以乳清为原料采用混菌发酵技术促进Nisin生产的研究

采用了一种不用外加碱或移除乳酸来达到控制发酵液中乳酸的新技术,即采用乳酸菌和酵母菌的混和发酵来控制pH值以促进乳酸链球菌素(nisin)的生产。其原理是乳酸菌在发酵生产nisin的过程中所产生的乳酸可以被酵母菌利用,从而达到控制发酵液pH值的目的;同时,混菌培养选用的酵母菌不能利用乳清培养基中的乳糖,从而避免了2株菌之间对碳源的竞争,创造了一个互利共生的微生态环境,从而达到促进nisin生产的目的。实验表明,乳酸根的积累对nisin发酵生产具有抑制作用;混合发酵的酵母菌利用了发酵液中的乳酸,促进了乳酸菌生长和nisin生产;酵母菌较乳酸菌提前3h接入到发酵培养基中时,能更好地控制pH值;最佳的接种比例为乳酸菌3%,酵母菌5%。     

与酵母菌具有共生作用乳酸菌LC5培养基的优化

用对酵母菌(YE4)有促进作用的乳酸菌(LC5)为研究对象,以OD值和活菌数为测定指标筛选合适的培养基。通过对5种乳酸菌常用培养基的筛选,确定M17培养基中LC5生长良好。以M17培养基作为基础培养基,进一步利用单因素试验和正交试验的设计方法对其进行优化,得出改良M17培养基的最佳成分为:大豆蛋白胨为0.6%,胰蛋白胨为0.6%,牛肉膏为0.4%,酵母浸提粉为0.5%,K2HPO4为0.7%,盐液A为0.4%,甘油为0.8%,乳糖为0.6%。优化后的培养基乳酸菌LC5的活菌数可达到1.89×109cfu/mL,是基础培养基的5.9倍,OD值是M17基础培养基的1.22倍。     

具有共生关系的乳酸菌和酵母菌的筛选

对分离自酸马奶酒中的9株乳酸菌和8株酵母菌存在共生作用的菌株进行筛选。分别于各自培养基中加入相应的代谢产物,以等量的生理盐水作为对照,测定其稳定期的浊度值和干重值。结果表明:乳酸菌的代谢产物促进的酵母菌生长有12个组合,而接近一半的酵母菌代谢产物对乳酸菌有促进作用。有7个组合的乳酸菌和酵母菌存在共生作用。结论是7个共生组合分别是:LAB2YST4、LAB4YST2、LAB5YST2、LAB7YST2、LAB7YST4、LAB8YST2、LAB9YST2。7个组合中酵母菌YST2和除LAB2外其余的乳酸菌均存在不同程度的共生,共生原因还待进一步研究。     

酸马奶酒中乳酸菌和酵母菌互作关系的研究

选取来自酸马奶酒中的2株乳酸菌(坚强肠球菌W1和粪肠球菌W2)和2株酵母菌(厚壁酵母属J1和娄德酵母属J2)进行互作关系的研究,采用固体平板涂布法的定性研究法和液体培养测定OD值、pH、滴定酸度的定量研究法。结果表明:2株乳酸菌发酵液都对酵母菌菌落生长有促进作用,发酵液经110℃、7min灭菌后,并不影响其促进作用;2株酵母菌发酵液对坚强肠球菌W1培养过程中的OD值、pH和滴定酸度无明显影响,而对粪肠球菌W2的生长及其产酸能力具有明显地促进作用;随着发酵液的添加量增加,促进作用更为明显。     

藏灵菇耐胃肠道环境酵母菌和乳酸菌的筛选

以藏灵菇发酵乳液为原料,分别利用改良的马铃薯乳糖培养基和改良MRS培养基分离出8株酵母菌和10株乳酸菌。以pH值为3.0,质量分数为0.3%牛胆盐的培养液模拟胃肠道环境作为初筛条件,以不同pH值和牛胆盐质量分数作为耐酸和耐胆盐的复筛条件,筛选得到耐受性好活力高的两株酵母菌C3和C8,两株乳酸菌M1和M3。     

发酵米浆中高发酵性能酵母菌和乳酸菌的筛选和鉴定

为了实现传统米发糕的规范化工业生产,从传统米发糕的发酵米浆中筛选发酵性能较好的菌株,并进行种属鉴定。首先从发酵米浆中分离出40 株酵母菌的疑似菌和30 株乳酸菌的疑似菌。经杜氏小管产气、发酵液特性和生长曲线分析三级筛选,最终得到1 株高发酵性能的酵母菌。经革兰氏染色等初筛、pH 值和生长曲线分析,最终得到1 株高发酵性能的乳酸菌。最后对两株菌株进行菌落形态观察和生理生化种属鉴定,确定分别为卡斯特酒香酵母和植物乳杆菌。     

影响鲜湿米粉食味品质微生物的分离与鉴定

从鲜湿米粉发酵液中分离并鉴定影响其食味品质的微生物菌株,以制备发酵剂推进米粉生产的工业化进程。首先从传统米粉发酵液中,分离得41株酵母菌和60株乳酸菌(LAB),并对其胞外酶活性(过氧化氢酶、β-葡萄糖苷酶、脂肪酶、α-淀粉酶、蛋白水解酶)、不同pH值和温度下的生长和酸化能力等技术特征进行分析,共筛选出11株酵母菌株和19株乳酸菌。然后通过对酵母菌的内转录间隔区序列和乳酸菌的16SrRNA基因鉴定,经基因型和系统发育树分析表明:3株酵母菌和4株乳酸菌。采用这7株菌对大米进行控制发酵制备鲜湿米粉,结果表明,乳酸菌中L.plantarum CSL 23发酵制备的米粉拉伸强度、硬度、内聚性和吸水率最高,黏附性和蒸煮损失最低;酵母中S.cerevisiae CSY 13发酵的米粉品质更好。因此,2株菌具有用来制备鲜湿米粉专用发酵剂的潜力,为规模化控制发酵生产鲜湿米粉提供试验依据。     

云南杨梅发酵液中酵母菌筛选及耐受性研究

以云南杨梅为材料进行自然发酵,发酵液经酵母浸出粉胨葡萄糖(YPD)培养基和麦芽汁培养基分离纯化获得酵母菌。对菌株进行产酯、产乙醇及产H₂S能力测定,结合杜氏小管发酵实验筛选,然后对筛选酵母菌进行耐受性测试,最后基于26S r DNA基因片段序列对筛选酵母菌进行分子生物学鉴定。结果表明,从云南杨梅发酵液中分离纯化获得59株酵母菌,筛选到3株优良酵母菌,编号为JYR-5、JYR-14、JYR-17,3株菌在温度40℃、乙醇体积分数5%、pH 2.5、葡萄糖质量浓度300 mg/L、SO₂质量浓度400 mg/L的条件下仍能正常生长。经鉴定,菌株JYR-5、JYR-14为酿酒酵母(Saccharomyces cerevisiae),菌株JYR-17为库德里阿兹威毕赤酵母(Pichia kudriavzevii)。因此,从云南杨梅发酵液中筛选的3株酵母菌为杨梅酒酿造微生物资源开发利用提供一定参考价值。     

耐盐乳酸菌与酵母菌在酱醪汁中协同作用的研究

采用透析培养的方法,研究了添加耐盐乳酸菌与酱醪中酵母菌的协同作用以及发酵工艺的控制和对酱油主要风味的影响.结果表明:在酱醪发酵过程中,添加耐盐乳酸菌明显促进T酵母哮菌的生长,而与S酵母菌存在着相互促进的关系.在酱醪发酵前期,同时添加104个/mL耐盐乳酸菌与T酵母菌时对发酵酱醪中的pH值和总酸起到稳定作用,提高了还原糖的产生速度,对中后期酱油主体风味的形成和提高具有良好的效果.     

分离自传统自然发酵食品中降胆固醇乳酸菌的筛选与评价

为了筛选优良的降胆固醇乳酸菌,本分析了从传统自然发酵食品中分离的30株乳酸菌,从中筛选出WS1、LP2、LP3、LP4、LP5和LP6 6株具有较强降胆固醇能力的乳酸菌,并通过对其胆盐耐受性及在含胆盐MRS培养基中的降胆固醇效率的分析,复筛出其中的WS1、LP2、LP3和LP6 4株乳酸菌。对4株乳酸菌的生长特征、产酸特征、不同生长阶段的种子液对胆固醇的降低效率的影响、耐酸特征及人工胃酸的耐受性进行分析和评价。结果显示,4株优选的乳酸菌降胆固醇能力较好,在胆盐中具有一定的存活力。胆盐的存在对降胆固醇能力具有一定的促进作用,4株菌在常规MRS培养基中生长和产酸特征稳定。不同生长时期的种子液对乳酸菌的降胆固醇能力有一定影响,对数期优于稳定期。耐酸性分析表明, pH越高,乳酸菌耐受性越好。pH 2.5,处理2h,乳酸菌依然具有一定的存活力;pH 3.5,乳酸菌存活力和存活时间都有所提高,pH 4.5时的存活力基本和自然pH的保持一致。在人工胃酸条件下,4株复筛乳酸菌在pH 2的条件下,存活时间超过4 h,随着pH的升高,存活率和存活时间都有所提高。本试验结果表明,4株乳酸菌均具有降胆固醇的特征,为后续优良菌株的降胆固醇机理研究打下了基础。     

酸马乳发酵过程中乳酸菌与酵母菌生长的相互影响

研究新疆酸马乳中乳酸乳球菌WLB5、干酪乳杆菌MLS5与马克思克鲁维酵母菌WWMJ1间的相互作用。结果表明：在酸马乳发酵过程中,马克思克鲁维酵母菌WWMJ1可以促进干酪乳杆菌MLS5的生长,干酪乳杆菌MLS5对马克思克鲁维酵母菌WWMJ1的生长有抑制作用,乳酸乳球菌WLB5能促进马克思克鲁维酵母菌WWMJ1的生长。乳酸乳球菌WLB5和干酪乳杆菌MLS5混合发酵有助于提高酸马乳中乳酸菌总活菌数。本研究可为酵母菌在发酵乳制品中的应用及开发新型乳制品提供一定参考。     

Fermentation and microflora of plaa-som, a thai fermented fish product prepared with different salt concentrations

Plaa-som is a Thai fermented fish product prepared from snakehead fish, salt, palm syrup and sometimes roasted rice. We studied the effects of different salt concentrations on decrease in pH and on microflora composition during fermentation. Two low-salt batches were prepared, containing 6% and 7% salt (w/w) as well as two high-salt batches, containing 9% and 11% salt. pH decreased rapidly from 6 to 4.5 in low-salt batches, whereas in high-salt batches, a slow or no decrease in pH was found. Lactic acid bacteria (LAB) and yeasts were isolated as the dominant microorganisms during fermentation. LAB counts increased to 10(8)-10(9) cfu g(-1) and yeast counts to 10(7)-5 x 10(7) cfu g(-1) in all batches, except in the 11% salt batch, where counts were 1-2 log lower. Phenotypic tests, ITS-PCR, carbohydrate fermentations and 16S rRNA gene sequencing identified LAB isolates as Pediococcus pentosaceus, Lactobacillus alimentarius/farciminis, Weisella confusa, L. plantarum and Lactococcus garviae. The latter species was only isolated from high-salt batches. Phenotypic characteristics, ITS-PCR and carbohydrate assimilation identified 95% of the yeasts as Zygosaccharomyces rouxii. It is concluded that the fermentation of plaa-som is delayed by a salt-level of 9% due to an inhibition of LAB growth. The growth of Z. rouxii has no influence on the fermentation rate, but may contribute positively to the flavour development of the product.     

Effect of oenological practices on microbial populations using culture-independent techniques

Sulphur dioxide (SO(2)) addition and yeast inoculation are well-established practices in winemaking for restricting the growth of indigenous yeasts and bacterial populations. The effect of these oenological practices on wine microbial populations has been evaluated using culture-independent methods. These are quantitative PCR (qPCR) for the enumeration of yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB), and PCR-DGGE to determine the yeast and bacteria species diversity. The PCR-DGGE method detected a low yeast and bacteria species diversity. On the contrary, the specificity of the primers designed for the qPCR allowed that minor microbial groups such as Hanseniaspora were accurately quantified regardless of a large presence of other microbial groups such as Saccharomyces. From an oenological point of view, inoculation increased the proportion of Saccharomyces vs. non-Saccharomyces in a shorter time. Hanseniaspora increased during the first phase and decreased during the latter phases of the process, especially in the sulphited fermentations. Both yeast inoculation and SO(2) kept the LAB populations at very low level, while the AAB populations were hardly affected by these two practices.     

Overview of Sourdough Technology: from Production to Marketing

Sourdough is a type of dough fermented by yeast and lactic acid bacteria (LAB) used as sponge dough in bread making. Sourdough may have four classifications according to the fermentation type and the technological process used. On sourdough type I, the fermentation of yeasts and LAB present in the flour occurs spontaneously whereas in type II, fermentation occurs after the inoculation of a starter culture. Type III is simply type II sourdough dehydrated and type IV is a mixture of type I and type II sourdough, produced on laboratory scale. With LAB’s growth during fermentation, pH decreases with subsequent hydrolysis of starch and protein, favoring the growth of yeasts. The yeast in turn, releases amino acids during autolysis, contributing to the growth of LAB. Due to this synergistic growth of LAB and yeasts, the products obtained from sourdough show more concentration of flavor, more elastic dough, and a longer shelf life than bread fermented only by yeast. Although, the benefits of this technology, using sourdough, is present just in a few countries, where it is also possible to obtain the dehydrated sourdough starters, for domestic use, in local supermarkets. Due to its fermentative process complexity, this paper presents updated information about the fabrication process of sourdough, main factors affecting this process, benefits of sourdough use for bakery products, and the varieties of dehydrated sourdough commercially available throughout the world.     

Investigation of Microbial Communities of Chinese Sourdoughs Using Culture‐Dependent and DGGE Approaches

Sourdough is typically characterized by the complex microbial communities mainly comprising of yeasts and lactic acid bacteria (LAB). The objective of this study was to explore the microbiota of Chinese traditional sourdoughs collected from different areas of China using culture‐dependent and denaturing gradient gel electrophoresis (DGGE) methods. A total of 131 yeasts, 2 molds, and 106 LAB strains were isolated and identified. Based on the culture‐dependent analysis, the populations of yeasts and LAB were at the level of 10⁵ to 10⁷ and 10⁶ to 10⁷ cfu/g, respectively. Similarly, the results of RT‐qPCR showed that the values of total yeasts and LAB populations were in the range of 10⁶ to 10⁷ and 10⁷ to 10⁸ copies/g, respectively. Using culture‐dependent method, a total of 7 yeasts, 2 molds and 7 LAB species were identified. Results showed that Saccharomyces cerevisiae and Lactobacillus plantarum were the predominant species among the yeasts and LAB microflora. Similarly, using PCR‐DGGE approach, 7 yeasts, 1 mold and 9 LAB species were detected. The yeast, S. cerevisiae, represented the predominant, while the yeast Candida tropicalis represented the subdominant species of the yeast community. Among the LAB community, Lactobacillus sanfranciscensis was the predominant species, while Lactococcus qarvieae, Enterococcus faecium, Lactobacillus delbrueckii and Enterococcus cecorum were among the less dominant species.     

Preservation of acidified cucumbers with a combination of fumaric acid and cinnamaldehyde that target lactic acid bacteria and yeasts

The naturally occurring compound, fumaric acid, was evaluated as a potential preservative for the long-term storage of cucumbers. Fumaric acid inhibited growth of lactic acid bacteria (LAB) in an acidified cucumber juice medium model system resembling conditions that could allow preservation of cucumbers in the presence of sodium benzoate. Forty millimolars of fumaric acid were required to inhibit growth of an extremely aciduric Lactobacillus plantarum LA0445 strain at pH 3.8. Half of this concentration was required to achieve inhibition of L. plantarum LA0445 at pH 3.5. As expected growth of the spoilage yeasts Zygosaccharomyces globiformis and Z. bailii was not inhibited by fumaric acid at near saturation concentrations in the same cucumber juice medium. To usefully apply fumaric acid as a preservative in acidified foods it will be necessary to combine it with a food grade yeast inhibitor. The antimicrobial agent, cinnamaldehyde (3.8 mM) prevented growth of Z. globiformis as well as the yeasts that were present on fresh cucumbers. Acidified cucumbers were successfully preserved, as indicated by lack of yeasts or LAB growth and microbial lactic acid or ethanol production by a combination of fumaric acid and cinnamaldehyde during storage at 30 °C for 2 mo. This combination of 2 naturally occurring preservative compounds may serve as an alternative approach to the use of sodium benzoate, potassium sorbate, or sodium metabisulfite for preservation of acidified vegetables without a thermal process. PRACTICAL APPLICATION: This study evaluates the potential application of alternative preservatives for the long-term storage of cucumbers in a reduced NaCl cover brine solution and without a thermal process.     

Optimum conditions for combined application of Leuconostoc sp. and Saccharomyces sp. to sourdough

Investigation of the optimum conditions for the combined application of Saccharomyces sp. (yeast) and Leuconostoc sp. (lactic acid bacteria, LAB) isolated in a previous study to the development of novel sourdough was carried out by response surface analysis. First, the cell growth conditions were analyzed. LAB showed good proliferation under conditions of 30–35°C, low pH, and high acidity, whereas the growth of yeast was inhibited. The growth of yeast was optimum at 25°C for 24 h. Based on these results, analysis of sourdough was carried out by varying the LAB population, temperature, and time after fixing the number of yeast. It was determined by response surface analysis that the optimal conditions for fermentation are LAB population of 10⁵ CFU/mL, temperature of 25°C, and reaction time of 24 h. From these results, the growth of LAB should be constantly maintained, and an appropriate pH that does not inhibit the growth of yeast due to the presence of generated organic acid is required to allow for the unique properties of sourdough. This study could give useful information for the development of novel sourdough.     

Pickling and storage of caperberries (Capparis spp.)

Caperberries (0.7–1.9?cm diameter), collected in June 1996, were pickled for 1 month in 5% and 10% brine. Some physical, chemical and microbiological analyses were made during their fermentation and storage. While lactic acid bacteria (LAB) increased during fermentation in 5% brine, they did not increase after 15 days in 10% brine. In some cases, total bacterial (TB) growth decreased. In both brines, yeasts/moulds and coliforms (except for 5% brine on day 15) were not present after 15 and 20 days, respectively. For both caper species, the most suitable salt concentration, for LAB activity was 5–10% NaCl, and the best length of fermentation with respect to product colour, flavour, acidity, pH and LAB activity in brine was 20–25 days. The texture was maintained well during storage. Acidity and pH were markedly affected by the concentration of brine. The quality of all samples was maintained with a 15% concentration of old or fresh brine during storage. The colour of the samples, especially those of C. ovata, stored with old brine was darker that of fresh berries. As no "off" flavour was observed, it was concluded that pickled caper berries can be stored in fresh brine with a 15% NaCl concentration.     

The Occurrence of Beer Spoilage Lactic Acid Bacteria in Craft Beer Production

Beer is one of the world's most ancient and widely consumed fermented alcoholic beverages produced with water, malted cereal grains (generally barley and wheat), hops, and yeast. Beer is considered an unfavorable substrate of growth for many microorganisms, however, there are a limited number of bacteria and yeasts, which are capable of growth and may spoil beer especially if it is not pasteurized or sterile‐filtered as craft beer. The aim of this research study was to track beer spoilage lactic acid bacteria (LAB) inside a brewery and during the craft beer production process. To that end, indoor air and work surface samples, collected in the brewery under study, together with commercial active dry yeasts, exhausted yeasts, yeast pellet (obtained after mature beer centrifugation), and spoiled beers were analyzed through culture‐dependent methods and PCR‐DGGE in order to identify the contaminant LAB species and the source of contamination. Lactobacillus brevis was detected in a spoiled beer and in a commercial active dry yeast. Other LAB species and bacteria ascribed to Staphylococcus sp., Enterobaceriaceae, and Acetobacter sp. were found in the brewery. In conclusion, the PCR‐DGGE technique coupled with the culture‐dependent method was found to be a useful tool for identifying the beer spoilage bacteria and the source of contamination. The analyses carried out on raw materials, by‐products, final products, and the brewery were useful for implementing a sanitization plan to be adopted in the production plant.