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

Enzyme Description

Species

Spirochaeta thermophila DSM 6192 ↗

Extremophile

Yes thermophilic organism

EC Number

3.2.1.40 ↗

Sequence

Length: 895 amino acids

MVHVYDVMCEGQRFPLGLSTARPRFSWRIGAAEGVGDLFQEAFQIQVALDEGFTSPVWDSGMVHEGRSQWVDYGGPELSPCTRYYVRVRVKPIGGELSPWSDATWFETGLPEWTAPFVSPEPVERVGRSSRPRYVRREFSIDRSVAGARLYATALGLYELSINGRRVGDSFFTPGWTSYRRLLYQTYDVTDYLRPGRNVVAGIVANGWYRGELTWFLERNLYGREIALSVDLRWKEEGGEGRLVTDSTWRSSDGPLVYAELYHGEVYDSRLEVPGWDLPGFDDGEWEECRVLEISGRAVEAQEAPPVRIHERLAPVRIITTPRGEKVLDFGQNLTGWVRFRVRGRSGDRVRLRHAEILDKEGNFYTDNLRKARNIVEYVLKGEGEEEFSPRFSFQGFRYVCVDEWPGEINPSDFTAEVLHSDMEPLLEFRCSHEPLNRLHHNIQWSWKGNSLAIPTDCPQRDERLGWTGDAAMFVGTATYLSDARSFLLQWLVDMTLDQRRDGAVPFVVPDVLSLLEQQDPNFREPPFGSAGWGDAITICPWTLYRRYGDRRILERLYPAMQRWVEYMRAQAREGLVWERGFHFGDWVALDAEEGSYVGLTPLPLIATAFYALSTEILARTAGVLGLEEDERRYKDLHKRIAAAWCREFFTGEGRLRVETQTGYTLGLAFGLVPESAREEAAARLADLVREKGHLTTGFLGTPFLLRVLSDHGYTDLAYTLLLRDEYPSWLYEVKAGATTIWEHWDGIKPDGTMWSPQMNSFNHYAYGAVGEWMYAVMGGIRAEDGGPGFSSFSLEPLPGGDVRECSVHYRSPYGLIASSWTIEDGRMHWVFEVPPNTKATVRLWGVESRSVEGAAKLGLQDRIEDGGPCAAAQVGSGRYELICPVPVRRFRSMT

Chen-Mu Luo et al. (2024) — Efficient biosynthesis of prunin in methanol cosolvent system by an organic solvent-tolerant α-L-rhamnosidase from Spirochaeta thermophila

Enzyme and Microbial Technology · doi:10.1016/j.enzmictec.2024.110410 ↗ · Activity - Classical Activity - Michaelis-Menten Stability - Incubation Activity + Stability - Conversion

46 measurements ▼

Database ID

NCBI: ADN01795.1 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (46 measurements)

46 measurements — page 3 of 3

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (1 hour at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose , Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (1 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	20%	100.0	44.9	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose , Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (1 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	10%	100.0	89.8	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose , Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (0.5 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	30%	100.0	60.6	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose , Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (0.5 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	20%	100.0	98.7	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose , Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (0.5 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	10%	100.0	102.6	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose , Prunin	—	—	Non-incubated control (in %), assay in aqueous phase

Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Benzene	20%	100.0	86.1	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Benzene	10%	100.0	102.0	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Acetonitrile	30%	100.0	17.9	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Acetonitrile	20%	100.0	40.2	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Acetonitrile	10%	100.0	81.1	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100.0	6.8	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100.0	29.8	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	66.7	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Ethanol	30%	100.0	2.8	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Ethanol	20%	100.0	44.7	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Ethanol	10%	100.0	80.6	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Methanol	20%	100.0	109.8	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Methanol	10%	100.0	123.2	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Cyclohexane	30%	100.0	95.7	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Cyclohexane	20%	100.0	96.7	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Cyclohexane	10%	100.0	103.5	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Petroleum Ether	30%	100.0	71.7	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Petroleum Ether	20%	100.0	88.6	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Petroleum Ether	10%	100.0	103.3	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Benzene	30%	100.0	89.1	%	50 mM phosphate buffer	6	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Classical aqueous control (in %)
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	65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	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	65°C	Naringin	L-Rhamnose, Prunin	—	—	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	65°C	Naringin	L-Rhamnose, Prunin	—	—	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	65°C	Naringin	L-Rhamnose, Prunin	—	—	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	65°C	5 g/L Naringin	L-Rhamnose, Prunin	—	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	65°C	5 g/L Naringin	L-Rhamnose, Prunin	—	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	65°C	5 g/L Naringin	L-Rhamnose, Prunin	—	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	65°C	5 g/L Naringin	L-Rhamnose, Prunin	—	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	65°C	5 g/L Naringin	L-Rhamnose, Prunin	—	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	65°C	5 g/L Naringin	L-Rhamnose, Prunin	—	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	65°C	5 g/L Naringin	L-Rhamnose, Prunin	—	200 rpm	Classical aqueous control (in %)
Stability - Incubation	Activity after incubation (3 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (3 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (4 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (4 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (4 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (3 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (2 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (2 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (2 hours at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (1 hour at 65°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: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (1 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	20%	100.0	44.9	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (1 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	10%	100.0	89.8	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (0.5 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	30%	100.0	60.6	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (0.5 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	20%	100.0	98.7	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase
Stability - Incubation	Activity after incubation (0.5 hour at 65°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	10%	100.0	102.6	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6, Assay: 6	Incubation: 65°C, Assay: 65°C	1 mM Naringin	L-Rhamnose, Prunin	—	—	Non-incubated control (in %), assay in aqueous phase

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Visualization : Activity — Classical

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

Visualization : Activity — Michaelis-Menten

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

Visualization : Stability — Incubation

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

Visualization : Activity + Stability — Conversion

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

Structure

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

Enzyme Description

Sequence

Experimental Data (46 measurements)

Visualization : Activity — Classical

Visualization : Activity — Michaelis-Menten

Visualization : Stability — Incubation

Visualization : Activity + Stability — Conversion

Structure

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