Purification and characterisation of Aspergillus sojae naringinase: The production of prunin exhibiting markedly enhanced solubility with in vitro inhibition of HMG-CoA reductase

Chang, Heon-Young; Lee, Yoon Bok; Bae, Hyun Ah; Huh, Ji Young; Nam, So Hyun; Sohn, Hosik; Lee, Hyong Joo; Lee, Soo Bok

doi:10.1016/j.foodchem.2010.06.024

Cited by 43 publications

(57 citation statements)

References 28 publications

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“…The purified enzyme migrated as a single band on the 12% SDS-PAGE gel (12) with an apparent molecular mass of about 66 kDa (Fig. 1A), which was similar to those for the previously reported ␣-L-rhamnosidase from Curvularia lunata (6) and naringinase from Aspergillus sojae (5). The 3-step-purified enzyme gave one major peak by high-performance liquid chromatography (HPLC) analysis (Tosoh TSKgel G2000SW; ⌽, 7.8 mm by 300 mm), which indicates that it was almost pure protein.…”

supporting

confidence: 87%

“…␣ -L-Rhamnosidase (EC 3.2.1.40) cleaves terminal ␣-L-rhamnose from a large number of natural products, such as flavonoids, saponins, and many other natural glycosides, and this enzyme has been used in the food industry for eliminating the naringin bitterness of citrus juices (4,20,25,26), removing hesperidin crystals from orange juices and releasing 7-O-␤-D-glucoside-hesperetin, an important precursor in sweetener production (17,24), and enhancing wine aromas (7,6,22). In addition, the deglycosylation of flavonoids in vitro by ␣-L-rhamnosidase has ushered in a new era of drug development (5,8,19,27). Hitherto, ␣-L-rhamnosidases from fungi, bacteria, and animals have been purified and characterized (29), and several genes encoding ␣-Lrhamnosidases have also been cloned (1, 2, 3, 9, 16).…”

mentioning

confidence: 99%

“…In addition, the deglycosylation of flavonoids in vitro by ␣-L-rhamnosidase has ushered in a new era of drug development (5,8,19,27). Hitherto, ␣-L-rhamnosidases from fungi, bacteria, and animals have been purified and characterized (29), and several genes encoding ␣-Lrhamnosidases have also been cloned (1,2,3,9,16).…”

mentioning

confidence: 99%

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Aspergillus niger DLFCC-90 Rhamnoside Hydrolase, a New Type of Flavonoid Glycoside Hydrolase

Liu

Zhang

et al. 2012

Appl Environ Microbiol

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A novel rutin-␣-L-rhamnosidase hydrolyzing ␣-L-rhamnoside of rutin, naringin, and hesperidin was purified and characterized from Aspergillus niger DLFCC-90, and the gene encoding this enzyme, which is highly homologous to the ␣-amylase gene, was cloned and expressed in Pichia pastoris GS115. The novel enzyme was classified in glycoside-hydrolase (GH) family 13. ␣ -L-Rhamnosidase (EC 3.2.1.40) cleaves terminal ␣-L-rhamnose from a large number of natural products, such as flavonoids, saponins, and many other natural glycosides, and this enzyme has been used in the food industry for eliminating the naringin bitterness of citrus juices (4,20,25,26), removing hesperidin crystals from orange juices and releasing 7-O-␤-D-glucoside-hesperetin, an important precursor in sweetener production (17,24), and enhancing wine aromas (7,6,22). In addition, the deglycosylation of flavonoids in vitro by ␣-L-rhamnosidase has ushered in a new era of drug development (5,8,19,27). Hitherto, ␣-L-rhamnosidases from fungi, bacteria, and animals have been purified and characterized (29), and several genes encoding ␣-Lrhamnosidases have also been cloned (1, 2, 3, 9, 16). Thus far, the ␣-L-rhamnosidases are categorized into four glycoside-hydrolase (GH) families, 28, 78, 106, and NC (nonclassified), in the CAZy (carbohydrate-active enzymes) database based on amino acid sequence similarities (10, 11).In this paper, a novel rutin-␣-L-rhamnosidase was purified and characterized from Aspergillus niger DLFCC-90, and the gene (rhaR) encoding this enzyme was also cloned and expressed in Pichia pastoris GS115.The strain A. niger DLFCC-90, obtained from the Culture Collection of Biotechnology Engineering of Dalian Polytechnic University (Dalian, China), was cultured with shaking at 28 to 30°C for 72 to 96 h in a wort medium of 5.0 Baume degrees containing 2% extract from flowers of Sophora japonica (Huai Hua in Chinese). To obtain pure enzyme, the cell-free culture was treated by a 3-step method, i.e., ammonium sulfate precipitation (75% saturation), a Sephadex G-75 column (Amersham Pharmacia) eluted with 0.02 M acetate buffer (pH 5.0), and a DEAE 52-cellulose column (Amersham Pharmacia) eluted with a linear gradient of KCl (0.0 to 0.5 M) in 0.02 M acetate buffer (pH 5.0). The determination of the protein concentration was performed as described in reference 13. The enzymatic activity was measured using 2.0 mg/ml rutin (Sigma) in 0.02 M acetate buffer (pH 5.0) as a substrate after a reaction at 50°C for 18 h, and products of rhamnose and isoquercitrin from the enzyme reaction were detected to calculate the enzyme activity (18, 28). After the above-described 3 steps of purification, the enzyme's specific activity was increased 5.3-fold. The purified enzyme migrated as a single band on the 12% SDS-PAGE gel (12) with an apparent molecular mass of about 66 kDa (Fig. 1A), which was similar to those for the previously reported ␣-L-rhamnosidase from Curvularia lunata (6) and naringinase from Aspergillus sojae (5). The 3-step-purified enzyme gave one major p...

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supporting

confidence: 87%

mentioning

confidence: 99%

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Aspergillus niger DLFCC-90 Rhamnoside Hydrolase, a New Type of Flavonoid Glycoside Hydrolase

Liu

Zhang

et al. 2012

Appl Environ Microbiol

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“…Recently, it has been shown that the rutinoside, O-a-L-rhamnosyl-(1 ? 2)-glucose of naringin is hydrolysed by the naringinase produced by an Aspergillus sojae strain isolated from a traditional Korean fermented soybean, to preferably yield a rhamnose and naringenin-7-O-glucoside, prunin (Chang et al, 2011). Prunin is considered to have a potential use as a sweetening agent in diabetes therapy without concurrent undesirable bitterness, and it possesses many biological activities, such as antiviral, anti-inflammatory, hypoglycemic, and hypocholesterolemic effects (Choi, Yokozawa, & Oura, 1991;Jeon, Park, & Choi, 2004;Kaul, Middleton, & Ogra, 1985).…”

Section: Introductionmentioning

confidence: 99%

Enzymatic bioconversion of citrus hesperidin by Aspergillus sojae naringinase: Enhanced solubility of hesperetin-7-O-glucoside with in vitro inhibition of human intestinal maltase, HMG-CoA reductase, and growth of Helicobacter pylori

et al. 2012

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“…FIC 1 , FIC 7 and FIC 8 were isolated from the buried citrus fruits and peels. Strains FIC 1 , FIC 6 and FIC 8 were isolated from the soil that was collected around the buried citrus fruits.…”

Section: Isolation Of Strainmentioning

confidence: 99%

Characterization of best naringinase producing fungus isolated from the citrus fruits

Srikantha

Ranganathan²,

Seevaratnam³

2016

IJBR

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Naringinase enzyme has potential application in food and pharmaceutical industry. Naringin and limonin are principle bitter components in the citrus fruit. The microorganisms that associate with citrus fruit may have the ability to degrade the naringin by extracellular naringinase enzymes that are produced by microorganisms. The objective of the study is to isolate naringinase producing fungus from the citrus fruit to debitter the citrus juice and to characeterize the fungus. Citrus fruits were allowed to spoil under the air and soil and the lesion was used to streak on fresh PDA plates. Out of the eight strains isolated from citrus fruits, five were positive for naringinase enzyme. When all the naringinase producing fungi were subjected to liquid fermentation medium for eight days at room temperature at 200 rpm and the crude enzyme was tested for naringinase enzyme at pH 5 and 50 ºC for 10 minutes, one strain showed the best naringenase activity (1.92 µmol/ml/min). This strain was identified as Aspergillus flavus based on the macroscopic, microscopic and biochemical tests. The culture conditions were optimized to increase the naringinase production via solid state fermentation system using paddy husk as the support. Though naringinase activity of Aspergillus flavus has started on the 2nd day, the highest activity (449.58Ug -1 Dry Matter) was obtained on the 8th day. Thereafter the naringinase activity has started to decline. Solid state fermentation using paddy husk as support could be used for large scale naringinase enzyme production.

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Purification and characterisation of Aspergillus sojae naringinase: The production of prunin exhibiting markedly enhanced solubility with in vitro inhibition of HMG-CoA reductase

Cited by 43 publications

References 28 publications

Aspergillus niger DLFCC-90 Rhamnoside Hydrolase, a New Type of Flavonoid Glycoside Hydrolase

Aspergillus niger DLFCC-90 Rhamnoside Hydrolase, a New Type of Flavonoid Glycoside Hydrolase

Enzymatic bioconversion of citrus hesperidin by Aspergillus sojae naringinase: Enhanced solubility of hesperetin-7-O-glucoside with in vitro inhibition of human intestinal maltase, HMG-CoA reductase, and growth of Helicobacter pylori

Characterization of best naringinase producing fungus isolated from the citrus fruits

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