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Abstract By using an extension to group N streptococci of a contransformation procedure we have introduced 4 different-sized cryptic plasmids for Streptococcus lactis into the plasmid-free S. lactis IL1403. A mixture of 4 cryptic plasmids with an indicator plasmid (pHV1301) conferring erythromycin resistance was used for IL1403 protoplast transformation. Under such conditions, 41.5% of the erythromycin-resistant transformants were contransformed with one of the cryptic plasmids in addition to pHV1301. Indicator plasmid pHV1301 was later spontaneously segregated from doubly transformed cells. This protocol should be very useful for constructing lactic streptococcal strains bearing any phenotypically cryptic plasmid. 相似文献
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本研究通过对乳酸链球菌亚种N8的Nisinz结构基因下游基因区的分子克隆、序列分析和ORF的检测,揭示出一个属于细菌性双成份调节子系统的基因,nisK,其起始区略与nisR末端重叠,并发现NisR和nisK同被转录一个多顺反子mRNA。在nisK编码区后,有一个ρ非依赖性终止子。 相似文献
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Cloning and nucleotide sequence analysis of the Lactobacillus delbrueckii ssp. lactis DSM7290 cysteine aminopeptidase gene pepC 总被引:1,自引:0,他引:1
Abstract A genomic library of Lactobacillus delbrueckii ssp. lactis DSM7290 in the low copy number vector pLG339, was screened for the presence of peptidase genes. Using the chromogenic substrate gly-ala-β-naphthylamide, which is not a substrate for any of the recently cloned peptidases of DSM7290, and the multiple peptidase deficient Escherichia coli strain CM89, allowed the isolation of clones, which contained the equivalent hydrolytic activity. To identify genes encoding the conserved catalytic active site of cysteine proteases, partial nucleotide sequencing with a degenerate oligonucleotide was performed on recombinant plasmids isolated from such clones. This allowed to identify two out of nine clones to carry the Lactobacillus pepC gene. A total of 2026 nucleotides were determined, and sequence analysis revealed a gene with strong homology to the recently cloned Lb. helveticus (73.2%) and Lactococcus lactis (51.03%) pepC genes, and the derived protein showed homology with the active site of a large number of cysteine proteases. The predicted open reading frame consists of 449 codons, coding for a protein of 50 909 Da. The enzyme is functional and extremely overexpressed in E. coli . 相似文献
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乳链菌肽的分离纯化和部分生物学性质 总被引:1,自引:0,他引:1
用乳酸链球菌SM526进行乳链菌肽的发酵生产,产量为40~50mg/l.经中空纤维超滤器超滤,非极性大孔吸附树脂XAD-2层析,CM-SephadexC-25层析和SephadexG-50层析纯化了该肽。SDS-PAGE表明达均一,RP-HPLC表明其纯度不低于95%。SDS-PAGE测其Mr约为3600,用IEF测其等电点为9.5.酸性条件下稳定且抗热;对胰蛋白酶、胃蛋白酶和木瓜蛋白酶不敏感,但对α-胰凝乳酶和蛋白酶K敏感。乳链菌肽对多种革兰氏阳性菌有强烈的抑制作用;以枯草杆菌和金黄色葡萄球菌为指示菌,其作用方式是杀菌。 相似文献
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Abstract Plasmid pIP501 was transferred by conjugation from Lactococcus lactis to Lactobacillus delbrückii subsp. bulgaricus and Lactobacillus helveticus . Only Lb. delbrückii subsp. bulgaricus transconjugants could act as a donor in crosses with Lc. lactis . No Lactobacillus transconjugants were detected after inter- or intra-species Lactobacillus crosses. Plasmid pIP501 has undergone no detectable deletion or rearrangement during transfer from Lc. lactis to Lactobacillus strains. 相似文献
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Alexander Akermann Jens Weiermüller Jens Christmann La Guirande Gregor Glaser Annette Knaus Roland Ulber 《Engineering in Life Science》2020,20(5-6):168-180
Brewers’ spent grain (BSG) is a low‐cost by‐product of the brewing process. BSG liquor names the liquid components of BSG, mainly glucose, maltose, and long‐chain α‐1,4‐glycosidic bond glucose oligomers. These substances should be separated in existing BSG biorefineries, as they might lead to an increased formation of microbe‐inhibiting compounds in well‐established hydrothermal/enzymatic saccharification processes. In most cases, this liquid fraction is discarded. The present study presents for the first time an optimized process with BSG liquor for the purpose of producing bulk chemicals (e.g., lactate) in relevant concentrations. The process comprises the application of yeast extract, produced from own brewing processes, as the sole supplemented complex constituent in a simultaneous fermentation and saccharification process. Kinetic parameters for the final optimized process conditions with the organism Lactobacillus delbrueckii subsp. lactis were: maximum specific growth rate µmax = 0.47 h?1, maximum lactate concentration cLac, max = 79.06 g L?1, process yield YPS = 0.89 gLac gSugar?1, lactate production rate qP = 4.18 gLac gCDW?1 h?1, and productivity P 15 h = 4.93 gLac L?1 h?1. BSG liquor, linked with yeast extract from Brewers’ yeast, can be a promising substrate for further bioprocess engineering tasks and contribute to a holistic and sustainable usage of Brewers’ spent grain. 相似文献
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