Purification and Characterization of RNase from Momordicacharantia L. - Details

author：

Lin, D. (Lin, D..) ^[1] | Wu, W. (Wu, W..) ^[2] | Ke, L. (Ke, L..) ^[3] | Zhou, J. (Zhou, J..) ^[4] | Rao, P. (Rao, P..) ^[5]

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Abstract：

Objective:　To　isolate　and　purify　a　ribonuclease　from　Momordicacharantia　L.　and　study　the　enzymatic　and　biological　characterizations　of　the　ribonuclease.　Methods:　Ethanol　precipitation,　C18　reversed　phase　high　performance　liquid　chromatography　(RP-HPLC),　Tricine-SDS-polyacrylamide　gel　electrophoresis　and　isoelectric　focusing　were　used　for　isolation,　purication　and　identification　of　the　ribonuclease.　The　enzyme　activity　of　ribonuclease　was　detected　using　tRNA　from　yeast　as　substrate.　Results:　A　ribonuclease,　designated　MC　-RNase,　showed　a　high　purity　by　Tricine-SDS-PAGE　and　RP-HPLC.　Its　TV-terminal　sequence　was　determined　as　VNEFDYYQVVLQWQPAT.　The　molecular　weight　and　isoelectric　point,　was　estimated　to　be　14　ku　and　8.1.　Enzymic　experiment　showed　the　optimum　temperature　is　50℃,　and　optimum　pH　is　5,　Pb2+　can　increase　the　enzyme　activity,　while　Zn2+,　Fe2+,　Fe3+　and　Mg2+　inhibited　the　enzyme　activity　of　MC-RNase.　The　result　of　cytotoxicity　test　showed　MC-RNasehad　no　effect　on　the　proliferation　of　L-02　cells　and　Hep-G2　cells　in　vitro.　Conclusion:　Isolation　and　identification　of　a　novel　ribonuclease　from　fresh　Momordicacharantia　L.　©　2017,　Editorial　Office　of　Journal　of　CIFST.　All　right　reserved.

Keyword：

Bitter gourd; Characterization; Purification; Ribonuclease

Community：

[ 1 ] [Lin, D.]College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, 310018, China
[ 2 ] [Wu, W.]Institute of Biotechnology, Fuzhou University, Fuzhou, 350002, China
[ 3 ] [Ke, L.]College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, 310018, China
[ 4 ] [Zhou, J.]College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, 310018, China
[ 5 ] [Rao, P.]College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, 310018, China

Reprint 's Address：

[Zhou, J.]College of Food Science and Biotechnology, Zhejiang Gongshang UniversityChina

Email：

jianwuzhou@zjgsu.edu.cn

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Source ：

Journal of Chinese Institute of Food Science and Technology

ISSN： 1009-7848

Year： 2017

Issue： 1

Volume： 17

Page： 148-153

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