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首页> 外文期刊>Applied Microbiology >Genomic Sequence of Bacteriophage ATCC 8074-B1 and Activity of Its Endolysin and Engineered Variants against Clostridium sporogenes
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Genomic Sequence of Bacteriophage ATCC 8074-B1 and Activity of Its Endolysin and Engineered Variants against Clostridium sporogenes

机译：噬菌体ATCC 8074-B1的基因组序列及其内溶素活性和工程化变体对产孢梭状芽孢杆菌的影响

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Lytic bacteriophage ATCC 8074-B1 produces large plaques on its host Clostridium sporogenes . Sequencing of the 47,595-bp genome allowed the identification of 82 putative open reading frames, including those encoding proteins for head and tail morphogenesis and lysis. However, sequences commonly associated with lysogeny were absent. ORF 22 encodes an endolysin, CS74L, that shows homology to N -acetylmuramoyl-l-alanine amidases, and when expressed in Escherichia coli , the protein causes effective lysis of C. sporogenes cells when added externally. CS74L was also active on Clostridium tyrobutyricum and Clostridium acetobutylicum . The catalytic domain expressed alone (CS74L_(1–177)) exhibited a similar activity and the same host range as the full-length endolysin. A chimeric endolysin consisting of the CS74L catalytic domain fused to the C-terminal domain of endolysin CD27L, derived from Clostridium difficile bacteriophage ΦCD27, was produced. This chimera (CSCD) lysed C. sporogenes cells with an activity equivalent to that of the catalytic domain alone. In contrast, the CD27L C-terminal domain reduced the efficacy of the CS74L catalytic domain when tested against C. tyrobutyricum . The addition of the CD27L C-terminal domain did not enable the lysin to target C. difficile or other CD27L-sensitive bacteria.
机译：溶菌性噬菌体ATCC 8074-B1在其宿主梭状芽孢杆菌中产生大噬斑。对47,595-bp基因组进行测序可鉴定出82个假定的开放阅读框，包括编码头和尾形态发生和裂解的蛋白质的那些。但是，通常没有与溶源性相关的序列。 ORF 22编码一种内溶素CS74L，与N-乙酰基村酰-1-丙氨酸酰胺酶具有同源性，当在大肠杆菌中表达时，该蛋白质在外部添加时可有效裂解孢子囊梭菌细胞。 CS74L对酪丁酸梭菌和丙酮丁醇梭菌也有活性。单独表达的催化结构域（CS74L_（1-177））的活性与全长溶血素的相似，且具有相同的宿主范围。产生了由CS74L催化结构域组成的嵌合内溶素，所述CS74L催化结构域与难溶梭状芽胞杆菌ΦCD27衍生的内溶素CD27L的C-末端结构域融合。此嵌合体（CSCD）裂解了C. sporogenes细胞，其活性与单独的催化结构域相当。相反，当针对酪丁酸梭菌进行测试时，CD27L C末端结构域降低了CS74L催化结构域的功效。 CD27L C末端域的添加不能使溶素靶向艰难梭菌或其他CD27L敏感细菌。

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