CAJ | 학술논문

以坛紫菜丝状体为材料,采用RACE方法获得坛紫菜碳酸酐酶(CA)基因的全长cDNA。该cDNA全长1081 bp,具有一个825 bp的开放阅读框,可编码274个氨基酸。序列同源性分析显示该cDNA序列推导的氨基酸序列与其他物种的碳酸酐酶具有较高的一致性,其中与条斑紫菜的一致性达到96%。氨基酸序列分析表明该蛋白为β-CA,含有两个CA活性位点,无跨膜结构,可能存在一个信号肽将其定位到叶绿体中,与藻类和细菌聚类。原核诱导表达得到一个72 kDa左右的融合蛋白,酶活测定结果显示此蛋白具有碳酸酐酶活性。该实验对进一步深入研究坛紫菜 CA 的功能及坛紫菜碳代谢、光合作用等生理过程具有重要的参考价值。
이단자채사상체위재료,채용RACE방법획득단자채탄산항매(CA)기인적전장cDNA。해cDNA전장1081 bp,구유일개825 bp적개방열독광,가편마274개안기산。서렬동원성분석현시해cDNA서렬추도적안기산서렬여기타물충적탄산항매구유교고적일치성,기중여조반자채적일치성체도96%。안기산서렬분석표명해단백위β-CA,함유량개CA활성위점,무과막결구,가능존재일개신호태장기정위도협록체중,여조류화세균취류。원핵유도표체득도일개72 kDa좌우적융합단백,매활측정결과현시차단백구유탄산항매활성。해실험대진일보심입연구단자채 CA 적공능급단자채탄대사、광합작용등생리과정구유중요적삼고개치。
Carbonic anhydrase (CA), a zinc-containing enzyme is widespread in living organisms, catalyses the reversible hydration of CO2 and 3HCO-. In this study, a full-length cDNA of CA was isolated from Porphyra haitanensis with rapid amplification of cDNA ends (RACE). This sequence was 1 081 bp in length and encodes a polypeptide of 274 amino acid residues. The deduced polypeptide showed high identities with the CA genes ranging from unicellular algae and bacteria to green plant. Phylogenetic tree analysis showed that the CA gene from P. haitanensis was more closely assembled with algae and bacteria. A ~72 kDa fused protein was obtained by the recombinant prokaryotic expression and the enzyme activity analysis showed that it had the activity of CA.