System , 2014, ACS synthetic biology.

[75]  M. Gutarra,et al.  Adding value to a toxic residue from the biodiesel industry: production of two distinct pool of lipases from Penicillium simplicissimum in castor bean waste , 2011, Journal of Industrial Microbiology & Biotechnology.

[76]  Continuous colorimetric screening assays for the detection of specific l- or d-α-amino acid transaminases in enzyme libraries , 2015, Applied Microbiology and Biotechnology.

[77]  Peter Chen,et al.  Electrospray Ionization Tandem Mass Spectrometry in High‐Throughput Screening of Homogeneous Catalysts. , 2003 .

[78]  K. Walker,et al.  Automated high-throughput protein purification using an ÄKTApurifier and a CETAC autosampler. , 2014, Journal of chromatography. A.

[79]  J. Silberring,et al.  Plasma-based ambient ionization mass spectrometry in bioanalytical sciences. , 2016, Mass spectrometry reviews.

[80]  F. Tse,et al.  Use of atmospheric pressure ionization mass spectrometry in enantioselective liquid chromatography. , 2001, Chirality.

[81]  Miguel Medina,et al.  Evidence for Irreversible Inhibition of Glycogen Synthase Kinase-3β by Tideglusib* , 2011, The Journal of Biological Chemistry.

[82]  Dirk Weuster-Botz,et al.  A new microfluidic concept for parallel operated milliliter‐scale stirred tank bioreactors , 2011, Biotechnology progress.

[83]  Jay D. Keasling,et al.  Versatile synthesis of probes for high-throughput enzyme activity screening , 2013, Analytical and Bioanalytical Chemistry.

[84]  M. Hann,et al.  Direct Measurement of Intracellular Compound Concentration by RapidFire Mass Spectrometry Offers Insights into Cell Permeability , 2016, Journal of biomolecular screening.

[85]  Christian Fonade,et al.  Why and How Membrane Bioreactors with Unsteady Filtration Conditions Can Improve the Efficiency of Biological Processes , 2003, Annals of the New York Academy of Sciences.

[86]  C. Criddle,et al.  Optimization of Methanotrophic Growth and Production of Poly(3-Hydroxybutyrate) in a High-Throughput Microbioreactor System , 2015, Applied and Environmental Microbiology.

[87]  Jörn Piel,et al.  A polyketide synthase-peptide synthetase gene cluster from an uncultured bacterial symbiont of Paederus beetles , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[88]  P. Dhulster,et al.  Recent Trends in Membrane Bioreactors , 2017 .

[89]  Jens Abildskov,et al.  Computer-aided solvent screening for biocatalysis , 2013 .

[90]  S. Trimpin.  "Magic" Ionization Mass Spectrometry , 2015, Journal of The American Society for Mass Spectrometry.

[91]  Hiroyuki Nakamura,et al.  Facile Preparation of an Enzyme‐Immobilized Microreactor using a Cross‐Linking Enzyme Membrane on a Microchannel Surface , 2006 .

[92]  C. Easley,et al.  Self-regulated, droplet-based sample chopper for microfluidic absorbance detection. , 2012, Analytical chemistry.

[93]  Z. Ouyang,et al.  Ambient ionization and miniature mass spectrometry system for chemical and biological analysis. , 2016, Trends in analytical chemistry : TRAC.

[94]  Michael L Shuler,et al.  Mini‐scale bioprocessing systems for highly parallel animal cell cultures , 2012, Biotechnology progress.

[95]  D. Paslier,et al.  Metabolic diversity among main microorganisms inside an arsenic-rich ecosystem revealed by meta- and proteo-genomics , 2011, The ISME Journal.

[96]  Y. Dufresne,et al.  Norine: A powerful resource for novel nonribosomal peptide discovery , 2015, Synthetic and systems biotechnology.

[97]  A. deMello.  Control and detection of chemical reactions in microfluidic systems , 2006, Nature.

[98]  Chapter 13 – Industrial Enzymes , 2015 .

[99]  A. deMello,et al.  Ultrafast surface enhanced resonance Raman scattering detection in droplet-based microfluidic systems. , 2011, Analytical chemistry.

[100]  P. Garstecki,et al.  Functionalization of polycarbonate with proteins; open-tubular enzymatic microreactors. , 2012, Lab on a chip.

[101]  J. Büchs,et al.  Screening of cellulases for biofuel production: online monitoring of the enzymatic hydrolysis of insoluble cellulose using high-throughput scattered light detection. , 2011, Biotechnology journal.

[102]  S. Leite,et al.  Palm oil hydrolysis catalyzed by lipases under ultrasound irradiation--the use of experimental design as a tool for variables evaluation. , 2012, Ultrasonics sonochemistry.

[103]  Manfred T Reetz,et al.  A Method for High-Throughput Screening of Enantioselective Catalysts. , 1999, Angewandte Chemie.

[104]  Shiyou Zhu,et al.  High‐throughput screens in mammalian cells using the CRISPR‐Cas9 system , 2015, The FEBS journal.

[105]  J. Wölcke,et al.  Miniaturized HTS technologies - uHTS. , 2001, Drug discovery today.

[106]  Kristie L. Rose,et al.  MALDI imaging mass spectrometry of Pacific White Shrimp L. vannamei and identification of abdominal muscle proteins , 2016, Proteomics.

[107]  Pedro Fernandes,et al.  High throughput in biotechnology: from shake-flasks to fully instrumented microfermentors. , 2009, Recent patents on biotechnology.

[108]  Li Zhang,et al.  Genome shuffling of Bacillus amyloliquefaciens for improving antimicrobial lipopeptide production and an analysis of relative gene expression using FQ RT-PCR , 2012, Journal of Industrial Microbiology & Biotechnology.

[109]  M. Welker.  Proteomics for routine identification of microorganisms , 2011, Proteomics.

[110]  George Georgiou,et al.  Directed Evolution of Highly Selective Proteases by Using a Novel FACS‐Based Screen that Capitalizes on the p53 Regulator MDM2 , 2012, Chembiochem : a European journal of chemical biology.

[111]  Milan Mrksich,et al.  Carbohydrate arrays for the evaluation of protein binding and enzymatic modification. , 2002, Chemistry & biology.

[112]  G. Collins.  The next generation. , 2006, Scientific American.

[113]  M. Palcic,et al.  High-throughput screening for enzyme inhibitors using frontal affinity chromatography with liquid chromatography and mass spectrometry. , 2005, Analytical Chemistry.

[114]  Dustin J Maly,et al.  Peptide microarrays for the determination of protease substrate specificity. , 2002, Journal of the American Chemical Society.

[115]  R. Aebersold,et al.  Crosslinking and Mass Spectrometry: An Integrated Technology to Understand the Structure and Function of Molecular Machines. , 2016, Trends in biochemical sciences.

[116]  Michelle L. Reyzer,et al.  MALDI imaging mass spectrometry: molecular snapshots of biochemical systems , 2007, Nature Methods.

[117]  Ronny Martínez,et al.  A Flow Cytometry–Based Screening System for Directed Evolution of Proteases , 2011, Journal of biomolecular screening.

[118]  J. Kinney,et al.  Discovery of cancer drug targets by CRISPR-Cas9 screening of protein domains , 2015, Nature Biotechnology.

[119]  H. Munier-Lehmann,et al.  Multiple Microfermentor Battery: a Versatile Tool for Use with Automated Parallel Cultures of Microorganisms Producing Recombinant Proteins and for Optimization of Cultivation Protocols , 2006, Applied and Environmental Microbiology.

[120]  Tilmann Weber,et al.  The secondary metabolite bioinformatics portal: Computational tools to facilitate synthetic biology of secondary metabolite production , 2016, Synthetic and systems biotechnology.

[121]  Marco Oldiges,et al.  An automated workflow for enhancing microbial bioprocess optimization on a novel microbioreactor platform , 2012, Microbial Cell Factories.

[122]  P. Dugourd,et al.  Chirality-dependent structuration of protonated or sodiated polyphenylalanines: IRMPD and ion mobility studies. , 2016, Physical chemistry chemical physics : PCCP.

[123]  Anna Bierczynska-Krzysik,et al.  Methods for samples preparation in proteomic research. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[124]  K. Hampel,et al.  Rapid steps in the glmS ribozyme catalytic pathway: cation and ligand requirements. , 2011, Biochemistry.

[125]  S. Mugo,et al.  The development of a capillary microreactor for transesterification reactions using lipase immobilized onto a silica monolith , 2013 .

[126]  Jonathan I. Betts,et al.  Miniature bioreactors: current practices and future opportunities , 2006, Microbial cell factories.

[127]  R. Aebersold,et al.  Analysis of protein complexes using mass spectrometry , 2007, Nature Reviews Molecular Cell Biology.

[128]  Christian Blesken,et al.  The microfluidic bioreactor for a new era of bioprocess development , 2016 .

[129]  L. Marri,et al.  A Chromogenic Assay Suitable for High-Throughput Determination of Limit Dextrinase Activity in Barley Malt Extracts. , 2015, Journal of agricultural and food chemistry.

[130]  Kai Blin,et al.  CRISPy-web: An online resource to design sgRNAs for CRISPR applications , 2016, Synthetic and systems biotechnology.

[131]  David L. Hawksworth,et al.  The fungal dimension of biodiversity: magnitude, significance, and conservation , 1991 .

[132]  William Stafford Noble,et al.  Technical advances in proteomics: new developments in data-independent acquisition , 2016, F1000Research.

[133]  Jan Brezovsky,et al.  Computational tools for designing and engineering enzymes. , 2014, Current opinion in chemical biology.

[134]  Jochen Büchs,et al.  Microscale and miniscale fermentation and screening. , 2015, Current opinion in biotechnology.

[135]  G. Haran,et al.  Vacuum Rabi splitting in a plasmonic cavity at the single quantum emitter limit , 2015, Nature Communications.

[136]  Karol M. Łącki,et al.  High throughput process development in biomanufacturing , 2014 .

[137]  R. C. Kuhad,et al.  Revisiting cellulase production and redefining current strategies based on major challenges. , 2016 .

[138]  M. Devignes,et al.  Prediction of Monomer Isomery in Florine: A Workflow Dedicated to Nonribosomal Peptide Discovery , 2014, PloS one.

[139]  James E. DiCarlo,et al.  RNA-Guided Human Genome Engineering via Cas9 , 2013, Science.

[140]  Ying Ge,et al.  Online Hydrophobic Interaction Chromatography-Mass Spectrometry for Top-Down Proteomics. , 2016, Analytical chemistry.

[141]  P. Dhulster,et al.  Facile immobilization of enzyme by entrapment using a plasma-deposited organosilicon thin film , 2014 .

[142]  D. Clark,et al.  Sol-gel encapsulated enzyme arrays for high-throughput screening of biocatalytic activity. , 2002, Biotechnology and bioengineering.

[143]  T. Northen,et al.  High-throughput platforms for metabolomics. , 2016, Current opinion in chemical biology.

[144]  Guoan Zheng,et al.  On-chip continuous monitoring of motile microorganisms on an ePetri platform. , 2012, Lab on a chip.

[145]  Pavel A. Pevzner,et al.  De Novo Peptide Sequencing via Tandem Mass Spectrometry , 1999, J. Comput. Biol..

[146]  B. Bocquet,et al.  Diffusion based kinetic selectivity modulation of enzymatic proteolysis in a microfluidic reactor: experimental analysis and stochastic modeling , 2014 .

[147]  A. Merrill,et al.  Sphingolipidomics: high-throughput, structure-specific, and quantitative analysis of sphingolipids by liquid chromatography tandem mass spectrometry. , 2005, Methods.

[148]  G. Antranikian,et al.  Characterization of a heat-active archaeal β-glucosidase from a hydrothermal spring metagenome. , 2014, Enzyme and microbial technology.

[149]  Rob Knight,et al.  The 'rare biosphere': a reality check , 2009, Nature Methods.

[150]  John M. Ward,et al.  Non-linear kinetic modelling of reversible bioconversions: Application to the transaminase catalyzed synthesis of chiral amino-alcohols , 2013 .

[151]  Brian Hubbard,et al.  Downstream processing of monoclonal antibodies--application of platform approaches. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[152]  Modeling of permeabilization process in Pseudomonas putida G7 for enhanced limonin bioconversion , 2012, Applied Microbiology and Biotechnology.

[153]  Shirly Pinto,et al.  Identification of DGAT2 Inhibitors Using Mass Spectrometry , 2016, Journal of biomolecular screening.

[154]  Kyle R. Conway,et al.  ClusterMine360: a database of microbial PKS/NRPS biosynthesis , 2012, Nucleic Acids Res..

[155]  W. Wiechert,et al.  Rapid assessment of oxygen transfer impact for Corynebacterium glutamicum , 2014, Bioprocess and Biosystems Engineering.

[156]  J. Zucko,et al.  ClustScan: an integrated program package for the semi-automatic annotation of modular biosynthetic gene clusters and in silico prediction of novel chemical structures , 2008, Nucleic acids research.

[157]  Robert Huber,et al.  Microbial Cell Factories Robo-lector – a Novel Platform for Automated High-throughput Cultivations in Microtiter Plates with High Information Content , 2022 .

[158]  Thomas Letzel,et al.  On-line coupling of high-performance liquid chromatography to a continuous-flow enzyme assay based on electrospray ionization mass spectrometry. , 2004, Analytical chemistry.

[159]  K. Schug.  Solution phase enantioselective recognition and discrimination by electrospray ionization--mass spectrometry: state-of-the-art, methods, and an eye towards increased throughput measurements. , 2007, Combinatorial chemistry & high throughput screening.

[160]  Paul Watts,et al.  The application of micro reactors to synthetic chemistry , 2001 .

[161]  Krist V Gernaey,et al.  Microscale technology and biocatalytic processes: opportunities and challenges for synthesis. , 2015, Trends in biotechnology.

[162]  M. Mazutti,et al.  Enzymatic Synthesis of Ascorbyl Palmitate in Organic Solvents: Process Optimization and Kinetic Evaluation , 2012, Food and Bioprocess Technology.

[163]  D. Volmer,et al.  Structural analysis of small to medium-sized molecules by mass spectrometry after electron-ion fragmentation (ExD) reactions. , 2016, The Analyst.

[164]  Mário A P Nunes,et al.  Microtiter plates versus stirred mini-bioreactors in biocatalysis: a scalable approach. , 2013, Bioresource technology.

[165]  Kai Blin,et al.  antiSMASH 3.0—a comprehensive resource for the genome mining of biosynthetic gene clusters , 2015, Nucleic Acids Res..

[166]  Clifford A. Meyer,et al.  Sequence determinants of improved CRISPR sgRNA design , 2015, Genome research.

[167]  Dirk Weuster-Botz,et al.  Miniature bioreactors for automated high‐throughput bioprocess design (HTBD): reproducibility of parallel fed‐batch cultivations with Escherichia coli , 2005, Biotechnology and applied biochemistry.

[168]  Bimo Ario Tejo,et al.  Optimized enzymatic synthesis of levulinate ester in solvent-free system , 2010 .

[169]  S. Leite,et al.  Epoxidation of oleic acid catalyzed by PSCI-Amano lipase optimized by experimental design , 2012 .

[170]  M. Lesaicherre,et al.  Enzymatic profiling system in a small-molecule microarray. , 2003, Organic letters.

[171]  G. Guiochon,et al.  Separation science is the key to successful biopharmaceuticals. , 2011, Journal of chromatography. A.

[172]  A. Abate,et al.  Ultrahigh-throughput screening in drop-based microfluidics for directed evolution , 2010, Proceedings of the National Academy of Sciences.

[173]  Wim J. Quax,et al.  High-Throughput Screening in Protein Engineering: Recent Advances and Future Perspectives , 2015, International journal of molecular sciences.

[174]  E. Claas,et al.  High-Throughput Identification of Bacteria and Yeast by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Conventional Medical Microbiology Laboratories , 2010, Journal of Clinical Microbiology.

[175]  P. Skagerlind,et al.  Industrial enzymes. , 2007, Advances in biochemical engineering/biotechnology.

[176]  M. Reetz,et al.  Second-generation MS-based high-throughput screening system for enantioselective catalysts and biocatalysts , 2002 .

[177]  Jochen Büchs,et al.  High-throughput screening for ionic liquids dissolving (ligno-)cellulose. , 2009, Bioresource technology.

[178]  William C. Skarnes,et al.  WGE: a CRISPR database for genome engineering , 2015, Bioinform..

[179]  C. Dumon,et al.  White biotechnology: State of the art strategies for the development of biocatalysts for biorefining. , 2015, Biotechnology advances.

[180]  Nicolas Szita,et al.  Oxygen Transfer Characteristics of Miniaturized Bioreactor Systems , 2013, Biotechnology and bioengineering.

[181]  T. Sandrin,et al.  MALDI TOF MS profiling of bacteria at the strain level: a review. , 2013, Mass spectrometry reviews.

[182]  Shinichiro Nakamura,et al.  Diastereomeric resolution directed towards chirality determination focussing on gas-phase energetics of coordinated sodium dissociation , 2016, Scientific Reports.

[183]  Timothy K Lu,et al.  Sequence-specific antimicrobials using efficiently delivered RNA-guided nucleases , 2014, Nature Biotechnology.