柠条幼苗苯丙氨酸解氨酶的纯化
Pur if ica tion of Phenyla lan ine Ammon ia-lya se in Ca ragana korsh inskii Juven ile
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摘要: 用离子交换层析和分子筛层析分离纯化了真叶期柠条根、速生期柠条叶、茎中的苯丙氨酸解氨酶( PAL) ,并测定了PAL全酶的分子量。用SDS-PAGE鉴定了全酶的组成,测定了各亚基的分子量。结果表明:柠条叶中存在306KD和222KD两种PAL,前者是由82KD和74KD亚基组成的异聚体,后者是由74KD亚基组成的同聚体;而柠条根和茎中仅存在74KD亚基组成的222KD全酶。该研究为pal基因的克隆及应用奠定了基础。Abstract: Phenylalanine ammonia-lyase ( PAL; EC 4. 3. 1. 5) of root in leaf stage, leaf and branch in rap id growthstage of Ca ragana korshinskii,was purified by ion exchange chromatography and gel filtration chromatography. Thesubunits of PAL by SDS2PAGE were analyzed. The results showed that when purified by ion exchangechromatography and gel filtration chromatography, there were two kinds of PAL , oneMW was 306KD and the otherwas 222KD, from leaf and one PAL whichMW was 222 KD from stem and root. By SDS-PAGE, it could concludethat the PAL, MW 306KD, was composed of two polypep tides which MW might be 82KD and two polypep tideswhich MW might 74 KD, and the 222KD-MW PAL was composed of four homology subunits, which MWwas 74KD.
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Key words:
- Caragana korsh insk ii
- / phenylalanine ammonia-lyase
- / purify
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