A thermostable Alpha-L-Rhamnosidase (Naringinase, RhamA) that catalyzes the cleavage of the bond between terminal L(+)-rhamnose and the aglycone of rhamnose-containing glycosides. The enzyme is very active on naringin but has also substantial activity with hesperidin as substrate.
Available as 100 units.
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Product : Rham142 | Naringinase 100 Units | Price : 280.00 EUR
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Enzyme activity: Rham142 Rhamnosidase A (naringinase, alpha-L-rhamnosidase) catalyzes the cleavage of the bond between terminal L(+)-rhamnose and the aglycone of rhamnose-containing glycosides. Hydrolysis of terminal non-reducing α-L-rhamnose residues in α-L-rhamnosides: naringin, hesperidin and rutin
Activity determination: Rhaminosidase activity was routinely determined in a 100mM KPO4 pH 7.5 buffer for 10 minutes with 2.0mM final conc. of p-nitro-phenyl-α-L-rhamnopyranoside.
Unit definition: One unit (U) of enzyme activity is the amount that leads to the release of 1 µmol of p-nitro-phenyl-α-L-rhamnopyranoside (pnpR) per minute
Synonyms: glycoside hydrolase; RhamA; naringinase; hesperidinase; α-L-rhamnosidase A; α-L-rhamnosidase N; α-L-rhamnoside rhamnohydrolase
Protein family: Glycosyl hydrolase family 78 ( GH78 ) – GH-H clan – CAZy database GH78 family
Enzyme classification: EC 22.214.171.124
Source: Bacteria; Thermomicrobia strain PRI-1686
Protein sequence: NBCI protein entry
Structural information: The crystal structure of alpha-L-rhamnosidase from Bacillus sp. GL1, sharing 52% sequence identity with ThermoactiveTM Rhamnosidase A, has been determined to a resolution of 1.9 Å ( Cui et al. 2007 ). –Protein Data Bank entry 2OKX
Substrates: Alpha-L-rhamnosidasaes catalyse the release of terminal rhamnose residues from polysaccharides and glycosides. Of the many natural compounds that contain terminal alpha-L-rhamnose, the flavonoids naringin, hesperidin, rutin and quercitrin have been the main natural test-substrates for alpha-L-rhamnosidases. Of these compounds, ThermoactiveTM Rhamnosidase A was found to be most active on Naringin as shown in Figure 1 (Birgisson et al 2004). The structure of naringin (4′,5,7-trihydroxyflavanone-7-α-L-rhamnopyranoside-(1,2)-β-D-glucopyranoside) and the hydrolysis by rhamnosidase is shown in Figure 2.
Figure 1. Substrate specificity: hydrolysis of terminal non-reducing α-L-rhamnose residues in the flovonoids naringin, hesperdin, rutin and quercitrin
Figure 2. Structure of naringin, with rhamnose 1,2 linked to a glucose residue linked to the flavonoid skeleton, and the hydrolysis of the terminal α-L-rhamnose residues by rhamnosidase
Applications: Naringin is a source of bitter flavor in fruit juice and rhamnosidases with naringinase activity are frequently used for debittering citrus juice. Other biotechnological applications include manufacture of prunin; manufacture of alpha-L-rhamnosidese fom natural glycosides; clarification of juices; enhancement of wine aromas by hydrolysis of terpenyl glycosides; conversion of chloropolysporin B to chloropolysporin C and production of pharmaceutically important compounds by removal of rhamnose residues from steriods such as diosgene, desglucoruscin and ginsenosides-Rg2 (Yadav et al. 2010). Beta-glucosidases may be used in combination with alpha-L-rhamnosidases for removal of glucose from the flavonoid skeleton. ThermoactiveTM Rhamnosidase A has been successfully demonstrated for use in production of rhamnose from narigin in a bioreactor containing immobilized E. coli cells expressing the gene for the enzyme (Birgisson et al 2007). L-Rhamnose or its derivatives are suitable chiral structural component and can be used for the synthesis of pharmaceutical products, plant protection agents and the preparation of fragrances in the foodstuffs and perfume industries.
Properties of ThermoActive™ Rhamnosidase A:
Figure 3. Temperature spetrum: The enzyme in relatively active in a rather broad temperature range (45-75°C)with optimum around 65°C
Figure 4. pH spectrum: pH range is about 4.5-9 with optimum about pH 7.5
Birgisson H, Hreggvidsson GO, Fridjónsson OH, Mort A, Kristjánsson JK, Mattiasson B (2004) Two new thermostable alpha-L-rhamnosidases from a novel thermophilic bacterium. Enzyme Microb. Technol. 34: 561-571
Birgisson H, Wheat JO, Hreggvidsson GO, Kristjánsson JK, Mattiasson B. (2007) Immobilization of a recombinant Escherichia coli producing a thermostable α-l-rhamnosidase: Creation of a bioreactor for hydrolyses of naringin. Enzyme and Microbial Technology 40:1181-1187
Cui Z, Maruyama Y, Mikami B, Hashimoto W and Murata K (2007) Crystal Structure of Glycoside Hydrolase Family 78 α-L-Rhamnosidase from Bacillus sp. GL. J Mol Biol 374: 384–398
Yadav V, Yadav PK, Yadav S & Yadav KDS (2010) α-L-Rhamnosidase: A review. Process Biochem 45:1226–1235