Purification and characterization of baicalin-β-d-glucuronidase hydrolyzing baicalin to baicalein from fresh roots of Scutellaria viscidula Bge

Zhang, Chengqi; Zhang, Yufei; Chen, Jiping; Liang, Xinmiao

doi:10.1016/j.procbio.2004.07.003

Cited by 30 publications

(16 citation statements)

References 18 publications

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“…PGUS‐E and PGUS‐P were strongly inhibited by Cu 2+ as β‐D‐glucuronidases from Scutellaria root and Streptococcus LJ‐22 were inhibited by Cu 2+ . The non‐glycosylated PGUS‐E exhibited less sensitivity to metal ions and more sensitivity to chatropic agents compared with PGUS‐P (Table ).…”

Section: Discussionmentioning

confidence: 99%

“…The variations in molecular weights of β‐glucuronidases have already been reported in organisms such as Eubacterium sp . GLH (43 kDa), Scutellaria root (55–58 kDa) and Pomacea sp . (90 kDa) .…”

Section: Discussionmentioning

confidence: 99%

“…β‐glucuronidases (EC 3.2.1.31) are enzymes that catalyse hydrolysis of β‐linked glucuronides to yield their various derivatives and free glucuronic acid. There is a great variety of β‐glucuronidases and they have been used extensively for the structural investigations of proteoglycans and also as positive selection markers for transformed plants, bacteria and fungi . Recently, much attention has been paid to their exploitation as biocatalysts for the biotransformation of valuable products.…”

Section: Introductionmentioning

confidence: 99%

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Purification, characterization and comparison of Penicillium purpurogenum β‐glucuronidases expressed in Escherichia coli and Pichia pastoris

Zou

Guo

Kaleem

et al. 2013

J of Chemical Tech & Biotech

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Background To investigate the effects of post‐translational modifications in different recombinant expression systems on the catalytic properties of recombinant β‐glucuronidase. The β‐glucuronidase (GUS) gene from Penicillium purpurogenum Li‐3 was cloned and successfully expressed in Escherichia coli BL21 and Pichia pastoris G115. Results The recombinant E. coli produced a 15‐fold increased level of β‐glucuronidase while the recombinant P. pastoris strain produced a 6.9‐fold increased level of β‐glucuronidase compared with their parent strains. The β‐glucuronidases from recombinant E. coli (PGUS‐E) and P. pastors (PGUS‐P) were purified to 35.9‐ and 47.4‐fold, respectively, through affinity, ion exchange and gel filtration chromatography. PGUS‐E from E. coli was a non‐glycosylated protein with an apparent molecular mass of 72.43 kDa, while PGUS‐P from P. pastors was appropriately glycosylated with a molecular mass of 78.83 kDa measured by MALDI/TOF‐MS. Although both recombinant β‐glucuronidases exhibited similar pH optima, the glycosylated PGUS‐P showed a significantly higher thermal stability and less sensitivity to metal ions compared with the non‐glycosylated PGUS‐E. The glycosylated PGUS‐P also displayed lower Km values, and higher kcat/Km ratios than the non‐glycosylated enzyme towards glycyrrhizin. Conclusion These results revealed the key role of post‐translational modifications in the P. pastors expression system on the catalytic properties of β‐glucuronidase and its potential stability over the prokaryotic expression system which could be applied as an important tool for the functional enhancement of industrial enzymes. © 2013 Society of Chemical Industry

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Purification, characterization and comparison of Penicillium purpurogenum β‐glucuronidases expressed in Escherichia coli and Pichia pastoris

Zou

Guo

Kaleem

et al. 2013

J of Chemical Tech & Biotech

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“…8‐ O ‐Methyl transferase is probably involved in the formation of wogonin from nor‐wogonin . The data about transformations of the flavones themselves exist as well, in particular, according to the glucuronides‐aglycones pathway, where the endogenous β ‐glucuronidase and UDP glycosyltransferases participate …”

Section: Introductionmentioning

confidence: 99%

The Relationship Between Endogenous β‐Glucuronidase Activity and Biologically Active Flavones‐Aglycone Contents in Hairy Roots of Baikal Skullcap

Dikaya

Solovyeva

Sidorov

et al. 2018

Chemistry & Biodiversity

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Here, we examine the relationship between contents of principal flavones in hairy roots of Scutellaria baicalensis with the activity of the β-glucuronidase (sGUS) enzyme during a culturing cycle. Using RP-HPLC, we show that the highest contents of aglycones, baicalin and wogonin is observed at the growth days 8, 14, and 71 and reach 45, 41, and 62% (based on the total weight of hairy roots of the Baikal skullcap), correspondingly. Their accumulation is accompanied by increase of the sGUS activity, which we determined fluorometrically. Moreover, the enzyme activity is characterized by significant and reasonable correlation only with the wogonin contents. Our results confirm a significant role of sGUS at the final steps of the metabolism in root-specific flavones of Baikal skullcap and suggest how one can optimize the conditions of culturing the hairy roots for biotechnological production of individual flavonoids. For example, at the culturing day 71 wogonin constituted over 80% of all flavones extracted from cells.

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“…Moreover, the endogenous enzyme seems to be less sensitive to heat treatment than the commercial GUS and seems to be independent of the pH value of the reaction. Zhang, Zhang, Chen, and Liang (2005) have purified and characterized the baicalin-β-D-glucuronidase from fresh roots of Scutellaria viscidula Bge, which can hydrolyze the β-D-glucuronide bond of baicalin to form baicalein. However, little information is available on the utilizing endogenous enzyme hydrolysis to enhance the extraction of target compounds from higher plants.…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

Enhanced extraction of isoflavonoids from Radix Astragali by incubation pretreatment combined with negative pressure cavitation and its antioxidant activity

Zhao

Wei

et al. 2011

Innovative Food Science & Emerging Technologies

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Purification and characterization of baicalin-β-d-glucuronidase hydrolyzing baicalin to baicalein from fresh roots of Scutellaria viscidula Bge

Cited by 30 publications

References 18 publications

Purification, characterization and comparison of Penicillium purpurogenum β‐glucuronidases expressed in Escherichia coli and Pichia pastoris

Purification, characterization and comparison of Penicillium purpurogenum β‐glucuronidases expressed in Escherichia coli and Pichia pastoris

The Relationship Between Endogenous β‐Glucuronidase Activity and Biologically Active Flavones‐Aglycone Contents in Hairy Roots of Baikal Skullcap

Enhanced extraction of isoflavonoids from Radix Astragali by incubation pretreatment combined with negative pressure cavitation and its antioxidant activity

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