Evidence for Production of a New Lantibiotic (Butyrivibriocin OR79A) by the Ruminal Anaerobe Butyrivibrio fibrisolvensOR79: Characterization of the Structural Gene Encoding Butyrivibriocin OR79A
Evidence for Production of a New Lantibiotic (Butyrivibriocin OR79A) by the Ruminal Anaerobe Butyrivibrio fibrisolvensOR79: Characterization of the Structural Gene Encoding Butyrivibriocin OR79A
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通过瘤胃厌氧菌 Butyrivibrio fibrisolvensOR79 生产新羊毛硫抗生素（Butyrivibriocin OR79A）的证据：编码 Butyrivibriocin OR79A 的结构基因的表征
DOI：
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发表时间：
1999
影响因子：
4.4
通讯作者：
R. Teather
中科院分区：
文献类型：
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作者：
M. Kalmokoff;D. Lu;M. Whitford;R. Teather
摘要溶纤丁酸弧菌OR 79是一种具有广谱抗菌活性的瘤胃厌氧菌。通过硫酸铵和酸性沉淀、反相色谱和高分辨率凝胶过滤的组合从用过的培养液中分离抑制剂。对分离的抑制剂的N-末端分析得到15个氨基酸的序列（G-N/Q-G/P-V-I-L-X-I-X-H-E-X-S-M-N）。在N末端的第二和第三位置检测到两个不同的氨基酸残基，表明存在两种不同的肽。克隆与确定的N-末端序列的一种组合具有显著同源性的基因，并通过北方印迹证实该基因的表达。该基因（bvi 79 A）编码47个氨基酸的前肽和25个氨基酸的成熟肽，丁酸弧菌素OR 79 A。显着的序列同源性，发现这个肽和以前报道的羊毛硫抗生素含有双甘氨酸前导肽酶加工位点之间。bvi 79 A的紧下游是第二个部分开放的阅读框，其编码与被认为参与羊毛硫氨酸残基合成的蛋白质具有显著同源性的肽。这些发现表明，分离的抑制肽代表新的羊毛硫抗生素。从总的和N-末端氨基酸测序的结果表明，第二个肽是相同的丁酸弧菌素OR 79 A，除了在位置2和3的成熟lantibiotic的氨基酸取代。当用基于bvi 79 A 5 '区域的寡核苷酸或基于第二个肽的预测序列的简并寡核苷酸探测总DNA的限制性酶消化物时，仅检测到单个编码区。
ABSTRACT The ruminal anaerobe Butyrivibrio fibrisolvens OR79 produces a bacteriocin-like activity demonstrating a very broad spectrum of activity. An inhibitor was isolated from spent culture fluid by a combination of ammonium sulfate and acidic precipitations, reverse-phase chromatography, and high-resolution gel filtration. N-terminal analysis of the isolated inhibitor yielded a 15-amino-acid sequence (G-N/Q-G/P-V-I-L-X-I-X-H-E-X-S-M-N). Two different amino acid residues were detected in the second and third positions from the N terminus, indicating the presence of two distinct peptides. A gene with significant homology to one combination of the determined N-terminal sequence was cloned, and expression of the gene was confirmed by Northern blotting. The gene (bvi79A) encoded a prepeptide of 47 amino acids and a mature peptide, butyrivibriocin OR79A, of 25 amino acids. Significant sequence homology was found between this peptide and previously reported lantibiotics containing the double-glycine leader peptidase processing site. Immediately downstream of bvi79Awas a second, partial open reading frame encoding a peptide with significant homology to proteins which are believed to be involved in the synthesis of lanthionine residues. These findings indicate that the isolated inhibitory peptides represent new lantibiotics. Results from both total and N-terminal amino acid sequencing indicated that the second peptide was identical to butyrivibriocin OR79A except for amino acid substitutions in positions 2 and 3 of the mature lantibiotic. Only a single coding region was detected when restriction enzyme digests of total DNA were probed either with an oligonucleotide based on the 5′ region of bvi79A or with degenerate oligonucleotides based on the predicted sequence of the second peptide.
影响因子：
3.5
作者：
Wendy A. Woodruff;Jan Novak;P. Caufield
通讯作者：
Wendy A. Woodruff;Jan Novak;P. Caufield
优化 Northern 印迹程序。
影响因子：
2.7
作者：
Rosen,KM;Lamperti,ED;Villa-Komaroff,L
通讯作者：
Villa-Komaroff,L