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Enhanced production of ginsenoside Rh2(S) from PPD-type major ginsenosides using BglSk cloned from Saccharibacillus kuerlensis together with two glycosidase in series

BACKGROUND: Ginsenoside Rh2(S) is a promising compound for the prevention of various kinds of cancers, inflammation, and diabetes. However, due to its low concentration (<0.02%), researchers are still trying to find an efficient glycoside hydrolase for the scaled-up production of Rh2(S). METHOD:...

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Autores principales: Siddiqi, Muhammad Zubair, Ximenes, Hipolito Amaral, Song, Bong-Kyu, Park, Hye Yoon, Lee, Woong Hee, Han, Hyosang, Im, Wan-Taek
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8324944/
https://www.ncbi.nlm.nih.gov/pubmed/34354454
http://dx.doi.org/10.1016/j.sjbs.2021.04.079
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author Siddiqi, Muhammad Zubair
Ximenes, Hipolito Amaral
Song, Bong-Kyu
Park, Hye Yoon
Lee, Woong Hee
Han, Hyosang
Im, Wan-Taek
author_facet Siddiqi, Muhammad Zubair
Ximenes, Hipolito Amaral
Song, Bong-Kyu
Park, Hye Yoon
Lee, Woong Hee
Han, Hyosang
Im, Wan-Taek
author_sort Siddiqi, Muhammad Zubair
collection PubMed
description BACKGROUND: Ginsenoside Rh2(S) is a promising compound for the prevention of various kinds of cancers, inflammation, and diabetes. However, due to its low concentration (<0.02%), researchers are still trying to find an efficient glycoside hydrolase for the scaled-up production of Rh2(S). METHOD: Three glycoside hydrolases (BglBX10, Abf22-3, and BglSk) were cloned in Escherichia coli BL21 (DE3) and the expressed recombinant enzyme was used for the scaled-up production of Rh2(S) through the conversion of PPD-type (protopanaxadiol) major ginsenosides (Rb1, Rc, and Rd, except Rb2) extracted from Korean red ginseng. Specific and specialized bioconversion pathways were designed that evolved the initial bioconversion of PPD-mix → Rg3(S) → Rh2(S). The reaction was started with 50 mg/mL of PPD-mix, 20 mg/mL of BglBX10, Abf22-3, and BglSk in series, respectively. The process was completed in a 10 L jar fermenter with a 5 L working volume at 37 °C for 48 hrs. RESULTS: The designed bioconversion pathways show that Abf22-3 and BglBX10 were responsible for the conversion of Rb1, Rc and Rd → Rg3(S), and then Rg3(S) was completely transformed to Rh2(S) by BglSk. As a result, 15.1 g of ginsenoside Rh2(S) with 98.0 ± 0.2% purity was obtained after strict purification using the Prep-HPLC system with a 100 φ diameter column. Additionally, BglSk was also investigated for its production activity with seven different kinds of PPD-mix type ginsenosides. CONCLUSION: Our pilot data demonstrate that BglSk is a suitable enzyme for the gram unit production of ginsenoside Rh2(S) at the industrial level.
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spelling pubmed-83249442021-08-04 Enhanced production of ginsenoside Rh2(S) from PPD-type major ginsenosides using BglSk cloned from Saccharibacillus kuerlensis together with two glycosidase in series Siddiqi, Muhammad Zubair Ximenes, Hipolito Amaral Song, Bong-Kyu Park, Hye Yoon Lee, Woong Hee Han, Hyosang Im, Wan-Taek Saudi J Biol Sci Original Article BACKGROUND: Ginsenoside Rh2(S) is a promising compound for the prevention of various kinds of cancers, inflammation, and diabetes. However, due to its low concentration (<0.02%), researchers are still trying to find an efficient glycoside hydrolase for the scaled-up production of Rh2(S). METHOD: Three glycoside hydrolases (BglBX10, Abf22-3, and BglSk) were cloned in Escherichia coli BL21 (DE3) and the expressed recombinant enzyme was used for the scaled-up production of Rh2(S) through the conversion of PPD-type (protopanaxadiol) major ginsenosides (Rb1, Rc, and Rd, except Rb2) extracted from Korean red ginseng. Specific and specialized bioconversion pathways were designed that evolved the initial bioconversion of PPD-mix → Rg3(S) → Rh2(S). The reaction was started with 50 mg/mL of PPD-mix, 20 mg/mL of BglBX10, Abf22-3, and BglSk in series, respectively. The process was completed in a 10 L jar fermenter with a 5 L working volume at 37 °C for 48 hrs. RESULTS: The designed bioconversion pathways show that Abf22-3 and BglBX10 were responsible for the conversion of Rb1, Rc and Rd → Rg3(S), and then Rg3(S) was completely transformed to Rh2(S) by BglSk. As a result, 15.1 g of ginsenoside Rh2(S) with 98.0 ± 0.2% purity was obtained after strict purification using the Prep-HPLC system with a 100 φ diameter column. Additionally, BglSk was also investigated for its production activity with seven different kinds of PPD-mix type ginsenosides. CONCLUSION: Our pilot data demonstrate that BglSk is a suitable enzyme for the gram unit production of ginsenoside Rh2(S) at the industrial level. Elsevier 2021-08 2021-05-01 /pmc/articles/PMC8324944/ /pubmed/34354454 http://dx.doi.org/10.1016/j.sjbs.2021.04.079 Text en © 2021 Published by Elsevier B.V. on behalf of King Saud University. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Siddiqi, Muhammad Zubair
Ximenes, Hipolito Amaral
Song, Bong-Kyu
Park, Hye Yoon
Lee, Woong Hee
Han, Hyosang
Im, Wan-Taek
Enhanced production of ginsenoside Rh2(S) from PPD-type major ginsenosides using BglSk cloned from Saccharibacillus kuerlensis together with two glycosidase in series
title Enhanced production of ginsenoside Rh2(S) from PPD-type major ginsenosides using BglSk cloned from Saccharibacillus kuerlensis together with two glycosidase in series
title_full Enhanced production of ginsenoside Rh2(S) from PPD-type major ginsenosides using BglSk cloned from Saccharibacillus kuerlensis together with two glycosidase in series
title_fullStr Enhanced production of ginsenoside Rh2(S) from PPD-type major ginsenosides using BglSk cloned from Saccharibacillus kuerlensis together with two glycosidase in series
title_full_unstemmed Enhanced production of ginsenoside Rh2(S) from PPD-type major ginsenosides using BglSk cloned from Saccharibacillus kuerlensis together with two glycosidase in series
title_short Enhanced production of ginsenoside Rh2(S) from PPD-type major ginsenosides using BglSk cloned from Saccharibacillus kuerlensis together with two glycosidase in series
title_sort enhanced production of ginsenoside rh2(s) from ppd-type major ginsenosides using bglsk cloned from saccharibacillus kuerlensis together with two glycosidase in series
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8324944/
https://www.ncbi.nlm.nih.gov/pubmed/34354454
http://dx.doi.org/10.1016/j.sjbs.2021.04.079
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