西藏“灵菇”乳的发酵与抑菌机制研究（简报）

根据发酵过程中菌粒中主要微生物数量与发酵乳中主要营养物和产物含量间的关系,分析探讨了西藏"灵菇"乳的发酵机制;考察了不同发酵时期所得发酵乳对大肠杆菌、金黄色葡萄球菌、沙门氏菌的抑菌活性,结合化学物质模拟试验揭示了发酵乳的抑菌机制。结果显示,发酵前期,菌粒中的各微生物由于向乳中扩散及生长延滞,数量稍有减少;随着发酵的进行,菌粒中乳酸菌和酵母菌的数量一直处于相对稳定的水平,但醋酸菌含量呈明显的先升后降趋势;在整个发酵过程中,菌粒中的乳酸菌数量一直处于最高水平,其次为酵母菌,醋醋菌数量最少;乳酸是西藏“灵菇”发酵乳中的主要抑菌活性物质;乙酸和乙醇的存在对低浓度乳酸的抑菌活性有增效作用,但当乳酸浓度较高时,这种增效作用减弱。西藏“灵菇”发酵乳的总抑菌效果是微生物与其代谢产物的共同作用结果。     

西藏"灵菇"乳的发酵与抑菌机制研究

根据发酵过程中菌粒中主要微生物数量与发酵乳中主要营养物和产物含量间的关系,分析探讨了西藏"灵菇"乳的发酵机制:考察了不同发酵时期所得发酵乳对人肠杆菌、金黄色葡萄球菌、沙门氏菌的抑菌活性,结合化学物质模拟试验揭示了发酵乳的抑菌机制.结果显示,发酵前期,菌粒中的各微生物由于向乳中扩散及生长延滞,数量稍有减少;随着发酵的进行,菌粒中乳酸菌和酵母菌的数量一直处于相对稳定的水平,但醋酸菌含量呈明显的先升后降趋势;在整个发酵过程中,菌粒中的乳酸菌数量一直处于最高水平,其次为酵母菌,醋醋菌数量最少:乳酸是西藏"灵菇"发酵乳中的主要抑菌活性物质;乙酸和乙醇的存在对低浓度乳酸的抑菌活性有增效作用,但当乳酸浓度较高时,这种增效作用减弱.西藏"灵菇"发酵乳的总抑菌效果是微生物与其代谢产物的共同作用结果.     

预发酵方式对餐厨垃圾酸化抑菌及甲烷发酵的影响

餐厨垃圾水分和有机物含量高,在收集储运过程中易酸腐变臭,影响其资源化利用。该文在新鲜餐厨垃圾中分别接种酵母菌和乙酸菌进行乙醇和乙酸预发酵,预发酵后的餐厨垃圾与酒糟混合进行甲烷发酵,并与不接任何菌种的对照组比较,考察2种预发酵方式对餐厨垃圾酸化抑菌及其与酒糟混合甲烷发酵的影响。研究结果表明,乙酸和乙醇预发酵对餐厨垃圾均具有良好的酸化抑菌效果,其大肠杆菌和金黄葡萄球菌数均比对照组降低了2个数量级以上;累积甲烷产量由高到低的顺序是:乙醇预发酵对照乙酸预发酵组,前者累积产甲烷量比对照组和乙酸预发酵组分别提高21.3%和49.8%;乙醇预发酵组乙醇浓度最高,而乙酸和丙酸浓度均最低;相反,乙酸预发酵组挥发性脂肪酸浓度积累导致产甲烷菌活性较低,其产气效率最低。因此,乙醇预发酵是一种既能实现餐厨垃圾酸化抑菌,又能维持发酵系统的稳定性,提高其后续产甲烷效率的有效预发酵方式。     

蚯蚓粪中乳酸菌的分离及其在大肠杆菌O157：H7中的抗菌活性

本研究利用SL培养基从蚯蚓粪中分离到54株具有产酸性能的菌株,并以E.coli O157：H7（EDL933株）作为指示菌株,采用点种法检测分离菌株的抑菌活性。结果表明其中6个菌株对指示菌具有拮抗作用,通过形态特征,结合16S rDNA序列分析,初步鉴定该6个菌株分别为食物魏斯特菌（Listeria welshimeri）、乳酸片球菌（Pediococcus acidilactici）、短乳杆菌（Lactobacillus brevis）和格氏乳球菌（Lactococcus garvieae）。分离到的乳酸菌对E.coli O157：H7（EDL933株）具有显著的抑制作用,发酵温度和初始pH值影响发酵液的抑菌作用,优化环境因子可以促进拮抗菌对E.coli O157：H7的抑制作用。本研究为进一步分离抗菌产物用于人畜共患病的预防和治疗提供了理论依据。     

乙醇预发酵对餐厨垃圾与酒糟混合甲烷发酵的影响

为了解决餐厨垃圾两相干式厌氧发酵产甲烷过程中的挥发酸抑制问题,该试验在酸发酵阶段添加产乙醇菌分别进行12、24和48 h的预发酵,然后在相同条件下进行甲烷发酵,考察不同乙醇预发酵时间对餐厨垃圾与酒糟混合发酵产甲烷的影响,考察其甲烷发酵过程中的pH值、总挥发性有机酸(total volatile fatty acid,TVFA)、乙酸、丙酸、乙醇等参数的变化,并与未经乙醇预发酵的对照组比较,以期为相关领域的理论与实践研究提供参考经验。结果表明:各组乙醇浓度和pH值从高到低的顺序依次为:预发酵48 h组24 h组12 h组对照组,而乙酸和TVFA则呈现相反的顺序;由于乙醇为中性物质,且比丙酸容易转化为乙酸,故可缓解由有机酸累积而导致的酸抑制,使乙醇预发酵12、24、48 h组的累积产甲烷量分别比未经预发酵对照组高9.1%、31.6%、19.9%左右,其中,乙醇预发酵24h时厌氧消化效果较好。研究为厌氧发酵产甲烷过程中的解决酸抑制问题提供参考。     

蒲公英不同部位抑菌作用及代谢物差异的UPLC-MS/MS分析

为了研究蒲公英不同部位抑菌作用和代谢物成分的差异,本物采用打孔法和超高效液相色谱串联质谱(UPLC-MS/MS)方法,研究了蒲公英根、茎叶和花的水提取物和醇提取物对5种致病菌的抑菌作用,并对不同部位的代谢物成分进行了定性和定量分析。结果表明,蒲公英不同部位初提取物对变形杆菌、嗜根考克氏菌和枯草芽孢杆菌均有抑菌作用,而对大肠杆菌和铜绿假单胞菌均无抑菌作用;醇提法的抑菌效果优于水提法,花提取物的抑菌活性最强,根最弱。蒲公英的不同部位共鉴定出449种代谢物,根、茎叶和花中相对含量最高的物质分别为反油酸(脂类)、γ-亚麻酸(脂类)和木犀草素(黄酮类);且根、茎叶和花中相对含量排名前20的物质中黄酮类分别有0种、3种和8种,表明黄酮类物质可能是蒲公英中的主要抑菌活性物质。正交偏最小二乘法-判别分析(OPLS-DA)结果显示,木犀草素-7-O-葡萄糖苷在花中上调幅度最大,橙皮素C-丙二酰己糖苷为根中特有物质。本研究结果为利用蒲公英不同部位研制和开发新型药物提供了理论依据。     

挥发性有机酸对产沼气效果的模拟试验

两相厌氧生物处理工艺在废弃物资源化利用中得到广泛应用。为了摸清产酸相的主要产物在产气过程中的效应,该文利用甲酸、乙酸、丙酸和丁酸等4种有机酸模拟产酸过程中的主要酸化产物,以活性污泥为接种物,设计单因素试验和正交试验,研究了在中温条件下,4种有机酸单一作用条件下和混合作用条件下,甲烷日产气量和累积产气量的变化。结果表明：单一有机酸作底物时,存在一个浓度阈值,高于此值会抑制甲烷产生,低于此值,产气效果随着酸浓度增大而提高;有机酸共同存在时,可以产生协同优势,其中乙酸的产气优势高于丁酸,丙酸具有抑制作用;4种单一酸中,单位浓度下乙酸产气效果最好,甲酸产气量最大,甲酸积累不易对发酵过程产生抑制作用,而丙酸累积最易造成抑制。因此,建议在沼气工程中适当调整产甲酸、乙酸和丁酸较多的发酵物料浓度,从而提高产气效率。     

菌酶协同处理提高脱脂薏米水提取液营养价值

为了高效利用脱脂薏米制备水提取液,该研究采用酸性蛋白酶、木瓜蛋白酶、中性蛋白酶、碱性蛋白酶和风味蛋白酶协同干酪乳杆菌发酵脱脂薏米,考察了菌酶协同处理对薏米水提取液可滴定酸、还原糖、总酚、多肽、氨基酸、抗氧化活性及酪氨酸酶和黄嘌呤氧化酶抑制活性的影响。与干酪乳杆菌单独发酵相比,菌酶协同可促进发酵薏米的蛋白质降解,显著提高水提取液中多肽、还原糖和总酚浓度（P<0.05）。菌酶协同处理后水提取液中小分子多肽比例显著增加（P<0.05）,游离氨基酸含量显著提高（P<0.05）,必需氨基酸与非必需氨基酸比值达到0.73～1.17。此外,菌酶协同处理后水提取液的铁离子还原能力和ABTS阳离子自由基清除能力较干酪乳杆菌单独发酵显著增强（P<0.05）,酪氨酸酶和黄嘌呤氧化酶抑制活性分别达到了52.3%～58.1%和36.7%～41.5%,显著高于干酪乳杆菌单独发酵（P<0.05）。结果表明,菌酶协同可显著提高脱脂薏米水提取液的营养品质,综合比较后木瓜蛋白酶的增强效果最佳。研究结果可为利用薏米提油后的副产物脱脂薏米开发高附加值薏米水提取液功能食品和化妆品提供参考。     

降解亚硝酸盐和生物胺乳杆菌筛选及其改善鱼肉香肠品质效果

为了筛选出优良亚硝酸盐和生物胺的降解植物乳杆菌,并研究在鱼肉发酵香肠中的应用。该文主要考察了19 株植物乳杆菌亚硝酸盐和生物胺的降解活性及相关基因的携带情况,从中筛选出优良菌株,并研究其对鱼肉香肠发酵过程中各种亚硝胺、生物胺、腐败微生物的抑制作用和对其他理化指标的影响。试验结果表明,L. plantarum CP3对亚硝酸盐和生物胺的降解能力最强,分别达到95%和77%,PCR结果显示此菌株中含有亚硝酸盐降解（假定蛋白（hypothetical protein,HP）、谷胱甘肽还原酶（glutathione reductase,GR）、一氧化还原酶（Oxidoreductase,ORD））和生物胺降解的相关基因（multicopper oxidases,suf I）,且不含生物胺生成相关基因（hdc,tdc,odc）。GC-MS测定结果显示L. plantarum CP3对发酵香肠中4种亚硝胺-二乙基亚硝胺（N-nitrosodiethylamine,NDEA）,二甲基亚硝胺（N-nitrosodimethylamine,NDMA）,亚硝基吗啉（N-nitrosomorpholine,NMOR）和甲基乙基亚硝胺（N-nitrosomethylamine,NMEA）的生成都具有显著的抑制作用,联合接种L.saki M4可以协同提高亚硝胺的降解能力。并且,与对照组相比,接种L. plantarum CP3+L.saki M4的鱼肉香肠,发酵48 h后腐胺、尸胺和酪胺的含量分别降低了76.83%,93.33%和88%。另外,接种乳酸菌处理组香肠中的肠杆菌、假单胞菌等腐败菌及pH值、硫代巴比妥酸（thiobarbituric acid,TBA）和挥发性盐基氮（total volatile basic nitrogen,TVB-N）的含量显著低于对照组。该研究结果体现了L. plantarum CP3作为香肠等肉制品发酵剂的良好开发应用前景。     

不同生物源助冻剂结合渗透脱水处理对蓝莓冻融品质的影响

为探究植物乳杆菌PC11（Lactobacillus plantarum PC11）发酵代谢物、葡聚糖和冬小麦蛋白组合糖液浸渍处理对蓝莓冻融品质的影响。该研究以冷冻和糖液浸渍（65 g/mL）冷冻为对照组,以PC11发酵菌代谢物（10 g/mL）、葡聚糖（1 g/mL）和冬小麦蛋白（0.1 g/mL）结合糖液浸渍预处理后冷冻为试验组,比较分析蓝莓的抗氧化能力、活性物质含量、质构特性、冷冻特性、汁液流失率、色差和细胞膜透性等指标。结果表明：单一糖液浸渍处理不如添加助冻剂结合糖液浸渍的效果理想,其中添加PC11发酵菌代谢物（10 g/mL）的处理组可以有效维持蓝莓冻融后的各项指标,此处理组胶着性、硬度、咀嚼性和弹性分别为0.89 N、1.22 N、0.56 N和0.65 mm,总花色苷和原花青素含量与其他组相比具有显著差异（P<0.05）,含量分别为2.59和0.54 mg/g,总花色苷的保留率比糖液浸渍组提高了1.78倍。该文提出的冷冻预处理办法为针对蓝莓原料不同加工方式的冷冻贮藏技术的发展提供了新思路。     

饼肥中的有机营养物质及其在发酵过程中的变化

分析测定了在恒温、恒湿条件下 4种植物油饼发酵过程中有机物质的变化。结果表明 ,由于不同植物油饼化学组成不同在发酵过程中所产生有机营养物质的成分和数量存在较大差异。发酵过程所产生的高级脂肪酸以C18、C16酸为主 ,并且不饱和脂肪酸的含量明显高于饱和脂肪酸 ;有机酸则以乳酸、苹果酸、柠檬酸、琥珀酸、酒石酸、乙酸含量较高 ,草酸含量较低 ;氨基酸则以谷氨酸含量最高 ,天门冬氨酸、亮氨酸和精氨酸含量次之。有机酸和高级脂肪酸含量随发酵时间的增加而增加 ,腐熟到一定程度时达到最高 ;而饼肥发酵 70d时各种氨基酸含量明显降低。 4种饼肥相比 ,豆饼和花生饼在发酵过程中氨基酸产生量大于芝麻饼和菜籽饼。从速度来看 ,芝麻饼腐熟最快 ,其次是豆饼 ,再次是花生饼 ,菜籽饼腐熟最慢。     

Wine volatile and amino acid composition after malolactic fermentation: effect of Oenococcus oeni and Lactobacillus plantarum starter cultures

Red wine amino acids and volatile compounds were analyzed before and after malolactic fermentation carried out by four different starter cultures of the species Oenococcus oeni and Lactobacillus plantarum. The purpose of this study was to determine whether differences can be attributed to the lactic acid bacteria strain used in this important step of the wine-making process. The malolactic cultures selected for this study were indigenous wine lactic acid bacteria strains. The data were evaluated using different multivariate analysis techniques. Results showed different malolactic behaviors for O. oeni and L. plantarum and significant metabolic differences between both species. A degree of diversity was found within each lactic acid bacteria group, since wines presented specific characteristics depending on the lactic acid bacteria strain used. In all cases, malolactic fermentation seemed to modify the amino acid and volatile composition of the wine.     

Characterization of Organic Acids Recovered from Rhizosphere of Corn Grown on Biosolids‐Treated Medium

Abstract

Organic acid production by plants and microorganisms was quantified in sand media amended with biosolids in the presence and absence of corn (Zea mays L.) in a sand‐culture hydroponic medium. Total quantities of organic acids were greatest in treatments containing both plants and biosolids, with lesser amounts in treatments with plants alone, biosolids‐treated media alone, and a nutrient solution–irrigated blank medium. Biosolids enhanced organic acid production in the rhizosphere and influenced the composition of organic acid mixtures. Only lactic, acetic, butyric, and oxalic acids were detected in media without plants. When the medium was planted, additional organic acids were recovered including tartaric, maleic, succinic, valeric, glutaric, pyruvic, and propionic. Lactic, acetic, and butyric acids were predominant in solutions recovered from the planted media and collectively accounted for 0.65 to 0.75 of the COO^? mole fraction. Oxalic, maleic, and tartaric acids were the second most abundant and varied from 0.05 to 0.1 of the mole fraction, followed by succinic, valeric, glutaric, propionic, and pyruvic acids, comprising ≤0.05 of the mole fraction. Plant growth stage had no effect on relative proportions of organic acids but did influence the total quantities of organic acids recovered. Biosolids sources did not have a significant effect on either the quantity or composition of organic acids in any media. The predominance of organic acids that are microbial fermentation products suggests that the carbon contained in root exudates and biosolid amendments was transformed into a mixture of various fermentation products that accumulated in the rhizosphere solution and sand medium as a result of microbial growth and activity.     

Changes in the pH of Two Different Composts Are Dependent on the Production of Organic Acids

The effects of pH in two different composts were studied. Yamagata compost was made under aerobic conditions, while Shimoduma compost was under semi-aerobic conditions. The change in pH of Yamagata compost was very limited, 7.38–8.97, and it started to increase only after 40-th d of composting. The pH increase from 40 to 70 d coincided with an increase in ammonium content, while a decrease after 70 d was probably as a result of nitrification. The concentration of organic acids (formic, acetic, propionic, iso-butyric and iso-valeric) produced in the first 14 d was high and after that, acetic, propionic and iso-valeric acids decreased quickly. In the Shimoduma compost, however, the range of pH change was wider (3.95–6.65) and increased almost linearly. The quantity of organic acids (citric, lactic, formic and acetic acid) produced depended on the periodic manual stirring. Immediately after stirring the amount of acids decreased, but it increased during the period without stirring.     

自动化机械发酵食醋过程中品质变化规律

为全面了解转鼓式反应器固态发酵食醋过程中主要成分和风味物质的动态变化规律,该文对反应器发酵食醋过程进行跟踪取样,利用高效液相色谱、固相微萃取技术、气相色谱-质谱联用技术,对酒精、总酸、还原糖、氨基态氮、有机酸、挥发性风味物质含量变化进行实时监测,结合主成分分析方法,探究食醋不同发酵阶段的风味物质差异。结果表明,酒精含量在发酵0～4 d内迅速增加,随后逐渐下降至零;总酸含量呈现先快速上升后缓慢上升的趋势;还原糖含量呈先快速下降后逐渐上升,最后逐渐下降的趋势;氨基态氮呈现先快速上升后缓慢上升的趋势;乙酸和乳酸是主要的有机酸,整个发酵过程中,乙酸含量持续增加,乳酸含量呈先上升后下降趋势,其他有机酸含量较少,发酵期间变化波动相对较小;共检测出64种挥发性风味物质,包括酯类25种,醇类12种,酸类6种,酚类5种,醛类5种,酮类6种,杂环类5种;发酵前、中、后期的重要挥发性物质分别是醇类化合物、酯类和醛类化合物、酸类化合物。该结果为推进转鼓式固态发酵食醋反应器的实际生产应用提供了理论基础和数据参考。     

腌制叶用芥菜发酵菌分离鉴定及应用研究

为获得发酵性能优良的菌株,生产优质的发酵叶用芥菜,本研究从自然发酵的芥菜中分离发酵用菌株,通过测定乳酸菌的生长、产酸和亚硝酸盐降解能力以及酵母菌的产气、产酯能力和蛋白酶活性等发酵特性进行筛选,并对筛选获得的优良发酵菌加以应用。结果表明,筛选获得的2株乳酸菌L8短乳杆菌(Lactobacillus brevis)、L9植物乳杆菌(Lactobacillus plantarum)和1株酵母菌Y9酿酒酵母菌(Saccharomyces cerevisiae)在腌制叶用芥菜中的发酵性能优良。与自然发酵的叶用芥菜相比,接菌发酵叶用芥菜的pH值和亚硝酸盐含量分别减少了2.22%和88.13%;总酸含量、氨基酸态氮含量分别增加了9.02%和28.09%。接菌发酵提高了腌制芥菜的营养品质和安全性。本研究丰富了叶用芥菜的发酵菌种,为蔬菜发酵加工产业奠定了技术基础。     

Evaluation of vinification lees as a general medium for lactobacillus strains

Lactobacillus species present high nutritional requirements, so it is necessary to find new low-cost nutrient components for fermentation media. This work compares the utilization of vinification lees (an important residue of wineries) from red and white winemaking technology, distilled or not. An amount of 20 g of lees/L was used as the unique nutrient to obtain lactic acid from glucose using Lactobacillus strains with different properties: L. plantarum CECT-221, L. pentosus CECT-4023, L. casei CECT-5275, and L. coryniformis subsp. torquens CECT-25600. Only L. casei using distilled lees showed values (Pmax = 92.1 g/L and Y(P/S) = 1.05 g/g) similar to those obtained with the MRS broth. The UV spectra of white and red lees, distilled or not, allowed an interpretation of the different phenolic compounds present and their influence on the fermentation. Their detoxification by extraction with organic compounds and fermentation with L. pentosus was also considered. Time courses of glucose and lactic acid were modeled according to reported models to obtain more information about the process.     

低分子量有机酸对砖红壤动电性质的影响

研究了 5种低分子量有机酸 ,即醋酸、柠檬酸、马来酸、苹果酸和草酸 ,对昆明和徐闻两种砖红壤动电性质的影响。结果表明 ,在试验条件下 ,两种砖红壤的 ζ电位随醋酸浓度的递增逐渐正移 ,而后 ζ电位正移速率显著增大 ,再后 ζ电位渐趋稳定 ;两者的 ζ电位随柠檬酸浓度的增加向更负的方向移动 ,在试验浓度范围 (0～ 0 .1 5mmolL-1 )内 ,负移的速率没有明显差别。在pH 3.2至 6 .0之间 ,含有不同有机酸的两种砖红壤悬液的ζ电位均具有如下顺序 :醋酸 >马来酸 >苹果酸 >柠檬酸 >草酸 ;含同一有机酸的悬液在相同pH下的 ζ电位 ,昆明砖红壤总是高于徐闻砖红壤。在不同有机酸存在下 ,两种砖红壤悬液等电点的顺序与上述的 ζ电位顺序相同 ;对于相同处理来说 ,昆明砖红壤的等电点高于徐闻砖红壤者 0 .5～ 1 .0pH单位。     

Reduction in Microbial Load of Wheat by Tempering with Organic Acid and Saline Solutions

Reducing microbial contamination in wheat is desirable to ensure consumer safety. In this study, the efficacy of adding organic acids and NaCl to tempering water to reduce microbial contamination in hard wheat was evaluated. Hard red winter wheat was tempered to 15.5% moisture by adding sterile distilled water (control) or tempering solutions containing organic acids (acetic, citric, lactic, or propionic; 1.0, 2.5, or 5.0 v/v), NaCl (26 or 52% w/v), or a combination of organic acid (acetic or lactic; 2.5 and 5.0% v/v) and NaCl (26 or 52% w/v). After tempering, the microbial load was significantly reduced by all the acid and NaCl treatments when compared with the control. Wheat tempered with 5% acetic, propionic, and lactic acids resulted in reductions of 1.7, 2.3, and 3.8 log colony forming units (CFU) per gram in aerobic plate count (APC), Enterobacteriaceae (Eb), and mold counts, respectively. The combination lactic acid (5.0%) and NaCl (52%) was the most effective against APC and Eb, with an average reduction of 4.3 and 4.7 log CFU/g, respectively. Tempering wheat with organic acids and saline solutions may provide milled products with improved microbiological quality when compared with the traditional tempering process using water.