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- 저자 : Changxia Liu (검색결과 2 건)
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Preparation and enzymatic activity of Fe3O4-IDA-Ni/NAD kinase magnetic catalyst
Changxia Liu,Yadi Yang,Huafeng Gao,Xiaoshuang Bai,Zheng-Jun Li 한국화학공학회 2020 Korean Journal of Chemical Engineering Vol.37 No.3
The use of oxidoreductases as biocatalysts for industrial production of valuable compounds has a strong demand for NADP. Herein, we prepared superparamagnetic NAD kinase catalyst to synthesize NADP in vitro. First, Fe3O4 particles were synthesized through a solvothermal method, followed by the chemical modification with epichlorohydrin, iminodiacetic acid, and Ni2+ to yield functional Fe3O4 sub-microspheres. Subsequently, NAD kinase of Escherichia coli was overexpressed and immobilized on to the surface of magnetic sub-microspheres. The immobilized NAD kinase was used to catalyze the conversion of NAD to NADP in a cell-free system. Under optimal condition, the conversion ratio of NAD reached 91.7% and remained at 86.3% after repeated use for five times. Our study revealed that the novel magnetic NAD kinase catalyst possessed favorable properties for magnetic manipulation and NADP production.
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Du Changxia,Chai Li’ang,Liu Chen,Yuyang Si,Fan Huaifu 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.1
Cucumber (Cucumis sativus) is one of the most important vegetable crop species in the world. As conventional breeding of cucumber is very challenging, genetic engineering is an alternative option for introducing important traits such as enhanced stress resistance and nutritional value. However, the efficiency of the transformation system depends on genotypes, transfor- mation conditions, etc. This study aims to optimize the transformation parameters of Agrobacterium-mediated transformation of cucumber. ‘Xintaimici’, which is a very popular and typical northern Chinese cucumber variety, was transformed with Agrobacterium GV3101. The strain carried the pCAMBIA2300s plasmid, which is a binary vector containing the marker gene neomycin phosphotransferase II (npt II). The results indicated that cefotaxime sodium was suitable for inhibiting Agrobacterium in the screening and bud elongation stages. Timentin was best used during the rooting stage. Furthermore, 25 mg/L kanamycin was used in the early stage of screening and increased to 50 mg/L for further screening. At the bud elongation and rooting stages, 75 and 100 mg/L kanamycin was used, respectively, to improve the screening efficiency. To obtain the highest regeneration frequency of resistant buds, 50, 150, and 100 μM acetosyringone was added to the preculture medium, infection solution, and coculture medium, respectively. To confirm the presence of the transgenes, DNA from npt II transformed cucumber plants was analyzed by polymerase chain reaction after transplanting resistant regenerated plants. We ultimately achieved an 8.1% transformation efficiency, which is among the highest values reported to date using the cucumber variety ‘Xintaimici’. Thus, an effective protocol for Agrobacterium tumefaciens-mediated genetic transformation of cucumber was optimized.
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