{"help":"Return the metadata of a dataset (package) and its resources. :param id: the id or name of the dataset :type id: string","success":true,"result":[{"id":"44edf99f-2fe2-4ce5-90cb-d24416802e3d","name":"plasma-driven-biocatalysis-using-cytochrome-p450-enzyme-cyp152bs\u03b2","title":"Plasma-driven biocatalysis using the cytochrome P450 enzyme CYP152BS\u03b2","author_email":"julia.bandow@rub.de","maintainer":"Research Data Repository","maintainer_email":"achim.vonkeudell@rub.de","license_title":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/","notes":"\u003Cp\u003EPlasma-driven biocatalysis utilizes in situ H2O2 production by atmospheric pressure plasmas to drive H2O2-dependent enzymatic reactions. Having previously established plasma-driven biocatalysis using recombinant unspecific peroxygenase from Agrocybe aegerita (rAaeUPO) to produce (R)-1-phenylethanol from ethylbenzene (ETBE), we here employed CYP152 from Bacillus subtilis (CYP152BS\u03b2). CYP152BS\u03b2 naturally hydroxylates medium and long-chain carboxylic acids, and, with short-chain carboxylic acids as decoy molecules, also converts non-natural substrates such as ETBE. To produce active CYP152BS\u03b2 overexpression and heme loading were optimized. The conversion of the non-natural substrates guaiacol and ABTS with heptanoic acid as decoy molecule and H2O2 from stock solution yielded 18.28 and 21.13 nmol product min-1 Embedded Image, respectively. These reactions also served to assess compatibility of CYP152BS\u03b2 with plasma-driven biocatalysis regarding temperature and H2O2 operating windows. To establish CYP152BS\u03b2-based plasma-driven biocatalysis, immobilized enzyme in a rotating bed reactor (5 ml reaction volume) was then supplied with H2O2 from a capillary plasma jet operated with 1280 ppm H2O in helium. After a 120 min run time a turnover number (TON) of 18.82 mol(R)-1-PhOl Embedded Image was reached. We conclude that plasma-driven biocatalysis can be extended to other H2O2-dependent enzymes. Future efforts will be directed at increasing the TON and product range.\u003C\/p\u003E\n","url":"https:\/\/rdpcidat.rub.de\/dataset\/plasma-driven-biocatalysis-using-cytochrome-p450-enzyme-cyp152bs%CE%B2","state":"Active","log_message":"Edited by kd.","private":true,"revision_timestamp":"Tue, 06\/10\/2025 - 13:29","metadata_created":"Tue, 02\/04\/2025 - 14:18","metadata_modified":"Tue, 06\/10\/2025 - 13:29","creator_user_id":"fbaa0bb1-6827-46c5-a646-bca2c4ce442a","type":"Dataset","resources":[{"id":"5cd83c15-b339-4a14-88aa-3641cb3e2b35","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/Fig.2_CypC_Spektren.xlsx","description":"\u003Cp\u003ESpectral analysis of purified CYP152BS\u03b2 before and after reconstitution.\u003Cbr \/\u003E\nAbsorption spectra were recorded using 12 \u00b5mol l-1 CYP152BS\u03b2 in potassium phosphate buffer (100 mmol l-1, pH 7). Buffer served as blank. R\/z values were calculated by relating absorption of the soret peak (at 420 nm) to absorption at 280 nm. Data shown are representative of three independent replicates.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:31","name":"Figure2","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"86.62 KB","created":"Tue, 02\/04\/2025 - 14:20","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:31"},{"id":"b0e19ee9-5b3d-4a47-a5e6-b3b38de22e5e","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/Fig3_ABTS.xlsx","description":"\u003Cp\u003EActivity of CYP152BS\u03b2 at different H2O2 concentrations using the non-natural substrates ABTS (a,b) and guaiacol (c,d).\u003Cbr \/\u003E\nActivity assays were performed at room temperature (approx. 22\u00b0C) using heptanoic acid as decoy molecule. Product conversion was measured photometrically at 405 nm (ABTS) or 470 nm (guaiacol) and product formation was calculated using Lambert-Beer law (\u03b5ABTS 36.8 mmol l-1 cm-1; \u03b5guaiacol 26.6 mmol l-1 cm-1). Specific activities were calculated based on the initial reaction velocities and given in mmol l-1 min-1 mg-1. Reactions were performed by adding up to 10 mM H2O2. For the conversion of ABTS, specific activity peaked at 1 mM, for guaiacol it peaked at 2.5 mM ((b,d) show blow-ups of the activity at low H2O2 concentrations, the dashed lines indicate the H2O2-dependent increase in specific activity). Means and standard deviations represent three independent experiments.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:27","name":"Figure3","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"85.02 KB","created":"Tue, 02\/04\/2025 - 14:21","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:27"},{"id":"c219a776-0b21-437e-82cf-7b748f36b0fb","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/Fig3_Guaiacol.xlsx","description":"\u003Cp\u003EActivity of CYP152BS\u03b2 at different H2O2 concentrations using the non-natural substrates ABTS (a,b) and guaiacol (c,d).\u003Cbr \/\u003E\nActivity assays were performed at room temperature (approx. 22\u00b0C) using heptanoic acid as decoy molecule. Product conversion was measured photometrically at 405 nm (ABTS) or 470 nm (guaiacol) and product formation was calculated using Lambert-Beer law (\u03b5ABTS 36.8 mmol l-1 cm-1; \u03b5guaiacol 26.6 mmol l-1 cm-1). Specific activities were calculated based on the initial reaction velocities and given in mmol l-1 min-1 mg-1. Reactions were performed by adding up to 10 mM H2O2. For the conversion of ABTS, specific activity peaked at 1 mM, for guaiacol it peaked at 2.5 mM ((b,d) show blow-ups of the activity at low H2O2 concentrations, the dashed lines indicate the H2O2-dependent increase in specific activity). Means and standard deviations represent three independent experiments.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:28","name":"Figure3Guaiacol","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"110.47 KB","created":"Tue, 02\/04\/2025 - 14:21","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:28"},{"id":"e4e0fa41-faec-4301-8c9a-9e3ac09fddf4","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/Fig4_Temps.xlsx","description":"\u003Cp\u003ETemperature dependency of CYP152BS\u03b2 using ABTS (a) and guaiacol (b) as non-natural substrates.\u003Cbr \/\u003E\nProduct conversion was measured photometrically at 405 nm or 470 nm, respectively. Activity assays were performed at different temperatures using a UV\/VIS spectrometer with an integrated Peltier element. All assay components were incubated at the respective temperatures for 5 min prior to starting the enzymatic reaction. Product formation was calculated using Lambert-Beer law (\u03b5ABTS 36.8 mmol l-1 cm-1; \u03b5guaiacol 26.6 mmol l-1 cm-1). Specific activities are given in mmol l-1 min-1 mg-1 and were calculated based on the initial velocities of the reactions. Means and standard deviations reflect three experiments.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:29","name":"Figure4","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"302.07 KB","created":"Tue, 02\/04\/2025 - 14:21","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:29"},{"id":"57bc97d5-e1b7-43ac-89ff-ed33aab06311","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/Fig5_20%25stacked.xlsx","description":"\u003Cp\u003EPlasma-driven biocatalysis (1280 ppm H2O in feed gas) with capillary plasma jet using CYP152BS\u03b2.\u003Cbr \/\u003E\nProduct conversion of the substrate ETBE using direct plasma treatment of CYP152BS\u03b2 immobilized on ReliZyme HA403 M beads. Reaction solution contained 5 ml potassium phosphate buffer (100 mmol l-1, pH 7) with 50 mmol l-1 ETBE and 20 mmol l-1 heptanoic acid (as decoy molecule). Plasma treatment was performed with a water concentration of 1280 ppm in the feed gas. Every 5 min, aliquots were withdrawn for product analysis by GC measurement and the reaction volume was topped up with reaction solution to 5 ml. Means and standard deviations reflect three experiments.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:30","name":"Figure5","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"59.94 KB","created":"Tue, 02\/04\/2025 - 14:22","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:30"},{"id":"bc1fb070-8096-41d0-9047-ef06f03915f5","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/Fig6_20%25total.xlsx","description":"\u003Cp\u003EPlasma-driven biocatalysis with the capillary plasma jet using CYP152BS\u03b2 using complete buffer exchange every 10 min.\u003Cbr \/\u003E\nThe reaction solution containing 5 ml potassium phosphate buffer (100 mmol l-1, pH 7) with 50 mmol l-1 ETBE and 20 mmol l-1 heptanoic acid (as decoy molecule) and was exchanged every 10 min. Product conversion of the substrate ETBE were evaluated every 10 min (a) and product accumulation (b) were evaluated with H2O2, which was supplied by plasma treatment of the reaction solution containing CYP152BS\u03b2 immobilized on ReliZyme HA403 M using a capillary plasma jet and 1280 ppm H2O in the feed gas by GC measurement. Plasma treatment was continued for a total of 120 min. Means and standard deviations reflect three experiments.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:30","name":"Figure6","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"67.47 KB","created":"Tue, 02\/04\/2025 - 14:22","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:30"},{"id":"f36d58a6-eea4-423a-9c38-4c2c1954dd60","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/SupplFig1_Spectra.xlsx","description":"\u003Cp\u003ESupplementary Figure 1: Spectral analysis of purified CYP152BSb overproduced in LB medium. For \u003Cem\u003EcypC\u003C\/em\u003E overexpression, cultures were inoculated to an OD600 of 0.05 and incubated at 37\u00b0C to an OD600 of 0.5-0.6, followed by an induction with IPTG (100 \u03bcmol l-1). After 4 h incubation at 30\u00b0C, the cells were harvested and used for protein purification. Absorption spectra were recorded using 12 \u03bcmol l- 1 CYP152BSb in potassium phosphate buffer (100 mmol l-1, pH 7). Buffer served as blank. R\/z value was calculated by relating absorption of the soret peak (at 420nm) to absorption at 280nm. Representative data of three independent replicates is displayed.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:31","name":"Supplementary Figure1","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"37.1 KB","created":"Tue, 02\/04\/2025 - 15:09","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:31"},{"id":"6099ab2b-be5e-4135-af5a-46554a4ad03d","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/SupplFig2_Chroma.xlsx","description":"\u003Cp\u003ESupplementary Figure 2: IMAC based purification of CYP152BSb. Lysate was loaded onto a HisTrap FF crude 5 ml column (GE Healthcare) and the His6-tagged CYP152BSb protein was purified with an \u00c4KTA pure25 system (GE Healthcare). Proteins were eluted with three stepwise increases in imidazole concentration (50 mmol l-1; 75 mmol l-1, 200 mmol l-1). The HisTrap FF crude 5 ml column was finally subjected to washing with 500 mmol l-1 imidazole. Absorption at 280 nm (general proteins) and 420 nm (heme containing protein) is displayed. Representative data of four independent replicates is displayed.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:39","name":"Supplementary Figure2","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"10.12 MB","created":"Tue, 02\/04\/2025 - 15:10","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:39"},{"id":"1c1453ce-82e8-4035-aaf6-80635330d8d6","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/SupplFig5.xlsx","description":"\u003Cp\u003ESupplementary Figure 5: Plasma-driven biocatalysis with capillary plasma jet using CYP152BSb and 6400 ppm H2O in feed gas. ETBE was hydroxylated using plasma treatment of a reaction solution containing CYP152BSb immobilized on ReliZyme HA403 M. The reaction solution contained 5 ml potassium phosphate buffer (100 mmol l-1, pH 7) with 50 mmol l-1 ETBE and 20 mmol l-1 heptanoic acid (as decoy molecule). Plasma treatment was performed with a water concentration of 6400 ppm in the feed gas. Every 5 min aliquots were withdrawn for product analysis by GC measurement. Means and standard deviations reflect three experiments.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:39","name":"Supplementary Figure5","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"33.37 KB","created":"Tue, 02\/04\/2025 - 15:11","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:39"},{"id":"0bf7d292-686f-4cea-9a5e-9346c5c6e168","revision_id":"","url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/SupplFig7.xlsx","description":"\u003Cp\u003ESupplementary Figure 7: Plasma-driven biocatalysis with capillary plasma jet using CYP152BSb and 640 ppm H2O in feed gas. ETBE was hydroxylated using plasma treatment of a reaction solution containing CYP152BSb immobilized on ReliZyme HA403 M. The reaction solution contained 5 ml potassium phosphate buffer (100 mmol l-1, pH 7) with 50 mmol l-1 ETBE and 20 mmol l-1 heptanoic acid (as decoy molecule). Plasma treatment was performed with a water concentration of 640 ppm in the feed gas. Every 5 min aliquots were withdrawn for product analysis by GC measurement. Means and standard deviations reflect three experiments.\u003C\/p\u003E\n","format":"xlsx","state":"Active","revision_timestamp":"Thu, 02\/06\/2025 - 10:39","name":"Supplementary Figure7","mimetype":"application\/vnd.openxmlformats-officedocument.spreadsheetml.sheet","size":"23.93 KB","created":"Tue, 02\/04\/2025 - 15:12","resource_group_id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","last_modified":"Date changed  Thu, 02\/06\/2025 - 10:39"}],"groups":[{"description":"","id":"a7cc37b6-5294-4469-8ad1-7e60df6ea28f","image_display_url":"https:\/\/rdpcidat.rub.de\/sites\/default\/files\/rublogoweiss_0_1.png","title":"Applied Microbiology","name":"group\/applied-microbiology"}]}]}