低胆固醇发酵牛肉香肠工艺参数的优化

为了降低牛肉香肠中的胆固醇含量,采用饱和D-最优试验设计方法优化香肠的工艺参数,以胆固醇降解率和感官评定为指标值,通过回归分析建立目标函数的数学模型,从而确定最优加工工艺参数为:啤酒酵母菌、植物乳杆菌配比为1︰2,接种量为0.9%,发酵温度为28℃,发酵时间为24h。在此条件下生产出的发酵牛肉香肠中胆固醇比原含量降低了34.98%。     

植物乳杆菌抑制黄曲霉活性代谢物的初步研究

植物乳杆菌（Lactobacillus plantarum）能够抑制黄曲霉生长,但起主要抑菌作用的物质尚未明确。该研究采用非靶向代谢组学技术比较分析了8株抑菌活性较好（S组）和8株抑菌活性较差（W组）的L. plantarum发酵上清液。结果显示,两组L. plantarum发酵上清液的代谢组存在显著差异（P<0.05）。通过数据库比对鉴定得到咪唑乙酸、酪氨酸等30个显著差异代谢物（P<0.05）,其中有机酸、脂肪酸等酸性物质较多为22个。通过与已报道的乳酸菌产生的抗真菌物质相比较,找到十八烷酸、吲哚乙酸等结构一致或结构类似物,表明上清液中酸性物质起主要的抑菌作用,且其抑菌活性依赖于低 pH 值的酸性环境。在L. plantarum产生的主要有机酸中,乳酸、乙酸、丙酸的抑菌活性良好,其抑制黄曲霉活性从大到小依次为丙酸、乙酸、乳酸。当乙酸浓度为2.64g/L、丙酸浓度为1.76 g/L时,可完全抑制浓度为106个/mL的黄曲霉孢子生长。综合表明,植物乳杆菌代谢物中有机酸和脂肪酸为主要抑菌物质,且抑菌活性随酸性物质浓度增大而增强。     

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。     

产β-甘露聚糖酶内生菌的诱变育种及产酶条件优化

以刺槐豆内生菌Paenibacillus sp.CH-3为出发菌株,采用紫外-硫酸二乙酯-亚硝酸盐对其进行复合诱变,并对菌株的发酵条件进行了优化。结果表明：经过紫外线-硫酸二乙酯-亚硝酸盐复合诱变,获得一株高产β-甘露聚糖酶的突变菌株Paenibacillus sp.CH3-05,其酶活力为160.2U/mL,较出发菌株（42U/mL）提高了281.4%。其最佳发酵培养基为：酵母膏0.25%,魔芋粉3%,磷酸氢二铵0.25%,氯化钠0.1%,硫酸镁0.3%,磷酸氢二钾0.2%,CaCl22 mmol/L。最佳发酵条件为：初始pH 6.5,接种量2%,培养温度35℃,转速200 r/min,培养时间78 h,在该条件下测得酶活为233.5 U/mL,较出发菌株提高456%。     

鱼菜共生系统中乳酸菌的筛选及其发酵矿化应用

针对鱼菜共生系统固体废弃物资源化利用效率低的问题,该研究旨在筛选出抗逆性好、矿化功能强的鱼源性乳酸菌,加强鱼粪残饵的发酵矿化性能。试验从鱼菜共生系统中的生物滤料和鱼体中分离乳酸菌,并通过抗逆性及发酵矿化性能检测,筛选出2株具有应用潜力的鱼粪残饵矿化菌株,经鉴定分别为乳酸乳球菌（Lactococcus lactis）L1和糊精乳杆菌（Lactobacillus dextrinicus）L2。耐温性、耐酸碱性和耐盐性检测结果显示,L1表现出较好的抗逆性。在 50 ℃时,L1存活率为96.60%,显著高于L2的存活率80.35%（P<0.05）;在pH值分别为5.0和9.0时,L1的存活率分别为65.43%和71.25%,高于L2的存活率31.10%和52.22%（P<0.05）;当盐浓度为60 g/L时,L1的存活率为37.33%,而L2无法存活。通过比较对照组（CK组,未添加乳酸菌）和乳酸乳球菌组（L组,添加乳酸乳球菌）发酵矿化过程中发酵液水质和矿物元素含量,结果显示 L组的有机物降解及矿化效果更好,除硫（S）元素以外,钾（K）、钙（Ca）、镁（Mg）、铁（Fe）、锰（Mn）和锌（Zn）元素的矿化率均在第3天达到最高（27.59%～94.67%）。综上所述,乳酸乳球菌（Lactococcus lactis）具有较强的抗逆性和显著的发酵矿化效果,且其最佳发酵矿化周期为3 d。该研究可为提高鱼菜共生系统固体废弃物资源化利用效率提供技术支持。     

复合乳酸菌接种发酵对浙东腌冬瓜发酵品质的影响

为缩短浙东传统腌冬瓜生产周期,并提高其质量品质,以自然腌制为对照,研究从浙东传统腌冬瓜分离获得的4株优势乳酸菌(植物乳杆菌、发酵乳杆菌、棒状乳杆菌和戊糖乳杆菌)按不同方式复配接种后对腌制冬瓜发酵品质的影响,并经室温发酵20d后检测产品的p H、总酸、亚硝酸盐含量、氨基酸含量、乳酸菌和细菌总数的变化情况及挥发性物质。结果表明,3种复合菌发酵方式的乳酸菌和细菌总数均高于对照,产酸迅速,亚硝酸盐含量显著降低。4种菌按相同比例复配发酵效果较好,与对照组比,其氨基酸总量和必需氨基酸含量分别提高32.02%和17.04%,而甲硫氨酸、苯丙氨酸、丝氨酸及精氨酸含量分别提高226.13%、178.00%、237.28%和95.52%;挥发性成分种类及含量与对照有较高的相似度,其中酯类和醛类物质相对含量分别提高10.00%和29.88%。综上,复合菌发酵改善了腌冬瓜的营养品质及风味。本研究结果为复合菌剂应用到发酵蔬菜中提供了一定的参考。     

两株蜡状芽孢杆菌对毒死蜱的降解动力学研究

利用微生物消除农药污染是一项安全、经济、有效的方法,降解动力学模型的构建有助于理解污染物的生物降解行为和估测系统中特征污染物的浓度变化,菌株对高浓度污染物的降解效果是降解菌在受污染水体中实际应用的关键。本研究采用基础培养基中定量添加毒死蜱和定时取样分析毒死蜱残留浓度的方法,探讨两株蜡状芽孢杆菌(HY-1和HY-2)的接种体培养时间、接种量和降解菌对毒死蜱的降解动力学,同时研究了降解菌对高浓度毒死蜱的降解率。结果表明:HY-1和HY-2最适接种体培养时间分别为10 h和19 h,接种体培养时间对菌株降解毒死蜱的影响较大。两菌株最适接菌量为8%(v/v),接种量从4%增至8%时,接种量对HY-1降解毒死蜱的影响大于HY-2。当毒死蜱的初始浓度为40 mg.L 1、80 mg.L 1、100 mg.L 1和120 mg.L 1时,一级动力学方程ln(C0/Ct)=kt可以用来拟合两菌株对毒死蜱的降解动力学及确定降解动力学参数,当毒死蜱初始浓度再次增加时,仅HY-2对毒死蜱的降解符合一级动力学方程。当毒死蜱初始浓度为40~120 mg.L 1时,菌株HY-1对毒死蜱的降解速率常数分布在0.013 5~0.015 7;当毒死蜱初始浓度为40~200 mg.L 1时,菌株HY-2的降解速率常数分布在0.008 0~0.015 3。菌株HY-2比HY-1可以在较高的毒死蜱浓度下发挥降解作用,且降解率较高。因此,两菌株在毒死蜱污染水体的净化去毒方面具有重要意义。     

尿嘧啶对植物乳植杆菌LIP-1冻干存活率的影响及其作用机制

为了研究在培养基中添加微量生长因子对菌株冷冻干燥存活率的影响,该研究以植物乳植杆菌LIP-1（Lactiplantibacillus plantarum LIP-1）为研究对象,探究培养基中添加尿嘧啶对菌株冷冻干燥存活率的影响及其作用机制。结果表明与未添加尿嘧啶的空白对照组相比,在培养基中添加0.05 g/L尿嘧啶能够显著提高植物乳植杆菌LIP-1的活菌数与冻干存活率（P＜0.05）。对其作用机制进行探究,发现尿嘧啶的加入能够促使菌株合成更多的尿苷二磷酸（P＜0.05）,从而使肽聚糖含量显著增加（P＜0.05）,进而提高细胞壁的稳定性;同时尿嘧啶的加入抑制了乳清酸向尿苷二磷酸的转化,使胞内的乳清酸含量显著升高（P＜0.05）,菌株利用乳清酸的抗氧化能力减轻了活性氧对细胞膜不饱和脂肪酸的氧化程度,进而减少了细胞膜的损伤程度。结果表明植物乳植杆菌LIP-1通过代谢尿嘧啶减轻了细胞壁与细胞膜受到的冻干损伤,提高了菌株的冻干存活率。研究结果为提高菌株冷冻干燥存活率提供了新的方法和思路。     

高活菌数复合益生菌发酵乳工艺优化

益生菌发酵乳中的活菌数是保证其功能特性的关键因素,为提高益生菌发酵乳中的活菌总数,以干酪乳杆菌和双歧杆菌复合菌种为试验对象,以发酵过程中的pH值和吸光度平均值值为试验指标,在单因素试验的基础上,利用 Box-Benhnken中心组合试验和响应面分析法研究了接种量、益生菌接种比例、发酵温度、葡萄糖添加量以及大豆多肽添加量对发酵乳pH值和活菌数的影响,并建立了复合益生菌发酵模型。响应面优化试验结果表明复合益生菌发酵的最佳工艺条件是脱脂乳乳固体质量分数为12%,接种量为6%,干酪乳杆菌：双歧杆菌接种体积比为3∶1,葡萄糖添加量为2.9%,大豆多肽添加量为0.8%,发酵温度为34℃,在此条件下预测值为1.076,验证试验得到实际值为1.087,与理论预测值相比,相对误差为1.0%。发酵乳最大活菌数为4.1×1011 cfu/mL,与已有研究相比,活菌数提高了1-2个对数级。在4℃条件下贮藏21 d后,发酵乳的活菌数仍然保持在4.7×1010 cfu/mL。研究结果为工业化生产高活菌数的益生菌饮料提供参考。     

优良乳酸菌的筛选及对腌制茭白品质的影响

为获得最适腌制茭白启动发酵剂,缩短腌制茭白生产周期、提高质量品质,从分离筛选保藏的4种乳酸菌菌株(植物乳杆菌、短乳杆菌、嗜柠檬酸明串球菌和戊糖片球菌)中优选出植物乳杆菌和戊糖片球菌2种优良乳酸菌,采用5种不同配比方式分别接种发酵,研究茭白腌制过程中p H、酸度、亚硝酸盐和微生物指标的变化,并对成品的色泽、质地和挥发性风味物质进行分析。结果表明,配比方式对茭白腌制过程中p H、酸度、乳酸菌数和细菌总数的影响不大;在不同配比方式接种中,接种1%植物乳杆菌和2%戊糖片球菌复合菌作为茭白腌制发酵剂,其腌制茭白的"亚硝峰"值最小,为1.62 mg·kg-1,色泽与鲜茭白最为接近,质地口感最好;不同配比方式接种茭白腌制共检测出挥发性风味物质118种,1%植物乳杆菌和2%戊糖片球菌腌制的茭白成品所含风味物质种类和相对含量均较高。因此,最终确定的最佳发酵剂为1%植物乳杆菌和2%戊糖片球菌。本研究结果为茭白的腌制及干制加工的工业化生产提供了理论依据。     

Combined use of starter cultures and preservatives to control production of biogenic amines and improve sensorial profile in low-acid salami

The combined effect of starter culture, nitrites, and nitrates has been studied in low-acidity salamis, typical products of northern Italy. Nine batches have been prepared, combining three different inoculations of starter cultures (control, Lactobacillus plantarum , and Lactobacillus plantarum together with Kocuria varians ) with three different preservatives (control, sodium nitrate, and sodium nitrite). All of the batches showed a good fermentation process with a proper pH decrease, which was quicker in batches inoculated with L. plantarum. The use of starter cultures and in particular the use of nitrites allowed the control of the proliferation of Enterobacteriaceae and enterococci. The accumulation of biogenic amines, especially putrescine, cadaverine, tryptamine, and tyramine, in salami ready for consumption (60 days of ripening) was strongly affected by the presence of Enterobacteriaceae and enterococci. Results obtained showed that the combined use of adequate preservatives and starter cultures allows the production of safer products with improved sensorial profile.     

Contribution of the microbial and meat endogenous enzymes to the free amino acid and amine contents of dry fermented sausages.

The role of the starter culture and meat endogenous enzymes on the free amino acid and amine contents of dry fermented sausages was studied. Five batches of sausages were prepared. The control batch was manufactured with aseptic ingredients without microbial inoculation. The other four experimental batches were manufactured with aseptic ingredients inoculated with Lactobacillus plantarum 4045 or Micrococcus-12 or L. plantarum 4045 and Micrococcus-12 or L. plantarum 4045 and Staphylococcus sp. Their effects on pH, a(w), myofibrillar proteins, and free amino acid and amine contents were studied. Sausages inoculated only with L. plantarum 4045 or with this starter combined with a Micrococcaceae had the lowest pH as a result of carbohydrate fermentation. In all batches similar patterns were observed for myofibrillar proteins and free amino acids which could indicate that meat endogenous proteases play an important role in proteolytic phenomena. No changes were observed in the amine fraction, indicating that the strains used as starter cultures did not show amino acid decarboxylase activity.     

Biogenic amines in low- and reduced-fat dry fermented sausages formulated with konjac gel

Biogenic amines in low- and reduced-fat dry fermented sausages made with konjac gel (KG) as pork backfat replacer were studied. An increase (P < 0.05) was observed in the microbial count during the fermentation process, reaching levels of over 8 Log cfu/g of total viable microorganisms and lactic acid bacteria. However, no significant differences were observed in the microbiota evolution as a function of the reformulation process (fat and konjac gel content). High levels of physiological amines (spermidine, spermine, and agmatine) were observed in the raw material. From day 2 of the fermentation process an increase (P < 0.05) was observed in tyramine and putrescine, which were the predominant amines at the end of the storage period. The increase in these amines was proportional to the presence of KG and fat reduction. This can also be seen for spermine, with agmatine showing the inverse. The biogenic amine levels in these products reformulated with KG are not considered to pose a health risk to consumers.     

Biogenic amine formation and nitrite reactions in meat batter as affected by high-pressure processing and chilled storage

Changes in biogenic amine formation and nitrite depletion in meat batters as affected by pressure-temperature combinations (300 MPa/30 min/7, 20, and 40 degrees C), cooking process (70 degrees C/30 min), and storage (54 days/2 degrees C) were studied. Changes in residual nitrite concentration in raw meat batters were conditioned by the temperature and not by the pressure applied. Cooking process decreased (P < 0.05) the residual nitrite concentration in all samples. High-pressure processing and cooking treatment increased (P < 0.05) the nitrate content. Whereas protein-bound nitrite concentration decreased with pressure processing, no effect was observed with the heating process of meat batters. High-pressure processing conditions had no effect on the rate of residual nitrite loss throughout the storage. The application of high pressure decreased (P < 0.05) the concentration of some biogenic amines (tyramine, agmatine, and spermine). Irrespective of the high processing conditions, generally, throughout storage biogenic amine levels did not change or increased, although quantitatively this effect was not very important.     

Quantitation of selected odor-active constituents in dry fermented sausages prepared with different curing salts

The odor-active compounds of dry-fermented sausages with added nitrite or nitrate as curing agents were identified by gas chromatography-olfactometry (GC-O) applying the detection frequency (DF) method. The quantification of these compounds in the sausage was determined by multiple headspace solid-phase microextraction (multiple HS-SPME). There were no specific odor-active compounds related to the use of nitrite or nitrate although there were differences in the DF value of several compounds. The nitrite-added sausages presented higher DF values for ethanol, 1-hexanol, propanoic acid, 2-heptenal, and nonanal while the nitrate-added sausages had higher DF values for phenylacetaldehyde and 3-methyl-butanal. Eighteen compounds were quantified by multiple HS-SPME. Most of them were above their air detection thresholds, but only hexanal, heptanal, and 1-octen-3-ol were in a concentration higher than their oil threshold values. These compounds would probably be the main contributors to the aroma of fermented sausages.     

Biogenic amines in wines from three Spanish regions

One hundred and sixty-three wines from La Rioja, Utiel-Requena, and Tarragona were analyzed to determine if there were any differences in the concentrations of six biogenic amines that are found in these three regions. The influence of grape variety, type of vinification, wine pH, malolactic fermentation, and storage in bottle on biogenic amine concentrations was studied. Results show important differences in putrescine and histamine concentrations among regions, varieties of grape, and type of wine; differences were less appreciable for the remaining biogenic amines studied. Low pH prevented biogenic amine formation. Malolactic fermentation and short storage periods in bottle (3-6 months) showed increases in histamine concentration, whereas longer periods of storage led to a general decrease in histamine. Several strains of lactic acid bacteria were isolated in this work, and their ability to form biogenic amines was assayed in synthetic media, grape must, and wine. Grape varieties, different types of winemaking, pH, and lactic acid bacteria may be responsible for the differences observed in the biogenic amine concentrations of the wines analyzed.     

High-level expression of recombinant beta-galactosidases in Lactobacillus plantarum and Lactobacillus sakei using a Sakacin P-based expression system

This work presents the cloning and expression of the genes encoding heterodimeric beta-galactosidases from Lactobacillus reuteri L103, Lactobacillus acidophilus R22, Lactobacillus plantarum WCFS1, and Lactobacillus sakei Lb790. These enzymes consist of two subunits of approximately 73 and 35 kDa, which are encoded by two overlapping genes, lacL and lacM, respectively. We have cloned these genes into the lactobacillal expression vectors pSIP403 and pSIP409, which are based on the sakacin P operon of L. sakei ( S?rvig et al. Microbiology 2005, 151, 2439- 2449 ), and expressed them in the host strains L. plantarum WCFS1 and L. sakei Lb790. Results varied considerably, ranging from 2.23 to 61.1 U/mg of beta-galactosidase activity, depending on the origin of the lacLM genes, the host strain, and the expression vector used. Highest expression levels were obtained in a laboratory cultivation of L. plantarum WCFS1 harboring the plasmid pEH3R containing the lacLM gene from L. reuteri L103. These cultivations yielded approximately 23 000 U of beta-galactosidase activity per liter, corresponding to the formation of roughly 100 mg of recombinant protein per liter of fermentation medium, and beta-galactosidase levels amounted to 55% of the total intracellular protein of the host organism. To further verify the suitability of this expression system, recombinant beta-galactosidase from L. reuteri was purified to apparent homogeneity. The properties of the purified enzyme were essentially identical with the properties of purified native beta-galactosidase from L. reuteri L103. The presented results lead the way to efficient overproduction of beta-galactosidase in a food-grade expression system, which is of high interest for applications in food industry.