Ξ±-L-rhamnosidase (gene St-Rha) from Spirochaeta thermophila DSM 6192
Enzyme Description
Extremophile
Yes
thermophilic organism
EC Number
Sequence
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
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Database ID
NCBI: ADN01795.1 β
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 | Extraction method | Data type | 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 % v/v | 100.0 | 94.9 | % | Digital | Continuous | 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 % v/v | 12.77 | 1.77 | mM | Direct | Continuous | 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 % v/v | 5.6 | 7.27 | 1/s | Direct | Continuous | 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 % v/v | 0.44 | 4.01 | 1/(s * mM) | Direct | Continuous | 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 % v/v | 22.0 | 62.0 | % | Digital | Continuous | 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 % v/v | 22.0 | 68.0 | % | Digital | Continuous | 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 % v/v | 22.0 | 63.0 | % | Digital | Continuous | 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 % v/v | 22.0 | 54.0 | % | Digital | Continuous | 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 % v/v | 22.0 | 51.0 | % | Digital | Continuous | 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 % v/v | 22.0 | 36.0 | % | Digital | Continuous | 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 % v/v | 22.0 | 30.0 | % | Digital | Continuous | 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 % v/v | 100.0 | 8.9 | % | Digital | Continuous | 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 % v/v | 100.0 | 0.0 | % | Digital | Continuous | 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 % v/v | 100.0 | 33.1 | % | Digital | Continuous | 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 % v/v | 100.0 | 0.0 | % | Digital | Continuous | 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 % v/v | 100.0 | 0.0 | % | Digital | Continuous | 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 % v/v | 100.0 | 51.2 | % | Digital | Continuous | 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 % v/v | 100.0 | 0.0 | % | Digital | Continuous | 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 % v/v | 100.0 | 7.2 | % | Digital | Continuous | 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 % v/v | 100.0 | 72.1 | % | Digital | Continuous | 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 |
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.