Biotransformation of the total flavonoid extract of epimedium into icaritin by two thermostable glycosidases from Dictyoglomus thermophilum DSM3960

Zhang, Shanshan; Luo, Jianianhua; Dong, Yurong; Wang, Zhenzhong; Xiao, Wei; Zhao, Linguo

doi:10.1016/j.procbio.2021.03.002

Cited by 19 publications

(22 citation statements)

References 32 publications

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“…The conversion rate of icariin by mutant H570Y was 4.25 times higher than that of the wild-type, reaching 18.41% ± 2.05%, but it was still lower than that of the mutant H570A (63.65% ± 3.96%). Based on these results, we proposed the hypothesis that DthRha also has the activity of hydrolyzing icariin, which was in agreement with results reported by Zhang et al (2021) .…”

Section: Discussionsupporting

confidence: 91%

“…This study aimed to develop an efficient α-L-rhamnosidase for hydrolyzing the rhamnosyl group of icariin to produce icariside I. TpeRha from T. petrophila DSM 13995 ( Xie et al, 2020 ) is one of the enzymes that has the hydrolytic activity for icariin to icariside I. Both Rhase-I from T. stollii CLY-6 ( Cheng et al, 2022 ) and DthRha from D. thermophilum DSM 3960 ( Zhang et al, 2021 ) have also been reported for the hydrolysis of the rhamnosyl group. However, the 3D structure of Rhase-I is not currently available, and homology modeling cannot be performed since the enzyme with the highest protein sequence identity with Rhase-I is only at 25.20% (PDB ID: 6q2f) ( Mensitieri et al, 2018 ).…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, in a study of structure determination by X-ray diffraction ( Guillotin et al, 2019 ), DthRha was not reported for its activity of hydrolyzing icariin to generate icariside I. This enzyme was later used in icaritin synthesis by Zhang et al (2021) . Therefore, TpeRha, which was first reported to have the activity of hydrolyzing icariin to generate icariside I, was selected as the target protein in this study.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, most of those reported α-L-rhamnosidases, such as AnRhaE from Aspergillus nidulans ( Lyu et al, 2019 ) and BtRha from Bacteroides thetaiotaomicron VPI-5482 ( Wu et al, 2018 ), could only hydrolyze the outer rhamnosidic bond. The inner rhamnosidic bond of epimedin C and icariin could only be hydrolyzed by three α-L-rhamnosidases up to now, TpeRha from T. petrophila DSM 13995 ( Xie et al, 2020 ), DthRha from Dicyolomus thermophilum DSM 3960 ( Zhang et al, 2021 ), and Rhase-Ⅰ from Talaromyces stollii CLY-6 ( Cheng et al, 2022 ). However, the catalytic efficiency of TpeRha and DthRha is insufficient, while the thermostability of Rhase-Ⅰ is poor.…”

Section: Introductionmentioning

confidence: 99%

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Computer-Aided Design of α-L-Rhamnosidase to Increase the Synthesis Efficiency of Icariside I

Huang

Hu²,

Lu³

et al. 2022

Front. Bioeng. Biotechnol.

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Icariside I, the glycosylation product of icaritin, is a novel effective anti-cancer agent with immunological anti-tumor activity. However, very limited natural icariside I content hinders its direct extraction from plants. Therefore, we employed a computer-aided protein design strategy to improve the catalytic efficiency and substrate specificity of the α-L-rhamnosidase from Thermotoga petrophila DSM 13995, to provide a highly-efficient preparation method. Several beneficial mutants were obtained by expanding the active cavity. The catalytic efficiencies of all mutants were improved 16–200-fold compared with the wild-type TpeRha. The double-point mutant DH was the best mutant and showed the highest catalytic efficiency (kcat/KM: 193.52 s−1 M−1) against icariin, which was a 209.76-fold increase compared with the wild-type TpeRha. Besides, the single-point mutant H570A showed higher substrate specificity than that of the wild-type TpeRha in hydrolysis of different substrates. This study provides enzyme design strategies and principles for the hydrolysis of rhamnosyl natural products.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Computer-Aided Design of α-L-Rhamnosidase to Increase the Synthesis Efficiency of Icariside I

Huang

Hu²,

Lu³

et al. 2022

Front. Bioeng. Biotechnol.

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“…Xie et al hydrolyzed epimedin C into icaritin using two recombinant enzymes of hyperthermophilic α-L-rhamnosidase and thermostable β-glucosidase [23], but the substrate was only epimedin C, and they did not use any high-content icariin, epimedin B or epimedin A from Epimedium herb. Zhang et al bioconverted the total flavonoid extract (containing icariin, and epimedin C, B and A) of Epimedium into icaritin with two thermostable glycosidases from Dictyoglomus thermophilum DSM3960, but the icaritin weight-yield low at 4%, i.e., only 0.20 g/l icaritin was obtained from 5 g/l of the total flavonoid extract of Epimedium [24].…”

Section: Introductionmentioning

confidence: 99%

Icaritin Preparation from Icariin by a Special Epimedium Flavonoid-Glycosidase from Aspergillus sp.y848 Strain

Wang

Liu

et al. 2022

J. Microbiol. Biotechnol.

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In this study, to obtain icaritin with high pharmacological activities from icariin, which has a content ratio of over 58% in the total flavonoids of Epimedium herb, a special Epimedium flavonoidglycosidase was produced, purified and characterized from Aspergillus sp.y848 strain. The optimal enzyme production was gained in a medium containing 5% (w/v) wheat bran extract and 0.7% (w/v) Epimedium leaf powder as the enzyme inducer, and strain culture at 30°C for 6-7 days. The molecular weight of the enzyme was approximately 73.2 kDa; the optimal pH and temperature were 5.0 and 40°C. The enzyme Km and Vmax values for icariin were 15.63 mM and 55.56 mM/h. Moreover, the enzyme hydrolyzed the 7-O-glucosides of icariin into icariside II, and finally hydrolyzed 3-Orhamnoside of icariside II into icaritin. The enzyme also hydrolyzed 7-O-glucosides of epimedin B to sagittatoside B, and then further hydrolyzed terminal 3-O-xyloside of sagittatoside B to icarisiede II, before finally hydrolyzing 3-O-rhamnoside of icarisiede II into icaritin. The enzyme only hydrolyzed 7-O-glucoside of epimedin A or epimedin C into sagittatoside A or sagittatoside C. It is possible to prepare icaritin from the high-content icariin in Epimedium herb using this enzyme. When 2.5% icariin was reacted at 40°C for 18-20 h by the low-cost crude enzyme, 5.04 g icaritin with 98% purity was obtained from 10 g icariin. Also, the icaritin molar yield was 92.5%. Our results showed icaritin was successfully produced via cost-effective and relatively simple methods from icariin by crude enzyme. Our results should be very useful for the development of medicines from Epimedium herb.

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α-l-rhamnosidase: production, properties, and applications

Pan

Zhang

et al. 2023

World J Microbiol Biotechnol

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Biotransformation of the total flavonoid extract of epimedium into icaritin by two thermostable glycosidases from Dictyoglomus thermophilum DSM3960

Cited by 19 publications

References 32 publications

Computer-Aided Design of α-L-Rhamnosidase to Increase the Synthesis Efficiency of Icariside I

Computer-Aided Design of α-L-Rhamnosidase to Increase the Synthesis Efficiency of Icariside I

Icaritin Preparation from Icariin by a Special Epimedium Flavonoid-Glycosidase from Aspergillus sp.y848 Strain

α-l-rhamnosidase: production, properties, and applications

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