植物乳杆菌发酵培养基的优化及其高密度培养技术

应用响应面法优化植物乳杆菌培养基配方以及利用中和法与指数流加法优化植物乳杆菌高密度培养的发酵条件。在单因素试验基础上,进一步采用SAS软件进行中心组合设计和响应面法优化发酵培养基。优化后的培养基配方为：葡萄糖质量分数5.43%、蛋白胨质量分数0.98%、K2HPO4质量分数0.59%。利用15L全自动发酵罐,在接种量3%、pH6.5、培养温度35℃的最佳条件下,采用氨水中和发酵培养基和指数流加碳、氮源,最终发酵液中植物乳杆菌菌体浓度达到9.3×109CFU/mL。     

基于pH下降速率的反馈补料策略高密度培养保加利亚乳杆菌NQ2508

保加利亚乳杆菌是酸奶发酵剂和益生菌制剂的重要菌种,其高密度培养对于提高菌种在应用产品生产中的效率具有重要的现实意义。葡萄糖浓度和发酵pH是影响保加利亚乳杆菌生长的关键因子,底物葡萄糖被转化为细胞菌体和主要代谢产物乳酸,探究葡萄糖消耗速率与表征乳酸生成的发酵pH下降速率的关系,是研究保加利亚乳杆菌代谢流方向的重要方法。该实验通过培养基组成和培养条件对保加利亚乳杆菌生长的影响进行研究,同时对葡萄糖消耗速率-pH降低速率相关模型研究,并基于底物质量守恒定律建立了一条基于pH下降速率的保加利亚乳杆菌高密度培养的反馈补料发酵策略。通过化学中和法控制发酵pH为6.0～5.0,基于pH下降速率进行反馈补料将葡萄糖质量浓度控制在8～2 g/L,发酵液的活菌数达到3.6×10⁹ CFU/mL,是批次分批培养的2.4倍。该策略的应用结果为提高保加利亚乳杆菌的工业化生产效率提供参考。     

培养条件对植物乳杆菌LIP-1抗冷冻活性的影响

以前期优化的MRS培养基为基础培养基,采用二次响应面分析法优化植物乳杆菌LIP-1的培养条件（温度、接种量、初始培养pH）,研究改变培养条件是否可提高植物乳杆菌 LIP-1高密度发酵培养后的活菌数以及对冷冻干燥后菌株的冻干抗性的影响。结果显示：最佳培养条件：培养pH6.7、培养温度36 ℃、接种量8.8%,在此培养条件下,植物乳杆菌LIP-1高密度发酵后活菌数为10.1855 lg（CFU/mL）,冷冻干燥后的存活率达83.40%,与未优化的培养条件相比,发酵培养后活菌数及冻干存活率显著提高（P<0.05）,分别提高0.2935 lg（CFU/mL）和8.46%。结轮：通过改变培养条件可显著提高植物乳杆菌LIP-1的高密度发酵活性和冷冻干燥存活率。     

植物乳杆菌高密度培养通气条件的研究

为研究通气条件对植物乳杆菌高密度生理活性、高密度培养及存活率的影响,在FUS-50-L (A)发酵罐中,分别采用4种通气策略进行发酵培养.结果表明,当通入空气时,植物乳杆菌生长速率较小,而通入氮气时,菌体的生理活性、比生长速率、营养物质消耗速率和细胞存活率最高,最大活菌数可达到1.8× 1010CFU/mL.说明植物乳杆菌在微量氧气短暂存在时能存活,欲实现发植物乳杆菌的高密度培养,必须通过通入氮气等惰性气体来保持发酵过程的厌氧状态.     

泡菜用乳酸菌富集培养的研究

泡菜用直投式乳酸菌发酵剂制作中,菌体富集和冻干浓缩是关键点。主要以实验室现有的3种菌株:植物乳杆菌、肠膜明串珠菌、短乳杆菌为出发菌株,进行富集培养的研究,采用缓冲盐法和化学中和法2种培养方式。通过缓冲盐法确定了植物乳杆菌、肠膜明串珠菌、短乳杆菌的最适pH均为7.0,最适培养温度分别为30℃、30℃、35℃;化学中和法采用控制pH为7.0,培养温度分别为30℃、30℃、35℃的培养条件。通过2种方法的比较,可知化学中和法得到的活菌数约为缓冲盐法的3～5倍。     

金银花乳杆菌发酵凉茶的研制

该研究的目的在于探究植物乳杆菌发酵金银花凉茶的工艺条件。以金银花为原料,通过单因素试验和正交试验,研究植物乳杆菌PMO接种量、发酵温度、发酵时间和糖添加量对金银花凉茶发酵过程的影响,并对发酵工艺条件进行优化。结果表明,植物乳杆菌PMO经过驯化过程,其在金银花凉茶中具有优良的增殖活性;金银花凉茶中绿原酸含量受发酵过程影响较小,始终稳定在29.20 mg/g左右;金银花凉茶最佳发酵工艺条件为植物乳杆菌PMO接种量3%、发酵温度37 ℃、发酵时间14 h、糖添加量7%。     

一株植物乳杆菌高密度培养的研究

通过单因素实验、Plackett-Burman法、最陡爬坡试验对分离自内蒙古传统发酵乳制品中高产γ-氨基丁酸的植物乳杆菌NDC75017培养基成分及培养条件进行优化研究。确定影响菌体生长的显著因素为葡萄糖、硫酸锰、柠檬酸氢二铵。利用Box-Behnken设计确定了植物乳杆菌NDC 75017的增殖培养基配方。结果表明,菌体在最优条件下进行培养,活菌数可达6.48×1010mL-1。细胞干重可达3.93 g/L。     

短乳杆菌的培养条件及高密度培养研究

对实验室自制泡菜中分离到的短乳杆菌进行生长特性研究,并采用响应面分析对影响短乳杆菌增殖的发酵培养基进行优化.首先采用PB实验筛选影响短乳杆菌增殖的显著因素,利用最陡爬坡实验逼近最大增殖区域,最后应用中心组和设计和响应面分析确定短乳杆菌高密度培养的最佳优化培养基成分为(%W/V):MRS基础培养基、乳糖0.2、大豆蛋白0.53、番茄汁0.4、胡萝卜汁1.04、醋酸钠0.05、碳酸钙0.125、磷酸氢二钠0.075,乳酸菌高密度培养活菌数可达5.21×109cfu/mL.     

发酵乳杆菌SS-31培养基及发酵条件的优化

对分离自广西柳州酸笋发酵液中具有优良益生特性的发酵乳杆菌SS-31进行增殖培养基优化,并对其高密度发酵条件进行探索。通过单因素实验、响应面优化等方法,以发酵乳杆菌SS-31的活菌数为主要参考指标,对其发酵增殖培养基成分和培养条件进行了优化探索。最终确定发酵乳杆菌SS-31最优培养基构成为：麦芽糖16.20 g/L、酵母浸粉20.16 g/L、磷酸氢二钾9.33 g/L、硫酸锰0.50 g/L、硫酸镁1.00 g/L、吐温80 1.00 g/L;最佳发酵条件为：SS-31接种体积分数为3%、初始pH为6.8,期间添加氨水保持发酵液pH稳定,在37 ℃下培养24 h后,活菌数可达到1.19×10¹⁰ CFU/mL,为其工业化生产奠定基础。     

低聚半乳糖对植物乳杆菌发酵乳特性及抗蜡样芽孢杆菌活性的改善

为探究低聚半乳糖对植物乳杆菌发酵乳特性及抗菌活性的影响,本文采用单因素法考察影响发酵乳特性的主要因素,并以响应面法优化发酵乳最佳发酵条件;以产肠毒素蜡样芽孢杆菌HN001为指示菌,探究低聚半乳糖的添加对植物乳杆菌ZDY2013发酵乳抑菌活性的改善作用。结果表明:植物乳杆菌能有效利用低聚半乳糖进行体外代谢,并抑制蜡样芽孢杆菌生长;牛奶中添加适量低聚半乳糖能够增加植物乳杆菌发酵乳中的活菌数、降低发酵乳的pH,并提高其持水力;响应面分析发现低聚半乳糖发酵乳的最佳制备条件为:2.0%的植物乳杆菌接种量、1.0%的低聚半乳糖添加量、发酵时间为24 h及发酵温度为42 ℃;添加低聚半乳糖的发酵乳能有效控制产肠毒素蜡样芽孢杆菌浓度在106 CFU/mL以下。该研究结果为低聚半乳糖及植物乳杆菌ZDY2013在发酵乳中的应用奠定了理论基础。     

一株产细菌素植物乳杆菌高密度发酵培养基的筛选及其在酸奶中的应用

目的:优化用于高密度发酵产细菌素的植物乳杆菌KLDS 1.0391的经济有效培养基,旨在生产具有益生作用的辅助发酵剂。方法:以发酵液中活菌数量和抑菌效果为指标,比较不同水解度的脱脂乳粉、乳清粉、麦麸,以及玉米浆粉的发酵效果,制备冻干形式的植物乳杆菌直投式发酵剂,将其添加至酸奶中,评价其对酸奶发酵时间、pH、滴定酸度、粘度和感官评价的影响,进而确定其在酸奶中应用的可行性。结果:确定脱脂乳粉、乳清粉和麦麸培养基的最佳水解度分别为15%、15%和20%;植物乳杆菌KLDS 1.0391在玉米浆粉培养基中获得的发酵效果最佳,活菌数量达到9.53 lg CFU/mL,抑菌圈直径达到12.69 mm。植物乳杆菌直投式菌种的添加对酸奶发酵时间和感官评价无显著影响(P>0.05),酸奶粘度下降,但可以缓解酸奶的后酸化程度。通过优化得到植物乳杆菌KLDS 1.0391高密度发酵的廉价培养基为:4%玉米浆粉,同时补充2%葡萄糖和0.1% Tween-80。结论:制得的植物乳杆菌直投式发酵剂可应用于酸奶生产中。     

Preparation of a Lactobacillus plantarum starter culture for cucumber fermentations that can meet kosher guidelines

A method is described for growth of a Lactobacillus plantarum starter culture in jars of commercially available pasteurized fresh-pack kosher dill cucumbers so that jars can be used to inoculate commercial scale cucumber fermentation tanks. A procedure is also described to transfer lactic acid bacteria from frozen storage in MRS broth into cucumber juice and commercial jars of kosher dill cucumbers so that a selected strain of lactic acid bacteria can be kosher certified for commercial fermentations in processing plants that operate under kosher certification. The strain of L. plantarum used in these experiments grew to maximum cell numbers in 4 d at 20 to 25 °C and then maintained viable cell numbers for 2 wk at >10(8) CFU/mL so the culture was suitable for inoculation of fermentation tanks. Refrigeration of jars of culture after they grow to maximum numbers minimizes die-off of cells sufficiently so that a pure culture can be maintained by aseptically transferring brine containing viable bacteria to a new pH-adjusted jar only once every 4 mo. PRACTICAL APPLICATION: This report describes a method to prepare a lactic acid bacteria starter culture suitable for kosher vegetable fermentations.     

植物乳杆菌最适生长底物解析及高密度培养工艺

为提高植物乳杆菌的增殖浓度,分别测定菌株在添加不同氮源、不同缓冲盐、不同浓度的MnSO4和不同促生长物质时菌株的生长浓度。结果表明,酵母类氮源是植物乳杆菌的最适氮源,缓冲盐在恒pH培养时对菌株生长无促进作用,锰浓度与最高活菌数呈正相关,在以酵母浸粉为氮源时植物乳杆菌培养不需要添加其他生长因子。进一步优化菌株的最适pH值和碳氮比,基于可耐受渗透压,优化恒pH培养和恒pH自动反馈补料培养基和培养工艺,得到各菌株的最适培养策略。3株菌的最适氮源添加量为40~45 g/L,MnSO4的最适添加量为0. 25 g/L,最适碳氮比为对数生长期生长速率被抑制时的碳氮消耗比。恒pH 5. 5自动反馈补料培养植物乳杆菌X1,活菌数达到4. 1×10^10CFU/mL;恒pH 5. 5分批培养植物乳杆菌N8,活菌数达到2. 9×10^10CFU/m L;恒pH 6. 0分批培养植物乳杆菌N9,活菌数达到6. 2×10^10CFU/mL。该研究结果的应用将显著提高植物乳杆菌的工业化生产效率。     

银耳多糖对冻干植物乳杆菌细胞膜及糖代谢酶的保护作用

为了探索银耳多糖作为益生菌冻干保护剂的潜在应用,该研究以植物乳杆菌为试验菌,通过测定植物乳杆菌冻干前后的活菌数、细胞膜性质以及糖代谢酶活力的变化,评价了银耳多糖复合酪蛋白酸钠对植物乳杆菌的冻干保护效果。结果表明：当银耳多糖与酪蛋白酸钠的复合比例为3:1时,菌的最大存活率达到55.39%,此时细胞内钙离子荧光强度显著降低,为29.98;DPH的荧光强度最低,为4.91;细胞内乳酸脱氢酶、钠钾ATP酶与钙镁ATP酶的活力最大,分别为0.32 U/mL、121.61 U/g与45.64 U/g。银耳多糖使膜受损的菌百分比从98.35%最大降低至37.90%。但冻干前后己糖激酶和丙酮酸激酶的活力无显著性变化。红外扫描结果表明银耳多糖与酪蛋白酸钠之间存在非共价相互作用;此外,银耳多糖的保护使菌粉表面的孔隙率降低,酪蛋白酸钠的复合则增加了菌粉的致密性。因此,银耳多糖在冷冻干燥过程中对植物乳杆菌细胞膜受到的损伤具有显著的抑制作用,对植物乳杆菌的乳酸脱氢酶、钠钾ATP酶及钙镁ATP酶具有明显的保护作用。     

Identification of potential probiotic starter cultures for Scandinavian-type fermented sausages

Potential probiotic cultures suitable as starter cultures for the Scandinavian-type fermented sausages were identified among strains well-adapted to fermented meats as well as strains originating from a culture collection. From 15 different fermented meat products, 22 strains were isolated as dominant non-starter lactic acid bacteria (NSLAB). The isolates were identified by RAPD, API and sequence analysis of 16S rRNA and showed to be five strains of Lactobacillus sakei, five strains of Lactobacillus farciminis, five strains belonging to the group of Lactobacillus plantarum/pentosus, four strains of Lactobacillus alimentarius, two strains of Lactobacillus brevis and one strain of Lactobacillus versmoldensis. Heterofermentative strains as well as strains not growing at 37 degrees C and not lowering pH below 5.1 in a meat model were excluded leaving 9 strains for further studies. These strains together with 19 strains from a culture collection were evaluated by in vitro methods including survival upon exposure to pH 2.5 or 0.3% oxgall and adhesion to the human colon adenocarcinoma cell line Caco-2 as well as antimicrobial activity against potential pathogens. Strains that fulfilled all the probiotic criteria and showed to be fast acid producers in a meat model included three strains belonging to the group of Lb. plantarum/pentosus (MF1291, MF1298, MF1300) which originated from the dominant NSLAB of fermented meat products. MF1291 and MF 1298 were further identified as Lb. plantarum and MF1300 as Lb. pentosus. The three strains were all successfully applied as starter cultures for the production of fermented sausage. The viable count at the end of the processing period reached high cell numbers (4.7x10(7)-2.9x10(8) cfu/g) and pH of the sausages decreased to pH 4.8-4.9 without any flavour deviation compared to sausage fermented by a commercial meat starter culture.     

植物乳杆菌B2纯种发酵萝卜泡菜的研究

以萝卜为原料,以自行分离的植物乳杆菌B2进行单一菌株发酵剂接种制作萝卜泡菜产品,与完全自然发酵产品比较,通过比较发酵周期、发酵酸度、pH、OD值等,并用固相微萃取法结合气-质联用技术分析产品的风味物质,同时感官评价。结果表明,以B2单一菌株发酵剂接种制作萝卜泡菜,各指标优于自然发酵泡菜。     

一株具降胆固醇功能的海洋源植物乳杆菌的筛选及其益生性能分析

本文采用邻苯二甲醛法从喙鲸内脏中筛选分离出一株高效降胆固醇能力的乳酸菌HJ-S2,考察并研究了乳酸菌HJ-S2的体外益生性能。通过耐酸、耐胆盐测定菌株的益生作用,研究了其对致病菌的抑菌作用及对人结肠癌细胞HT-29的粘附能力,利用形态学、API50CH和16S rDNA对菌株进行鉴定。乳酸菌HJ-S2体外降胆固醇能力达到48.82%,在p H值3的酸性环境中培养3h耐受率达到82.73%,在含0.3%的胆盐环境中培养3 h耐受率达80.62%。菌株代谢物对致病菌大肠杆菌、金黄色葡萄球菌和沙门氏菌均有一定的抑菌作用,且菌株对人结肠癌细胞HT-29很强的粘附能力,粘附细菌数为132.52,经鉴定为植物乳杆菌Lactobacillus plantarum,并命名为HJ-S2(CGMCCNO:17720)。筛选得到的植物乳杆菌HJ-S2体外具有高效降胆固醇能力,可进一步开发为功能性微生态制剂。     

分离自Kefir的植物乳杆菌体外降胆固醇作用的研究

对分离Kefir中的植物乳杆菌体外降胆固醇作用进行了研究。结果表明，植物乳杆菌在含胆盐的培养基中具有良好的降胆固醇能力，24h后培养基中的胆固醇可降低60％(0．18g／L)；该菌体外降胆固醇作用的机理主要是其产生的胆盐水解酶水解胆盐变为游离态的胆酸，与胆固醇形成复合物，在酸性pH值条件下发生共沉淀。同时，也发现该菌产生的胆盐水解酶对反应底物(各种胆盐)的特异性不同，以脱氧牛磺胆酸最佳。     

Application of central composite design and response surface methodology to the fermentation of olive juice by Lactobacillus plantarum and Debaryomyces hansenii

The objectives of this study were to investigate the effects of NaCl, calcium acetate and calcium lactate in concentrations corresponding to ionic strengths equivalent to 2-10%, w/v salt brines as well as the 50% replacement of NaCl contained in the above mixture by KCl. A central composite design and response surface methodology were used to optimize the maximum specific growth rate of Lactobacillus plantarum ATCC 8014 and Debaryomyces hansenii 2114. The fermentation was carried out in olive juice obtained from Kalamon black olives at 30 degrees C with initial pH 5.0. Mathematical models describing the combined effects of these factors on the maximum specific growth rate of L. plantarum or D. hansenii were established. Both strains in single cultures showed higher maximum specific growth rate in olive juice supplemented with NaCl/KCl, Ca-acetate and Ca-lactate. But in mixed culture fermentations of olive juice supplemented with NaCl, Ca-acetate and Ca-lactate, higher specific growth rates were obtained. Under the optimum growth conditions determined for the single culture fermentations, i.e. 378.4 mM NaCl, 34.1 mM Ca-acetate and 39.9 mM Ca-lactate, mixed culture fermentation was undertaken by varying the time of inoculation of the yeast strain. When D. hansenii was inoculated 48 h before L. plantarum the maximum specific growth rate of L. plantarum was increased to 0.247 per hour, which was significantly higher compared to L. plantarum alone (0.211 per hour). In mixed culture fermentation of olive juice supplemented with the mixture of NaCl/KCl under similar conditions as above, a maximum specific growth rate of L. plantarum of 0.218 per hour was determined. The optimum conditions determined for mixed culture fermentation are useful in fermentation of black olives Kalamon variety under lower salt content.     

植物乳杆菌的降胆固研究

对植物乳杆菌MA2的体外和体内降胆固醇活性进行了研究,结果显示:培养基中添加的胆固醇的初始浓度对植物乳杆菌MA2的胆固醇去除率影响显著,当胆固醇的终质量浓度为30mg/dL,不添加胆盐的条件下,菌株MA2的胆固醇去除率达到(55±4.5)%。死菌体和休眠菌体也可去除培养基中的胆固醇,且菌体细胞浓度的影响显著。添加胆盐对菌株MA2的胆固醇去除率影响显著。体外实验表明植物乳杆菌MA2通过吸附胆固醇,进而将胆固醇吸入菌体细胞中来降低培养基中的胆固醇。MA2菌株可显著降低大鼠血清总胆固醇、甘油三酯的水平,表明其作为降血脂的益生菌的应用潜力。