Bifidobacterium longum의 Sucrose 대사관련scr 유전자군의 특성 규명
Characterization of the scr Gene Cluster InvolvedIn Sucrose Utilization in Bifidobacterium longum.
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Bifidobacterium longum SJ32 균주로부터 cloning한 sucrose phosphorylase 유전자를 포함하는 8.6 kb의 EcoRI 단편의 염기서열을 결정하였다. 5개의 open reading frame이 존재하였고, 상동성 검색의 결과, sucrose 대사에 관여하는 sucrose phosphorylase (ScrP), sucrose transporter (ScrT), GalR-LacI-type transcriptional regulator (ScrR) 유전자의 존재를 확인하였다. SJ32 균주의 scrPTR 유전자군은 다른 B. longum 균주의 scrPTR과 배열이 동일하고, 각 유전자 산물이 아미노산 수준에서 94% 이상의 상동성을 가지고 있지만, 주변 유전자는 다르게 나타나 B. longum 균주 간의 scrPTR 유전자군의 horizontal transfer를 추정하게 한다. 대장균에서의 scrP와 scrT의 동시 발현은 세포 내로의 sucrose 유입을 증가시켜 sucrose phosphorylase의 활성 증가에 영향을 주는 것으로 나타나, scrT가 sucrose transporter 유전자임을 뒷받침한다. 기존에 보고된 B. longum NCC2705 균주의 유전체로부터 scrPTR 외에도 sucrose 대사에 관여하는 다양한 sucrose multiple transport system에 관여하는 유전자의 존재가 확인되어, B. longum이 다양한 sucrose 유입체계를 보유하고 있음이 추정된다.
Abstract
The nucleotide sequence of 8.6-kb EcoRI fragment containing sucrose phosphorylase gene isolated from Bifidobacterium longum SJ32 was determined. It was found that the fragment contained five open reading frames including the gene cluster for sucrose utilization such as a sucrose phosphorylase (ScrP), a sucrose transporter (ScrT), and a GalR-LacI-type transcriptional regulator (ScrR) identified by amino acid homology. Each gene showed over 94% amino acid homology among various B. longum strains. Genomic organization of the gene cluster is the same as those of other strains of B. longum but different from that of B. lactis. In spite of high homology of each gene among B. longum strains, the difference of flanking sequences of the gene cluster between strains SJ32 and NCC2705 insinuates the horizontal transfer of scrPTR between B. longum strains. The increase of sucrose phosphorylase activity in heterologous E. coli system by the co-expression of scrT with scrP against the single expression of scrP was measured. It seems to be the result of sucrose uptake increment by scrT in the host and is an indirect evidence that scrT is the gene for sucrose transport. The existence of multiple sucrose uptake systems in B. longum is supposed from the findings of several genes besides scrPTR involved in sucrose uptake in the genome of B. longum NCC2705.
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