A novel 7α-hydroxysteroid dehydrogenase: Magnesium ion significantly enhances its activity and thermostability

Ji, Shunlin; Pan, Yinping; Zhu, Lingyu; Tan, Jun; Tang, Shuang; Yang, Qiong; Zhi, Zhang; Lou, Deshuai; Wang, Bochu

doi:10.1016/j.ijbiomac.2021.02.082

Cited by 10 publications

(3 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7α-Hydroxysteroid dehydrogenase and 7β-HSDH belong to the same short-chain dehydrogenase/reductase (SDR) family with a molecular mass of approximately 30 kDa and generally exist in homodimeric or homotetrameric structures (Oppermann et al, 2003;Kavanagh et al, 2008). The first 7α-HSDHs were isolated from aerobic or anaerobic bacteria, and in recent years, a larger number of 7α-HSDHs have been identified from black bear feces by homologous alignment (Ji et al, 2014(Ji et al, , 2021Tang et al, 2019) or metagenomics methods (Song et al, 2017). The cofactor-dependent and enzymatic activities of the reported 7α-HSDHs are shown in Table 1.…”

Section: Cdca As a Substrate For Udca Synthesismentioning

confidence: 99%

Biological synthesis of ursodeoxycholic acid

et al. 2023

View full text Add to dashboard Cite

Ursodeoxycholic acid (UDCA) is a fundamental treatment drug for numerous hepatobiliary diseases that also has adjuvant therapeutic effects on certain cancers and neurological diseases. Chemical UDCA synthesis is environmentally unfriendly with low yields. Biological UDCA synthesis by free-enzyme catalysis or whole-cell synthesis using inexpensive and readily available chenodeoxycholic acid (CDCA), cholic acid (CA), or lithocholic acid (LCA) as substrates is being developed. The free enzyme-catalyzed one-pot, one-step/two-step method uses hydroxysteroid dehydrogenase (HSDH); whole-cell synthesis, mainly uses engineered bacteria (mainly Escherichia coli) expressing the relevant HSDHs. To further develop these methods, HSDHs with specific coenzyme dependence, high enzyme activity, good stability, and high substrate loading concentration, P450 monooxygenase with C-7 hydroxylation activity and engineered strain harboring HSDHs must be exploited.

show abstract

Section: Cdca As a Substrate For Udca Synthesismentioning

confidence: 99%

Biological synthesis of ursodeoxycholic acid

et al. 2023

View full text Add to dashboard Cite

show abstract

“…To evaluate the C-terminal truncation strategy for altering substrate selectivity, the same method was implemented in two other 7α-HSDHs (J-1-1 and S1-a-1). J-1-1 [41] and S1-a-1 [38] originated from the gut microbiota of black bears, which share 66.79%, 59.93% amino acid sequence identities with St-2-2, respectively. As shown in Figure 3A, J-1-1∆C6 completely lost catalytic activities on TCA and GCA but remained active on TCDCA, GCDCA of 18.45 U/mg and 15.32 U/mg, respectively.…”

Section: Verification Of the C-terminal Truncation Strategy On Other ...mentioning

confidence: 99%

A Novel NADP(H)-Dependent 7alpha-HSDH: Discovery and Construction of Substrate Selectivity Mutant by C-Terminal Truncation

et al. 2022

Self Cite

View full text Add to dashboard Cite

7α-Hydroxysteroid dehydrogenase (7α-HSDH) plays an important role in the biosynthesis of tauroursodeoxycholic acid (TUDCA) using complex substrate chicken bile powder as raw material. However, chicken bile powder contains 4.74% taurocholic acid (TCA), and a new by-product tauroursocholic acid (TUCA) will be produced, having the risk of causing colorectal cancer. Here, we obtained a novel NADP(H)-dependent 7α-HSDH with good thermostability from Ursus thibetanus gut microbiota (named St-2-2). St-2-2 could catalyze taurochenodeoxycholic acid (TCDCA) and TCA with the catalytic activity of 128.13 and 269.39 U/mg, respectively. Interestingly, by a structure-based C-terminal truncation strategy, St-2-2△C10 only remained catalytic activity on TCDCA (14.19 U/mg) and had no activity on TCA. As a result, it can selectively catalyze TCDCA in waste chicken bile powder. MD simulation and structural analysis indicated that enhanced surface hydrophilicity and improved C-terminal rigidity affected the entry and exit of substrates. Hydrogen bond interactions between different subunits and interaction changes in Phe249 of the C-terminal loop inverted the substrate catalytic activity. This is the first report on substrate selectivity of 7α-HSDH by C-terminal truncation strategy and it can be extended to other 7α-HSDHs (J-1-1, S1-a-1).

show abstract

“…Besides, there are some progresses of 7α-and 7β-HSDH in application. 7α-and 7β-HSDH have been successfully expressed in E. coli and purified using specific tags [17,21]. Furthermore, both free enzymes and immobilized enzymes are suitable for the two steps catalytic reactions.…”

Section: Introductionmentioning

confidence: 99%

The Impact of Bilirubin on 7α- and 7β- Hydroxysteroid Dehydrogenases: Spectra and Docking Analysis

Ji¹,

Chen²,

Zhu³

et al. 2023

Preprint

View full text Add to dashboard Cite

7α- and 7β-hydroxysteroid dehydrogenases (HSDH) are a pair of enzymes that can catalyze the isomerization of hydroxyl group at site 7 of bile acids. In previous study, we found that the activities of 7α- and 7β-HSDH can be inhibited by bilirubin. In order to clarify the impact, the effects of bilirubin on enzymes were studied by kinetics, spectrum and docking analysis. Relative activity of 7α-HSDH remained less than 40 % under 1 mM bilirubin, for 7β-HSDH, only 18 % activity left at the same condition. Using taurochenodeoxycholic acid (TCDCA) as substrate, the Km of 7α-HSDH was up to 0.63 mM from 0.24 mM after binding with bilirubin, the Km of 7β-HSDH rose from 1.14 mM to 1.87 mM for the catalysis of tauroursodeoxycholic acid (TUDCA). The affinity of 7α- and 7β-HSDH to substrate decreased with the effect of bilirubin. The binding of bilirubin with 7α- or 7β-HSDH were analyzed by UV–Vis spectra, Fluorescence spectra and Circular dichroism (CD) spectra. The results reflected that bilirubin caused a slight change in the secondary structure of 7α- or 7β-HSDH, and the changes were correlated with the ratio of bilirubin to enzyme. 10 candidate molecular docking results were presented to reflect the binding of bilirubin with 7α- or 7β-HSDH and to explore the inhibition mechanism. This research not only provides the more in-depth understanding of 7α- and 7β-HSDH, but also reminds us to avoid the inhibition of bilirubin on hydroxysteroid dehydrogenases.

show abstract

A novel 7α-hydroxysteroid dehydrogenase: Magnesium ion significantly enhances its activity and thermostability

Cited by 10 publications

References 36 publications

Biological synthesis of ursodeoxycholic acid

Biological synthesis of ursodeoxycholic acid

A Novel NADP(H)-Dependent 7alpha-HSDH: Discovery and Construction of Substrate Selectivity Mutant by C-Terminal Truncation

The Impact of Bilirubin on 7α- and 7β- Hydroxysteroid Dehydrogenases: Spectra and Docking Analysis

Contact Info

Product

Resources

About