正交旋转试验优化产Ⅱa类细菌素乳酸菌的扩大发酵条件

利用前期实验数据,通过五因子二次正交旋转试验对扩大发酵条件进行优化,考察了转速、装液量、接种量、温度和不同初始pH值对产Ⅱ a类细菌素屎肠球菌发酵的影响.得到发酵培养基最佳条件:转速150 r/min.接种量2%,装液量450mL,温度30℃,初始pH值6.5.     

产超氧化物歧化酶乳酸菌的筛选及发酵条件优化

从不同地区的传统发酵食品中筛选出104株菌落形态各异的乳酸菌,通过过氧化氢抗性试验筛选出20株具有抗氧化能力的乳酸菌,并测定其产SOD的能力。结果表明,20株乳酸菌中C8的SOD活性最高,达349.62 U/mg。通过革兰氏染色、过氧化氢酶试验和16S rRNA基因序列分析,构建系统发育树,根据同源性分析确定C8为乳酸乳球菌。为提高菌株产SOD的能力,通过单因素实验探究培养温度、初始接种量、初始pH值和灌装量对乳酸乳球菌产SOD的影响,并通过正交试验进一步优化菌株的发酵条件。最终确定最佳发酵条件为:发酵温度37℃,初始接种量3%,初始pH 7.5,灌装量75mL。在此培养条件下,乳酸乳球菌的SOD活性达93.95 U/mL,与优化前相比提高了2.2倍,为后期乳酸乳球菌在发酵产业的应用奠定基础。     

益生屎肠球菌RS3的分离鉴定及增殖条件的优化

为筛选具有益生作用的乳酸菌,该研究从泡菜中筛选到1株细菌菌株RS3,经菌落形态观察、生化试验和16 S rRNA序列分析确定为屎肠球菌。为高效获得屎肠球菌RS3菌体,对其最佳增殖条件进行优化研究。利用单因素筛选和中心复合响应面试验对屎肠球菌RS3菌体增殖用液体培养基和增殖条件进行研究,优化的最佳培养基为乳糖15 g/L、胰蛋白胨25 g/L、初始pH值为8.2、培养温度37℃。与优化前培养条件相比,优化后培养条件下菌体生物量提高了54.9%,乳酸产量提高了53%,为后期规模化培养和益生菌剂开发奠定基础。     

屎肠球菌HY07产细菌素发酵条件的优化

对屎肠球菌HY07产细菌素的培养基组成和发酵条件进行了研究.通过单因素水平试验和正交试验,确定产细菌素的最佳培养条件为37℃培养24h,培养基初始pH为6.最佳培养基组成为:葡萄糖1％,鱼粉蛋白胨1.5％,牛肉膏1.5％,磷酸氢二钾0.2％,柠檬酸二铵0.2％,乙酸钠0.5％、硫酸锰0.025％,吐温-800.4％.在此条件下培养屎肠球菌HY07,所产细菌素抑菌圈可达12.60mm.     

嗜酸乳杆菌细菌素Lactobacilin XH1发酵条件优化

以细菌素相对抑菌效价为考察指标,通过单因素和正交试验对嗜酸乳杆菌细菌素Lactobacillin XH1发酵条件进行优化,研究了培养基初始pH、培养温度、培养时间及接种量4个因素对细菌素产生的影响。结果表明,优化后最佳发酵条件为培养基初始pH 6.5、培养温度35℃、接种量为2.5%和培养时间28 h,在此条件下细菌素Lactobacillin XH1相对抑菌效价值达到247.49±1.05 AU/mL,比优化前提高了2倍。     

产细菌素屎肠球菌E6的特性分析及发酵条件优化

研究了屎肠球菌E6抑菌活性的生物学特性,并通过响应面法优化其发酵条件.结果表明,屎肠球菌E6产蛋白质类细菌素,能够抑制单增李斯特氏菌、大肠杆菌、金黄色葡萄球菌和枯草芽孢杆菌生长,在pH3.0～7.0条件下有明显抑菌活性,60～121℃热处理20min后仍具有抑菌活性.通过Box-Behnken实验设计优化屎肠球菌E6发酵条件为培养时间36.0h,培养温度31.0℃,培养基pH5.1.在此条件下,发酵上清液抑菌圈直径可达20.17mm,较优化前提高了27.2％.     

响应面法优化产细菌素屎肠球菌BC-3发酵培养基

响应面分析法优化屎肠球菌BC-3产细菌素的培养基。采用不同的培养基对屎肠球菌BC-3进行培养,筛选出最适培养基。结合单因素法和响应面分析法,优化出最适屎肠球菌BC-3产细菌素的发酵培养基配比。结果表明:屎肠球菌BC-3产细菌素的最佳培养基为蛋白胨27.11g/L,牛肉膏51.65g/L,磷酸氢二钾4.78g/L,在此条件下,细菌素的产量最高,抑菌圈达到最大,为13.21mm,比优化前提高了42.81%。     

一株产γ-氨基丁酸屎肠球菌的筛选和发酵条件优化及其益生特性分析

为拓宽产γ-氨基丁酸(γ-aminobutyric acid,GABA)微生物资源,以四川泡菜为分离源从中筛选高产GABA乳酸菌菌株,并对高产乳酸菌菌株进行生理生化、分子生物学鉴定,高产GABA发酵条件优化及其益生特性分析。结果表明:采用高效液相色谱对菌株产GABA能力分析发现菌株AB157的GABA产量最高为1.08 g/L。生理生化和16S rRNA鉴定菌株AB157为屎肠球菌(Enterococcus faecium)。通过单因素实验和响应面优化,确定屎肠球菌AB157的最佳培养条件为L-谷氨酸钠底物浓度5.2 g/L、发酵温度31 ℃、初始pH7、发酵时间70 h,在此条件下,屎肠球菌AB157的GABA产量达到1.60 g/L。屎肠球菌AB157的益生分析表明,其在pH2.0和3 g/L的胆盐环境中的存活率分别为39%和59%,胆固醇的脱除量为18.09 μg/mL,说明该菌株具有良好的耐酸和耐胆盐特性以及较强的降胆固醇能力,可作为潜在功能性乳酸菌资源进行后续研究开发。     

乳球菌CW22产细菌素发酵条件的优化

对乳球菌CW22产细菌素的发酵条件进行了优化,分别研究了培养时间、温度、接种量、培养基起始pH值、培养基碳源、氮源等因素对细菌素产生的影响,通过单因素水平试验和正交试验,确定产细菌素的最佳培养基组合和最佳发酵条件为:葡萄糖3%,胰蛋白胨1.5%,蛋白胨2%,酵母膏1.5%,硫酸镁0.087%,吐温-80.2%,磷酸氢二钾0.2%,柠檬酸铵0.2%,乙酸钠0.5%,硫酸锰0.025%,30℃培养28 h,培养基起始pH值为6.0,接种量2%.通过优化发酵条件,细菌素产量提高了313%.     

乳酶生片中乳酸菌的分离鉴定及其增殖培养条件优化

该研究采用传统培养分离法从乳酶生片中分离乳酸菌,通过形态观察、生理生化试验及分子生物技术对其进行鉴定,并采用单因素及响应面试验对其增殖培养条件及培养基进行优化。结果表明,从乳酶生片中分离得到一株乳酸菌Y1,经鉴定为屎肠球菌（Enterococcus faecium）,其最适培养条件为初始pH 8.0,生长温度37 ℃,接种量1%,最优增殖培养基为乳糖11 g/L,乳清粉10 g/L,酵母浸粉22 g/L,硫酸镁0.28 g/L,硫酸锰0.18 g/L。用此培养基培养所得E. faecium Y1的活菌数为2.90×109 CFU/mL,比优化前活菌数提高了80.12%。     

屎肠球菌BC-3产细菌素条件优化及其理化特性研究

从东北传统发酵酸菜中筛选到一株屎肠球菌BC-3,通过其代谢产生的细菌素来控制食品中的某些腐败菌和病原菌,开发其作为高效广谱食品生物防腐剂。通过滤纸片法对屎肠球菌BC-3产生细菌素发酵条件进行优化,结果表明,发酵时间为20 h、pH值为6.5、接种量为1.5%、温度为37.0℃时,抑菌圈直径最大,为17.87 mm。经硫酸铵粗提后,对肠球菌素E3生物学特性进行分析,结果表明其具有较高耐热性,在pH为2.0~10.0的范围内稳定性较好,可被蛋白酶降解,经有机溶剂处理后稳定性几乎不受影响。     

Enterocin A production by Enterococcus faecium FAIR-E 406 is characterised by a temperature- and pH-dependent switch-off mechanism when growth is limited due to nutrient depletion

The biokinetics of cell growth and bacteriocin production of Enterococcus faecium FAIR-E 406 was studied as a function of temperature (20-45 degrees C) and pH (5.5-8.5) using de Man-Rogosa-Sharpe medium. Growth of E. faecium FAIR-E 406 was characterized by three successive growth phases and was modelled with the mechanistic nutrient depletion model. Bacteriocin production showed primary metabolite kinetics but was limited to the early growth phase. The critical biomass for switching off bacteriocin production was dependent on medium pH and incubation temperature, and was inversely correlated with the specific bacteriocin production. Doubling the concentration of the nitrogen source as well as a step-wise pH increase shifted the bacteriocin production towards a higher switch-off cell density.     

植物乳杆菌(Lactobacillus plantarum)8-6产细菌素发酵条件的优化

对植物乳杆菌(Lactobacillus plantarum)8-6产细菌素的发酵条件进行了优化,分别研究了培养时间、温度、接种量、培养基起始pH值、培养基碳源、氮源等因素对细菌素产生的影响,通过单因素水平试验和正交试验,确定产细菌素的最佳培养基组合和最佳发酵条件为葡萄糖3%,胰蛋白胨2%,蛋白胨1%,酵母膏1%,硫酸镁0.058%,吐温-80 0.2%,30℃培养24h,培养基起始pH值为6.5,接种量2%。乳杆菌8-6优化后效价为1825.56IU/mL,比优化前提高了373.15%。     

产细菌素屎肠球菌的筛选鉴定及其抑菌特性

从采自四川红原的传统发酵酸乳中分离得到308 株疑似乳酸菌,通过96 孔板法初筛和平板打孔法复筛,筛选出1 株具有明显抑菌作用的菌株SC-Y112。通过形态学观察、16S rDNA序列同源性分析和ropA管家基因序列同源性分析结合生理生化实验,鉴定该菌株为屎肠球菌（Enterococcus faecium）。通过排酸及排除H2O2影响后,该菌株的发酵上清液仍具有明显抑菌活性,经蛋白酶处理后,抑菌效果明显消失或下降,说明发酵上清液中的抑菌成分具有蛋白质性质。进一步探究该菌株所产细菌素的生物学特性、遗传稳定性及抑菌特性表明,该细菌素在菌株接种后6 h产生,12 h达到最高;菌株在连续传代86 代内,产细菌素能力较为稳定;细菌素在 pH 3.0～7.0的条件下稳定,在100 ℃处理30 min后仍具有抑菌活性;SC-Y112所产细菌素具有广谱抗菌性,对单核细胞性李斯特菌有明显的抑菌活性。     

Enterococcus faecium RZS C5, an interesting bacteriocin producer to be used as a co-culture in food fermentation

Enterocins, bacteriocins produced by enterococci, are gaining interest because of their industrial potential. Due to its bacteriocin production, Enterococcus faecium RZS C5, a natural cheese isolate, has a strong activity towards Listeria monocytogenes. For this reason, the strain may be applicable as a bacteriocin-producing co-culture in food fermentation in order to reduce the risk on Listeria outgrowth. The strain displays remarkable bacteriocin production kinetics. Whereas most lactic acid bacteria produce bacteriocin in a growth-associated way until the beginning of the stationary phase, bacteriocin production by E. faecium RZS C5 in MRS broth at controlled pH values below 7.5 is characterised by a boost of bacteriocin activity levels in the very early growth phase. In addition, bacteriocin production kinetics are closely linked to the environmental and cultural conditions. However, no straightforward statement about the effect of environmental stress on bacteriocin production can be made since the effect is dependent on the type of stress applied. Kinetic experiments in milk and on pilot scale, applying Cheddar cheese-making conditions, have indicated that the strain may be effective as a bacteriocin-producing co-culture. Further research is needed to evaluate the use of E. faecium RZS C5 as a co-culture for the production of fermented sausage.     

香菇产抑菌活性物质液态发酵条件优化

以抑菌活性为评价指标,以接种量、初始pH值、摇床转速、发酵时间、发酵温度为考察因素,采用单因素及正交试验优选液态发酵条件.结果表明,产抑菌活性物质的最佳发酵条件是接种量7％,发酵液初始pH值为5,培养时间8d、培养温度为28℃.在此条件下,发酵液对黄曲霉的抑菌圈直径为8.9 cm.发酵液初始pH值和发酵时间对试验结果的影响显著.     

动物双歧杆菌细菌素bifidocin A群体感应合成调控行为分析

bifidocin A是由Bifidobacterium animalis BB04代谢合成的一种新型广谱高效细菌素。为探讨该细菌素的群体感应合成调控行为,本研究通过监测发酵过程中菌体生长及细菌素抑菌活性变化规律,分析菌体密度和细菌素合成的相关性;基于低产细菌素培养模型体系的构建,检测发酵上清液中是否存在群体感应自诱导肽,判断是否存在细菌素合成相关群体感应系统;并通过对发酵上清液中自诱导肽进行提纯和分子质量测定,初步明确其分子特性。结果表明,当菌体细胞达到活菌数为7.31（lg（CFU/mL））时,细菌素才开始合成;发酵过程中,菌体密度与细菌素的合成呈现正相关性;成功构建了低产细菌素培养模型体系,其培养条件为培养温度37?℃、培养基起始pH?5、培养基浓度1/10改良MRS（Man Rogosa Sharpe）培养基、接种量1%、培养时间24?h;基于此模型体系,确定发酵上清液中确实存在可以诱导细菌素合成的自诱导肽,细菌素bifidocin?A的合成是受群体感应系统调控的;通过对发酵上清液进行超滤管筛分、葡聚糖凝胶柱层析及高效液相色谱层析提纯,获得了高纯度自诱导肽样品;采用基质辅助激光解析电离飞行质谱测定其分子质量为3?587.253?Da。     

环境因素对屎肠球菌产苯乙胺和酪胺的影响

为揭示环境因素对屎肠球菌产苯乙胺和酪胺的影响,研究了NaCl、糖的添加量、pH、含氧量、温度环境因素以及产胺菌之间对生物胺的影响。结果表明,当pH为5.00时,屎肠球菌的生长和产生物胺能力都受到抑制。当pH为6.00时,产胺菌的产胺能力在有氧条件下显著低于厌氧条件下。通过4因素二次回归方程分析pH、温度、加糖量和加食盐量对生物胺产生的影响,结果表明,pH、糖、盐和温度对屎肠球菌生长有显著的影响,其中受pH和温度影响较大。pH和温度对苯乙胺和酪胺产生有显著的交互影响,并且这种交互作用随着温度升高显著增强。     

The stimulating effect of a harsh environment on the bacteriocin activity by Enterococcus faecium RZS C5 and dependency on the environmental stress factor used

Bacteriocin production by Enterococcus faecium RZS C5 occurs in a growth-associated way but is generally switched off in the very early growth phase. The influence of environmental stress on the bacteriocin production kinetics by E. faecium RZS C5 was analysed at a controlled temperature of 35 degrees C and constant pH 6.5. The effect of environmental stress on bacteriocin production was depending on the type of stress applied. Oxidative stress did not interfere with cell growth or bacteriocin activity. In contrast, salt stress decreased both the cell growth and the specific bacteriocin production. Nevertheless, moderate levels of sodium chloride improved bacteriocin activity because they increased the biomass concentration at which bacteriocin production was switched off. Environmental stress due to limitations in sugar or complex nutrients did not affect the early shut-off mechanism or the specific bacteriocin production. However, bacteriocin stability decreased or increased at low levels of sugar or complex nutrients, respectively.