[논문]Bioconversion of Cyanidin-3-Rutinoside to Cyanidin-3-Glucoside in Black Raspberry by Crude α-ʟ-Rhamnosidase from Aspergillus Species

Lim, Taehwan; Jung, Hana; Hwang, Keum Taek

doi:10.4014/jmb.1503.03098

Bioconversion of Cyanidin-3-Rutinoside to Cyanidin-3-Glucoside in Black Raspberry by Crude α-ʟ-Rhamnosidase from Aspergillus Species 원문보기

Journal of microbiology and biotechnology, v.25 no.11, 2015년, pp.1842 - 1848

Lim, Taehwan (Department of Food and Nutrition, and Research Institute of Human Ecology, Seoul National University) , Jung, Hana (Department of Food and Nutrition, and Research Institute of Human Ecology, Seoul National University) , Hwang, Keum Taek (Department of Food and Nutrition, and Research Institute of Human Ecology, Seoul National University)

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Cyanidin-3-glucoside (C3G) has been known to be more bioavailable than cyanidin-3-rutinoside (C3R), the most abundant anthocyanin in black raspberry (Rubus occidentalis). The aim of this study was to enhance the bioavailability of anthocyanins in black raspberry by cleaving ʟ-rhamnose in C3R using crude enzyme extracts (CEEs) from Aspergillus usamii KCTC 6956, A. awamori KCTC 60380, A. niger KCCM 11724, A. oryzae KCCM 12698, and A. kawachii KCCM 32819. The enzyme activities of the CEEs were determined by a spectrophotometric method using ρ-nitrophenyl-rhamnopyranoside and ρ-nitrophenyl-glucopyranoside. The CEE from A. usamii had the highest α-ʟ-rhamnosidase activity with 2.73 U/ml at 60℃, followed by those from A. awamori and A. niger. When bioconversion of C3R to C3G in black raspberry was analyzed by HPLC-DAD, the CEEs from A. usamii and A. awamori hydrolyzed 95.7% and 95.6% of C3R to C3G, respectively, after 2 h incubation. The CEEs from A. kawachii and A. oryzae did not convert C3R to C3G in black raspberry.

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The most effective CEE of each species was mixed with black raspberry juice (BRJ) to examine the bioconversion of C3R to C3G in the food matrix. To the best of our knowledge, this study is the first trial to compare the bioconversion of an anthocyanin in black raspberry using the five Aspergillus species.

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A. usamii KCTC 6956 was purchased from Korean Collection for Type Cultures (Daejeon, Korea). A.

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8. The protein concentration was estimated by the Bradford method [2] using bovine serum albumin as the standard.

성능/효과

1 and 2) and presented in Table 2. The anthocyanin fraction from black raspberry mainly consisted of C3XR, C3G, and C3R (Fig. 3).
3). The bioconversion of C3R to C3G in black raspberry resulted in a decreased C3R peak area and an increased C3G peak area (Fig. 3).
The effects of the temperature on α-ʟ-rhamnosidase and β-D-glucosidase activities of the CEEs obtained on the 7th day were measured at 30℃, 40℃, 50℃, and 60℃ (Fig. 2).
Concentrations of ʟ-rhamnose have also been found to affect α-ʟ-rhamnosidase activity [11,28]. The results of the previous studies imply that the quantity of carbon source remaining in the medium during incubation might importantly affect the enzyme secretion of microorganisms. Generally, Aspergillus species have a strong activity of β-D-glucosidase, which converts a glucoside form to an aglycone form, as well as α-ʟ-rhamnosidase activity [3].
kawachii and A. oryzae had no effects on the bioconversion of the anthocyanin in black raspberry.
niger were found to be effective α-ʟ-rhamnosidase sources as screened by a spectrophotometric method using pNPR, and A. usamii could be the most effective source for the bioconversion of C3R to C3G in black raspberry. The results of the present study suggest that the hydrolytic properties of the food-grade α-ʟ-rhamnosidase from A.
The CEE from A. usamii that had the highest α-ʟ-rhamnosidase activity for pNPR was also the most effective on the bioconversion in black raspberry, hydrolyzing 95.7% of C3R to C3G. The CEEs from A.

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Bioconversion of Cyanidin-3-Rutinoside to Cyanidin-3-Glucoside in Black Raspberry by Crude α-ʟ-Rhamnosidase from Aspergillus Species 원문보기

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Bioconversion of Cyanidin-3-Rutinoside to Cyanidin-3-Glucoside in Black Raspberry by Crude α-ʟ-Rhamnosidase from Aspergillus Species 원문보기

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