Welcome to Acta Agronomica Sinica，

Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (10): 1569-1574.doi: 10.3724/SP.J.1006.2016.01569

• RESEARCH NOTES • Previous Articles    

Cloning, Expression, and Enzymatic Characteristics of Betaine Aldehyde De-hydrogenase Gene in Sesuvium portulacastrum L.

YU Shi-Zhou1,4,YANG Cheng-Long2,*,GUO Jian-Chun3,DUAN Rui-Jun3   

  1. 1 Guizhou Rape Institute, Guiyang 550008, China; 2 Guizhou Institute of Subtropical Crops, Xingyi 562400, China; 3 Institute of Tropical Bioscience and Biotechnology / Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture / Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China; 4 Guizhou Hemufu Seed Co. Ltd, Guiyang 550008, China
  • Received:2016-02-19 Revised:2016-07-11 Online:2016-10-12 Published:2016-07-28
  • Contact: 杨成龙,E-mail:yangchenglong208@163.com,Tel:0859-3911295 E-mail:yushizhou2008@163.com
  • Supported by:

    ThisstudywassupportedbyNationalAgriculturalScienceandTechnologyAchievementsTransformationFundProject(2014GB2F200240),theSpecialProjectofIndependentInnovationProject[(2014)014],theProjectofGuizhouProvinceforServiceEnterprisebyScientificResearchInstitution[(2015)4001],theKeySpecialProjectforGuizhouScienceandCooperation[(2013)6005],theSpecialProjectofGuizhouAcademyofAgriculturalSciences[(2015)16],GuizhouTop100TalentsHierarchyProjectofScienceandTechnologyDepartmentofGuizhouProvince[(2015)4018],andtheTalentTeamProjectofGuizhouAcademyofAgriculturalSciences[CR(2014)64].

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Abstract:

Among many osmotic materials, glycine betaine is a best organic micro-molecular, and functionally works for osmotic regulation in plants, which is non-toxic to plant growth. A lot of glycine betaine accumulated in plant can enhance the resistance of plants to environmental stresses. In the study, a full-length sequence of betaine aldehyde dehydrogenase gene from Sesuvium portulacastrum was ligated with the vector pET-[28a](+), named pET-SpBADH, and successfully transformed into BL21(DE3) to obtain the corresponding recombinant engineering bacteria, which could highly express 55 kD protein induced by IPTG, with the expression level to 301 μg mL–1. The purified protein was obtained, showing the optimum pH value of 7.2, and maintain high catalytic activity the enzyme under slightly alkaline conditions. SpBADH protein very sensitive to high temperature effected the enzyme activity, with the optimum temperature to 37℃. The enzyme activity was only 20% when temperature was over 55℃. The small organic molecules of the reveral compounds of alcohol had a protective effect on the catalytic activity of the enzyme. The microenvironment of catalytic activity could be maintained by its own characteristics.

Key words: SesuviumportulacastrumL., Betainealdehydedehydrogenase, Prokaryoticexpression, Enzymeactivityassay

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