Christoph Syldatk

Karlsruhe Institute of Technology, Carlsruhe, Baden-Württemberg, Germany

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Publications (192)336.43 Total impact

  • Julia Andre · David Saleh · Christoph Syldatk · Rudolf Hausmann
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    ABSTRACT: A thorough comparison of spacer-mediated covalent and non-covalent immobilization of trypsin on micro-magnetic particles was accomplished in the present study. Trypsin was coupled via diaminoalkanes, aminoalkanoic acids, bovine serum albumin (BSA), and biotin-derivate spacers onto magnetic particles. A comparison of resulting synthetic and hydrolytic activities after immobilization was performed. Whereas hydrolytic trypsin activity was measured employing N-α-Benzoyl-DL-arginine 4-nitroanilide (BAPNA) assay, synthetic trypsin activity was measured employing a dipeptide synthesis assay. Within spacer-mediated trypsin immobilization, diaminoalkanes, aminoalkanoic acids and biotin spacers showed an up to 40% increased synthetic specific activity of trypsin compared to the spacer-free method. Within the hydrolytic reaction type, coupling of trypsin via diaminoalkanes and biotin spacers resulted in a specific activity increase of up to 30%. BSA-bound trypsin displayed only minor increasing effects on both activities of trypsin. Furthermore, protein loading-dependent specific synthetic and hydrolytic activities were evaluated for 8-aminooctanoic acid, 12-aminododecanoic acid and 1,12-diamonododecane as spacers and compared to the direct covalent binding method. The protein binding capacity of spacer-modified particles was lower compared to the direct binding method. Synthetic activity of 8-aminooctanoic acid-bound trypsin was higher than in the case of the spacer-free method over a broad protein loading range with an up to 10-fold increase. The hydrolytic activity was increased by 64% using 8-aminooctanoic acid as spacer within a lower protein loading range. Spacer-bound trypsin showed a slightly lowered reusability over ten sequential cycles compared to spacer-free covalent binding method.
    No preview · Article · Jan 2016 · Journal of Molecular Catalysis B Enzymatic
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    ABSTRACT: Microorganisms produce a great variety of secondary metabolites that feature surface active and bioactive properties. Those possessing an amphiphilc molecular structure are also termed biosurfactant and are of great interest due to their often unique properties. Rouxiella sp. DSM 100043 is a gram negative enterobacter isolated from peat-bog soil and described as a new biosurfactant producing species in this study. Rouxiella sp. produces glycolipids, biosurfactants with a carbohydrate moiety in its structure. This study characterizes the composition of glycolipids with different hydrophobicities that have been produced during cultivation in a bioreactor and been extracted and purified from separated foam. Using two dimensional nuclear magnetic resonance spectroscopy, the hydrophilic moieties are elucidated as glucose with various acylation sites and as talose within the most polar glycolipids. The presence of 3' hydroxy lauroleic acid as well as myristic and myristoleic acid has been detected.
    Full-text · Article · Dec 2015 · AMB Express
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    ABSTRACT: Surfactin is one of the most promising biosurfactants due to its extraordinary surface activity. Commonly, the well-established Cooper medium, a glucose-based mineral salt medium, is utilized for the microbial production of Surfactin. The current study investigated the enhancement of Surfactin yields by analyzing the effects of different glucose concentrations, next to the introduction of an alternative chelating agent and nitrogen source. The utilization of 8 g/L glucose, 0.008 mM Na3citrate and 50 mM (NH4)2SO4 increased Surfactin yields from 0.7 to 1.1 g/L during shake flask experiments applying Bacillus subtilis DSM10T. Consequentially conducted shake flask experiments, employing five other Surfactin producer strains during cultivation in the former and enhanced version of the Cooper medium, suggest a general enhancement of Surfactin yields during application of the enhanced version of the Cooper medium. The enhancement of the medium composition is therefore most likely independent from the employed producer strain. The following utilization of the enhanced medium composition during fed-batch fermentation with integrated foam fractionation yielded 30 % more Surfactin in comparison to batch fermentations with integrated foam fractionation employing the former version of the Cooper medium. Electronic supplementary material The online version of this article (doi:10.1186/s13568-015-0145-0) contains supplementary material, which is available to authorized users.
    Preview · Article · Dec 2015 · AMB Express
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    ABSTRACT: A novel substrate, 6-(4-nitrophenyl)dihydropyrimidine-2,4(1H,3H)-dione (pNO 2 PheDU), was chemically synthesized and analytically verified for the potential biocatalytic synthesis of enantiopure β-amino acids. The hydantoinase (EC 3.5.2.2) from Arthrobacter crystallopoietes DSM20117 was chosen to prove the enzymatic hydrolysis of this substrate, since previous investigations showed activities of this enzyme toward 6-monosubstituted dihydrouracils. Whole cell biotransformations with recombinant Escherichia coli expressing the hydantoinase showed degradation of pNO 2 PheDU. Additionally, the corresponding N-carbamoyl-β-amino acid (NCarbpNO 2 βPhe) was chemically synthesized, an HPLC-method with chiral stationary phases for detection of this product was established and thus (S)-enantioselectivity toward pNO 2 PheDU has been shown. Consequently this novel substrate is a potential precursor for the enantiopure β-amino acid para-nitro-β-phenylalanine (pNO 2 βPhe).
    Full-text · Article · Dec 2015 · AMB Express
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    ABSTRACT: Surfactin is one of the most popular biosurfactants due to its numerous potential applications. The usually aerobic production via fermentation of Bacillus subtilis is accompanied by vigorous foaming which leads to complex constructions and great expense. Therefore it is reasonable to search for alternative foam-free production processes. The current study introduces a novel approach to produce Surfactin in a foam-free process applying a strictly anaerobic bioreactor cultivation. The process was performed several times with different glucose concentrations in mineral salt medium. The fermentations were analyzed regarding specific (qSurfactin, vol. qSurfactin) and overall product yields (YP/X, YP/S) as well as substrate utilization (YX/S). Fermentations in which 2.5 g/L glucose were employed proofed to be the most effective, reaching product yields of YP/X = 0.278 g/g. Most interesting, the product yields exceeded classical aerobic fermentations, in which foam fractionation was applied. Additionally, values for specific production rate qSurfactin (0.005 g/(g∙h)) and product yield per consumed substrate (YP/S = 0.033 g/g) surpass results of comparable foam-free processes. The current study introduces an alternative to produce a biosurfactant that overcomes the challenges of severe foaming and need for additional constructions. Electronic supplementary material The online version of this article (doi:10.1186/s13568-015-0107-6) contains supplementary material, which is available to authorized users.
    Preview · Article · Dec 2015 · AMB Express
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    ABSTRACT: Immobilization of enzymes onto different carriers increases enzyme's stability and reusability within biotechnological and pharmaceutical applications. However, some immobilization techniques are associated with loss of enzymatic specificity and/or activity. Possible reasons for this loss are mass transport limitations or structural changes. For this reason an immobilization method must be selected depending on immobilisate's demands. In this work different immobilization media were compared towards the synthetic and hydrolytic activities of immobilized trypsin as model enzyme on magnetic micro-particles. Porcine trypsin immobilization was carried out in organic and aqueous media with magnetic microparticles. The immobilization conditions in organic solvent were optimized for a peptide synthesis reaction. The highest carrier activity was achieved at 1 % of water (v/v) in dioxane. The resulting immobilizate could be used over ten cycles with activity retention of 90 % in peptide synthesis reaction in 80 % (v/v) ethanol and in hydrolysis reaction with activity retention of 87 % in buffered aqueous solution. Further, the optimized method was applied in peptide synthesis and hydrolysis reactions in comparison to an aqueous immobilization method varying the protein input. The dioxane immobilization method showed a higher activity coupling yield by factor 2 in peptide synthesis with a maximum activity coupling yield of 19.2 % compared to aqueous immobilization. The hydrolysis activity coupling yield displayed a maximum value of 20.4 % in dioxane immobilization method while the aqueous method achieved a maximum value of 38.5 %. Comparing the specific activity yields of the tested immobilization methods revealed maximum values of 5.2 % and 100 % in peptide synthesis and 33.3 % and 87.5 % in hydrolysis reaction for the dioxane and aqueous method, respectively. By immobilizing trypsin in dioxane, a beneficial effect on the synthetic trypsin activity resilience compared to aqueous immobilization medium was shown. The results indicate a substantial potential of the micro-aqueous organic protease immobilization method for preservation of enzymatic activity during enzyme coupling step. These results may be of substantial interest for enzymatic peptide synthesis reactions at mild conditions with high selectivity in industrial drug production.
    Preview · Article · Dec 2015 · BMC Biotechnology
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    ABSTRACT: The risk of hydrocarbon contamination in marine polar areas is constantly increasing. Autochthonous bacteria, due to their ability to cope and survive under extreme environmental conditions, can play a fundamental role in the hydrocarbon degradation. The degradation process is often enhanced by the production of biosurfactant molecules. The present study reports for the first time on the isolation of biosurfactant-producing bacteria from Arctic and Antarctic shoreline sediments. A total of 199 psychrotolerant bacterial isolates were obtained from hydrocarbon-amended (with crude or diesel oil) microcosms. A total of 18 isolates were selected for their ability to grow in the presence of crude oil and produce biosurfactants, as it was revealed by the production of good E24 values (≥50 %) and/or reduction in the surface tension (under 30 mN/m). The positive response of the isolates to both tests suggests a possible production of biosurfactants with emulsifying and interfacial activities. Biosurfactant-producing isolates were mainly affiliated to the genera Rhodococcus (14 isolates), followed by Pseudomonas (two isolates), Pseudoalteromonas (one isolate) and Idiomarina (one isolate). Thin-layer chromatography of biosurfactant crude extracts revealed that the majority of the selected isolates were able to produce glycolipidic surfactants. Our results enlarge the knowledge, which is still poor and fragmentary, on biosurfactant producers from polar areas and indicate marine polar sediments as a source of bacteria with potential applications in the remediation of hydrocarbon-contaminated cold environments.
    No preview · Article · Oct 2015 · Polar Biology
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    ABSTRACT: Biosurfactants are surface-active agents produced by microorganisms and show increasing significance in various industrial applications. A great variety of these secondary metabolites are described to occur within actinomycetes, amongst trehalose lipids and oligosaccharide lipids produced by the family Tsukamurellaceae. This study reports on the production of not yet described compounds with surface active behavior by non-pathogenic Tsukamurella pseudospumae and T. spumae during growth on hydrophobic carbon sources. Extracts of the purified compounds differ in terms of structure and performance properties to other biosurfactants described within their family. Infrared and nuclear magnetic resonance spectroscopic analysis revealed the presence of aromatic moieties within the surfactant produced, which to date is only known to occur within phenolic glycolipids of some mycobateria. Copyright © 2015 Elsevier B.V. All rights reserved.
    No preview · Article · Jul 2015 · Journal of Biotechnology
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    ABSTRACT: The effect of heavy metals on the activity of biosurfactants produced by Joostella strain A8 from the polychaete Megalomma claparedei was investigated. Biosurfactant activity was first improved by evaluating the influence of abiotic parameters. Higher E24 indices were achieved at 25 °C in mineral salt medium supplemented with 2 % glucose, 3 % sodium chloride (w/v) and 0.1 % ammonium chloride (w/v). Considerable surface tension reduction was never recorded. Heavy metal tolerance was preliminarily assayed by plate diffusion method resulting in the order of toxicity Cd > Cu > Zn. The activity of biosurfactants was then evaluated in the presence of heavy metals at different concentrations in liquid cultures that were incubated under optimal conditions for biosurfactant activity. The production of stable emulsions resulted generally higher in the presence of metals. These findings suggest that biosurfactant production could represent a bacterial adaptive strategy to defend cells from a stress condition derived from heavy metals in the bulk environment.
    No preview · Article · Jun 2015 · Ecotoxicology
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    ABSTRACT: Globally the change towards the establishment of a bio-based economy has resulted in an increased need for bio-based applications. This, in turn, has served as a driving force for the discovery and application of novel biosurfactants. The class Actinobacteria represents a vast group of microorganisms with the ability to produce a diverse range of secondary metabolites, including surfactants. Understanding the extensive nature of the biosurfactants produced by actinobacterial strains can assist in finding novel biosurfactants with new potential applications. This review therefore presents a comprehensive overview of the knowledge available on actinobacterial surfactants, the chemical structures that have been completely or partly elucidated, as well as the identity of the biosurfactant-producing strains. Producer strains of not yet elucidated compounds are discussed, as well as the original habitats of all the producer strains, which seems to indicate that biosurfactant production is environmentally driven. Methodology applied in the isolation, purification and structural elucidation of the different types of surface active compounds, as well as surfactant activity tests, are also discussed. Overall, actinobacterial surfactants can be summarized to include the dominantly occurring trehalose-comprising surfactants, other non-trehalose containing glycolipids, lipopeptides and the more rare actinobacterial surfactants. The lack of structural information on a large proportion of actinobacterial surfactants should be considered as a driving force to further explore the abundance and diversity of these compounds. This would allow for a better understanding of actinobacterial surface active compounds and their potential for biotechnological application.
    Full-text · Article · Mar 2015 · Frontiers in Microbiology
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    ABSTRACT: Bioprocess engineering is a highly interdisciplinary field of study which is strongly benefited by practical courses where students can actively experience the interconnection between biology, engineering, and physical sciences. This work describes a lab course developed for 2nd year undergraduate students of bioprocess engineering and related disciplines, where students are challenged with a real-life bioprocess-engineering application, the production of recombinant protein in a fed-batch process. The lab course was designed to introduce students to the subject of operating and supervising an experiment in a bioreactor, along with the analysis of collected data and a final critical evaluation of the experiment. To provide visual feedback of the experimental outcome, the organism used during class was Escherichia coli which carried a plasmid to recombinantly produce enhanced green fluorescent protein (eGFP) upon induction. This can easily be visualized in both the bioreactor and samples by using ultraviolet light. The lab course is performed with bioreactors of the simplest design, and is therefore highly flexible, robust and easy to reproduce. As part of this work the implementation and framework, the results, the evaluation and assessment of student learning combined with opinion surveys are presented, which provides a basis for instructors intending to implement a similar lab course at their respective institution.
    No preview · Article · Mar 2015 · Biochemistry and Molecular Biology Education
  • Siegmund Lang · Christoph Syldatk · Udo Rau
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    ABSTRACT: A review on the chemo-enzymic synthesis of glycolipids using lipases, glycolipidases and proteases, and the enzymic modification of naturally occurring glycolipids. Important advantages of the biocatalysts are mild reaction conditions, their regioselectivities and, in some cases, their stereoselectivities. These methods are well established for preparative use, and may also have a significant impact on the industrial prodn. of glycolipids for use as surfactants, sweeteners, food ingredients, cosmetics, and pharmaceuticals. Some recommendations to overcome the limitations assocd. with these methods are given. Novel methods such as the lipase-catalyzed solid-phase synthesis, including addn. of adjuvants, which can successfully be applied at the preparative scale, may not easily be used on an industrial scale. [on SciFinder(R)]
    No preview · Conference Paper · Feb 2015
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    ABSTRACT: Globally, the drive towards the establishment of a bio-based economy has resulted in an increased need for bio-based applications. This, in turn, has served as a driving force for the discovery and application of novel biosurfactants. The class Actinobacteria represents a vast group of microorganisms with the ability to produce a diverse range of secondary metabolites, including surfactants. Understanding the extensive nature of the biosurfactants produced by actinobacterial strains can assist in finding novel biosurfactants with new potential applications. This review therefore presents a comprehensive overview of the knowledge available on actinobacterial surfactants, the chemical structures that have been completely or partly elucidated, as well as the identity of the biosurfactant-producing strains. Producer strains of not yet elucidated compounds are discussed, as well as the original habitats of all the producer strains, which seems to indicate that biosurfactant production is environmentally driven. Methodology applied in the isolation, purification and structural elucidation of the different types of surface active compounds, as well as surfactant activity tests, are also discussed. Overall, actinobacterial surfactants can be summarized to include the dominantly occurring trehalose-comprising surfactants, other non-trehalose containing glycolipids, lipopeptides and the more rare actinobacterial surfactants. The lack of structural information on a large proportion of actinobacterial surfactants should be considered as a driving force to further explore the abundance and diversity of these compounds. This would allow for a better understanding of actinobacterial surface active compounds and their potential for biotechnological application.
    Full-text · Article · Jan 2015
  • Christin Slomka · Ulrike Engel · Christoph Syldatk · Jens Rudat

    No preview · Chapter · Jan 2015
  • Martin Pöhnlein · Christoph Syldatk · Siegmund Lang · Rudolf hausmann

    No preview · Article · Jan 2015 · European Journal of Lipid Science and Technology
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    Martin Pöhnlein · Tim Finkbeiner · Christoph Syldatk · Rudolf Hausmann
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    ABSTRACT: A microtiter plate-based assay was developed to evaluate the ability of lipases to perform transesterifications when employed in different organic solvents. A 4-nitrophenol assay was carried out employing seven different lipase formulations and two fatty acid methyl esters with different chain lengths in a total of six organic solvents with logP values approximately between 1 and -1. This assay delivered results within comparatively short times measured by a color reaction and thus facilitates the choice of an enzyme-solvent combination for the synthesis of glycolipids. To validate the findings, glycolipid syntheses were performed using the same lipase formulation in the same solvents. When comparing the results obtained using the microtiter plate-based assay to the results of the glycolipid syntheses using the same lipases and solvents, matching results were obtained.
    Preview · Article · Nov 2014 · Biotechnology Letters
  • Janina Beuker · Christoph Syldatk · Rudolf Hausmann

    No preview · Chapter · Nov 2014
  • Rudolf Hausmann · Christoph Syldatk

    No preview · Chapter · Nov 2014
  • Marius Henkel · Christoph Syldatk · Rudolf Hausmann

    No preview · Chapter · Nov 2014
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    ABSTRACT: Enzymatic synthesis of sugar fatty acid esters in organic solvents is a well-described procedure to synthesize glycolipids. This study aims at replacing these solvents with deep eutectic solvents (DES), a group of solvents that gained more and more interest during the last years, since they can be easily produced from non-toxic resources. Enzymatic glycolipid synthesis in deep eutectic solvents was investigated, employing Candida antarctica lipase B (Novozyme 435) in various deep eutectic solvents. A successful lipase-catalyzed synthesis of glucose fatty acid esters gave proof of this concept, while using the two deep eutectic solvents consisting of choline chloride and urea (CC : U) and choline chloride and glucose (CC : Glc). Additionally the DES consisting of choline chloride and glucose was observed to act as solvent and substrate for the synthesis at the same time.Practical application:Glycolipids find applications in many every-day products like cosmetic and pharmaceutical formulations, food and classic cleaning products, utilizing their good detergent or emulsification properties. Glycolipids can, among other routes, be synthesized via lipase-catalyzed reactions, which are often carried out in organic solvents. By replacing these organic solvents with more ecologically friendly solvents like deep eutectic solvents, the reaction might be improved and the amount of waste produced could be reduced.
    No preview · Article · Nov 2014 · European Journal of Lipid Science and Technology

Publication Stats

3k Citations
336.43 Total Impact Points

Institutions

  • 2004-2016
    • Karlsruhe Institute of Technology
      • • Institute of Process Engineering in Life Sciences
      • • Chair of Technical Biology
      • • Engler Bunte Institute
      Carlsruhe, Baden-Württemberg, Germany
  • 2010
    • Klinikum Stuttgart
      Stuttgart, Baden-Württemberg, Germany
  • 2006-2010
    • BIOLOG Life Science Institute
      Bremen, Bremen, Germany
  • 1994-2008
    • Universität Stuttgart
      • • Institute for Biochemical Engineering
      • • Institute of Industrial Genetics
      Stuttgart, Baden-Württemberg, Germany
  • 1986-2006
    • Technische Universität Braunschweig
      • Institute of technical chemistry
      Brunswyck, Lower Saxony, Germany