食品生物防腐剂产生菌门间远缘细胞融合研究

乳酸链球菌素(Nisin)和ε-聚赖氨酸是由微生物产生的安全的天然防腐剂,分别由乳酸链球菌和白色链霉菌产生。这两种生物防腐剂的抑菌效果不同,存在互补性。为了获得具有更广谱抑菌作用的优良性状的菌株,探索远缘细胞融合的可行性,以自主分离的ε-聚赖氨酸产生菌白色链霉菌YB12和Nisin产生菌NZ9700作为亲本菌株,通过原生质体融合技术将两个菌株进行门间的细胞融合;通过抗药性筛选、形态学观察、SRAP-PCR分子鉴定,成功获得两株稳定遗传的融合菌株。同时对融合菌株的抑菌活性进行了检测。     

西藏灵菇中产胞外多糖嗜热链球菌的分离筛选及其发酵性能测定

从西藏灵菇中分离筛选、鉴定高产胞外多糖的乳酸菌菌株,并对其发酵性能和流变学参数进行测试。筛选的9株菌,菌种鉴定结果均为嗜热链球菌,其中菌株KC1、KC2、KC6、KC17产胞外多糖,菌株KC5、KC7、KC15、KC16、KC22不产胞外多糖。产胞外多糖菌株发酵乳发酵性能、黏度和黏附性指标均明显高于不产胞外多糖的嗜热链球菌,进一步证实了嗜热链球菌产生的胞外多糖可以赋予发酵乳良好的质地。     

富硒乳酸菌的分离、筛选、驯化及富硒研究

从酸奶中分离纯化出保加利亚杆菌(LactobacillusBulgaria,LB)和嗜热链球菌(StreptocociusThermopiles,ST),通过耐硒筛选,确定嗜热链球菌为耐硒菌种,以浓度梯度法对其驯化,从而得到富硒菌株,并进行富硒实验。初步确定富硒条件为:采用蕃茄酵母培养基,接种量10%,培养温度39℃,pH7,第6小时时加硒,硒浓度为15μg/mL,培养11h,富硒率为72.8%。     

Diversity of Streptococcus thermophilus phages in a large-production cheese factory in Argentina

Phage infections still represent a serious risk to the dairy industry, in which Streptococcus thermophilus is used in starter cultures for the manufacture of yogurt and cheese. The goal of the present study was to analyze the biodiversity of the virulent S. thermophilus phage population in one Argentinean cheese plant. Ten distinct S. thermophilus phages were isolated from cheese whey samples collected in a 2-mo survey. They were then characterized by their morphology, host range, and restriction patterns. These phages were also classified within the 2 main groups of S. thermophilus phages (cos- and pac-type) using a newly adapted multiplex PCR method. Six phages were classified as cos-type phages, whereas the 4 others belonged to the pac-type group. This study illustrates the phage diversity that can be found in one factory that rotates several cultures of S. thermophilus. Limiting the number of starter cultures is likely to reduce phage biodiversity within a fermentation facility.     

培养条件对Streptococcus equisimilis合成透明质酸相对分子质量的影响

作者比较了不同培养条件对Streptococcus equisimilis合成透明质酸相对分子质量的影响。结果表明:高质量浓度葡萄糖有利于长链透明质酸的合成,当发酵体系葡萄糖初始质量浓度从20g/L上升至80g/L,相对分子质量从1.24×106增大到2.02×106,提高了62.9%。葡萄糖补料培养不利于高相对分子质量透明质酸的合成,利用葡萄糖间歇和连续补料培养的方式得到透明质酸相对分子质量分别为1.53×106和1.42×106,比分批培养下降了19.9%和25.7%。在33~39℃范围内,较低温度有利于高相对分子质量透明质酸的合成,在33℃培养条件下透明质酸相对分子质量最高可达2.54×106。发酵液p H显著影响透明质酸的相对分子质量,在p H8时,达到了最高的2.38×106。较高溶氧水平有利于高相对分子质量透明质酸的合成,在0~45%溶氧浓度范围内,相对分子质量随溶氧水平的增加从1.16×106提升至2.43×106,增长了109.4%。本研究结果为后续高相对分子质量透明质酸的生产提供有用的实验依据。     

Molecular Epidemiology and Population Structure of Bovine Streptococcus uberis

The molecular epidemiology and population structure of 30 bovine subclinical mastitis field isolates of Streptococcus uberis, collected from 6 Portuguese herds (among 12 farms screened) during 2002 and 2003, were examined by using pulsed-field gel electrophoresis (PFGE) for clustering of the isolates and multilocus sequence typing (MLST) to assess the relationship between PFGE patterns and to identify genetic lineages. The 30 isolates were clustered into 18 PFGE types, using a similarity cutoff of 80%, and 3 PFGE types accounted for almost half of the isolates (46.6%). These major types were herd specific, suggesting either cow-to-cow transmission or infection with isolates from the same environmental reservoirs. The remaining unrelated PFGE types of isolates were from different herds strongly suggesting environmental sources of Strep. uberis infection. All 30 isolates were analyzed by MLST and clustered into 14 sequence types (ST). These ST were found to be novel, either with 10 new alleles of 6 housekeeping genes or with different combinations of previously assigned alleles. Five of these ST were clustered into 3 clonal complexes (lineages), ST-143, ST-86, and ST-5, known to include bovine isolates from several geographic locations (Australia, New Zealand, United Kingdom, Sweden, and Denmark) and 9 singletons. To our knowledge, this is the first report that documents molecular typing studies of bovine isolates of Strep. uberis from Portugal, which were shown to represent novel genomic backgrounds of this pathogen.     

Cloning and Characterization of Surface-Localized α-Enolase of Streptococcus iniae,an Effective Protective Antigen in Mice

Streptococcus iniae is a major fish pathogen that can also cause human bacteremia, cellulitis and meningitis. Screening for and identification of protective antigens plays an important role in developing therapies against S. iniae infections. In this study, we indicated that the α-enolase of S. iniae was not only distributed in the cytoplasm and associated to cell walls, but was also secreted to the bacterial cell surface. The functional identity of the purified recombinant α-enolase protein was verified by its ability to catalyze the conversion of 2-phosphoglycerate (2-PGE) to phosphoenolpyruvate (PEP), and both the recombinant and native proteins interacted with human plasminogen. The rabbit anti-rENO serum blockade assay shows that α-enolase participates in S. iniae adhesion to and invasion of BHK-21 cells. In addition, the recombinant α-enolase can confer effective protection against S. iniae infection in mice, which suggests that α-enolase has potential as a vaccine candidate in mammals. We conclude that S. iniae α-enolase is a moonlighting protein that also associates with the bacterial outer surface and functions as a protective antigen in mice.     

搅拌与混合对兽疫链球菌发酵生产透明质酸的影响

研究了透明质酸的流变学特性以及搅拌和混合对兽疫链球菌发酵生产透明质酸的影响。结果表明：透明质酸溶液属于典型的非牛顿凯松流体;搅拌转速和透明质酸浓度对气液传氧速率有很大影响;在较高的搅拌速率下发酵时可以得到较高的透明质酸产量,650 r·min^-1时透明质酸产量为4.1 g·L^-1,产率系数为0.08 g·g^-1;用有效搅拌模型分析发酵过程发现,只有在高搅拌转速时发酵体系才处于全部混合状态。     

酶法转化制备L-瓜氨酸

利用粪链球菌精氨酸脱亚胺酶转化L-精氨酸制备L-瓜氨酸。考察了菌龄、转化温度等多种因素对精氨酸脱亚胺酶活力的影响。酶法制备L-瓜氨酸的最适工艺条件是:菌体发酵时间20 h,转化温度37℃,转化液起始pH为6.0,ρ〔十六烷基三甲基溴化铵(CTAB)〕=0.3 g/L使酶活提高了414%。c(Co2+)=10-3mol/L使酶活增高约50%,c(Cu2+)=10-2mol/L或c(Zn2+)=10-2mol/L使酶活下降约50%。Ca2+、Mg2+和Mn2+对酶活影响较小。     

SARS-CoV-2 Spike Protein and Mouse Coronavirus Inhibit Biofilm Formation by Streptococcus pneumoniae and Staphylococcus aureus

The presence of co-infections or superinfections with bacterial pathogens in COVID-19 patients is associated with poor outcomes, including increased morbidity and mortality. We hypothesized that SARS-CoV-2 and its components interact with the biofilms generated by commensal bacteria, which may contribute to co-infections. This study employed crystal violet staining and particle-tracking microrheology to characterize the formation of biofilms by Streptococcus pneumoniae and Staphylococcus aureus that commonly cause secondary bacterial pneumonia. Microrheology analyses suggested that these biofilms were inhomogeneous soft solids, consistent with their dynamic characteristics. Biofilm formation by both bacteria was significantly inhibited by co-incubation with recombinant SARS-CoV-2 spike S1 subunit and both S1 + S2 subunits, but not with S2 extracellular domain nor nucleocapsid protein. Addition of spike S1 and S2 antibodies to spike protein could partially restore bacterial biofilm production. Furthermore, biofilm formation in vitro was also compromised by live murine hepatitis virus, a related beta-coronavirus. Supporting data from LC-MS-based proteomics of spike–biofilm interactions revealed differential expression of proteins involved in quorum sensing and biofilm maturation, such as the AI-2E family transporter and LuxS, a key enzyme for AI-2 biosynthesis. Our findings suggest that these opportunistic pathogens may egress from biofilms to resume a more virulent planktonic lifestyle during coronavirus infections. The dispersion of pathogens from biofilms may culminate in potentially severe secondary infections with poor prognosis. Further detailed investigations are warranted to establish bacterial biofilms as risk factors for secondary pneumonia in COVID-19 patients.