摘要
根据Gen Bank中的枯草芽孢杆菌(Bacillus subtilis)168菌株假定的赖氨酸脱羧酶基因(LDC)序列设计同源引物,提取枯草芽孢杆菌BJ3-2基因组DNA进行PCR扩增,并克隆至p GEM-T载体后测序。序列分析结果表明:Bacillus subtilis BJ3-2 yaa O基因开放阅读框(ORF)长为1 443 bp,获得基因登录号为KJ561349,BLAST比对与已报道枯草芽孢杆菌的yaa O基因核苷酸序列及氨基酸序列同源性均高达90%以上,对赖氨酸脱羧酶氨基酸序列构建系统进化树分析,发现Bacillus subtilis BJ3-2的LDC与B.subtilis JS亲缘关系最近,SWISS-MODEL预测该蛋白的3D结构与鸟氨酸/赖氨酸/精氨酸脱羧酶蛋白相似性为39.22%,属于典型的III型磷酸吡哆醛(PLP)依赖型鸟氨酸/赖氨酸/精氨酸脱羧酶家族成员。yaa O基因序列分析及氨基酸保守结构域的分析,为有效控制水豆豉产品中尸胺含量过高的研究提供了理论基础。
According to the Bacillus subtilis 168 putative lysine decarboxylase gene sequence registered in GenBank,analogy primers were designed using software Primer Premier.The yaaO was amplified by PCR from genomic DNA of the B.subtilis B J3-2,and cloned to the pGEM-T vector for sequencing.Sequence analysis showed that the open reading frame length of B.subtilis B J3-2 yaaO gene was 1443 bp,which achieved the GenBank accession number KJ561349.After blasting with NCBI Database,the results revealed that the identities between yaaO gene nucleotide sequence of B.subtilis B J3-2 and the other similar nucleotide and amino acid sequences were beyond 90%.Phylogenetic tree indicated that B.subtilis B J3-2 LDC was the nearest relative to B.subtilis JS.The similarity of three-dimensional structure between lysine decarboxylase predicted by SWISS-MODEL and omithine/lysine/arginine decarboxylase was 39.22%,which belonged to the classical type Ⅲ PLP-dependent ornithine decarboxy-lase/lysine decarboxylase/arginine decarboxylase family.The gene sequence and conserved structure domain analysis of yaaO would bring some theoretical basis for further exploration of controlling the cadaverine content of the fermented soybeans.
出处
《中国酿造》
CAS
2014年第12期116-120,共5页
China Brewing
基金
国家自然科学基金项目(31260394/C2007)
