Reference This study Noh et al, 2009 Wang et al, 2011 Hong et al, 2012 An et al, 2011 doi:10.1371/journal.pone.0085727.t003 7 Characterization of a Novel b-glucosidase moieties at the C3 position of aglycon. BglPm hydrolyzes outer glucoses of C3 and C20 position of ginsenosides, similar to the b-glucosidase from Sphingomonas sp. 2F2, but BglPm 80-49-9 cost cannot hydrolyze inner glucose moiety of ginsenosides at C3 and C20. A few ginsenoside hydrolyzing recombinant enzymes have been characterized to convert major ginsenosides to F2. A bglucosidase from Sulfolobus solfataricus can hydrolyze Rb1 and Rd into F2, but this enzyme can continuously hydrolyze F2 into C-K. A BglSp from Sphingomonas sp. 2F2 showed F2 production abilities from ginsenosides Rb1, Rb2, Rd, and Rc but the low conversion activity limited its application for F2 production from PPDGM. Until now, the low transformation efficiencies and unconformable conversion 22948146 pathways of recombinant glycosidase hydrolases limited the applications for the production F2 as gramscale from PPDGM. PPDGM as the substrate with a concentration of 50 mg/ml as the final concentration in 5 L in order to produce F2. As shown in Fig. 7, the ginsenoside Rc was completely converted to Rd 3.6. Optimization of PPDGM concentration F2 production efficiency of crude recombinant BglPm were tested with four PPDGM concentration in order to determine the appropriate substrate concentration for the decreasing reactor AZ-876 volume 25837696 and economical enzyme concentration for reducing production costs. The time course of the ginsenosides on the process was determined via HPLC Microcystin-LR analyses and the abundance of ginsenoside F2 was calculated. In the test condition of a high substrate concentration, the ginsenosides mixture of Rb1 and Rd were completely converted ginsenoside F2 within 7 hours. The higher reaction condition did not complete the conversion within 40 hours. Endowing the advantage to the smaller reactor size, the condition of a high concentration substrate was adopted for the next scaled-up biotransformation step. Thus, these three reaction conditions were excluded in the next step. 3.7. Scaled-up production of ginsenosides F2 PPDGM was used as substrate for the mass production of ginsenoside F2 with high purity because it is relatively abundant and can be efficiently separated in specialized ways from crude American ginseng extracts. The enzyme reaction occurred using the crude recombinant Abf22-3 followed by BglPm with Characterization of a Novel b-glucosidase within six hours after the crude Abf22-3 was applied to the PPDGM. Thus, at this point, the crude lyophilized BglPm was applied to convert both the intact ginsenoside Rb1 and Rd in the PPDGM and transformed Rd via Abf22-3 to F2. The ginsenoside F2 was purchase Rubusoside produced consecutively up to 7 hours after initiation until ginsenosides Rb1 and Rd were exhausted. The reaction sample of each point were withdrawn and analyzed via HPLC, of which the chromatography images are shown in Fig. 8. It was demonstrated that when the bioconversion rate was nearly complete for ginsenoside Rb1, Rc and Rd which were not detected by the HPLC analysis. Among the PPDGM, ginsenosides Rb2 and Rb3 occupying approximately 7.6% were not transformed by Abf223, but can be converted by BglPm, and metabolites C-O and CMx1 were remained in the solution. The research team behind this paper has searched the ginsenoside hydrolyzing bacteria and constructed several ginsenoside-hydrolyzing recombinant enzymes through the.Reference This study Noh et al, 2009 Wang et al, 2011 Hong et al, 2012 An et al, 2011 doi:10.1371/journal.pone.0085727.t003 7 Characterization of a Novel b-glucosidase moieties at the C3 position of aglycon. BglPm hydrolyzes outer glucoses of C3 and C20 position of ginsenosides, similar to the b-glucosidase from Sphingomonas sp. 2F2, but BglPm cannot hydrolyze inner glucose moiety of ginsenosides at C3 and C20. A few ginsenoside hydrolyzing recombinant enzymes have been characterized to convert major ginsenosides to F2. A bglucosidase from Sulfolobus solfataricus can hydrolyze Rb1 and Rd into F2, but this enzyme can continuously hydrolyze F2 into C-K. A BglSp from Sphingomonas sp. 2F2 showed F2 production abilities from ginsenosides Rb1, Rb2, Rd, and Rc but the low conversion activity limited its application for F2 production from PPDGM. Until now, the low transformation efficiencies and unconformable conversion 22948146 pathways of recombinant glycosidase hydrolases limited the applications for the production F2 as gramscale from PPDGM. PPDGM as the substrate with a concentration of 50 mg/ml as the final concentration in 5 L in order to produce F2. As shown in Fig. 7, the ginsenoside Rc was completely converted to Rd 3.6. Optimization of PPDGM concentration F2 production efficiency of crude recombinant BglPm were tested with four PPDGM concentration in order to determine the appropriate substrate concentration for the decreasing reactor volume 25837696 and economical enzyme concentration for reducing production costs. The time course of the ginsenosides on the process was determined via HPLC analyses and the abundance of ginsenoside F2 was calculated. In the test condition of a high substrate concentration, the ginsenosides mixture of Rb1 and Rd were completely converted ginsenoside F2 within 7 hours. The higher reaction condition did not complete the conversion within 40 hours. Endowing the advantage to the smaller reactor size, the condition of a high concentration substrate was adopted for the next scaled-up biotransformation step. Thus, these three reaction conditions were excluded in the next step. 3.7. Scaled-up production of ginsenosides F2 PPDGM was used as substrate for the mass production of ginsenoside F2 with high purity because it is relatively abundant and can be efficiently separated in specialized ways from crude American ginseng extracts. The enzyme reaction occurred using the crude recombinant Abf22-3 followed by BglPm with Characterization of a Novel b-glucosidase within six hours after the crude Abf22-3 was applied to the PPDGM. Thus, at this point, the crude lyophilized BglPm was applied to convert both the intact ginsenoside Rb1 and Rd in the PPDGM and transformed Rd via Abf22-3 to F2. The ginsenoside F2 was produced consecutively up to 7 hours after initiation until ginsenosides Rb1 and Rd were exhausted. The reaction sample of each point were withdrawn and analyzed via HPLC, of which the chromatography images are shown in Fig. 8. It was demonstrated that when the bioconversion rate was nearly complete for ginsenoside Rb1, Rc and Rd which were not detected by the HPLC analysis. Among the PPDGM, ginsenosides Rb2 and Rb3 occupying approximately 7.6% were not transformed by Abf223, but can be converted by BglPm, and metabolites C-O and CMx1 were remained in the solution. The research team behind this paper has searched the ginsenoside hydrolyzing bacteria and constructed several ginsenoside-hydrolyzing recombinant enzymes through the.Reference This study Noh et al, 2009 Wang et al, 2011 Hong et al, 2012 An et al, 2011 doi:10.1371/journal.pone.0085727.t003 7 Characterization of a Novel b-glucosidase moieties at the C3 position of aglycon. BglPm hydrolyzes outer glucoses of C3 and C20 position of ginsenosides, similar to the b-glucosidase from Sphingomonas sp. 2F2, but BglPm cannot hydrolyze inner glucose moiety of ginsenosides at C3 and C20. A few ginsenoside hydrolyzing recombinant enzymes have been characterized to convert major ginsenosides to F2. A bglucosidase from Sulfolobus solfataricus can hydrolyze Rb1 and Rd into F2, but this enzyme can continuously hydrolyze F2 into C-K. A BglSp from Sphingomonas sp. 2F2 showed F2 production abilities from ginsenosides Rb1, Rb2, Rd, and Rc but the low conversion activity limited its application for F2 production from PPDGM. Until now, the low transformation efficiencies and unconformable conversion 22948146 pathways of recombinant glycosidase hydrolases limited the applications for the production F2 as gramscale from PPDGM. PPDGM as the substrate with a concentration of 50 mg/ml as the final concentration in 5 L in order to produce F2. As shown in Fig. 7, the ginsenoside Rc was completely converted to Rd 3.6. Optimization of PPDGM concentration F2 production efficiency of crude recombinant BglPm were tested with four PPDGM concentration in order to determine the appropriate substrate concentration for the decreasing reactor volume 25837696 and economical enzyme concentration for reducing production costs. The time course of the ginsenosides on the process was determined via HPLC analyses and the abundance of ginsenoside F2 was calculated. In the test condition of a high substrate concentration, the ginsenosides mixture of Rb1 and Rd were completely converted ginsenoside F2 within 7 hours. The higher reaction condition did not complete the conversion within 40 hours. Endowing the advantage to the smaller reactor size, the condition of a high concentration substrate was adopted for the next scaled-up biotransformation step. Thus, these three reaction conditions were excluded in the next step. 3.7. Scaled-up production of ginsenosides F2 PPDGM was used as substrate for the mass production of ginsenoside F2 with high purity because it is relatively abundant and can be efficiently separated in specialized ways from crude American ginseng extracts. The enzyme reaction occurred using the crude recombinant Abf22-3 followed by BglPm with Characterization of a Novel b-glucosidase within six hours after the crude Abf22-3 was applied to the PPDGM. Thus, at this point, the crude lyophilized BglPm was applied to convert both the intact ginsenoside Rb1 and Rd in the PPDGM and transformed Rd via Abf22-3 to F2. The ginsenoside F2 was produced consecutively up to 7 hours after initiation until ginsenosides Rb1 and Rd were exhausted. The reaction sample of each point were withdrawn and analyzed via HPLC, of which the chromatography images are shown in Fig. 8. It was demonstrated that when the bioconversion rate was nearly complete for ginsenoside Rb1, Rc and Rd which were not detected by the HPLC analysis. Among the PPDGM, ginsenosides Rb2 and Rb3 occupying approximately 7.6% were not transformed by Abf223, but can be converted by BglPm, and metabolites C-O and CMx1 were remained in the solution. The research team behind this paper has searched the ginsenoside hydrolyzing bacteria and constructed several ginsenoside-hydrolyzing recombinant enzymes through the.Reference This study Noh et al, 2009 Wang et al, 2011 Hong et al, 2012 An et al, 2011 doi:10.1371/journal.pone.0085727.t003 7 Characterization of a Novel b-glucosidase moieties at the C3 position of aglycon. BglPm hydrolyzes outer glucoses of C3 and C20 position of ginsenosides, similar to the b-glucosidase from Sphingomonas sp. 2F2, but BglPm cannot hydrolyze inner glucose moiety of ginsenosides at C3 and C20. A few ginsenoside hydrolyzing recombinant enzymes have been characterized to convert major ginsenosides to F2. A bglucosidase from Sulfolobus solfataricus can hydrolyze Rb1 and Rd into F2, but this enzyme can continuously hydrolyze F2 into C-K. A BglSp from Sphingomonas sp. 2F2 showed F2 production abilities from ginsenosides Rb1, Rb2, Rd, and Rc but the low conversion activity limited its application for F2 production from PPDGM. Until now, the low transformation efficiencies and unconformable conversion 22948146 pathways of recombinant glycosidase hydrolases limited the applications for the production F2 as gramscale from PPDGM. PPDGM as the substrate with a concentration of 50 mg/ml as the final concentration in 5 L in order to produce F2. As shown in Fig. 7, the ginsenoside Rc was completely converted to Rd 3.6. Optimization of PPDGM concentration F2 production efficiency of crude recombinant BglPm were tested with four PPDGM concentration in order to determine the appropriate substrate concentration for the decreasing reactor volume 25837696 and economical enzyme concentration for reducing production costs. The time course of the ginsenosides on the process was determined via HPLC analyses and the abundance of ginsenoside F2 was calculated. In the test condition of a high substrate concentration, the ginsenosides mixture of Rb1 and Rd were completely converted ginsenoside F2 within 7 hours. The higher reaction condition did not complete the conversion within 40 hours. Endowing the advantage to the smaller reactor size, the condition of a high concentration substrate was adopted for the next scaled-up biotransformation step. Thus, these three reaction conditions were excluded in the next step. 3.7. Scaled-up production of ginsenosides F2 PPDGM was used as substrate for the mass production of ginsenoside F2 with high purity because it is relatively abundant and can be efficiently separated in specialized ways from crude American ginseng extracts. The enzyme reaction occurred using the crude recombinant Abf22-3 followed by BglPm with Characterization of a Novel b-glucosidase within six hours after the crude Abf22-3 was applied to the PPDGM. Thus, at this point, the crude lyophilized BglPm was applied to convert both the intact ginsenoside Rb1 and Rd in the PPDGM and transformed Rd via Abf22-3 to F2. The ginsenoside F2 was produced consecutively up to 7 hours after initiation until ginsenosides Rb1 and Rd were exhausted. The reaction sample of each point were withdrawn and analyzed via HPLC, of which the chromatography images are shown in Fig. 8. It was demonstrated that when the bioconversion rate was nearly complete for ginsenoside Rb1, Rc and Rd which were not detected by the HPLC analysis. Among the PPDGM, ginsenosides Rb2 and Rb3 occupying approximately 7.6% were not transformed by Abf223, but can be converted by BglPm, and metabolites C-O and CMx1 were remained in the solution. The research team behind this paper has searched the ginsenoside hydrolyzing bacteria and constructed several ginsenoside-hydrolyzing recombinant enzymes through the.
