产广谱细菌素米酒乳杆菌C2发酵培养基的研究

为了提高米酒乳杆菌C2发酵产细菌素的水平,对发酵培养基的组成进行研究。采用琼脂扩散法,以金黄色葡萄球菌为指示菌,对发酵的基础培养基、碳源、氮源、无机盐和氨基酸对细菌素产生的影响进行研究,并确定最佳的培养基组成。实验结果表明：米酒乳杆菌C 2发酵产细菌素的最佳基础培养基为SL培养基,培养基中的碳源、氮源、无机盐及补加氨基酸对细菌素产生的影响显著。当培养基的组成为：葡萄糖20 g·L^-1,酵母浸粉25 g·L^-1,MgSO4.7H2O 0.38 g·L^-1,L-半胱氨酸1 g·L^-1,柠檬酸三铵2g·L^-1,乙酸钠2.5 g·L^-1,吐温80 1 ml.L-1时细菌素的产量最大,抑菌圈直径可达28.97 mm。     

细菌素Nisin延长低温熟肉制品货架期的效果

在低温肉制品加工过程中注入细菌素（Nisin）,观察了其感官指标和细菌数的变化。结果表明,细菌素（Nisin）在低温内制品中使用能有效抑制细菌繁殖速度,从而延长肉制品的货架期,较好地解决低温肉制品货架期短的问题。     

嗜酸乳杆菌La-1产细菌素发酵条件的研究

合适的发酵条件能够有效地提高嗜酸乳杆菌La-1细菌素的产量。在已经确定碳源、氮源和复合维生素的基础上,采用正交试验的方法研究了以MRS为基础的发酵培养基中各营养成分的相互协同作用,10L发酵罐试验证实,优化后的培养基和各项优化技术参数能使细菌总数提高了53.6%,细菌素抗菌活性提高了35.3%,抑菌圈达到了25mm以上。     

Interplay between gut microbiota and antimicrobial peptides

The gut microbiota is comprised of a diverse array of microorganisms that interact with immune system and exert crucial roles for the health. Changes in the gut microbiota composition and functionality are associated with multiple diseases. As such, mobilizing a rapid and appropriate antimicrobial response depending on the nature of each stimulus is crucial for maintaining the balance between homeostasis and inflammation in the gut. Major players in this scenario are antimicrobial peptides (AMP), which belong to an ancient defense system found in all organisms and participate in a preservative co-evolution with a complex microbiome. Particularly increasing interactions between AMP and microbiota have been found in the gut. Here, we focus on the mechanisms by which AMP help to maintain a balanced microbiota and advancing our understanding of the circumstances of such balanced interactions between gut microbiota and host AMP. This review aims to provide a comprehensive overview on the interplay of diverse antimicrobial responses with enteric pathogens and the gut microbiota, which should have therapeutic implications for different intestinal disorders.     

Necrotic enteritis-producing strains of Clostridium perfringens displace non-necrotic enteritis strains from the gut of chicks

We inoculated broiler chicks with mixtures of Clostridium perfringens strains to investigate the single strain dominance observed in natural cases of necrotic enteritis (NE) [Nauerby, B., Pedersen, K., Madsen, M., 2003. Analysis by pulsed-field gel electrophoresis of the genetic diversity among Clostridium perfringens isolates from chickens. Vet. Microbiol. 94, 257-266]. Pre-inoculation bacteriologic culture of chick intestines yielded up to six pulsed-field gel electrophoresis (PFGE) types of C. perfringens. Birds developed typical NE lesions in response to administration (2x per day for 4 days) of a combined inoculum comprising one NE strain (JGS4143, PFGE pattern 8) and four non-NE strains (from piglet necrotizing enteritis, chicken normal flora, human gas gangrene, and bovine neonatal enteritis). After inoculation commenced, only the NE strain was recovered through the first post-inoculation day, in spite of intense efforts to recover pre-challenge flora strains and the other challenge strains. Thereafter, pre-inoculation and previously undetected PFGE types were found, and JGS4143 became undetectable. Birds inoculated simultaneously with five NE strains (from disease in chickens or turkeys, and including JGS4143) also developed lesions, but again only JGS4143 was recovered through the 1st day post-challenge. At that time, birds began to be repopulated with pre-challenge PFGE types. Two NE strains (JGS4143 and JGS4064) produced bacteriocins, which inhibited each other and normal flora strains (n=17), while normal flora strains inhibited neither NE strains nor each other. Thus, it appears that naturally occurring dominance of the gut by NE strains can be reproduced experimentally. Bacteriocins directed against normal flora could possibly provide the necessary advantage, although inhibition of one NE strain by another suggests that other factors may be partially or completely responsible for the dominance.     

一株产细菌素乳酸菌的筛选及细菌素特性研究

[目的]研究从酸马奶酒中筛选出的1株产细菌素的乳酸菌的细菌素特性.[方法]从内蒙古地区酸马奶酒中分离到19株乳酸菌,采用牛津杯双层平板扩散法,排除有机酸、过氧化氢的干扰后,经蛋白酶敏感性试验,筛选出1株产细菌素的乳酸菌菌株XMD6.[结果]通过形态学特征、生理生化试验及16S rRNA基因序列分析,鉴定为坚强肠球菌(E.durans).其所产细菌素在80℃对热稳定,显示抑菌活性pH范围为2～7.抑菌谱试验表明,该菌能抑制食品中常见的革兰氏阳性、革兰氏阴性的致病菌及腐败菌,具有较广的抑菌谱.[结论]研究可为今后开发利用新型细菌素防腐剂提供理论依据.     

细菌素Bacteriocin E50-52(H)基因设计 及其毕赤酵母表达载体构建

设计及合成细菌素Bacteriocin E50-52(H)基因,克隆到组成型分泌表达质粒pGAPZαA中构建重组质粒pGAPZα-Bacteriocin E(H),经PCR、测序验证正确后,电转化整合到毕赤酵母基因组,基因组PCR、测序验证结果表明成功构建了细菌素组成型重组表达载体,为在毕赤酵母中表达奠定了基础。     

产广谱细菌素乳酸菌CW3的筛选和初步鉴定

[目的]对CW3菌株进行筛选,并且进行初步鉴定。[方法]首先采用牛津杯双层平板法进行产广谱细菌素菌株的初筛,再将初筛得到的菌株进行排除酸和过氧化氢干扰,并检测抑菌物质的蛋白质性质,最终对复筛得到的菌株进行鉴定。[结果]试验得出,排除有机酸、过氧化氢等干扰因素后,发酵液仍有抑菌作用;用胰蛋白酶和胃蛋白酶处理后,发酵液抑菌活性急剧下降,确定产生的抑菌物质具有蛋白质性质,是一类细菌素。经过生理生化试验初步鉴定菌株CW3为植物乳杆菌。[结论]菌株CW3是一种能产广谱细菌素的植物乳杆菌。     

乳链球菌P-99发酵乳清生产乳链菌肽的研究

研究了乳清培养基中乳链球菌P-99的细胞生长和乳链菌肽合成特性,采用正交试验对乳清发酵培养基中的碳源、氮源和磷源进行了优化。结果表明,乳链球菌P-99能够在没有添加物的乳清培养基中生长并合成乳链菌肽,其细胞生长和产物合成可以分别用Monod方程和Luedeking-Piret方程拟合。培养基优化试验结果表明,在乳清发酵培养基中添加19g/L乳糖、0.8g/L甘氨酸、0.45g/LKH2PO4时,乳链菌肽的合成量提高了31.7%。     

青枯假单胞杆菌产生细菌素的研究

测定来自全国不同寄主和地区的211个青枯病细菌(Pseudomonassolanacearum)菌株产生细菌素的能力,结果表明,有80％分属于不同小种和生物型的8个指示菌能产生细菌素,其中能产生细菌素的桑菌株占其总数的94.2％。 用7个拮抗作用强而稳定的菌株测定对211个供试菌株的拮抗作用,结果表明,不同菌株产生细菌素的能力和拮抗范围有差异。其中油橄榄菌株(OE-104)和桑菌株(MA-1、MA-701)拮抗能力最强,拮抗范围最广。不同小种、生物型及寄主来源的菌株间的桔抗作用大于来源相同菌株间的拮抗作用。此外,试验证明培养温度、培养基成分和培养时间对菌株产生抑制圈的大小有影响。这些结果对筛选青枯病细菌的产细菌素拮抗菌有参考价值。