Ξ±-L-rhamnosidase (gene St-Rha) from Spirochaeta thermophila DSM 6192

Enzyme Description

Extremophile
Yes thermophilic organism
EC Number

Sequence

Length: 895 amino acids
MVHVYDVMCEGQRFPLGLSTARPRFSWRIGAAEGVGDLFQEAFQIQVALDEGFTSPVWDSGMVHEGRSQWVDYGGPELSPCTRYYVRVRVKPIGGELSPWSDATWFETGLPEWTAPFVSPEPVERVGRSSRPRYVRREFSIDRSVAGARLYATALGLYELSINGRRVGDSFFTPGWTSYRRLLYQTYDVTDYLRPGRNVVAGIVANGWYRGELTWFLERNLYGREIALSVDLRWKEEGGEGRLVTDSTWRSSDGPLVYAELYHGEVYDSRLEVPGWDLPGFDDGEWEECRVLEISGRAVEAQEAPPVRIHERLAPVRIITTPRGEKVLDFGQNLTGWVRFRVRGRSGDRVRLRHAEILDKEGNFYTDNLRKARNIVEYVLKGEGEEEFSPRFSFQGFRYVCVDEWPGEINPSDFTAEVLHSDMEPLLEFRCSHEPLNRLHHNIQWSWKGNSLAIPTDCPQRDERLGWTGDAAMFVGTATYLSDARSFLLQWLVDMTLDQRRDGAVPFVVPDVLSLLEQQDPNFREPPFGSAGWGDAITICPWTLYRRYGDRRILERLYPAMQRWVEYMRAQAREGLVWERGFHFGDWVALDAEEGSYVGLTPLPLIATAFYALSTEILARTAGVLGLEEDERRYKDLHKRIAAAWCREFFTGEGRLRVETQTGYTLGLAFGLVPESAREEAAARLADLVREKGHLTTGFLGTPFLLRVLSDHGYTDLAYTLLLRDEYPSWLYEVKAGATTIWEHWDGIKPDGTMWSPQMNSFNHYAYGAVGEWMYAVMGGIRAEDGGPGFSSFSLEPLPGGDVRECSVHYRSPYGLIASSWTIEDGRMHWVFEVPPNTKATVRLWGVESRSVEGAAKLGLQDRIEDGGPCAAAQVGSGRYELICPVPVRRFRSMT
Chen-Mu Luo et al. (2024) β€” Efficient biosynthesis of prunin in methanol cosolvent system by an organic solvent-tolerant Ξ±-L-rhamnosidase from Spirochaeta thermophila
Enzyme and Microbial Technology  Β· doi:10.1016/j.enzmictec.2024.110410 β†—  Β· Activity - Classical Activity - Michaelis-Menten Stability - Incubation Activity + Stability - Conversion
46 measurements
Database ID
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (46 measurements)

46 measurements β€” page 2 of 3
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Methanol 30% 100.0 94.9 % 50 mM phosphate buffer 6 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Classical aqueous control (in %)
Activity - Michaelis-Menten Km (mM), measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Methanol 20% 12.77 1.77 mM 50 mM phosphate buffer 6 65Β°C Naringin L-Rhamnose , Prunin β€” β€” Classical aqueous control (in mM)
Activity - Michaelis-Menten kcat/Km (1/(s*mM)), measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Methanol 20% 5.6 7.27 1/s 50 mM phosphate buffer 6 65Β°C Naringin L-Rhamnose , Prunin β€” β€” Classical aqueous control (in 1/s)
Activity - Michaelis-Menten kcat (1/s), measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Methanol 20% 0.44 4.01 1/(s * mM) 50 mM phosphate buffer 6 65Β°C Naringin L-Rhamnose , Prunin β€” β€” Classical aqueous control (in 1/(s * mM))
Activity + Stability - Conversion Conversion measured by HPLC (naringin and prunin concenrations) in the presence of organic solvent Methanol 25% 22.0 62.0 % 50 mM phosphate buffer 6 65Β°C 5 g/L Naringin L-Rhamnose , Prunin β€” 200 rpm Classical aqueous control (in %)
Activity + Stability - Conversion Conversion measured by HPLC (naringin and prunin concenrations) in the presence of organic solvent Methanol 20% 22.0 68.0 % 50 mM phosphate buffer 6 65Β°C 5 g/L Naringin L-Rhamnose , Prunin β€” 200 rpm Classical aqueous control (in %)
Activity + Stability - Conversion Conversion measured by HPLC (naringin and prunin concenrations) in the presence of organic solvent Methanol 15% 22.0 63.0 % 50 mM phosphate buffer 6 65Β°C 5 g/L Naringin L-Rhamnose , Prunin β€” 200 rpm Classical aqueous control (in %)
Activity + Stability - Conversion Conversion measured by HPLC (naringin and prunin concenrations) in the presence of organic solvent Methanol 10% 22.0 54.0 % 50 mM phosphate buffer 6 65Β°C 5 g/L Naringin L-Rhamnose , Prunin β€” 200 rpm Classical aqueous control (in %)
Activity + Stability - Conversion Conversion measured by HPLC (naringin and prunin concenrations) in the presence of organic solvent Methanol 30% 22.0 51.0 % 50 mM phosphate buffer 6 65Β°C 5 g/L Naringin L-Rhamnose , Prunin β€” 200 rpm Classical aqueous control (in %)
Activity + Stability - Conversion Conversion measured by HPLC (naringin and prunin concenrations) in the presence of organic solvent Methanol 35% 22.0 36.0 % 50 mM phosphate buffer 6 65Β°C 5 g/L Naringin L-Rhamnose , Prunin β€” 200 rpm Classical aqueous control (in %)
Activity + Stability - Conversion Conversion measured by HPLC (naringin and prunin concenrations) in the presence of organic solvent Methanol 40% 22.0 30.0 % 50 mM phosphate buffer 6 65Β°C 5 g/L Naringin L-Rhamnose , Prunin β€” 200 rpm Classical aqueous control (in %)
Stability - Incubation Activity after incubation (3 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 20% 100.0 8.9 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
Stability - Incubation Activity after incubation (3 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 30% 100.0 0.0 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
Stability - Incubation Activity after incubation (4 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 10% 100.0 33.1 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
Stability - Incubation Activity after incubation (4 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 20% 100.0 0.0 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
Stability - Incubation Activity after incubation (4 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 30% 100.0 0.0 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
Stability - Incubation Activity after incubation (3 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 10% 100.0 51.2 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
Stability - Incubation Activity after incubation (2 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 30% 100.0 0.0 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
Stability - Incubation Activity after incubation (2 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 20% 100.0 7.2 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
Stability - Incubation Activity after incubation (2 hours at 65Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Methanol 10% 100.0 72.1 % Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer Incubation: 6, Assay: 6 Incubation: 65Β°C, Assay: 65Β°C 1 mM Naringin L-Rhamnose , Prunin β€” β€” Non-incubated control (in %), assay in aqueous phase
1 2 3

Visualization : Activity β€” Classical

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Activity β€” Michaelis-Menten

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability β€” Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Activity + Stability β€” Conversion

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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