CAJ | 학술논문

NCIMB 10467是一株木质素降解菌,根据其16S rDNA序列将其重新分类为Burkholderia菌属.研究显示,在NCIMB 10467菌株中,不同的底物可以诱导该菌株对于原儿茶酸的多种代谢形式.根据克隆到的一段原儿茶酸邻位开环酶,即原儿茶酸3,4-双加氧酶(P34D;EC 1.13.11.3)α-亚基的保守序列,通过染色体步移的方法,得到一段9505bp的DNA片段.序列分析显示,在这段9.5 kb的DNA片段中,两个可能的开放阅读框pcaG和pcaH分别编码P34D的α-亚基和β-亚基.将pcaGH克隆并在大肠杆菌中进行表达后,可以检测到P34D的活性.而pcaH在NCIMB 10467菌株中的敲除则使该菌完全丧失了代谢原儿茶酸的能力.由此证实,克隆到的pcaGH基因确实编码原儿茶酸3,4-双加氧酶,并且对于NCIMB 10467菌株对原儿茶酸的代谢是必需的.
NCIMB 10467시일주목질소강해균,근거기16S rDNA서렬장기중신분류위Burkholderia균속.연구현시,재NCIMB 10467균주중,불동적저물가이유도해균주대우원인다산적다충대사형식.근거극륭도적일단원인다산린위개배매,즉원인다산3,4-쌍가양매(P34D;EC 1.13.11.3)α-아기적보수서렬,통과염색체보이적방법,득도일단9505bp적DNA편단.서렬분석현시,재저단9.5 kb적DNA편단중,량개가능적개방열독광pcaG화pcaH분별편마P34D적α-아기화β-아기.장pcaGH극륭병재대장간균중진행표체후,가이검측도P34D적활성.이pcaH재NCIMB 10467균주중적고제칙사해균완전상실료대사원인다산적능력.유차증실,극륭도적pcaGH기인학실편마원인다산3,4-쌍가양매,병차대우NCIMB 10467균주대원인다산적대사시필수적.
Strain NCIMB 10467, a lignin degrader, was reclassified as genus Burkholderia according to its 16S rDNA sequence. It seems that the metabolism of protocatechuate by this strain is diverse under the induction of various substrates. A 9505-bp DNA fragment extending from α conserved region of the gene, which encodes β subunit of orthocleavage protocatechuate 3, 4-dioxygenase(P34D; EC 1.13.11.3),was obtained by genome walking. Sequence analysis revealed two deduced open reading frames, pcaG and pcaH, encoding the α and β subunits of P34D respectively in this fragment. The P34D activity could be detected when pcaGH were expressed in E. Coli and the disruption of pcaH in strain NCIMB 10467 has lead to loss of its ability to catabolize protocatechuate. It was proved that the cloned pcaGH were encoding a functional protocatechuate 3, 4-dioxygenase which was necessary for the protocatechuate metabolism in this strain.